Abstract

Mg is a material of choice for biodegradable implants. The main challenge for using Mg in temporary implants is to provide protective surfaces that mitigate its rapid degradation in biological fluids and also confer sufficient cytocompatibility and bacterial resistance to Mg-coated surfaces. Even though carbonate mineralization is the most important source of biominerals, such as the skeletons and shells of many marine organisms, there has been little success in the controlled growth of carbonate layers by synthetic processes. We present here the formation mechanism, antibacterial activity, and cell viability of magnesian calcite biomimetic coatings grown on biodegradable Mg via a green, one-step route. Cell compatibility assessment showed cell viability higher than 80% after 72 h using fibroblast cells (NCTC, clone L929) and higher than 60% after 72 h using human osteoblast-like cells (SaOS-2); the cells displayed a normal appearance and a density similar to the control sample. Antimicrobial potential evaluation against both Gram-positive (Staphylococcus aureus (ATCC 25923)) and Gram-negative (Pseudomonas aeruginosa (ATCC 27853)) strains demonstrated that the coated samples significantly inhibited bacterial adhesion and biofilm formation compared to the untreated control. Calcite coatings grown on biodegradable Mg by a single coating process showed the necessary properties of cell compatibility and bacterial resistance for application in surface-modified Mg biomaterials for temporary implants.

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Background: The force a muscle exerts is partly determined by anatomical parameters, such as its physiological cross-section. The temporal muscle is structurally heterogeneous. To the authors' knowledge, the ultrastructure of this muscle has been poorly specifically studied.Methods: Five adult Wistar rats weighting 350-400 g were used as temporal muscle donors. Tissues were specifically processed and studied under transmission electron microscope.Results: On ultrathin cuts, the general ultrastructural pattern of striated muscles was observed. Moreover, pennate sarcomeres were identified, sharing a one-end insertion on the same Z-disc. Bipennate morphologies resulted when two neighbor sarcomeres, attached on different neighbor Z-discs and separated at that end by a triad, converged to the same Z-disc at the opposite ends, thus building a thicker myofibril distinctively flanked by triads. Tripennate morphologies were identified when sarcomeres from three different Z-discs converged to the same Z-disc at the opposite ends.Conclusions: These results support recent evidence of sarcomeres branching gathered in mice. Adequate identification of the sites of excitation-contraction coupling should be on both sides of a myofibril, on bidimensional ultrathin cuts, to avoid false positive results due to putative longitudinal folds of myofibrils.& COPY; 2023 Elsevier GmbH. All rights reserved.

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The bioactivity of the versatile biodegradable biopolymer poly(lactic acid) (PLA) can be obtained by combining it with natural or synthetic compounds. This paper deals with the preparation of bioactive formulations involving the melt processing of PLA loaded with a medicinal plant (sage) and an edible oil (coconut oil), together with an organomodifed montmorillonite nanoclay, and an assessment of the resulting structural, surface, morphological, mechanical, and biological properties of the biocomposites. By modulating the components, the prepared biocomposites show flexibility, both antioxidant and antimicrobial activity, as well as a high degree of cytocompatibility, being capable to induce the cell adherence and proliferation on their surface. Overall, the obtained results suggest that the developed PLA-based biocomposites could potentially be used as bioactive materials in medical applications.

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In this paper, the extract from the leaves of Hedera helix L.was subjected to isolation and fractionation of saponins, in order to determine their in vitro antiproliferative activity. The ethanolic extract was obtained by maceration from H.helix L.leaves, grown in Romania, and the saponins were isolated and fractionated using column chromatography. The total saponin content in the extract and the isolated saponins was determined. Three saponins of interest, hederacoside C, a-hederin and hederagenin were identified by LC/MS analysis. Four selected fractions rich in these saponins tested in vitro, showed antiproliferative effects at a concentration of 200 mg/mL in a culture of human cervix carcinoma Hep-2 cells. The tested fractions were non-cytotoxic in the range of concentration 2-400 mg/mL on normal fibroblasts cells from NCTC cell line. Our results demonstrated the selected saponin fractions isolated from H.helix L.leaves may be considered as possible antitumor agents.

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This study aimed to evaluate the hypoglycemic potential, antioxidant activity and prebiotic activity of a hydroalcoholic extract of blueberry pomace (BP), an aqueous extract of chia seeds (CS) and a novel combination of BP-CS extracts (BCM) for further use as ingredient of functional food. Spectrometric and HPLC analyses were used to characterize the total phenolic and flavonoid content and composition of BP, while CS was analyzed for total carbohydrate content. Data showed that the BCM mixture exerted an inhibition of alpha-amylase activity, which was 1.36 times higher than that of BP and 1.25 higher than CS extract. The mixture also showed better scavenging activity of free DPPH radicals than individual extracts, and had an IC50 value of 603.12 mu g/mL. In vitro testing indicated that both serum- and colon-reaching products of simulated intestinal digestion of BCM presented the capacity to protect Caco-2 intestinal cells against oxidative stress by inhibition of reactive oxygen species production. In addition, the colon-reaching product of BCM digestion had the capacity to significantly (p < 0.05) stimulate the growth of Lactobacillus rhamnosus and Lactobacillus acidophilus, revealing a prebiotic potential. All these results indicated that improved biological activity of the novel combination of BP and CS extracts could be due to the synergistic action of constituents. The combination is recommended for further testing and the development of novel functional food for controlling type 2 diabetes and gastrointestinal conditions.

Abstract

Polyphenolic extracts from natural sources have received great interest due to their beneficial properties for human health. A method to reduce their variability is to use the design of experiments which allows a limited number of experiments to be performed while exploring the experimental space. Firstly, a 2(3)-full factorial model was used to investigate the polyphenols extraction from wild bilberry leaves. Spectrophotometric data (the content of polyphenols, flavonoids, chlorophyll and radical scavenger activity) and extraction yield were used as responses, and six statistical models were determined depending on the two numerical factors (temperature and alcohol % of ethanol-water mixture) being significant (p < 0.05) in all cases. Numerical optimisation performed by Design Expert 13 software correlates well with the chemical profile determined by high-performance liquid chromatography and the amount of the polyphenol. Afterwards, under the optimised conditions, an extract was prepared in three extraction steps for which composition, chemical stability and antimicrobial properties were evaluated. The antimicrobial potential of the extract was compared with that of the standard compounds (rutin and chlorogenic acid), and the results supported a synergistic effect of the extract components.

Abstract

The present research aims to describe a new methodology to obtain biocompatible hydrogels based on Aloe vera used for wound healing applications. The properties of two hydrogels (differing in Aloe vera concentration, AV5 and AV10) prepared by an all-green synthesis method from raw, natural, renewable and bioavailable materials such as salicylic acid, allantoin and xanthan gum were investigated. The morphology of the Aloe vera based hydrogel biomaterials was studied by SEM analysis. The rheological properties of the hydrogels, as well as their cell viability, biocompatibility and cytotoxicity, were determined. The antibacterial activity of Aloe vera based hydrogels was evaluated both on Gram-positive, Staphylococcus aureus and on Gram-negative, Pseudomonas aeruginosa strains. The obtained novel green Aloe vera based hydrogels showed good antibacterial properties. In vitro scratch assay demonstrated the capacity of both AV5 and AV10 hydrogels to accelerate cell proliferation and migration and induce closure of a wounded area. A corroboration of all morphological, rheological, cytocompatibility and cell viability results indicates that this Aloe vera based hydrogel may be suitable for wound healing applications.

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Featured Application: Nutritional Profiling for subsequent dietary supplements/nutraceuticals development and formulation as well as by-products valorification. In recent years, the scientific community has made significant progress in understanding nutrition, leading consumers to shift their preferences away from animal-based protein products and towards natural, plant-based protein sources. This study aimed to determine the nutritional value, in vitro cytotoxicity and antioxidant activity for different sources of high protein content products (pea, yeast, almond, spirulina and Pleurotus spp.) with potential usage as raw materials for dietary supplements, especially since these products do not benefit from stricter regulation requirements regarding their actual health benefits. The characterization of raw materials consisted in evaluation of their nutritional profile (by addressing moisture content, crude protein content, extractable fat, ash, carbohydrates) and microbial contamination (TAMC, TYMC, Enterobacteriaceae and beta-glucuronidase positive Escherichia coli), total content of free amino acids, soluble proteins, phenols and flavonoids, as well as antioxidant activity through chemical assays. We used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) release to evaluate the potential cytotoxicity of selected raw materials. Results obtained indicate high percentages of proteins for the pea powder (77.96%) and Spirulina powder (64.79%), Pleurotus spp. flour had strong antioxidant activity, while the highest contamination values were registered for Pleurotus spp. powder (4.6 +/- 10(5) CFU/g or 5.66 log CFU/g). Cytotoxicity results demonstrate that tested ingredients have an impact on the metabolic activity of cells, affecting cellular integrity and provoking leakage of DNA at several concentrations. While plant-based protein supplementation may appear to be a promising solution to balance our busy lives, there are several advantages and disadvantages associated with them, including issues related to their absorption rate, bioavailability, cytotoxicity and actual nutritional benefits.

Abstract

Lonicera caerulea L., (blue honeysuckle, blue honeyberry or haskap) is an Eastern Siberian shrub with fruits known to have potential medicinal applications. The aim of this paper was to investigate the physico-chemical properties, composition and the correlation to the total antioxidant capacity of some fluid extracts of haskap berries for their use as nutraceuticals. The dried fruits were collected from Moldavia region in North-Eastern Romania and the extracts were prepared at a concentration of 10% (w/v) in ethanol concentrations of 50:50 (v:v), 70:30 (v:v) and 96:4 (v:v), respectively. The total content of main bioactive compounds, such as polyphenols, flavonoids, tannins, anthocyanins, carotenoids, lycopene, lutein, vitamin C and carbohydrates was determined by UV-Vis spectrophotometric assays. Total antioxidant capacity of extracts was assessed by photochemiluminescence method, in comparison to Trolox (R), an analogue of vitamin E, used as antioxidant standard. The obtained results emphasized that the hydroalcoholic extract in 70:30 (v:v) ethanol had significantly (p<0.01) higher content of vitamin C, carbohydrates and carotenoids than the other two extracts, and similar quantities of total phenolics, flavonoids, lycopene and lutein, In accordance, the same extract had the highest total antioxidant activity and close to the values of resveratrol, a known antioxidant polyphenolic compound. In conclusion, hydroalcoholic extracts of Lonicera caerulea L. berries could be used as valuable products for the formulation of novel nutraceuticals with significant antioxidant activity.

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Lonicera caerulaea L. and Aronia melanocarpa (Michx.) Elliot fruits are frequently used for their health benefits as they are rich in bioactive compounds. They are recognized as a source of natural and valuable phytonutrients, which makes them a superfood. L. caerulea presents antioxidant activity three to five times higher than other berries which are more commonly consumed, such as blackberries or strawberries. In addition, their ascorbic acid level is the highest among fruits. The species A. melanocarpa is considered one of the richest known sources of antioxidants, surpassing currants, cranberries, blueberries, elderberries, and gooseberries, and contains one of the highest amounts of sorbitol. The non-edible leaves of genus Aronia became more extensively analyzed as a byproduct or waste material due to their high polyphenol, flavonoid, and phenolic acid content, along with a small amount of anthocyanins, which are used as ingredients in nutraceuticals, herbal teas, bio-cosmetics, cosmeceuticals, food and by the pharmaceutical industry. These plants are a rich source of vitamins, tocopherols, folic acid, and carotenoids. However, they remain outside of mainstream fruit consumption, being well known only to a small audience. This review aims to shed light on L. caerulaea and A. melanocarpa and their bioactive compounds as healthy superfoods with antioxidant, anti-inflammatory, antitumor, antimicrobial, and anti-diabetic effects, and hepato-, cardio-, and neuro-protective potential. In this view, we hope to promote their cultivation and processing, increase their commercial availability, and also highlight the ability of these species to be used as potential nutraceutical sources, helpful for human health.

Abstract

The richness and structure of symbiont assemblages are shaped by many factors acting at different spatial and temporal scales. Among them, host phylogeny and geographic distance play essential roles. To explore drivers of richness and structure of symbiont assemblages, feather mites and seabirds are an attractive model due to their peculiar traits. Feather mites are permanent ectosymbionts and considered highly host-specific with limited dispersal abilities. Seabirds harbour species-rich feather mite communities and their colonial breeding provides opportunities for symbionts to exploit several host species. To unravel the richness and test the influence of host phylogeny and geographic distance on mite communities, we collected feather mites from 11 seabird species breeding across the Atlantic Ocean and Mediterranean Sea. Using morphological criteria, we identified 33 mite species, of which 17 were new or recently described species. Based on community similarity analyses, mite communities were clearly structured by host genera, while the effect of geography within host genera or species was weak and sometimes negligible. We found a weak but significant effect of geographic distance on similarity patterns in mite communities for Cory's shearwaters Calonectris borealis. Feather mite specificity mainly occurred at the host-genus rather than at host-species level, suggesting that previously inferred host species-specificity may have resulted from poorly sampling closely related host species. Overall, our results show that host phylogeny plays a greater role than geography in determining the composition and structure of mite assemblages and pinpoints the importance of sampling mites from closely-related host species before describing mite specificity patterns.

Abstract

The richness and structure of symbiont assemblages are shaped by many factors acting at different spatial and temporal scales. Among them, host phylogeny and geographic distance play essential roles. To explore drivers of richness and structure of symbiont assemblages, feather mites and seabirds are an attractive model due to their peculiar traits. Feather mites are permanent ectosymbionts and considered highly host-specific with limited dispersal abilities. Seabirds harbour species-rich feather mite communities and their colonial breeding provides opportunities for symbionts to exploit several host species. To unravel the richness and test the influence of host phylogeny and geographic distance on mite communities, we collected feather mites from 11 seabird species breeding across the Atlantic Ocean and Mediterranean Sea. Using morphological criteria, we identified 33 mite species, of which 17 were new or recently described species. Based on community similarity analyses, mite communities were clearly structured by host genera, while the effect of geography within host genera or species was weak and sometimes negligible. We found a weak but significant effect of geographic distance on similarity patterns in mite communities for Cory’s shearwaters Calonectris borealis. Feather mite specificity mainly occurred at the host-genus rather than at host-species level, suggesting that previously inferred host species-specificity may have resulted from poorly sampling closely related host species. Overall, our results show that host phylogeny plays a greater role than geography in determining the composition and structure of mite assemblages and pinpoints the importance of sampling mites from closely-related host species before describing mite specificity patterns. © 2023, The Author(s).

Abstract

Polyphenolic extracts from natural sources have received great interest due to their beneficial properties for human health. A method to reduce their variability is to use the design of experiments which allows a limited number of experiments to be performed while exploring the experimental space. Firstly, a 23-full factorial model was used to investigate the polyphenols extraction from wild bilberry leaves. Spectrophotometric data (the content of polyphenols, flavonoids, chlorophyll and radical scavenger activity) and extraction yield were used as responses, and six statistical models were determined depending on the two numerical factors (temperature and alcohol % of ethanol–water mixture) being significant (p &lt; 0.05) in all cases. Numerical optimisation performed by Design Expert 13 software correlates well with the chemical profile determined by high-performance liquid chromatography and the amount of the polyphenol. Afterwards, under the optimised conditions, an extract was prepared in three extraction steps for which composition, chemical stability and antimicrobial properties were evaluated. The antimicrobial potential of the extract was compared with that of the standard compounds (rutin and chlorogenic acid), and the results supported a synergistic effect of the extract components. © 2023 by the authors.

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Osteochondral structure reconstruction by tissue engineering, a challenge in regenerative medicine, requires a scaffold that ensures both articular cartilage and subchondral bone remodeling. Functional hydrogels and scaffolds present a strategy for the controlled delivery of signaling molecules (growth factors and therapeutic drugs) and are considered a promising therapeutic approach. Icariin is a pharmacologically-active small molecule of prenylated flavonol glycoside and the main bioactive flavonoid isolated from Epimedium spp. The in vitro and in vivo testing of icariin showed chondrogenic and ostseoinductive effects, comparable to bone morphogenetic proteins, and suggested its use as an alternative to growth factors, representing a low-cost, promising approach for osteochondral regeneration. This paper reviews the complex structure of the osteochondral tissue, underlining the main aspects of osteochondral defects and those specifically occurring in osteoarthritis. The significance of icariin's structure and the extraction methods were emphasized. Studies revealing the valuable chondrogenic and osteogenic effects of icariin for osteochondral restoration were also reviewed. The review highlighted th recent state-of-the-art related to hydrogels and scaffolds enriched with icariin developed as biocompatible materials for osteochondral regeneration strategies.

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The aim of this study was to investigate the main volatiles and polyphenolic compounds in hydrosols extracted from three Romanian Lamiaceae plants, sage (Salvia officinalis), rosemary (Rosmarinus officinalis) and lavender (Lavandula angustifolia) by steam distillation. The antioxidant and acetylcholinesterase inhibition capacity of hydrosols were assessed for their further use as biopesticides. The results have shown that sage and rosemary hydrosols presented better antioxidant activity than lavender hydrosol in all free radical scavenging and metal chelating assays. A strong relationship between the antioxidant activity of hydrosols' and their total phenolic and flavonoid content was found based on Pearson correlation coefficients. Sage and rosemary hydrosols exerted higher acetylcholinesterase inhibition than lavender hydrosol, but this activity was moderate correlated to phenolic content and low correlated to flavonoid content. These results suggested that volatiles and other compounds present in hydrosols were also involved in this process. In conclusion, Lamiaceae plant hydrosols could be further tested, as such or mixed, as natural products for the control of pests in eco-agrosystems.

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This study aimed to investigate the free radicals scavenging, inhibition of acetylhydrolase and antimicrobial activity of five mixtures of hydrolates obtained by hydrodistillation of Lamiaceae plants, Rosmarinus officinalis, Salvia officinalis and Lavandula angustifolia. Chemical composition of hydrolates was analysed by gas chromatography coupled with mass spectrometry for volatiles and UV-VIS spectrophotometry for total phenolic and flavonoid content. The results of free radicals scavenging and acetylhydrolase inhibition activity have shown improved effect of selected hydrolates mixtures, compared to individual hydrolates. The effect was in accordance to their polyphenolic content, indicating synergistic action of the combined biologic active compounds from different extracts of rosemary, sage and lavender. In turn, the antimicrobial activity of hydrolates mixtures was similar to that of individual hydrolates in Staphylococcus aureus, Pseudomonas aeruginosa and Rhizoctonia solani cultures, revealing no synergistic effect of volatiles, as mainly responsible compounds. In conclusion, selected mixtures of aromatic plant hydrolates can provide added value to these by-products through further development of natural pesticide and/or fungicide products useful for the ecological crop protection.

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The aim of this study was to investigate the melissopalynology, physicochemical characteristics, antioxidant and antibacterial activity of seven honey samples harvested from different geographical regions and climates of Romania. The melissopalynological analysis revealed that monofloral and multifloral samples contained a wide diversity of minor pollen types from Romanian flora. The moisture, pH and free acidity values were within international limit. HPLC analysis indicated high content of fructose and glucose and low content of sucrose. Bioactive compounds including proteins, phenolics, flavonoids and ascorbic acid were present in variable quantities, according to the botanical origin and geographical area. The highest phenolics and ascorbic acid content was in multifloral honeys from Crisana mountain and meadow and the extrafloral honeydew honey. The same honey samples have exerted free radical scavenging and antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa. The free radical scavenging activity was strongly correlated to phenolics and ascorbic acid content, while the antimicrobial activity was medium correlated only to phenolics content. In conclusion, the selected Romanian honey samples with best antioxidant and antimicrobial activity could be further tested for the development of novel biomedical products.

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Research background. Various protocols for enzymatic hydrolysis of fish by-products are increasingly tested to ensure value-added products with functional and biological properties important for food, cosmetic and medical applications. In addition, they attempt to minimize waste from industrial processing and environmental requirements. This study aims to establish an efficient protocol based on two-step enzymatic hydrolysis of freshwater fish skin and to evaluate the effect of resulting bioactive peptides on free radical scavenging, redox balance and regulation of fibroblast proliferation and migration. Experimental approach. Pepsin-soluble collagen extracted from silver carp (Hypophthalmichthys molitrix) skin was hydrolyzed by proteinase K at specific sites under controlled conditions. The molecular mass of ultrafiltration permeate was determined by gradient electrophoresis and gel filtration chromatography. The biological activity of intermediate and small size bioactive peptides was evaluated in experimental models in vitro mimicking oxidative stress and skin wound conditions. Results and conclusions. Extracted fish collagen was hydrolysed using proteinase K, the most efficient enzyme for the cleavage of the primary structure of the molecule, as previously found in silico. Established optimal conditions increased the enzyme specificity and the process yield. Bioactive peptides exerted significantly higher scavenging activity on free stable radicals and hydroxyl radicals often found in vivo, compared to fish collagen. They stimulated fibroblast metabolism in a dose-dependent manner and up-regulated cell migration in a scratch wound model. Pretreatment of fibroblasts with induced oxidative stress using optimal concentrations of fish peptides prevented the increase of reactive oxygen species production. In conclusion, bioactive peptides from carp skin demonstrated valuable properties of maintaining redox balance and skin wound healing process improvement, which indicated further potential applications in the development of pharmaceutical and nutraceutical formulations. Novelty and scientific contribution. In this study the enzymatic hydrolysis was applied to isolated protein, in contrast to previous studies using waste tissue with variable composition. Recovered bioactive peptides acted not only as antioxidant agents, but also as regulators of oxidative stress and wound healing processes in skin cell models. Their nutritional and cosmetic application is recommended in novel formulations fighting skin ageing phenomena.

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The aim of this study was to obtain biocomposites consisting of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), bacterial cellulose (BC) and alpha-tocopherol by a melt processing technique for potential use in biomedical applications. The melt processing and roughness of biocomposites were evaluated and compared to sample without BC. The degradation rate of PHBV/BC biocomposites was measured in phosphate buffer saline (PBS) by determining the mass variation and evidencing of thermal and structural changes by differential scanning calorimetry (DSC) and attenuated total reflectance-Fourier transformed infrared spectrometry (ATR-FTIR). The cell viability, cell morphology, cell cycle distribution and total collagen content were investigated on murine NCTC fibroblasts. Overall, the adding of BC to polyester matrix led to an adequate melt processing of biocomposites and increased surface roughness and cytocompatibility, allowing the cells to secrete the extracellular matrix (collagen) and stimulate cell proliferation. Results showed that the PHBV/BC biocomposites were favorable for long-term degradation and could be used for the design of medical devices with controlled degradability.

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Finding innovative solutions to improve the lives of people affected by trauma, bone disease, or aging continues to be a challenge worldwide. Tissue engineering is the most rapidly growing area in the domain of biomaterials. Cerium-containing MBG-derived biomaterials scaffolds were synthesized using polymethyl methacrylate (PMMA) as a sacrificial template. The obtained scaffolds were characterized by X-ray powder diffraction (XRPD), infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The Ce4+/Ce3+ ratio in the scaffolds was estimated. In vitro testing revealed good cytocompatibility of the investigated scaffolds in mouse fibroblast cell line (NCTC clone L929). The results obtained regarding bioactivity, antibacterial activity, and controlled drug delivery functions recommend these scaffolds as potential candidates for bone tissue engineering applications.

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Corrosion protective coatings were grown on pure magnesium in naturally carbonated mineral water containing Ca2+ using a simple, green conversion method. Dissolution of the native corrosion product, Mg(OH)2, mediates the surface nucleation of hydroxycarbonates. The pH increase due to CO2 degassing to atmospheric pressure leads to calcium carbonate nucleation and lateral growth by the incorporation of nanoparticles in a continuous layer of calcite -Ca(Mg)CO3- microcrystals. The coated surfaces have a much improved corrosion resistance in physiological fluids, measured by EIS, weight loss, corrosion rates and hydrogen release. Cell viability/ morphology assessment demonstrates that the coating is non-toxic and promotes the proliferation of osteoblastic cells.

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A red-emitting fluorescent Riboflavin (RF)/Polyvinylpyrrolidone (PVP)-coated silver nanoparticles system, lambda(em) = 527 nm, phi = 0.242, with a diameter of the metallic core of 27.33 nm and a zeta potential of - 25.05 mV was prepared and investigated regarding its biological activity. We found that PVP has a key role in RF adsorption around the SNPs surface leading to an enhancement of antioxidant properties (similar to 70%), low cytotoxicity (> 90% cell viability, at 50 mu L/mL, after 48 h of incubation) as well as to an efficient process of its cellular uptake (similar to 60%, after 24 h of incubation) in L929 cells. The results are relevant concerning the involvement of RF and its coenzymes forms in SNPs - based systems, in cellular respiration as well as for future studies as antioxidant marker system on tumoral cells for viewing and monitoring them, by cellular imaging.

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A facile, green synthesis methodology to obtain zinc oxide nanoparticles using three polysaccharide gums (Acacia gum, Guar gum and Xanthan gum) of biological origin was developed. Subsequently, biosynthesized zinc oxide nanoparticles were incorporated into a sustainable chitosan hydrogel matrix functionalized with propolis extract. This study has revealed that the selected polysaccharides as chelates represents a suitable approach to synthesize ZnO nanoparticles of particular interest with controlled morphology. The formation of ZnO nanoparticles using polysaccharide gums was confirmed by FTIR, XRD, UV-Vis spectroscopy, thermal analysis, SEM, Raman and photoluminescence spectroscopies. The rheological behaviour of obtained hydrogels was evaluated. The AFM studies demonstrate that all synthesized chitosan incorporated ZnO composites hydrogels functionalized with propolis extract exhibit corrugated topographies. The present study highlights the possible incorporation of various guest molecules into hydrogel matrix due to its tuneable morphologies. The obtained hydrogel composites were cytocompatible in L929 fibroblast cell culture, in a range of concentrations between 50 and 1000 mu g/mL, as assessed by MTT, LDH and Live/Dead double staining assays. By enhancing the biological properties, these novel green hydrogels show attractive superior performance in a wide concentration range to develop future in vivo suitable natural platforms as effective delivery systems of pharmacologic agents for biomedical applications.

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The marine algal ecosystem is characterized by a rich ecological biodiversity and can be considered as an unexploited resource for the discovery and isolation of novel bioactive compounds. In recent years, marine macroalgae have begun to be explored for their valuable composition in bioactive compounds and opportunity to obtain different nutraceuticals. In comparison with their terrestrial counterparts, Black Sea macroalgae are potentially good sources of bioactive compounds with specific and unique biological activities, insufficiently used. Macroalgae present in different marine environments contain several biologically active metabolites, including polysaccharides, oligosaccharides, polyunsaturated fatty acids, sterols, proteins polyphenols, carotenoids, vitamins, and minerals. As a result, they have received huge interest given their promising potentialities in supporting antitumoral, antimicrobial, anti-inflammatory, immunomodulatory, antiangiogenic, antidiabetic, and neuroprotective properties. An additional advantage of ulvans, fucoidans and carrageenans is the biocompatibility and limited or no toxicity. This therapeutic potential is a great natural treasure to be exploited for the development of novel drug delivery systems in both preventive and therapeutic approaches. This overview aims to provide an insight into current knowledge focused on specific bioactive compounds, which represent each class of macroalgae e.g., ulvans, fucoidans and carrageenans, respectively, as valuable potential players in the development of innovative drug delivery systems.

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Lately, in the world of medicine, the use of polymers for the development of innovative therapies seems to be a major concern among researchers. In our case, as a continuation of the research that has been developed so far regarding obtaining new isoniazid (INH) derivatives for tuberculosis treatment, this work aimed to test the ability of the encapsulation method to reduce the toxicity of the drug, isoniazid and its new derivatives. To achieve this goal, the following methods were applied: a structural confirmation of isoniazid derivatives using LC-HRMS/MS; the obtaining of microparticles based on polymeric support; the determination of their loading and biodegradation capacities; in vitro biocompatibility using MTT cell viability assays; and, last but not least, in vivo toxicological screening for the determination of chronic toxicity in laboratory mice, including the performance of a histopathological study and testing for liver enzymes. The results showed a significant reduction in tissue alterations, the disappearance of cell necrosis and microvesicular steatosis areas and lower values of the liver enzymes TGO, TGP and alkaline phosphatase when using encapsulated forms of drugs. In conclusion, the encapsulation of INH and INH derivatives with chitosan had beneficial effects, suggesting a reduction in hepatotoxicity and, therefore, the achievement of the aim of this paper.

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Natural compounds are an important source of beneficial components that could be used in cancer therapy along with well-known cytostatic agents to enhance the therapeutic effect while targeting tumoral tissues. Therefore, nanoplatforms containing mesoporous silica and a natural polysaccharide, ulvan, extracted from Ulva Lactuca seaweed, were developed for irinotecan. Either mesoporous silica-ulvan nanoplatforms or irinotecan-loaded materials were structurally and morphologically characterized. In vitro drug release experiments in phosphate buffer solution with a pH of 7.6 emphasized the complete recovery of irinotecan in 8 h. Slower kinetics were obtained for the nanoplatforms with a higher amount of natural polysaccharide. Ulvan extract proved to be biocompatible up to 2 mg/mL on fibroblasts L929 cell line. The irinotecan-loaded nanoplatforms exhibited better anticancer activity than that of the drug alone on human colorectal adenocarcinoma cells (HT-29), reducing their viability to 60% after 24 h. Moreover, the cell cycle analysis proved that the irinotecan loading onto developed nanoplatforms caused an increase in the cell number trapped at G0/G1 phase and influenced the development of the tumoral cells.

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The aim of this study was to establish the best ultrasound assisted extraction (UAE) conditions of saponins from Hedera helix L. leaves and to evaluate the in vitro biocompatibility of the extracts richest in saponins. Different parameters, such as extraction time, temperature, ultrasound power, solvent to plant material ratio, and solvent concentration, were investigated. The most efficient extraction conditions were a temperature of 50 degrees C, an ultrasound amplitude of 40%, an extraction time of 60 min, a plant material to solvent ratio of 1:20 (w:v), and 80% ethanol as solvent. In vitro cytotoxicity of the extracts richest in saponins and their influence on the DNA content of L929 (NCTC) fibroblasts were tested. Until 200 mu g/mL, the studied extracts were cytocompatible with L929 fibroblast cell lines at 48 h of treatment. These in vitro cell culture results provide useful information for further applications of Hedera helix extracts in a pharmaceutical field.

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Background and Objectives: The dental pulp stem cells are highly proliferative and can differentiate into various cell types, including endothelial cells. We aimed to evaluate the ultrastructural characteristics of the human dental pulp cells of the permanent frontal teeth. Materials and Methods: Human adult bioptic dental pulp was collected from n = 10 healthy frontal teeth of five adult patients, prior to prosthetic treatments for aesthetic purposes. Tissues were examined under transmission electron microscopy. Results: We identified cells with a peculiar trait: giant nucleoli resembling intranuclear endoplasmic reticulum, which mimicked extrusion towards the cytoplasm. These were either partly embedded within the nuclei, the case in which their adnuclear side was coated by marginal heterochromatin and the abnuclear side was coated by a thin rim of ribosomes, or were apparently isolated from the nuclei, while still being covered by ribosomes. Conclusions: Similar electron microscopy features were previously reported in the human endometrium, as nucleolar channel system; or R-Rings induced by Nopp140. To our knowledge, this is the first report of extruded nucleolar structure in the dental pulp. Moreover, the aspect of giant extruded nucleoli was not previously reported in any human cell type, although similar evidence was gathered in other species as well as in plants.

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Three feather mite species of the genus Brephosceles Hull, 1934 (Alloptidae: Alloptinae) were found on the European storm petrel, Hydrobates pelagicus (Procellariiformes: Hydrobatidae), in the Mediterranean Sea and North-east Atlantic Ocean. Brephosceles holoplax sp. n. is described as a new species; B. pelagicus (Vitzthum, 1921) and B. longirostris (Vitzthum, 1921) comb. n. formerly known only from females are redescribed from both sexes. The latter species has been transferred from the genus Alloptes Canestrini, 1879. All descriptions are supported by DNA barcode data.

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Over the past years, research attention has been focusing more on waste-derived, naturally derived, and renewable materials, in the view of a more sustainable economy. In this work, different topical formulations were obtained from the valorization of marine and agro-industrial by-products and the use of Carbopol 940 as gelling agent. In particular, the combination of extracts obtained from the marine snail, Rapanosa venosa, with Cladophora vagabunda and grape pomace extracts, was investigated for wound healing purposes. Rapana venosa has demonstrated wound healing properties and antioxidant activity. Similarly, grape pomace extracts have been shown to accelerate the healing process. However, their synergic use has not been explored yet. To this aim, four different formulations were produced. Three formulations differed for the presence of a different extract of Rapana venosa: marine collagen, marine gelatin, and collagen hydrolysate, while another formulation used mammalian gelatin as further control. Physico-chemical properties of the extracts as well as of the formulations were analyzed. Furthermore, thermal stability was evaluated by thermogravimetric analysis. Antioxidant capacity and biological behavior, in terms of cytocompatibility, wound healing, and antimicrobial potential, were assessed. The results highlighted for all the formulations (i) a good conservation and thermal stability in time, (ii) a neutralizing activity against free radicals, (iii) and high degree of cytocompatibility and tissue regeneration potential. In particular, collagen, gelatin, and collagen hydrolysate obtained from the Rapana venosa marine snail represent an important, valuable alternative to mammalian products.

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A new series of pyrazolo-benzimidazole hybrid Mannich bases were synthesized, characterized by H-1-NMR, C-13-NMR, IR, UV-Vis, MS, and elemental analysis. In vitro cytotoxicity of the new compounds studied on fibroblast cells showed that the newly synthesized pyrazolo-benzimidazole hybrid derivatives were noncytotoxic until the concentration of 1 mu M and two compounds presented a high degree of biocompatibility. The antibacterial and antibiofilm activity of the newly synthesized compounds was assayed on Gram-positive Staphylococcus aureus ATCC25923, Enterococcus faecalis ATCC29212, and Gram-negative Pseudomonas aeruginosa ATCC27853, Escherichia coli ATCC25922 strains. All synthesized compounds 5a-g are more active against all three tested bacterial strains Staphylococcus aureus ATCC25923, Enterococcus faecalis ATCC29212, and Escherichia coli ATCC25922 than reference drugs (Metronidazole, Nitrofurantoin), with the exception of compounds 5d and 5g, which are less active compared to Nitrofurantoin, and all synthesized compounds 5a-g are more active against Pseudomonas aeruginosa ATCC27853 compared to reference drugs (Metronidazole, Nitrofurantoin). Compound 5f showed the best activity against Staphylococcus aureus ATCC 25923, with a MIC of 150 mu g/mL and has also inhibited the biofilm formed by all the bacterial strains, having an MBIC of 310 mu g/mL compared to the reference drugs (Metronidazole, Nitrofurantoin).

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Bioactive collagen-chitosan-lemongrass (COL-CS-LG) membranes were prepared by casting method and analyzed for potential biomedical applications. For COL-CS-LG membranes, LG essential oil release, antioxidant properties, in vitro cytotoxicity and antimicrobial assessments were conducted, as well as free radical determination after gamma irradiation by chemiluminescence, and structural characteristics analysis through Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy (ATR-FTIR) and Differential Scanning Calorimetry (DSC). The evaluation of non-isothermal chemiluminescence after gamma radiation exposure to COL-CS-LG membranes revealed a slowing down of the oxidation process at temperatures exceeding 200 degrees C, in correlation with antioxidant activity. Antimicrobial properties and minimum inhibitory concentrations were found to be in correlation with cytotoxicity limits, offering the optimum composition for designing new biomaterials.

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Background: New materials are currently designed for efficient treatment of oral tissue lesions by guided tissue regeneration. The aim of this study was to develop a multifunctional 3D hybrid biomaterial consisting of extracellular matrix components, collagen, chondroitin 4-sulfate and fibronectin, functionalised with silver nanoparticles, intended to improve periodontitis treatment protocols. Methods: Structural observations were performed by autometallography, scanning and transmission electron microscopy. In vitro tests of 3D constructs of embedded gingival fibroblasts within hybrid biomaterial were performed by MTS and Live/Dead assays. Genotoxicity was assessed by comet assay. In vivo experiments using chick embryo chorioallantoic membrane (CAM) assay analysed the degradation and nanoparticles release, but also angiogenesis, new tissue formation in 3D constructs and the regenerative potential of the hybrid material. Biological activity was investigated in experimental models of inflamed THP-1 macrophages and oral specific bacterial cultures. Results: Light micrographs showed distribution of silver nanoparticles on collagen fibrils. Scanning electron micrographs revealed a microstructure with interconnected pores, which favoured cell adhesion and infiltration. Cell viability and proliferation were significantly higher within the 3D hybrid biomaterial than in 2D culture conditions, while absence of the hybrid material?s genotoxic effect was found. In vivo experiments showed that the hybrid material was colonised by cells and blood vessels, initiating synthesis of new extracellular matrix. Besides the known effect of chondroitin sulfate, incorporated silver nanoparticles increased the antiinflammatory activity of the hybrid biomaterial. The silver nanoparticles maintained their antibacterial activity even after embedding in the polymeric scaffold and inhibited the growth of F. nucleatum and P. gingivalis. Conclusion: The novel biomimetic scaffold functionalised with silver nanoparticles presented regenerative, antiinflammatory and antimicrobial potential for oral cavity lesions repair.

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Tomato peels are used as a valuable material to extract lycopene-rich oleoresins by supercritical CO2 extraction. The extraction involves continuous circling of CO2 to the extractor after removing the solute in the separators, S40 and S45, where the solvent power of the CO2 is reduced by reducing pressure down to 20 MPa in S40 and 5 MPa in S45, respectively, leading to two extracts. Lycopene is found to be the major compound, representing 93% and 76% of the total carotenoids in S40 and S45 extracts, respectively. The two extracts are microencapsulated in whey protein concentrate and acacia gum by complex coacervation and freeze-drying, leading to corresponding P40 and P45 powders, with antioxidant activity of 8.57 +/- 0.74 and 9.37 +/- 0.48 mMol TEAC/g DW in P40 and P45, respectively. Different structural and morphological patterns are observed, with finer microparticles of 1-2 mu m in P45. Both powders show dose and time-dependent antiproliferative activity. The half-maximal inhibitory concentration values are 100 mu g/mL for P40 and 750 mu g/mL for P45 sample, indicating a higher antiproliferative effect of P40 over P45 in HT-29 cell culture. The powders have an extended range of cytocompatibility, up to 1000 mu g/mL, in L929 normal cells, stimulating the cell growth. Lycopene retention is tested, and values of 48% and 29% in P40 and P45 are found after 21 days at 25 degrees C, with the degradation rate in P45 significantly higher, due to the higher content of the surface lycopene, which favored its degradation.

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Significant quantities of onion are cultivated annually, such that industrial processing leads to an appreciable amount of by-products, estimated at around 500,000 tons. Onion skins are considered an important source of naturally occurring antioxidant compounds, particularly flavonoid compounds. Our study follows the development of a sustainable solution in order to manage the by-products of yellow onion skins by designing ingredients with multifunctional activities. A green solvent aqueous extraction of flavonoids was applied to obtain a safe, flavonoid-enriched extract, yielding a total flavonoid content of 50.21 +/- 0.09 mg quercetin equivalent (QE)/g dry weight (DW), and an antioxidant activity of 250.81 +/- 6.76 mM Trolox/g DW. Complex biopolymeric matrices consisting of whey protein isolates, whey protein hydrolysates, maltodextrin, and pectin were further dissolved in the flavonoid-enriched aqueous extract, followed by freeze-drying. Two powders were obtained, both showing satisfactory phytochemical content and good stability during storage. The application of confocal microscopy revealed that the microscopic structure of the powders have a distribution of the bioactive compounds within the biopolymeric matrices. The in vitro digestion suggested remarkable stability in the gastric tract and a flavonoid-controlled release in the intestinal phase. A significant compatibility range of up to 1 mg/mL for both powders was found, whereas concentrations between 10 and 250 mu g/mL stimulated cell proliferation after 24 h of cultivation. The powders showed satisfactory thermal and pH stability, which favors their addition to different food matrices.

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Severe skin injuries, including burns, represent a real concern for the global health-care system and therefore, there is an increased interest in developing wound dressings, in order to stimulate and enhance skin tissue repair. The aim of this study was to design novel hybrid materials, biomimetic to skin extracellular matrix and enriched with silver nanoparticles (nAg), in order to provide both dermal tissue regeneration and antimicrobial activity. Two material variants (variant A and variant B) consisting of type I collagen (COL), chondroitin sulfate (CS) and k-elastin peptides (EL) enriched with positively-charged nAg, were conditioned as membranes. UV exposure ensured both sterilisation and cross-linking of the materials. Physico-chemical characterization of the hybrid biomaterials showed values of density and swelling degree higher than those of COL membrane, while the process of in vitro degradation followed a similar pattern. Infrared spectroscopy and x-ray diffraction indicated alterations of the characteristic structural features and crystallinity of COL after blending with CS and EL and nAg embedding. Scanning electron microscopy observations revealed different surface morphologies of the hybrid membranes, according to their composition. In vitro studies on L929 fibroblasts and HaCaT keratinocytes showed that both hybrid membranes exhibited good cytocompatibility and promoted higher cell proliferation compared to COL sample, as evaluated by MTT and Live/Dead assays. The presence of actin filaments highlighted by fluorescent labelling confirmed the fibroblast and keratinocyte adhesion onto the surface of hybrid membranes. Most importantly, both materials showed an increased wound healing ability in an in vitro scratch assay model, stimulating cell migration at 24 h post-seeding. In addition, good antimicrobial activity was recorded, especially against Gram-positive bacterial strain. Altogether, our findings recommend COL-CS-EL-nAg hybrid membranes as good candidates for wound healing acceleration and bioengineering of skin tissue.

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Osteoarthritis (OA) is a degenerative joint disease. An objective of the nanomedicine and drug delivery systems field is to design suitable pharmaceutical nanocarriers with controllable properties for drug delivery and site-specific targeting, in order to achieve greater efficacy and minimal toxicity, compared to the conventional drugs. The aim of this review is to present recent data on natural bioactive compounds with anti-inflammatory properties and efficacy in the treatment of OA, their formulation in lipid nanostructured carriers, mainly liposomes, as controlled release systems and the possibility to be intra-articularly (IA) administered. The literature regarding glycosaminoglycans, proteins, polyphenols and their ability to modify the cell response and mechanisms of action in different models of inflammation are reviewed. The advantages and limits of using lipid nanoformulations as drug delivery systems in OA treatment and the suitable route of administration are also discussed. Liposomes containing glycosaminoglycans presented good biocompatibility, lack of immune system activation, targeted delivery of bioactive compounds to the site of action, protection and efficiency of the encapsulated material, and prolonged duration of action, being highly recommended as controlled delivery systems in OA therapy through IA administration. Lipid nanoformulations of polyphenols were tested both in vivo and in vitro models that mimic OA conditions after IA or other routes of administration, recommending their clinical application.

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The present study describes a comprehensive investigation of the spectroscopic characteristics, stability and in vitro antioxidant and cytotoxic properties of the Flavin MonoNucleotide (FMN) and Flavin Adenine Dinucleotide (FAD) in Dextran70 (Dx70) and Dx70/phospatidylcholine (PC) biomimetic systems by means of the UV-Vis absorption, fluorescence spectroscopy, chemiluminescence and Neutral Red assay. The affinity of FMN, FAD and the precursor riboflavin (RF) to an unsaturated phospholipid bilayer model as well as the location of the probes within the lipid bilayer were assessed from united-atom molecular dynamics simulations carried out on an unsaturated phospholipid bilayer model system, and the theoretical and experimental characterization of the two probes within biomembranes was complemented with the light microscopy survey of the cell morphology of L929 fibroblast cells cultivated in the presence of various dosage of FAD/FMN. In lipid bilayers, FMN/FAD resulted in a noticeable improvement of the antioxidant activity (the scavenging of reactive oxygen species up to 40%) and a significant effect on cellular viability in the L929 fibroblast cells. The results are important in the oxidative stress process concerning the redox reactions of flavins in humans as well as in further studies on different systems belonging to the category of flavoenzymes/flavoproteins, required for cellular respiration. (C) 2020 Elsevier B.V. All rights reserved.

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The paper presents some aspects regarding the obtaining of new ecological fertilizers for organic and classical agriculture by exploiting valuable residual bioresources, such as marine biomass and waste from viticulture and secondary vinification process. Biomass composed of marine green and brown algae, invertebrate shells, fermented and unfermented grape marc and chopped woody shoots of vines were combined in different proportions, resulting in eight experimental variants of fertilizer, which were incorporated into steppe chernozemic mollisols, specific to the Murfatlar area of Constan.a County, Romania. In 2019, in the proposed mixtures of soil - residual biomass, under greenhouse conditions, different types of vegetal species were cultivated, namely, Vitis vinifera L. (Pinot Gris variety), Lolium perene L., Sinapis alba L., and Trifolium repens L. The comparative statistical interpretation of the obtained results indicated a significant increase of essential nutrients ( phosphorus and potassium), classifying the fertilized soils into the category of those with very good nutrients supply. An improvement of the soil organic matter (humus) was observed, the increase compared to the control being of 40.9% in the case of the V5 variant. A moderate increase of the 3% CaCO3 content was also observed. Considering the experimental results obtained under greenhouse conditions, we will continue the study on experimental lots cultivated with different white and black varieties of Vitis vinifera L, representative for the Murfatlar vineyard.

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In our study a Hedera helix L. leaves extract was fractioned by flash chromatography. The extract and its nine fractions were tested for biocompatibility on NCTC normal fibroblasts, and antiproliferative activity on Hep-2 epithelial tumor cells. In vitro tests revealed that the samples were biocompatible on NCTC cells up to 200 mu g/mL. Fractions 7 and 8 manifested strong antiproliferative effect on Hep-2 cells, and were analyzed versus ivy extract to determine the content in phytochemical constituents and antioxidant activity. The obtained results highlighted that the high content of saponins is responsible for the fractions 7 and 8 antiproliferative activity.

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In recent years, the rising number of bone diseases which affect millions of people worldwide has led to an increased demand for materials with restoring and augmentation properties that can be used in therapies for bone pathologies. In this work, PMMA- MBG composite scaffolds containing ceria (0, 1, 3 mol%) were obtained by the phase separation method. The obtained composite scaffolds were characterized by X-ray diffraction, infrared spectroscopy, and scanning electron microscopy. UV-Vis measurement and EDX analysis confirmed the presence of cerium ions in the composite scaffolds. Evaluation of the in-vitro biocompatibility using MTT assay showed that composite scaffold containing 1 mol% of ceria presented higher viability than control cells (100%) for concentrations ranging between 5 and 50% after 96 h of incubation.

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The aim of this paper was to evaluate the total phenolic content and comparative antioxidant activity by three methods, photo chemiluminescence FRAP, DPPH of some grape pomace polyphenolic extracts from white and red varieties of Vitif vinifera (L)., obtained by maceration for 12 days, at room temperature in 40%, 70% aqueous ethanol solution or ethanol. The grape pomace was obtained after the secondary winemaking process of five grape varieties Vitis vinifera (L.) for white wines, Sauvignon Blanc, Muscat Ottonel, and red wines, Feteasca Neagra, Cabernet Sauvignon and Mamaia, created at Research Station for Viticulture and Oenology of Murfatlar, Constanta County, Romania. The results emphasize that an increased phenolic content and antioxidant capacity were obtained for the white varieties, Muscat Ottonel and for the red ones, Cabernet Sauvignon. Regarding the extraction efficiency, for all analyzed grape pomace 70% hydroalcoholic extracts present an increased activity. Based on our preliminary results regarding the radical scavenger activity of grape marc hydroalcoholic extracts, we can consider these by product as a promising resource for dermato-cosmetics with anti-aging effect.

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In the present study, we evaluated for the first time the photoprotective effect of fish bone bioactive peptides (FBBP) preparation isolated from silver carp (Hypophthalmichthys molitrix) discarded tissue using in vitro experimental models of skin cells exposed to ultraviolet B (UVB) irradiation and stressing agents. FBBP preparation was obtained by papain treatment of minced bones and centrifugal ultrafiltration, and the molecular weight (MW) distribution was characterized by size exclusion and reversed-phase high performance liquid chromatography (RP-HPLC). In vitro assessment of the effect of FBBP pretreatment in UVB-irradiated L929 fibroblasts and HaCaT keratinocytes revealed their cytoprotective activity. Their capacity to efficiently reduce reactive oxygen species (ROS) production and lipid peroxidation varied in a dose-dependent manner, and it was greater in fibroblasts. A decrease of proinflammatory cytokines secretion, in particular of tumor necrosis factor alpha (TNF-alpha), was found after FBBP pretreatment of THP-1-derived inflamed macrophages. Melanin production and tyrosinase activity investigated in UVB-irradiated Mel-Juso cells were lowered in direct relation to FBBP concentrations. FBBP fractions with high radical scavenging activity were separated by ion exchange chromatography, and two collagenic sequences were identified. All these results offer new scientific data on aquaculture fish bone-derived peptides confirming their ability to control the antioxidant, anti-inflammatory and pigmentation processes developed during UV irradiation of skin cells and recommend their use as valuable natural ingredients of photoprotective cosmeceutical products.

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Current advances in computational modelling and simulation have led to the inclusion of computer scientists as partners in the process of engineering of new nanomaterials and nanodevices. This trend is now, more than ever, visible in the field of deoxyribonucleic acid (DNA)-based nanotechnology, as DNA's intrinsic principle of self-assembly has been proven to be highly algorithmic and programmable. As a raw material, DNA is a rather unremarkable fabric. However, as a way to achieve patterns, dynamic behavior, or nano-shape reconstruction, DNA has been proven to be one of the most functional nanomaterials. It would thus be of great potential to pair up DNA's highly functional assembly characteristics with the mechanic properties of other well-known bio-nanomaterials, such as graphene, cellulos, or fibroin. In the current study, we perform projections regarding the structural properties of a fibril mesh (or filter) for which assembly would be guided by the controlled aggregation of DNA scaffold subunits. The formation of such a 2D fibril mesh structure is ensured by the mechanistic assembly properties borrowed from the DNA assembly apparatus. For generating inexpensive pre-experimental assessments regarding the efficiency of various assembly strategies, we introduced in this study a computational model for the simulation of fibril mesh assembly dynamical systems. Our approach was based on providing solutions towards two main circumstances. First, we created a functional computational model that is restrictive enough to be able to numerically simulate the controlled aggregation of up to 1000s of elementary fibril elements yet rich enough to provide actionable insides on the structural characteristics for the generated assembly. Second, we used the provided numerical model in order to generate projections regarding effective ways of manipulating one of the the key structural properties of such generated filters, namely the average size of the openings (gaps) within these meshes, also known as the filter's aperture. This work is a continuation of Amarioarei et al., 2018, where a preliminary version of this research was discussed.

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Acid-soluble, undenatured, type I collagen (BSC) isolated, for the first time, from gilthead bream skin and the novel fabricated 3D porous wound dressing were analyzed for physicochemical and biological properties, in order to offer a safe alternative to commercial bovine collagen (BC) products. SDS-polyacrylamide analysis confirmed the purity of BSC preparation. The hydroxyproline content and temperature of denaturation of BSC were lower than those of BC, in accordance with the structural data recorded by FT-IR spectroscopy. However, certain concentrations of BSC stimulated the cell metabolism of L929 fibroblasts in a higher proportion than BC. The 3D wound dressing presented high porosity and low surface hydrophobicity that could help cell attachment and growth. The rapid biodegradation of BSC wound dressing could explain the improved in vitro cell migration and wound closure rate. In conclusion, the skin of gilthead bream from the Black Sea coast represented a valuable source for the biomedical industry, providing biocompatible, biodegradable collagen and 3D porous wound dressing, as novel material with enhanced wound healing activity.

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In this study, sodium alginate film (Alg) was coated with electrospun collagen glue (Col) extracted from rabbit skin waste, loaded with different commercial antimicrobial agents (chitosan, AG425K and ZnONPs) and investigated in terms of morphological, structural and biological properties. The coated nanostructures were characterized using scanning electron microscopy coupled with the energy-dispersive X-ray (SEM/EDS), Attenuated Total Reflectance Fourier-Transform Infrared spectroscopy (ATR FT-IR), and Atomic Force Microscopy (AFM) tests. The cytotoxicity was investigated on murine L929 fibroblasts using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide salt (MTT) and lactate dehydrogenase (LDH) assays. Microbiological tests were performed against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and Candida albicans ATCC 27853 standard strains. In vitro cell culture tests showed a good cytocompatibility of the coated nanostructured systems, except the sample loaded with ZnONPs, which exhibited a highly cytotoxic effect. Alg-Col-ZnONPs nanostructure inhibited the growth and multiplication of the Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 bacterial strains. The results of new coated nanostructures may be useful for the development of sustainable biomaterials in a circular economy, with bioactive properties for medical wound dressings.

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In this study, flavonoids extracted from yellow onion skins and Lactobacillus casei were encapsulated in a combination of whey protein isolate, inulin and maltodextrin with an encapsulation efficiency of 84.82 +/- 0.72% for flavonoids and 72.49 +/- 0.11% for lactic acid bacteria. The obtained powder showed a flavonoid content of 89.49 +/- 4.12 mg quercetin equivalents/g dry weight (DW) and an antioxidant activity of 39.27 +/- 0.45 mM Trolox/g DW. The powder presented a significant antidiabetic and anti-inflammatory potential, with an inhibitory effect on alpha-amylase, lipase and lipoxygenase of 76.40 +/- 2.30%, 82.58 +/- 3.36% and 49.01 +/- 0.62%, respectively. The results obtained for in vitro digestion showed that the coating materials have a protective effect on the flavonoids release. Cytotoxicity results indicated that the powder was cytocompatible up to a concentration of 500 mu g/mL. The functional potential of the powder was tested by adding in a selected food matrix, highlighting a good stability of the phytochemicals, whereas an increase with 1 log cell forming unit (CFU)/g DW was observed after 21 days of storage. The obtained results are promising in the valorization of natural antioxidants in combination with lactic acid bacteria in order to develop multifunctional ingredients with value-added for food and pharmaceutics applications.

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The main aim of this study was to transform bovine colostrum into a tribiotic product through a metabolic conversion with a microbial consortium based on artisanal and starter microorganisms. As such, the bovine colostrum was firstly transformed enzymatically for 48 h with a selected strain of a non-pathogenic Candida lipolytica strain, and then lactic acid fermentation was performed for 48 h, by using a co-culture of 25 g/L fresh kefir grains and a 10 g/L commercial FreshQ (R) culture (Chr. Hansen, Denmark). The obtained product is recommended to be used as an ingredient for the milk-derived nutraceuticals' formulation. As such, the product displayed increased antioxidant potential of 3.15 mM Trolox equivalent/g. Furthermore, the peptide fractions with a molecular weight lower than 3 kDa showed a notable in vitro ABTS radical scavenging activity, similar to a concentration of 2 nM captopril, hence proving a high potential in reducing the blood pressure.

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Bionanocomposites including poly(lactic acid) (PLA), collagen, and silver nanoparticles (AgNPs) were prepared as biocompatible and stable films. Thermal properties of the PLA-based bionanocomposites indicated an increase in the crystallinity of PLA plasticized due to a small quantity of AgNPs. The results on the stability study indicate the promising contribution of the AgNPs on the durability of PLA-based bionanocomposites. In vitro biocompatibility conducted on the mouse fibroblast cell line NCTC, clone 929, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed high values of cell viability (>80%) after cell cultivation in the presence of bionanocomposite formulations for 48 h, while the percentages of lactate dehydrogenase (LDH) released in the culture medium were reduced (<15%), indicating no damages of the cell membranes. In addition, cell cycle analysis assessed by flow cytometry indicated that all tested bionanocomposites did not affect cell proliferation and maintained the normal growth rate of cells. The obtained results recommend the potential use of PLA-based bionanocomposites for biomedical coatings.

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Fermented bovine colostrum rich in bioactive peptides was obtained using Candida lipolytica strains in co-culture with kefir grains. During fermentation, the pH and total titratable acidity of the product were daily measured. The hydrolysis degree and the electrophoretic pattern of water-soluble extracts (WSE) of fermented colostrum were analysed. Fractions enriched in peptides with a molecular weight lower than 10 kDa were separated by ultracentrifugation. They showed better modulation of angiotensin-converting enzyme (ACE) inhibition and cell proliferation, but lower radical scavenging capacity, compared to corresponding WSE.

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Lavender flowers were used in this study as a source of phytochemicals as naturally occurring antioxidants. Two different extraction techniques were applied, such as ultrasound-assisted (UAE) and supercritical fluids (SCE) methods. The comparative evaluation of the phytochemicals profile evidenced a higher content of chlorophyll a and b of 5.22 +/- 0.12 mg/g dry weight (D.W.) and 2.95 +/- 0.16 mg/g D.W, whereas the carotenoids content was 18.24 +/- 0.04 mg/g D.W. in the SCE extract. Seven main compounds were found in both extracts: beta-linalool, eucalyptol, linalool acetate, beta-trans-ocimene, and limonene in SCE and linalool acetate, beta-linalool, 6-methyl-2-(2-oxiranyl)-5-hepten-2-ol, linalool oxide, lavandulyl acetate and camphor in UAE. The (n-3) acids had a higher contribution in SCE. The extracts were microencapsulated in different combinations of wall materials based on polysaccharides and milk proteins. The four variants showed different phytochemical and morphological profiles, with a better encapsulating efficiency for proteins (up to 98%), but with a higher content of encapsulated carotenoids for polysaccharides, the latter showing remarkable antimicrobial activity against selected microorganisms. Carboxymethyl cellulose and whey proteins led to a double encapsulation of lipophilic compounds. The powders were tested in two food matrices as ingredients, with multiple targeted functions, such as flavoring, antimicrobial, antioxidant activity that can successfully replace synthetic additives.

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Phytochemicals are priceless sources of bioactive compounds with multiple health benefices. The main objective of the current investigation was to develop nanostructured herbal formulations conditioned as appropriate hydrogel (HG) conferring an enhanced transdermal absorption of bioactive compounds from selective extracts and vegetable oils. The direct impact of research is represented by the identification of prototype products which manifest an improved therapeutic response, by means of cumulative antioxidant, anti-inflammatory and anti acne actions, without causing any side health effects. The combinatorial effect of Carrot Extract (CE) and Marigold Extract (ME) Nanostructured Lipid Carriers (NLC) based on rosehip oil or black cumin oils was accompanied by a high biocompatibility and a significant ability to capture both short- and long-life free radicals. HG-NLC-ME-CE has been shown to be an efficient carrier with a differentiated potential for in vitro release of the two active principles, e.g. it delayed the release of carotenoids while the hydrophilic active (azelaic acid, AA) was faster released. The HG-NLC efficacy in skin inflammation treatment (demonstrated by in vitro and in vivo tests) revealed a reduced expression of inflammatory cytokines (IL-1 beta and TNF-alpha), more pronounced in the case of TNF-alpha. Moreover, a superior in vivo anti-inflammatory effect of HG-based NLC-CE/ME-AA as compared to that obtained for a commercial product was detected, i.e. after 3 h of HG-NLC treatment, a significant reduction of rat paw edema was quantified. In pre-clinical studies, the quantification of the hydration and elasticity effects in the viable epidermis provided the evidence of the high potential of developed prototypes, suitable for implementation in the market area. The degree of skin hydration and skin elasticity were remarkable enhanced after topical application of developed prototypes, a hydration effect up to 74% being determined and a skin elasticity reaching 90%. The knowledge acquired from this investigation could be utilized by the cosmetic industry to design novel topical products with improved quality and health benefices, endowed with antioxidant, anti-inflammatory and anti-acne actions and with desired hydration and elasticity profiles, in order to achieve better therapeutic efficacy and no drug toxicity.

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Two chitosan extracts were prepared by chemical and enzymatic treatment of Ganoderma lucidum mushroom, as an alternative source to crustacean shells. The molecular weight of the enzymatic extract was lower than that of the chemical one and of shrimp chitosan, as determined by viscosity measurements. Characteristic signals were identified in the H-1-NMR spectra and high deacetylation degree indicated good physico-chemical properties for both mushroom chitosan extracts. The scavenging capacity of mushroom chitosan extracts was moderate against the synthetic radicals of 2,2 '-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH), but higher values were observed for the enzymatic extract, compared to the chemical extract and shrimp chitosan. In vitro cytotoxicity was evaluated in L929 mouse fibroblast cell lines and the results of MTT assay showed good cytocompatibility in the tested range of concentrations. The growth of Gram-positive bacteria was inhibited more than Gram-negative bacteria in the presence of mushroom chitosan extracts, in particular by the chemical one, indicating their efficiency as antimicrobial agents. All these results strengthen the evidence of mushroom polysaccharide preparations availability for biomedical applications.

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This study focuses on combining different bioprocessing tools in order to develop an in-depth engineering approach for enhancing the biological properties of two valuable food by-products, namely fish waste and yellow onion skins, in a single new bioactive formulation. Bone tissue from phytophagous carp (Hypophthalmichthys molitrix) was used to obtain bioactive peptides through papain-assisted hydrolysis. The peptides with molecular weight lower than 3 kDa were characterized through MALDI-ToF/ToF mass spectrometry and bioinformatics tools. As a prerequisite for microencapsulation, the ability of these peptides to bind the flavonoids extracted from yellow onion skins was further tested through fluorescence quenching measurements. The results obtained demonstrate a considerable binding potency with a binding value of 10(6) and also the presence of one single or one class of binding site during the interaction process of flavonoids with peptides, in which the main forces involved are hydrogen bonds and van der Waals interactions. In the freeze-drying microencapsulation process, an efficiency for total flavonoids of 88.68 +/- 2.37% was obtained, considering the total flavonoids and total polyphenols from the powder of 75.72 +/- 2.58 quercetin equivalents/g dry weight (DW) and 97.32 +/- 2.80 gallic acid equivalents/g DW, respectively. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test on the L929 cell line cultivated in the presence of different concentrations of microencapsulated samples (0.05-1.5 mg/mL) proved no sign of cytotoxicity, the cell viability being over 80% for all the samples.

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The effects of two lyophilized extracts obtained from the aerial parts ofThymus marschallianusWilld. and harvested from wild flora (TMW) and obtained from culture (TMC) were evaluated in Wistar rats with experimentally induced hyperglycemia. The hyperglycemia was induced by streptozotocin (STZ) administration and the obtained results were evaluated in comparison for TMW and TMC. The polyphenolic composition of extracts was evaluated by spectrophotometrical and LC-MS methods.In vitroantioxidant capacity assays (DPPH, FRAP, EPR) were performed in order to preliminary establish the ability of tested samples to protect against free radical induced damage. Afterwards, the effects of these extracts were assessedin vivoon rats with experimental-induced hyperglycemia. Oxidative stress biomarkers (e.g. malondialdehyde-MDA), phosphorylated transcription factor subunit of nuclear kappaB (NF-kB) p65, methyl CpG binding protein (MECP) 2 and histone deacetylase 1 (HDAC1) expressions in hippocampus and frontal lobe were assessed. Open Field Test (OFT) and Elevated Plus Maze (EPM) were conducted on tested animals. Malondialdehyde (MDA) levels and HDAC1and MeCP2 expressions increased significantly in hippocampus (p<0.05) and frontal lobe (p<0.001) of diabetes group compared to the control group in parallel with decreasing of GSH/GSSG ratio. TMW and TMC administration reduced blood glucose levels and diminished lipid peroxidation, HDAC1 expression and enhanced antioxidant capacity in frontal lobe. TMW improved central locomotion of rats, increased phospho-NFkB p65 and diminished MECP2 expressions in hippocampus. Both tested samples exerted a beneficial effect by increasing the antioxidant defense. Our findings indicate that the administration of these extracts might represent a good option in the treatment of diabetes and its complications.

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The influence of spore concentration on the ability of a Trichoderma consortium to colonize the Passiflora caerulea phyllosphere was evaluated by determining the effects of foliar treatments with two spore concentrations, in two repeated treatments, on the morphological, physiological, and ultrastructural characteristics, and on the yield and quality of P. caerulea. The studied crop quality features were related to its nutraceutical use: the accumulation of polyphenols and flavonoids, antioxidant activity, and effects on mouse fibroblast L929 cells. The Trichoderma consortium consisted of two strains, T. asperellum T36b and T. harzianum Td50b, and the concentrations used were 10(6) colony forming units (cfu)/mL and 10(8) cfu/mL. As a reference treatment, a commercial product that was based on herbs and algal extracts was used. As compared to the negative control, the treatment with the Trichoderma consortium at 10(8) cfu/mL concentration determines the accumulation of higher level of polyphenols and flavonoids and increased antioxidant activity. This enhancement of P. caerulea quality characteristics after treatment with the higher concentration of Trichoderma consortium was associated with larger leaves, increased number and size of chloroplasts, improved plant physiology characteristics, and an increased yield. The treatment with high concentration of Trichoderma consortium spores promotes phyllosphere colonization and benefits both crop yield and quality.

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Concentrated collagen hydrolysate (HC10CC), rabbit collagen glue (RCG), and keratin hydrolysate (KH) were investigated in terms of their extraction from mammalian by-products and processing by electrospinning. The electrospun nanofibers were characterized by scanning electron microscopy coupled with the energy dispersive X-ray spectroscopy (SEM/EDS), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), differential scanning calorimetry (DSC), and indentation tests. The cytotoxicity of the electrospun nanofibers was conducted on L929 fibroblast cells using MTT and LDH assays and cell morphology observations. The electrospun RCG and KH nanofibers morphology showed an average size of nanofibers ranging between 44 and 410 nm, while the electrospun HC10CC nanofibers exhibited higher sizes. The ATR-FTIR spectra performed both on extracted proteins and electrospun nanofibers showed that the triple helix structure of collagen is partially preserved. The results were in agreement with the circular dichroism analysis for protein extracts. Furthermore, the viscoelastic properties of electrospun KH nanofibers were superior to those of electrospun RCG nanofibers. Based on both in vitro quantitative and qualitative analysis, the electrospun nanofibers were not cytotoxic, inducing a healthy cellular response. The results of new electrospun protein-based nanofibers may be useful for further research on bioactive properties of these nanofibers for tissue engineering.

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Angiogenesis is a physiological process involving the growth of new blood vessels, which provides oxygen and required nutrients for the development of various pathological conditions. In a tumor microenvironment, this process upregulates the growth and proliferation of tumor cells, thus any stage of angiogenesis can be a potential target for cancer therapies. In the present study, chitosan and his derivatives have been used to design novel polymer-based nanoparticles. The therapeutic potential of these newly designed nanoparticles has been evaluated. The antioxidant and MTT assays were performed to know the antioxidant properties and their biocompatibility. The in vivo antiangiogenic properties of the nanoparticles were evaluated by using a chick Chorioallantoic Membrane (CAM) model. The obtained results demonstrate that chitosan derivatives-based nanostructures strongly enhance the therapeutic effect compared to chitosan alone, which also correlates with antitumor activity, demonstrated by the in vitro MTT assay on human epithelial cervical Hep-2 tumor cells. This study opens up new direction for the use of the chitosan derivatives-based nanoparticles for designing of antiangiogenic nanostructured materials, for future cancer therapy. © 2020 by the authors.

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The main aim of this study was to transform bovine colostrum into a tribiotic product through a metabolic conversion with a microbial consortium based on artisanal and starter microorganisms. As such, the bovine colostrum was firstly transformed enzymatically for 48 h with a selected strain of a non-pathogenic Candida lipolytica strain, and then lactic acid fermentation was performed for 48 h, by using a co-culture of 25 g/L fresh kefir grains and a 10 g/L commercial FreshQ® culture (Chr. Hansen, Denmark). The obtained product is recommended to be used as an ingredient for the milk-derived nutraceuticals’ formulation. As such, the product displayed increased antioxidant potential of 3.15 mM Trolox equivalent/g. Furthermore, the peptide fractions with a molecular weight lower than 3 kDa showed a notable in vitro ABTS radical scavenging activity, similar to a concentration of 2 nM captopril, hence proving a high potential in reducing the blood pressure. © 2020, © 2020 The Author(s). Published with license by Taylor & Francis Group, LLC.

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Several therapeutic strategies are currently in development for severe periodontitis and other associated chronic inflammatory diseases. Guided tissue regeneration of the periodontium is based on surgical implantation of natural or synthetic polymers conditioned as membranes, injectable biomaterials (hydrogels), or three-dimensional (3D) matrices. Combinations of biomaterials with bioactive factors represent the next generation of regenerative strategy. Cell delivery strategy based on scaffold-cell constructs showed potential in periodontitis treatment. Bioengineering of periodontal tissues using cell sheets and genetically modified stem cells is currently proposed to complete existing (pre)clinical procedures for periodontal regeneration. 3D structures can be built using computer-assisted manufacturing technologies to improve the implant architecture effect on new tissue formation. The aim of this review was to summarize the advantages and drawbacks of biomimetic composite matrices used as biomaterials for periodontal tissue engineering. Their conditioning as two-dimensional or 3D scaffolds using conventional or emerging technologies was also discussed. Further biotechnologies are required for developing novel products tailored to stimulate periodontal regeneration. Additional preclinical studies will be useful to closely investigate the mechanisms and identify specific markers involved in cell-implant interactions, envisaging further clinical tests. Future therapeutic protocols will be developed based on these novel procedures and techniques.

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We report the encapsulation of two grape pomace polyphenolic extracts into mesoporous MCM-41-type silica matrices (pristine and Zn or Mg heteroatom modified) to reduce the extract sensitivity and enhance its stability, while preserving the radical scavenger activity. Various grapes marc (Cabernet Saugvinon and Feteasca Neagra from the Black Sea region and commercially available grape skins powder) were used to prepare ethanolic extracts either through conventional extraction, or microwave-assisted procedure. The polyphenolic extracts composition was analysed by reversed phase-high pressure liquid chromatography and spectrometric determination of total polyphenols and ascorbic acid (using Folin Ciocalteu reagent), total flavonoids (by AlCl3 cornplexation), as well as total anthocyanin monomeric pigments content. The encapsulated extract into MCM-41 silica, as well as Zn-MCM-41 and Mg-MCM-41 matrices showed an enhanced radical scavenger activity assessed by DPPH procedure developed for solid samples. The cytocompatibility tests performed on HaCaT keratinocyte human cells demonstrated a good cytocompatibility for the Cabernet Saugvinon and grape skins extracts free and encapsulated into MCM-41-type matrices.

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Eggplant peels (Solanum melongena L.) constitute a promising source of phenolic compounds, flavonoids, and anthocyanins that are beneficial for human health. The objectives of this study were to extract the polyphenolics from eggplant peels and microencapsulation in selected polymers, such as pectin, carboxymethylcellulose and whey proteins hydrolysates by freeze-drying. A combination of ethanolic with ultrasound-assisted methods was used for the extraction, leading to an extract with antioxidant activity of 157.82 +/- 9.46mmol Trolox/g dry weight. The next experiment was to obtain bioactive peptide, both by experimental and predictive methods, which were further used as coating materials. The bioinformatics tools were used for checking the susceptibility of alpha-lactalbumin and beta-lactoglobulin for digestion with thermolysin. The anthocyanins were encapsulated in different combinations of selected polymers, with encapsulation efficiency up to 77.60 +/- 1.92%, highlighting a higher microencapsulation efficiency of carboxymethylcellulose to incorporate anthocyanins. Our results suggested whey peptides had a role in regulating the microencapsulation patterns, whereas carboxymethylcellulose and pectin favored filamentous structuring and double encapsulation, respectively. The in vitro tests showed high biocompatibility of powders cultivated in a cell culture of murine fibroblasts. A significant protective effect in simulated digestion was observed, with a controlled release in the intestinal juice. Accelerated storage stability test showed an increase in antioxidant activity.

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Bone defects due to trauma or diseases causing bone loss have led to an increased demand for new materials that have applications in therapies for bone pathologies. Mesoporous bioactive glasses (MBGs), based on 70SiO(2)-(26-x)CaO-4P(2)O(5)-xCeO(2) (x = 0, 1 and 5 mol % for MBGS, MBGS1Ce and MBGS5Ce) were synthesized by evaporation-induced self-assembly (EISA) method. Ceria was selected due to its antimicrobial and antioxidant properties and positive impact on cells differentiation and mineralization. The pore size values of 2.8 nm confirmed the mesoporosity of the obtained bio-glasses. The biocompatibility tests on mouse fibroblast cells (NCTC clone L929) using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) activity assays revealed no cytotoxicity of Cerium containing MBGs. Formation of a hydroxycarbonate apatite (HCA) layer on MBGs surface, e.g. bioactivity of BMGs, was confirmed by XRD, FTIR, SEM and Raman Spectroscopy while presence of Ce3+/Ce4+ in the glasses before body simulated fluid (SBF) immersion was determined by XPS, UV-Vis measurements. The obtained results show that investigated MBGs are potential materials for applications in bone regeneration.

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In this study, three of the saponins present in leaves of Hedera helix L., alpha-hederin, hederagenin, and hederacoside C were studied for their antiproliferative activity. The three saponins were analyzed in different concentrations by in vitro tests on normal fibroblasts cells and cervix ephitelial tumor cells. Determination of cytotoxicity and antitumor effects was performed using the MTT method. From the tested saponins, a-hederin was biocompatible in normal fibroblasts cells at concentrations between 2-10 mu g/mL. Its antiproliferative activity was exerted in the concentration range of 10-400 mu g/mL in cervix ephitelial tumor cells. Similarly, hederagenin presented antiproliferative activity at concentrations between 25-400 mu g/mL. In turn, hederacoside C was shown to be noncytotoxic in normal fibroblasts and cervix ephitelial tumor cell culture at all the tested concentrations. The obtained experimental results were analyzed by Mixture design, a specialized form of the response surface method (RSM) provided by the Design Expert 11 software, and the optimal composition of obtained saponins mixture was selected and verified in vitro for antiproliferative activity. The results showed that an optimal saponins mixture has the potential to be used in pharmacological applications.

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In this study, we aimed to obtain gelatin from the marine snail Rapana venosa using acidic and enzymatic extraction methods and to characterize these natural products for cosmetic and pharmaceutical applications. Marine gelatins presented protein values and hydroxyproline content similar to those of commercial mammalian gelatin, but with higher melting temperatures. Their electrophoretic profile and Fourier transform infrared (FTIR) spectra revealed protein and absorption bands situated in the amide region, specific for gelatin molecule. Scanning electron microscopy (SEM) analysis showed significant differences in the structure of the lyophilized samples, depending on the type of gelatin. In vitro studies performed on human keratinocytes showed no cytotoxic effect of acid-extracted gelatin at all tested concentrations and moderate cytotoxicity of enzymatic extracted gelatin at concentrations higher than 0.5 mg/mL. Also, both marine gelatins favored keratinocyte cell adhesion. No irritant potential was recorded as the level of IL-1 alpha and IL-6 proinflammatory cytokines released by HaCaT cells cultivated in the presence of marine gelatins was significantly reduced. Together, these data suggest that marine snails are an alternative source of gelatins with potential use in pharmaceutical and skincare products.

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Layers of carbon nanowalls (CNW) were exposed to cold plasma to improve the attachment of fibroblast cells on surface. Plasma was generated at low pressure by a radio-frequency (RF) parallel-plate discharge in Ar/O-2 mixture. 10 min of exposure allowed the insertion of oxygen-containing groups and considerable enhancement in surface hydrophilicity while the surface morphology was not significantly changed. The cells were cultivated on CNW samples and the analyses were performed after 24 h culture. Significant differences in density and morphology of cell attached on tested surfaces were observed. The number of viable cells increased after plasma treatment.

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Hederagenin, a saponin known for its therapeutic effect was isolated from leaves of Hedera helix. Hederagenin was obtained by repeated maceration of ground plant material in 95% (v/v) ethanol. The ethanolic extract was subjected to acid hydrolysis and purification with acetonitrile. Isolated hederagenin (IHe) was analyzed by HPLC-MS/MS and compared to standard hederagenin. In vitro cytotoxicity of IHe was tested in a culture of fibroblast cells from NCTC clone L929 cell line by MTT assay, using dioscin as positive control. For the determination of IHe antitumor effect, in vitro tests were performed in a culture of human cervix carcinoma Hep-2 cells cultivated in the presence of different concentrations of sample for 48 h. IHe was biocompatible in the range of concentrations 2-200 mu g/mL, in NCTC cell culture. The compound showed cell cytotoxicity in the concentration range of 100-400 mu g/mL, in Hep-2 cell culture, revealing its antitumor activity. These results demonstrated the possible use of hederagenin isolated from H. helix extract as an antitumor agent.

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The aim of this study was to improve the bovine colostrum biological function through fermentation with kefir grains enhanced with selected yeasts, for developing new nutraceutical and cosmeceutical products. It was found that fermentations with co-culture of 2.5 g% artisanal kefir grains and selected yeast strains (10(6) CFU/100 mL) increased the functional quality of the fermented products compared to the product obtained only with kefir grains. Fresh fermented products obtained with a consortium based on kefir grains and Candida lipolytica MIUG D67 demonstrated an increased antioxidant activity of 2.69 mM Trolox Equivalent/g, after 48 h of fermentation. Instead, peptide fractions with MW<10 kDa isolated by membrane filtration from lyophilized fermented products, based on colostrum fermentation with kefir grains enhanced with Candida lipolytica MIUG D99 starter, presented markedly increase in vitro of ABTS radical scavenging activity, similar to a concentration of 2 nM captopril. These results indicated their possible application in enhance of the quality of the fermented products in order to increase the postbiotic composition with functional impact in vivo.

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Novel L-Arginine-Dextran70 based-silver nanoparticles (SNPs) (SNPs(Arg)), functionalized with Riboflavin (RF) and Human Serum Albumin (HSA) were characterized by UV-Vis absorption, Transmission Electron Microscopy (TEM), Atomic Force Microscopy (AFM), fluorescence and circular dichroism spectroscopy, chemiluminescence and Neutral red assays. TEM analysis showed the formed faceted particles, large clumps/fused aggregates, nano featured with the mean particle size of 41.60 nm. Chemiluminescence and Neutral red assays for in vitro antioxidant and cytotoxic activities of the SNPs(Arg)/RF/HSA systems have been studied. It was pointed out that SNPs(Arg) functionalized with RF and HSA resulted in a bio-nanosystem which leads to a high activity against oxygen free radicals, altered viability, morphology, apoptotic bodies and decreased cell density of L929 fibroblast cells. Results are relevant for understanding the redox properties of RF in the case of biological applications and especially for RF containing drugs.

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In the present work, we report the characterization of TiO2-hydroxyapatite (HA) nanocomposites obtained by a two-step sintering (TSS) process of a mixture of HA and titanium hydride (TiH2) powders. The reactions underwent by TiH2 in the presence of HA and hydrogen release, and subsequently, titanium oxidation was examined by thermal analysis. A longer holding time in the second sintering stage enabled obtaining a homogenous TiO2-HA (36% rutile) composite with a thermal expansion coefficient of 11.46 . 10(-6) C-1 in the 40-1000 degrees C range. Unconventional TSS process hinders HA decomposition to detrimental tricalcium phosphate (TCP). Wear rate of ceramics was determined by tribological measurements and the material biocompatibility was evaluated using MTT assay. Overall, cell viability results correlated with morphological observations indicated a good biocompatibility of HA-based composites at all tested concentrations. Incorporation of the TiO2 phase in HA by TSS process was found to be an efficient way to prepare bioceramics with improved performances. (C) 2017 Production and hosting by Elsevier B.V. on behalf of King Saud University.

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Silver nanoparticles (AgNP) suspensions were biosynthesized by silver ions reduction in the presence of collagen, a nontoxic, organic polymer, intending to improve their medical use in periodontitis treatment. Spectrophotometric measurements showed a time- and concentration-dependent increase of AgNP formation in each suspension variant. Transmission electron microscopy revealed spherical morphology of AgNP in collagen and their mean diameter size was around 30 nm. The particle size distribution and zeta potential values of AgNP in collagen were determined by dynamic light scattering measurements. The surface charge of AgNP in collagen was positive, while commercial AgNP stabilized in citrate had negative surface charge. In vitro cytotoxicity testing of AgNP in collagen showed that they were biocompatible with human gingival fibroblasts in a wider range of concentrations than commercial nanoparticles. The antibacterial activity of AgNP in collagen against two pathogenic strains present in the periodontal pocket was dose-dependent and higher than that of AgNP in citrate. All these results demonstrated that AgNP prepared in collagen gel had improved properties, like small diameter, positive surface charge, high biocompatibility in human gingival fibroblasts, efficiency against bacterial growth and, thus, better therapeutic potential in periodontal disease treatment.

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Bionanocomposites based on poly(lactic acid) (PLA), plasticized with commercial tributyl o-acetyl citrate (ATBC) and containing hydrolyzed collagen (HC) up to 10 wt% and silver nanoparticles (AgNPs), were prepared by a melt mixing procedure. The properties of antimicrobial PLA based formulations were investigated in terms of morphology (atomic force microscopy, AFM), mechanical, thermal (differential scanning calorimetry, DSC), spectral (by attenuated total reflectance, Fourier-transform infrared spectroscopy), cell proliferation (by flow cytometry) and immunohistochemical properties induced by collagen. The incorporation of HC into antimicrobial PLA biocomposites led to the slight reduction both in mechanical properties and the degree of crystallinity with respect to those of PLA/ATBC sample. These properties can be attributed to the smooth surface improvement of the bionanocomposite. In vitro testing using L929 fibroblasts in the presence of PLA-based bionanocomposites showed that all samples presented good biocompatibility, as it was indicated by the cell cycle distribution and DNA content analyses. Furthermore, these new biocomposites induced an increase of collagen production in vitro. Overall, PLA/HC5/AgNPs and PLA/HC10/AgNPs bionanocomposites showed very good in vitro biocompatibility, and therefore, could be considered as valuable materials for medical devices, such as tubes, catheters, drains or connectors, with a relatively long service life.

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The collagen hydrolysate (HC) and silver nanoparticles (AgNPs) effect on the mechanical, thermal, in vitro degradation, cytotoxicity and antifungal properties of polylactic acid (PLA) plasticized and compatibilized with a mixture of polyethylene glycol (PEG 4000) and Lapol 108 (MB) was studied. The presence of HC led to the decreased tensile properties and to the increased hydrophilicity of biocomposites surfaces. Tested biocomposites led to a very good cellular viability percentage, unaltered cell morphology and a modest antifungal activity. The extrusion trial showed that the PLA/MB/PEG/AgNPs bionanocomposite is a promising potential material for urinary drains. [GRAPHICS]

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Specific ultrastructural anatomy of masticatory muscles is commonly referred to a general pattern assigned to striated muscles. Junctional feet consisting of calcium channels of the sarcoplasmic reticulum (i.e. the ryanodine receptors, RyRs) physically connected to the calcium channels of the t-tubules build triads within striated muscles. Functional RyRs were demonstrated in the nuclear envelopes of pancreas and of a skeletal muscle derived cell line, but not in muscle in situ. It was hypothesized that ryanodine receptors (RyRs) could also exist in the nuclear envelope in the masseter muscle, thus aiming at studying this by transmission electron microscopy. There were identified paired and consistent subsarcolemmal clusters of mitochondria, appearing as outpockets of the muscle fibers, usually flanking an endomysial microvessel. It was observed on grazing longitudinal cuts that the I-band-limited mitochondria were not strictly located in a single intermyofibrillar space but continued transversally over the I-band to the next intermyofibrillar space. It appeared that the I-band-limited transverse mitochondria participate with the column-forming mitochondria in building a rather incomplete mitochondrial reticulum of the masseter muscle. Subsarcolemmal nuclei presented nuclear envelope-associated RyRs. Moreover, t-tubules were contacting the nuclear envelope and they were seemingly filled from the perinuclear space. This could suggest that nucleoplasmic calcium could contribute to balance the cytosolic concentration via pre-built anatomical routes: (i) indirectly, via the RyRs of the nuclear envelope and (ii) directly via the communication of t-tubules and sarcoplasmic reticulum through the perinuclear space. (C) 2019 Elsevier GmbH. All rights reserved.

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Tuberculostatic drugs are the most common drug groups with global hepatotoxicity. Awareness of potentially severe hepatotoxic reactions is vital, as hepatic impairment can be a devastating and often fatal condition. The treatment problems that may arise, within this class of medicines, are mainly of two types: adverse reactions (collateral, toxic or hypersensitive reactions) and the initial or acquired resistance of Mycobacterium tuberculosis to one or more antituberculosis drugs. Prevention of adverse reactions, increase treatment adherence and success rates, providing better control of tuberculosis (TB). In this regard, obtaining new drugs with low toxicity and high tuberculostatic potential is essential. Thus, in this work, we have designed or synthesized new derivatives of isoniazid (INH), such as new Isonicotinoylhydrazone (INH-a, INH-b and INH-c). These derivatives demonstrated good biocompatibility, antimicrobial property similar to that of parent isoniazid and last but not least, a significantly improved Pharmacotoxicological profile compared to that of isoniazid. © 2019 Elsevier B.V.

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In the present work, we report the characterization of TiO2-hydroxyapatite (HA) nanocomposites obtained by a two-step sintering (TSS) process of a mixture of HA and titanium hydride (TiH2) powders. The reactions underwent by TiH2 in the presence of HA and hydrogen release, and subsequently, titanium oxidation was examined by thermal analysis. A longer holding time in the second sintering stage enabled obtaining a homogenous TiO2-HA (36% rutile) composite with a thermal expansion coefficient of 11.46 · 10−6 C−1 in the 40–1000 °C range. Unconventional TSS process hinders HA decomposition to detrimental tricalcium phosphate (TCP). Wear rate of ceramics was determined by tribological measurements and the material biocompatibility was evaluated using MTT assay. Overall, cell viability results correlated with morphological observations indicated a good biocompatibility of HA-based composites at all tested concentrations. Incorporation of the TiO2 phase in HA by TSS process was found to be an efficient way to prepare bioceramics with improved performances. © 2017

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Skin wound dressings are commonly used to stimulate and enhance skin tissue repair. Even if wounds seem easy to repair for clinicians and to replicate in an in vitro set-up for scientists, chronic wounds remain currently an open challenge in skin tissue engineering for patients with complementary diseases. The seemingly simple process of skin healing hides a heterogenous sequence of events, specific timing, and high level of organization and coordination among the involved cell types. Taken together, all these aspects make wound healing a unique process, but we are not yet able to completely repair the chronic wounds or to reproduce them in vitro with high fidelity. This review highlights the main characteristics and properties of a natural polymer, which is widely used as biomaterial, namely collagen and of its denatured form, gelatin. Available wound dressings based on collagen/gelatin and proposed variants loaded with bioactive compounds derived from plants are presented. Applications of these composite biomaterials are discussed with emphasis on skin wound healing. A perspective on current issues is given in the light of future research. The emerging technologies support the development of innovative dressings based exclusively on natural constituents, either polymeric or bioactive compounds. (C) 2019 Elsevier B.V. All rights reserved.

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The aqueous anthocyanin-rich extract derived from black rice (Oryza sativa L.) was encapsulated by freeze drying using milk proteins and peptides as coating materials. The molecular modelling approach indicated that all major casein fractions and whey proteins were able to bind at least one anthocyanin molecule. The hydrophobic interactions and hydrogen bonding across the interfaces appeared to be mainly responsible for the stabilizations of the complexes formed between the coating material and bioactive compounds. Two dark purple colored powders, differentiated by the ratio of the encapsulation materials used, rich in phytochemicals were obtained, with an encapsulation efficiency of up to 99%. The powders were tested for antioxidant activity, cytocompatibility, and thermal stability. The morphological structure of the powders highlighted the presence of encapsulated anthocyanins. Both powders showed a remarkable antioxidant activity of about 46 mM Trolox/g D.W., and cytocompatibility on the L929 fibroblast culture. At certain concentrations, both powders stimulated cell proliferation. The powders showed a good thermal stability between 75 and 100 degrees C for 15 min. The powders were tested in a food model system and checked for stability of phytochemicals during storage. The added value of the powders was demonstrated throughout the antioxidant activity, which remained unchanged during storage.

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Composite materials containing natural biocompatible polymers, specific for the extracellular matrix present in the periodontal tissue, are prepared by mixing type I soluble collagen (COL) and fibronectin (FN) as fibrillar proteins and chondroitin sulfate (CS) as polysaccharidic component. The threedimensional porous structure is obtained by freeze-drying process and it is analyzed by scanning electron microscopy (SEM). Specific techniques of immunohistochemistry and transmission electron microscopy (TEM) are used to investigate the distribution and interaction between polymeric components within the scaffold. Light and electron microscopy observations are also performed on solutions of CS and FN to reveal their structural and morphological properties. The cytocompatibility of composite material is tested in gingival fibroblasts and osteoblast cell cultures using quantitative assays of cell viability. The results demonstrate that COL-CS-FN composite material has optimal characteristics for further testing as device for oral tissue wound healing.

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In this paper, we present the fabrication and characterization of new chitosan-based membranes while using a new biotechnology for immobilizing alkaline phosphatase (ALP). This technology involved metal ions incorporation to develop new biopolymeric supports. The chemical structure and morphological characteristics of proposed membranes were evaluated by infrared spectroscopy (FT-IR) and the scanning electron microscopy technique (SEM). The inductively coupled plasma mass spectrometry (ICP-MS) evidenced the metal ion release in time. Moreover, the effect of Mg2+ on the enzymatic activity and the antibacterial investigations while using Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus bacteria, hemolysis, and biocompatibility behavior were studied. Immobilizing ALP into the chitosan membranes composition followed by the incorporation of Mg2+ led to polymeric supports with enhanced cellular viability when comparing to chitosan-based membranes without Mg2+. The results obtained evidenced promising performance in biomedical applications for the new biopolymeric supports that are based on chitosan, ALP, and metal ions.

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Telocytes (TCs) are cells with long, thin and moniliform processes called telopodes. These cells have been found in numerous tissues, including the eye choroid and sclera. Lamina fusca (LF), an anatomical structure located at the sclera-choroid junction, has outer fibroblastic lamellae containing cells with long telopodes. The purpose of this study was to evaluate, via transmission electron microscopy, the LF for the presence of endothelial-specific ultrastructural features, such as Weibel-Palade bodies (WPBs), in the residing TCs. We found that the outer fibroblastic layer of LF lacked pigmented cells but contained numerous cells with telopodes. These cells had incomplete or absent basal laminae, were united by focal adhesions and close contacts, and displayed scarce caveolae and shedding vesicles. Within the stromal cells of LF, numerous WPBs in various stages of maturation and vesicular structures, as secretory pods that ensure the exocytosis of WPBs content, were observed. The WPBs content of the cells with telopodes in the LF could indicate either their involvement in vasculogenesis and/or lymphangiogenesis or that they are the P-selectin- and CD63-containing pools that play roles in sclera) or choroidal inflammation. (C) 2018 Elsevier GmbH. All rights reserved.

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Stromal cells/telocytes (SCs/TCs) were recently described in the human adult trigeminal ganglion (TG). As some markers are equally expressed in SCs/TCs and endothelial cells, we hypothesized that a subset of the TG SCs/TCs is in fact represented by endothelial progenitor cells of a myelomonocytic origin. This study aimed to evaluate whether the interstitial cells of the human adult TG correlate with the myelomonocytic lineage. We used primary antibodies for c-erbB2/HER-2, CD31, nestin, CD10, CD117/c-kit, von Willebrand factor (vWF), CD34, Stro-1, CD146, alpha-smooth muscle actin (alpha-SMA), CD68, VEGFR-2 and cytokeratin 7 (CK7). The TG pial mesothelium and subpial vascular microstroma expressed c-erbB2/HER-2, CK7 and VEGFR-2. SCs/TCs neighbouring the neuronoglial units (NGUs) also expressed HER-2, which suggests a pial origin. These cells were also positive for CD10, CD31, CD34, CD68 and nestin. Endothelial cells expressed CD10, CD31, CD34, CD146, nestin and vWF. We also found vasculogenic networks with spindle-shaped and stellate endothelial progenitors expressing CD10, CD3 1, CD34, CD68, CD146 and VEGFR-2. Isolated mesenchymal stromal cells expressed Stro-1, CD146, CK7, c-kit and nestin. Pericytes expressed a-SMA and CD146. Using transmission electron microscopy (TEM), we found endothelial specific Weibel-Palade bodies in spindle-shaped stromal progenitors. Our study supports the hypothesis that an intrinsic vasculogenic niche potentially involved in microvascular maintenance and repair might be present in the human adult trigeminal ganglion and that it might be supplied by either the pial mesothelium or the bone marrow niche. (C) 2018 Elsevier GmbH. All rights reserved.

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Kombucha is an oriental traditional beverage made usually of sweetened tea fermented by a symbiotic consortium of bacteria and yeast embedded within a cellulose membrane. The beverage has been characterized for health benefits related mainly to the tea itself but very few in relation to the microbial consortia. The objective of our work was to isolate lactic acid bacteria from a local kombucha source and characterize the strains for their probiotic potential. Five lactic acid bacteria have been isolated, characterized morphologically and by molecular tools; the results proved their belonging to the lactobacilli/lactococci group. Sequencing results lead to the conclusion that they all have 99% identity with strains of Pediococcus pentosaceus. The on plate screening for bacteriocin production have been expressed on synthetic media only for three of the total five isolates. To complete the probiotic protential profiles of the isolates, they have been tested for their resistance to bile salts; only one isolate finally proved to have clear probiotic potential.

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The present work explores new hybrid bioactive, enzymatically degradable hydrogel nanocomposites. They are composed by stimuli-sensitive semi-interpenetrating collagen/poly(N-isopropyl acrylamide) polymeric matrix, DelliteA (R) 67G and CloisiteA (R) 93A nanoclays, and hydroxyapatite particles. Morphology of the hybrid hydrogel nanocomposites was examined by near infrared chemical imaging, X-ray diffraction, scanning electron microscopy, and transmission electron microscopy techniques. It has been found that the porous hydrogel nanocomposites showed a mix between intercalated and exfoliated structure, with interconnected pores and the inorganic material was evenly distributed in the polymeric matrix because of specific interactions between components. The collagen resistance against enzymatic degradation and the thermal stability were improved due to the protein encapsulation in the synthetic polymer matrix. The hydrogel nanocomposites showed suitable swelling characteristics for tissue engineering. In vitro cytocompatibility and cell viability revealed that the hybrid nanocomposites were non-cytotoxic for rat osteoblasts. These hybrid nanocomposites were designed as potential synthetic bone tissue engineering.

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Telocytes (TCs) are stromal cells defined by the presence of long and slender prolongations (telopodes). They are a biologically and functionally heterogeneous population that has not been previously investigated in the sclera. The purpose of this study is to investigate the presence and characteristics of scleral telocytes through a combined immunohistochemical and transmission electron microscopy (TEM) study using samples from ten adult patients. Stromal cells with a TC-like morphology expressed CD34, CD45, CD105, vimentin and occasionally CD68 but were negative for collagen III, CD31, CD133, and CD146. Conjunctival epithelial cells expressed CD45, CD105, CD146, and vimentin. These phenotypes support a scleral niche with immune TCs and haematopoietic stem cells (HSCs). In TEM, we often found spindle shaped stromal cells projecting telopodes or filopodes, with extremely long nuclei extended even within those prolongations. We separated these cells into a light subtype, which contained a complete set of organelles, and a dark subtype, consisting of undifferentiated stem/progenitor cells. The light cells contained dense vesicles, Weibel-Palade bodies, and rounded beta-granule-like structures. These storage areas for the von Willebrand factor (vWF) are known to express selectins that are critically involved in HSC homing and could also indicate endothelial progenitors. The dark cells were scarcely myoid, populated the episcleral perivascular niches and the scleral stroma, and were equipped with lipid storage areas such as lamellar bodies and lipid droplets (LDs). Previously, unreported intranuclear LDs were found in these cells, which is characteristic of an HSC population. It appears that the human scleral stroma is a niche harbouring TC-like cells with immune and HSC phenotypes, and the mere presence or characteristics of telopodes are not enough to differentiate them. (C) 2018 Elsevier GmbH. All rights reserved.

Abstract

Berteroa incana is a wild herb widespread in temperate zones which was practically not studied for its biological effects. Methanolic and aqueous extracts of B. incana were assessed for the content in polyphenols and the related antioxidant and antimicrobial activities and the polysaccharide extract for the content in saccharides and the associated cytostatic effect. The results obtained highlighted that the methanolic extracts of B. incana contain moderate amounts of polyphenols, the most representative been isoquercitrin 4.41 +/- 0.02 mg100 g(-1)dry weight plant material (DW), quercetin 4.21 +/- 0.05, sinapic acid 5.23 +/- 0.12 and ferulic acid 5.05 +/- 0.12 mg 100 g(-1)DW, with correlated moderate antioxidant activities (IC (50) 13.40 +/- 0.01 mu g mL(-1)) and absent antibacterial activity. The polysaccharide fraction showed high content in saccharides, especially in arabinose (312.22 +/- 7.54 mg g(-1) polysaccharide extract) and glucose (279.22 +/- 5.59), and promising cytostatic effect. [GRAPHICS] .

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The aim of this study was to develop three-dimensional porous composites of collagen (Col) incorporating polyphenolic-rich wormwood extract and to investigate their interaction with human skin cells, in order to optimize wound healing treatments. The scaffolds' ultrastructure was observed by scanning electron microscopy, and biodegradability and bioactive compounds release were investigated in physiologic environment. Interaction of composites in direct and indirect contact with human skin cells was evaluated using two in vitro experimental models. ColWE scaffolds presented high porosity, swelling degree, and increased stability against enzymatic degradation, compared to Col scaffold. Composite scaffolds incorporating higher quantities of wormwood extract allowed better control of polyphenolics release. ColWE 0.5 variant favored the attachment and proliferation of human dermal fibroblasts and keratinocyte cells. In addition, the composite scaffold stimulated the synthesis of skin extracellular matrix components. All these results demonstrated that ColWE composites with improved physico-chemical and biological properties could be used in advanced wound healing applications.

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The bioavailability of pollen bioactive compounds for humans is limited. In this study, our aim was to enhance the health-related benefits of pollen by fermentation with a Kombucha/SCOBY (symbiotic culture of bacteria and yeasts) consortium. We performed the fermentation of pollen suspended from the beginning with SCOBY on sweetened green tea or on Kombucha vinegar, by adding pollen after 20 days of Kombucha fermentation. We analyzed: formation of bioactive compounds (anti-oxidant polyphenols, soluble silicon, hydroxy-acids, short chain fatty acids-SCFA); parameters related to Kombucha fermentation (dynamics of lactic acid bacteria-LAB, formation of organic acids, soluble sugar evolution on Kombucha vinegar); the influence of Kombucha fermentation on pollen morphology and ultrastructure; in vitro cytotoxic and antitumoral effects of the Kombucha fermented pollen. The pollen addition increases LAB proportion in the total number of SCOBY microbial strains. SEM images highlight the adhesion of the SCOBY bacteria to pollen. Ultrastructural analysis reveals the release of the pollen content. The content of bioactive compounds (polyphenols, soluble silicon species and SCFA) is higher in the fermented pollen and the product shows a moderate antitumoral effect on Caco-2 cells. The health benefits of pollen are enhanced by fermentation with a Kombucha consortium.

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A new series of substituted N,N-bis-[(1H-pyrazol-1-yl)methyl]-aminohexadecane Mannich bases were synthesized, characterized by IR, H-1 NMR C-13 NMR, UV-Vis, MS and elemental analysis, and tested for their biological activity. All the synthesized compounds were tested for in vitro antimicrobial activity against a panel of selected bacterial and fungal strains using erythromycin and clotrimazole as standards. Most of the synthesized compounds demonstrated very good activity at minimum inhibitory concentrations (MICs). Compound 3b with an halogen atom into the pharmacophore structure exhibited the most significant activity against Bacillus subtilis (MIC = 0.007 mu gmLL(-1)) versus erythromycin as standard. In vitro cytotoxicity of the new compounds was studied using MTT assay. The analysis of the test cells showed that the newly synthesized alkylaminopyrazoles derivatives were biocompatible until a concentration of 5 mu gmL(-1); two compounds presented a high degree of biocompatibility on the studied concentration range. (C) 2017 Elsevier B.V. All rights reserved.

Abstract

Stable Schiff bases containing a furoxan moiety are synthesized as single regioisomers by the reaction of 3-methyl-2-oxy-furazan-4-carbaldehydewith various amino compounds at room temperature. The structures of synthesized compounds were fully characterized by multinuclear NMR spectroscopy and X-ray crystallography. The effect of synthesized Schiff bases containing a furoxan moiety on biological generation of reactive oxygen species and nitric oxide in plant tissues was investigated for the first time by fluorescence microscopy and the released NO identified as nitrite with Griess reagent. There is a good correlation between the biological generation of NO determined by fluorescence microscopy and with Griess reagent. Some of the synthesized compounds exhibited both nitric oxide and reactive oxygen species generation abilities and represent potential NO donors in plant tissues.

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Telocytes (TCs) are cells with long, thin and moniliform processes called telopodes. These cells have been found in numerous tissues, including the eye choroid and sclera. Lamina fusca (LF), an anatomical structure located at the sclera-choroid junction, has outer fibroblastic lamellae containing cells with long telopodes. The purpose of this study was to evaluate, via transmission electron microscopy, the LF for the presence of endothelial-specific ultrastructural features, such as Weibel–Palade bodies (WPBs), in the residing TCs. We found that the outer fibroblastic layer of LF lacked pigmented cells but contained numerous cells with telopodes. These cells had incomplete or absent basal laminae, were united by focal adhesions and close contacts, and displayed scarce caveolae and shedding vesicles. Within the stromal cells of LF, numerous WPBs in various stages of maturation and vesicular structures, as secretory pods that ensure the exocytosis of WPBs content, were observed. The WPBs content of the cells with telopodes in the LF could indicate either their involvement in vasculogenesis and/or lymphangiogenesis or that they are the P-selectin- and CD63-containing pools that play roles in scleral or choroidal inflammation. © 2018 Elsevier GmbH

Abstract

The tympanic membrane (TM) integrity is of utmost importance for the sense of hearing. Therefore, the intrinsic potential of the TM to regenerate and repair deserves complete characterisation. Existing studies brought evidence on the epithelial stem niche of the TM. However, the stromal compartment was not evaluated for harbouring a distinctive stem, or progenitor, niche. We aimed doing this in transmission electron microscopy. We used TMs dissected out from 3 male Oryctolagus cuniculus rabbits. Evidence of stromal quiescent stem cells was gathered. Moreover, endothelial progenitor cells were found in the TM, being accurately identified by two specific ultrastructural markers of the endothelial lineage: the Weibel-Palade bodies and the stomatal diaphragms of the subplasmalemmal caveolae. The stromal stem niche of the TM appears to be a distinctive contributor during physiological and pathological processes of the TM, such as cholesteatoma formation, at least as a biological support for processes of vasculogenesis. However, further characterisation of the molecular pattern of the stromal stem niche of the TM is mandatory.

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Establishing the postmortem interval (PMI) is vital in legal medicine as it allows to retrospectively estimate the hour of death, which is essential for the police as a starting point for their inquiries (especially in violent deaths). Ultrastructure studies aimed specifically to detect autolytic changes are scarcely identified in the scientific literature. Moreover, they are performed in a variety of conditions (different temperatures, species, in vitro / in situ, and so on), making the results difficult to interpret for legal medicine purposes. The main aim of this review is to determine the potential usefulness of ultrastructure studies for the estimation of the postmortem interval and to provide a summary of relevant scientific literature in the area, which might be useful as a starting point for more specific and detailed studies in the field. We performed a search on the ISI Thomson Web of Knowledge database using a series of predefined keywords; the articles fulfilling the inclusion criteria were systematically analyzed to identify ultrastructure changes associated with autolysis. Our investigation revealed 20 relevant articles, which detailed ultrastructure changes in the brain, heart, liver, pancreas, kidney, bone, sweat glands, thyroid, skeletal muscle, cartilage and sweat glands. For each organ, we arranged systematically postmortem ultrastructure changes that were described by various authors. Ultrastructure changes appear early and may be useful in determining the time since death in the early postmortem interval. However, most studies published in this area followed methodologies that could not allow a proper reproducibility in forensic circumstances. Therefore, before using ultrastructure for estimating the PMI in practical environments, further studies are needed. They should be performed ideally on human samples, obtained at regular intervals after death, at variable, decreasing temperatures.

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Atorvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, is known to exert lipid-lowering, but also anti-inflammatory effects in extra-arterial locations. In order to avoid its toxicity associated with long-term oral treatment, in this study we have proposed novel lipid nanostructures containing atorvastatin to improve its efficiency and bioavailability after local application in periodontitis inflammation therapy. The physico-chemical characterization of the nanostructures was performed using dynamic light scattering technique and morphological observations were made by light microscopy. The encapsulation efficiency was determined by high performance liquid chromatography analysis of loaded atorvastatin. In vitro cytotoxicity and anti-inflammatory activity were evaluated in human premonocytic THP-1 cell line and a model of lipopolysaccharide-induced inflammation in macrophages, respectively. The results showed that the population of atorvastatin lipid nanostructures presented a mean diameter of 178 nm and a good homogeneity after sonication and extrusion treatments applying, as indicated by the low polydispersity index of 0.223. The efficiency of atorvastatin encapsulation was high (87.3%) and the nanostructures cytotoxicity was reduced for lipid concentrations ranging from 50 mu M to 500 mu M. Experiments in THP-1 cells differentiated to macrophages demonstrated that atorvastatin liposomal formulation led to a higher inhibition of lipopolysaccharide-induced proinflammatory cytokines (interleukin 6, tumor necrosis factor alpha and interleukin 8) release, compared to free drug. In conclusion, atorvastatin lipid nanostructures could be used to develop an efficient local treatment of periodontitis inflammation.

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Cytological responses in different organs of sentinel organisms have proven to be useful tools for characterizing the health status of those organisms and assessing the impact of environmental contaminants. Our study shows that nickel (II) accumulated in both germ cells (oogonia and developing oocytes) and somatic cells (muscle cells, follicle cells) in the Astacus leptodactylus ovary. Muscle cells from ovarian wall show disorganization and the disruption of cytoplasmic microtubules and pyknosis of the cell nucleus. Follicle cells, both those that surround the developing oocytes and also those that are not associated with the oocytes contained within the cytoplasm vacuoles of different sizes, degenerated mitochondria, myelin bodies, disorganized microtubules, and pyknotic nuclei. The most evident pathological phenomenon was the alteration and disorganization of the basal matrix, which separates the ovarian interstitium from ovarian follicles compartment. Exposure to nickel induces cytoplasmic vacuolation in oogonia and developing oocytes, structural alteration of the developing yolk granules and condensation of the nucleoli. Ultrastructural autometallography has shown grains of silver-enhanced nickel inside the cytoplasm of the muscle cells with altered morphology, including the cytoplasm, nucleus, and basal matrix of the follicle cells, and in intracisternal granules and developing yolk granules of the oocytes.

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Polysaccharides display several biological activities, such as antitumour, anti-oxidation, hypoglycaemic, antiviral and immunity enhancing properties. The polysaccharide fraction from Phemeranthus confertiflorus was extracted, hydrolysed and 8 monosaccharides were analysed and quantified by capillary electrophoresis. The polysaccharide fraction was further assessed for cytotoxicity on normal and tumour cell lines. The results obtained can promote Phemeranthus as an eligible plant with potential for human health, because of high monosaccharides content (predominant rhamnose) and promising cytostatic effect of polysaccharide fraction.

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The work aims to simulate in vitro, the antioxidant activity of riboflavin (RF) (Vitamin B-2) in Polyethylene Glycols (PEGs) (Tween20 and Myrj52)/Bovine Serum Albumin (BSA) systems, using the chemiluminescent system luminol-hydrogen peroxide, in alkaline solution, Tris-HCl buffer, pH 8.5. The tested concentrations were 2-20 mu M for RF and 0.06 to 0.6% for PEGs. It was found that 0.12% PEGs enhance the antioxidant activity of RF to higher and lower values, according to their molecular structure, Tween20 > Myrj52. RF has a pro-oxidant effect in the presence of BSA, while in the PEG/BSA systems, its antioxidant activity increases up to similar to 20%; a cross-linking of PEG to RF in the presence of BSA is considered. Additionally, studies by fluorescence spectroscopy on RF embedded in Tween20/BSA system-enhanced Tyrosine fluorescence contribution. In vitro cytotoxicity evaluation of RF in PEG/BSA systems was performed by Neutral Red assay and by monitoring lactate dehydrogenase release. PEG/BSA system offers great promise for enhancement of the RF antioxidant activity and biocompatibility, maintaining the integrity of the BSA, facts which are useful in the oxidative stress and drug delivery processes.

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Therapeutic benefits of phytochemicals are widespread, but the association of natural lipophilic and hydrophilic compounds in the same nanostructured delivery system represents a challenge that was not considered up to present. This work aimed to evaluate the effects of two plant-derived oils rich in omega-3 (rosehip oil Ro) and omega-6 (black caraway oil Bco) on the synthesis of marigold extract (ME) and azelaic acid (AzA) co -loaded nanostructured lipid carriers(NLC) and to determine its in vitro and in vivo anti-inflammatory action. The lipid core and surfactant shell guaranteed a high encapsulation efficiency of both water soluble -AM and lipid soluble -ME(93% entrapment for carotenoids and 83% for AM). A high degree of biocompatibility in the concentration range of 5-400 mu g/mL NLC-ME-AzA for 72 h of treatment was determined (i.e. at 400 mu g/mL the L929 cells displayed a proliferative effect with a cell viability of 110%). The co-presence of ME and AM in the same NLC was accompanied by a reduced expression of inflammatory IL-6 and IL-1 beta cytokines, as evidenced by ELISA method. The in vivo plestismometric method showed that paw edema was significantly reduced for 3 h following NLC-ME-AzA hydrogel treatment (0.006 mg ME and 26.2 mg AAz) in rats challenged with carrageenan.

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Portulaca pilosa L and Portulaca oleracea L were comparatively studied for the total content of polyphenols and flavonoids, antioxidant activity, individual polyphenols, short-chain organic acids, and saccharides in extracts of plants collected from Bucharest ''delta'' using spectrometry and capillary electrophoresis. The polysaccharide fractions were assessed for cytotoxicity on normal and tumor cell lines. The results obtained highlighted that Portulaca pilosa could be considered more valuable than Portulaca oleracea because of its higher content in important flavonoids (quercetin 101.70 +/- 2.68 mu g g(-1) dry weight plant material, rutin 96.24 +/- 0.74 mu g g(-1) DW plant material) and some phenolic acids (chlorogenic acid 161.33 +/- 0.67 mu g g(-1) DW plant material, p-coumaric acid 61.40 +/- 5.50 mu g g(-1) DW plant material) of the ethanolic extracts, for its lower content in oxalic acid that is considered anti-nutrient of the aqueous extracts, and for its higher content in saccharides, especially rhamnose and xylose, with consequently higher cytostatic effect of the polysaccharide fraction. The current study presented for the first time the content in polyphenols, short chain organic acids, and saccharides of Portulaca pilosa and proved that this species has noticeable antioxidant activity, low toxicity on normal cells, and high toxicity on tumor cells and could be considered important for health and food industry.

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Oxidative stress and inflammation can be involved in cognitive dysfunction associated with neurodegenerative disorders. Diazepam (DZP) administration has been chosen to simulate the memory impairment. The aim of this study was to evaluate the effects of curcumin (CUR) on spatial cognition, ambulatory activity, and blood and brain oxidative stress levels. The ERK/NF-kappa B signaling pathway and the histopathological changes in the hippocampus and frontal lobe, in diazepam-treated rats, were also analyzed. The animals were divided into 4 groups: control, carboxymethylcellulose (CMC) + CUR, CMC + DZP, and CUR + CMC+ DZP. CUR (150 mg/kg b.w.) was orally administered for 28 days. DZP (2 mg/kg b.w.) was intraperitoneally administered 20 minutes before the behavioral tests (open field test, Y-maze, and elevated plus maze). CUR improved the spontaneous alternation behavior, decreased the oxidative stress levels, both in the blood and in the hippocampus, and downregulated the extracellular signal-regulated kinase (ERK 1/2)/nuclear transcription factor-(NF-) kappa B/pNF-kappa B pathway in the hippocampus and the iNOS expression in the hippocampus and frontal lobe of the DZP-treated rats. Histopathologically, no microscopic changes were found. The immunohistochemical signal of iNOS decreased in the DZP and CUR-treated group. Thus, our findings suggest that curcumin administration may improve the cognitive performance and may also have an antioxidant effect.

Abstract

Telocytes (TCs) are morphologically defined as small-sized cells with long, thin, moniliform processes called telopodes (Tps). Numerous papers imply that TCs are a distinctive cell type, and that transmission electron microscopy (TEM) is the gold standard tool for their identification. We aimed to reproduce previous studies on myocardial TCs to check their validity. For this purpose we performed an immunohistochemical study on human cardiac samples from six autopsied donor cadavers, using antibodies against CD10, CD31, CD34, CD146, Ki67, alpha-smooth muscle actin (a-SMA), Platelet-Derived Growth Factor Receptor-alpha (PDGFRa) and laminin. Additionally we performed a TEM study on cardiac samples from three human autopsied donor cadavers and five adult Sprague-Dawley rats. We found endothelial cells (ECs), cords, and filopodia-projecting endothelial tip cells (ETCs) that expressed CD10, CD31, CD34, CD146, and PDGFR-a. Often, endothelial cells closely neighbored the sarcolemmal basal laminae. Endothelial progenitor cells, as well as nascent capillaries, were CD31+/CD34+. Proliferative endothelial cells expressed Ki67. In larger vessels we found pericytes that expressed CD146 and alpha-SMA; scarce alpha-SMA-expressing spindle-shaped cells lining cardiomyocytes were suggestive of a pericytic role in angiogenic sprout guidance. The TEM study showed that endothelial tubes are almost exclusively found in the narrow myocardial interstitia. ECs that built them up appeared identical to the cells that previous TEM studies have suggested to be myocardial telocytes. A subset of stromal cells with TC-like phenotype and telopodes-like processes actually seem to configure blood vessels, and therefore belong to the endothelial lineage. This study shows that data presented in previous studies on myocardial telocytes is not enough to allow the reproducibility of the results. At least a subset of cells considered to be TCs might belong to the endothelial lineage. (C) 2016 Elsevier GmbH. All rights reserved.

Abstract

Several 3,5-disubstituted isoxazoles were obtained in good yields by regiospecific 1,3-dipolar cycloaddition reactions between aromatic nitrile oxides, generated in situ from the corresponding hydroxyimidoyl chlorides, with non-symmetrical activated alkynes in the presence of catalytic amounts of copper(I) iodide. Effects of 3,5-disubstituted isoxazoles on nitric oxide and reactive oxygen species generation in Arabidopsis tissues was studied using specific diaminofluoresceine dyes as fluorescence indicators.

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The synthesis and physicochemical characterization of riboflavin (RF) loaded Myrj52-silver nanoparticles (SNPs) were performed and their cytotoxic properties were evaluated. Transmission electron microscopy analysis showed the formation of colloidal spherical SNPs with an average size of similar to 12 nm. The structure, stability, dynamics and conformation of human serum albumin (HSA) binding RF on SNPs have been studied. The influence of RF on the secondary structure of HSA in the presence of SNPs showed a predominant alpha-helix structure (61.7%). No changes induced by RF binding to HSA were observed. The cytotoxicity of RF and RF-HSA loaded Myrj52-SNPs was evaluated in vitro using the Neutral Red assay and cell morphology observations in a culture of L929 fibroblast cells. The results have relevance with regard to the RF loaded Myrj52-SNP formulation for pharmaceuticals.

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This study has been carried out to investigate a new method to improve metallic implant bioactivity for osseointegration by producing a calcium phosphate (Ca-P) surface layer using an immersion process in the Simulated Body Fluid (SBF) of Polypyrrole (PPy) and Polypyrrole/poly(sodium-4styrensulfonate) (NaPSS) conductive hybrid polymers electrochemically synthesized on Ti6A17Nb alloy. As formed PPy/Ca-P and PPy/ PSS-/Ca-P films were characterized electrochemically by Cyclic Voltammetry (CV), Tafel plots and spectroscopically using FT-IR technique. Also, in vitro cytotoxicity of these new organic/inorganic hybrids films was evaluated by yellow 3- (4,5-dimethylthiazolyI-2)-2,5-diphenyltetrazolium bromide (MTT) assay after cultivation in a primary culture of osteoblast cells. The PPy/Ca-P hybrid film showed the best electrochemical stability and the highest value of cell viability.

Abstract

The growing scientific interest in exploitation of vegetable bioactives has raised a number of questions regarding their imminent presence in pharmaceutical formulations. This study intends to demonstrate that a dual combination between vegetable oil (e.g. thistle oil, safflower oil, sea buckthorn oil) and a carrot extract represents an optimal approach to formulate safe carrier systems that manifest cell regeneration effect and promising antioxidant and anti-inflammatory activity. Inclusion of both natural actives into lipid carriers imparted a strong negative charge on the nanocarrier surface (up to -45 mV) and displayed average sizes of 70 nm to 140 nm. The entrapment efficiency of carrot extract into nanostructured carriers ranged between 78.3 and 88.3%. The in vitro release study has demonstrated that the entrapment of the extract represents a viable way for an equilibrated release of carotenoids. Besides the excellent antioxidant properties (e.g. scavenging up to 98% of the free oxygen radicals), the results of cellular integrity (e.g. cell viability of 133%) recommend these nanocarriers based on dual carrot extract-bioactive oil as a promising trend for the treatment of certain disorders in which oxidative stress plays a prominent role. In addition, the lipid nanocarriers based on safflower oil and sea buckthorn oil demonstrated an anti-inflammatory effect on LPS induced THP-1 macrophages, by inhibiting the secretion of two pro inflammatory cytokines, IL-6 and TNF-alpha. (C) 2016 Elsevier B.V. All rights reserved.

Abstract

Current procedures used for skin injury reepithelization have yet to improve. At present, stem cell-based therapies provide great potential in repairing different damaged tissues due to the multipotent character of these cells. This study aimed to investigate the in vitro differentiation capacity of human adipose-derived stem cells (ASCs) into epidermal cells and to evaluate their interaction with HaCaT keratinocytes in coculture experimental models that mimicked the living environment of human skin. The mesenchymal phenotype of isolated stem cells was indicated by their spindle-shaped morphology and the expression of specific surface markers CD73, CD90, and CD105. After 21 days of cultivation in specific induction medium, the epidermal keratinocyte phenotype of differentiated ASCs was demonstrated by the expression of cytokeratin 19 and involucrin markers, at both the protein and gene level. Significant (P < 0.05) proliferation of ASCs and HaCaT cells was recorded by an MTT assay in a Transwell system coculture as compared with control cells. Flow cytometry of HaCaT-ASC direct coculture showed a significant (P < 0.05) increase in DNA content in the G2/M and S phases and a decrease in the G0/G1 phase. All these data demonstrated that ASCs could be useful in future cell-based therapies for skin wound healing.

Abstract

We defined the somatic environment in which female germinal cells develop, and performed ultrastructural analyses of various somatic cell types, with particular reference to muscle cells and follicle cells, that reside within the ovary at different stages of oogenesis. Our findings show that ovarian wall of the crayfish is composed of long muscle cells, blood cells, blood vessels and hemal sinuses. The follicle and germinal cells lie within a common compartment of ovarian follicles that is defined by a continuous basal matrix. The follicle cells form branching cords and migrate to surround the developing oocytes. A thick basal matrix separates the ovarian interstitium from ovarian follicles compartment. Transmission electron microscopy shows that inner layer of basal matrix invaginates deeply into the ovarian compartment. Our results suggest that before being surrounded by follicle cells to form follicles, oogonia and early previtellogenic oocytes reside within a niche surrounded by a basal matrix that separates them from ovarian interstitium. We found coated pits and coated vesicles in the cortical cytoplasm of previtellogenic and vitellogenic oocytes, suggesting the receptor mediated endocytosis for transfer of material from the outside of the oocytes, via follicle cells. The interstitial compartment between the inner muscular layer of the ovarian wall and the basal matrix of the ovarian follicle compartment contains muscle cells, hemal sinuses, blood vessels and blood cells. Granular hemocytes, within and outside the vessels, were the most abundant cell population in the ovarian interstitium of crayfish after spawning and in the immature ovary. (C) 2015 Wiley Periodicals, Inc.

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The aim of this study was to develop a new variant of membranes based on collagen (COL), chitosan (CHI) and alkaline phosphatase (ALP) immobilized and cross-linking with glutaraldehyde (GA) at different concentrations. The biodegradation in the presence of collagenase was investigated. Biocompatibility was evaluated by MTT assay using a mouse fibroblast cell culture type NCTC (clone 929). Non-cross-linked samples were biocompatible and membranes cross-linked with low concentrations of GA (0.04, 0.08%) were also biocompatible. However, high concentrations of GA lead to a decreased biocompatibility. The adsorption behaviour of Ca2+ ions to all membranes were evaluated using the Freundlich isotherms. Haemolytic studies were performed in order to consider their applications in biomineralization process. By the addition of collagen and ALP to chitosan, the haemolytic index decreases, the COL-CHI-ALP membrane being in the non-haemolytic domain, while the COL-CHI-ALP-GA membrane has a haemolytic index greater than 2, and is slightly haemolytic.

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The authors present and evaluate an original product in the armamentarium of the preparations for burns and wounds treatment, made solely on the basis of medicinal plants and natural ingredients. The ointment formulation comprises olive oil extract from a mixture of nine medicinal plants with wound healing activity (Calendula officinalis L., Matricariachamomilla L., Symphytumofficinale L., Hypericumperforatum L., Achilleamillefolium L., Arctiumlappa L., Plantago major L., Althaea officinalis L., Quercusrobur L.), sea buckthorn oil, lavender essential oil and as thickening agents, coconut oil, beeswax and conifer resin. The LC-MS analyses of the ethanolic extracts from the plant mixture and of the ethanolic re-extracts from oil plant mixture extract and from ointment have evidenced high levels of polyphenols like caffeic, chlorogenic, gallic and ferulic acids, as well as quercetin and rutin, all of which being known compounds with good wound healing activity. The neutral red assay has shown no cytotoxic effect on fibroblast NCTC cell line exposed to herbal extracts. Finally, the wound healing action of the submitted ointment has been clinically confirmed and highlighted by some case reports.

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The Holy Quran, the sacred Muslim religious book, is also a sum of principles and teachings covering all aspects of human life. About 750 verses (of 6236) refer to various aspects of nature, and to the relationship between man and nature. The aim of this paper is to briefly analyse some of the many ecological verses. The article highlights the eco-educational dimension of the Holy Quran by means of a qualitative investigation. It addresses the ecological considerations on the rational use of natural resources, ethical interactions between man and nature, and the rights of plants, animals and of the physical elements of the environment.

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Neurons from sensory ganglia are exposed to oxidative attack in diabetes. Altered mitochondrial morphologies are due to impaired dynamics (fusion, fission) and to cristae remodeling. This study aimed to evaluate using transmission electron microscopy mitochondrial changes in diabetic trigeminal ganglia suggestive for ignition of apoptosis, in absence of classical morphological signs of apoptosis. We used samples of trigeminal ganglia (from six type 2 diabetes human donors and five streptozotocin (STZ)-induced diabetic rats). In human diabetic samples we found three main distributions of mitochondria: (a) small dark normal mitochondria, seemingly resulted from fission processes; (b) small dark damaged mitochondria, with side-vesiculations (single-and double-coated), large matrix vesicles and cytosolic leakage of reactive species, mixed with larger light mitochondria, swollen, and with crystolysis; (c) prevailing light mitochondria. In STZ-treated rats a type (c) distribution prevailed, except for nociceptive neurons where we found a different distribution: large and giant mitochondria, suggestive for impaired mitochondrial fission, mitochondrial fenestrations, matrix vesicles interconnected by lamellar cristae, and mitochondrial leakage into the cytosol. Thus, the ultrastructural pattern of mitochondria damage in diabetic samples of sensory neurons may provide clues on the initiation of intrinsic apoptosis, even if the classical morphological signs of apoptosis are not present. Further studies, combining use of biochemical and ultrastructural techniques, may allow a better quantification of the degree in which mitochondrial damage, with membrane alterations and cytosolic leaks, may be used as morphological signs suggesting the point-of-no return for apoptosis. (C) 2016 Wiley Periodicals, Inc.

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Chitosan is a non-toxic, biocompatible, biodegradable natural cationic polymer known for its low imunogenicity, antimicrobial, antioxidant effects and wound-healing activity. To improve its therapeutic potential, new chitosan-sulfonamide derivatives have been designed to develop new wound dressing biomaterials. The structural, morphological and physico-chemical properties of synthesized chitosan derivatives were analyzed by FT-IR, H-1 NMR spectroscopy, scanning electron microscopy, swelling ability and porosity. Antimicrobial, in vivo testing and biodegradation behavior have been also performed. The chitosan derivative membranes showed improved swelling and biodegradation rate, which are important characteristics required for the wound healing process. The antimicrobial assay evidenced that chitosan-based sulfadiazine, sulfadimethoxine and sulfamethoxazole derivatives were the most active. The MTT assay showed that some of chitosan derivatives are nontoxic. Furthermore, the in vivo study on burn wound model induced in Wistar rats demonstrated an improved healing effect and enhanced epithelialization of chitosan-sulfonamide derivatives compared to neat chitosan. The obtained results strongly recommend the use of some of the newly developed chitosan derivatives as antimicrobial wound dressing biomaterials. (C) 2016 Elsevier Ltd. All rights reserved.

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Chondroitin-4 sulfate (CS4) is the main glycosaminoglycan extracted from bovine trachea. CS4 play an important role in osteoarthritis treatment, anticoagulant activity, reduces the degradation of cartilage matrix components, reduces necrosis and apoptosis of chondrocytes and reduces the activity of collagenase. Chondroitin sulfate is also responsible for proteoglycans degradation. Chondroitin sulfate can bind calcium ions with different affinities, depending on their sulfation position. The purpose of this study was to determine the structural properties and the influence of Ca2+ cations. We carried out measurements on CS4 solutions and mixtures of liquid CS4 with Ca2+ by Small-Angle Neutron Scattering (SANS). CS4 have a mass fractal behavior and the addition of a salt (CaCl2) in CS4 solutions generates the appearance of a correlation peak due to local ordering between adjacent chains with inter-chain distances between 483 angstrom and 233 angstrom for a calcium concentration of 0.01% w/w.

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Telocytes (TCs) are typically defined as cells with telopodes by their ultrastructural features. Their presence was reported in various organs, however little is known about their presence in human trigeminal ganglion. To address this issue, samples of trigeminal ganglia were tested by immunocytochemistry for CD34 and examined by transmission electron microscopy (TEM). We found that TCs are CD34 positive and form networks within the ganglion in close vicinity to microvessels and nerve fibers around the neuronal-glial units (NGUs). TEM examination confirmed the existence of spindle-shaped and bipolar TCs with one or two telopodes measuring between 15 to 53 mu m. We propose that TCs are cells with stemness capacity which might contribute in regeneration and repair processes by: modulation of the stem cell activity or by acting as progenitors of other cells present in the normal tissue. In addition, further studies are needed to establish if they might influence the neuronal circuits.

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The present work reports on the biocompatibility of poly (3-hydroxybutyrate) and poly (3-hydroxybutyrateco-3-hydroxyvalerate) loaded with bacterial cellulose and microcrystalline cellulose via melt processing. Biocompatibility was tested by physico-chemical and in vitro methods. Physico-chemical tests of biocomposites, such as reducing substances, acidity, alkalinity, absorbance by UV/VIS, residue on evaporation were performed on aqueous extract. The cell viability was evaluated by MTT assay. Cell morphology evaluation of cell culture treated with composites was visualized by light microscopy. Also, thermal properties of biocomposites were investigated by DSC analysis. The obtained results have shown good biocompatibility of all biocomposites.

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Introduction: We have identified some critical aspects concerning umbilical cord tissue mesenchymal stem cells: the lack of standards for cell isolation, expansion and cryopreservation, the lack of unanimous opinions upon their multilineage differentiation potential and the existence of very few results related to the functional characterization of the cells isolated from cryopreserved umbilical cord tissue. Umbilical cord tissue cryopreservation appears to be the optimal solution for umbilical cord tissue mesenchymal stem cells storage for future clinical use. Umbilical cord tissue cryopreservation allows mesenchymal stem cells isolation before expected use, according with the specific clinical applications, by different customized isolation and expansion protocols agreed by cell therapy institutions. Methods: Using an optimized protocol for umbilical cord tissue cryopreservation in autologous cord blood plasma, isolation explant method and growth media supplemented with FBS or human serum, we performed comparative studies with respect to the characteristics of mesenchymal stem cells (MSC) isolated from different compartments of the same umbilical cord tissue such as Wharton's jelly, vein, arteries, before cryopreservation (pre freeze) and after cryopreservation (post thaw). Results: Expression of histochemical and immunohistochemical markers as well as electron microscopy observations revealed similar adipogenic, chondrogenic and osteogenic differentiation capacity for cells isolated from pre freeze and corresponding post thaw tissue fragments of Wharton's jelly, vein or arteries of the same umbilical cord tissue, regardless growth media used for cells isolation and expansion. Discussion: Our efficient umbilical cord tissue cryopreservation protocol is reliable for clinical applicability of mesenchymal stem cells that could next be isolated and expanded in compliance with future accepted standards. (C) 2015 Elsevier Ltd. All rights reserved.

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Intraluminal contents of benign and malignant prostatic tissue are associated with varying forms of acellular structures. These include corpora amylacea, prostatic calculi, and prostatic crystalloids. There are relatively few microscopy studies about the characterization of intraluminal structures from benign and malignant prostatic glands and little is known about their chemical composition. In the present study, we used a combination of special histochemical methods, immunohistochemistry, and transmission electron microscopy to characterize intraluminal contents of benign and malignant prostate glands. The study was done on 33 radical prostatectomy and four transurethral resections of prostate specimens. Histochemical methods such as von Kossa, autometallography (AMG), as well as PSA immunohistochemistry and transmission electron microscopy were performed to characterize intraluminal contents of benign and malignant prostate glands. Von Kossa staining was observed in acellular structures, corpora amylacea, prostatic calculi, and calcified blood vessels. AMG staining was observed in the lumen of small glands, in the epithelium lining prostate glands, and corpora amylacea. PSA staining showed prostatic glands with both positive and negative corpora amylacea and epithelial cells. Ultrastructural observation revealed the presence of a variety of highly heterogeneous aggregates composed of fibrillar elements that were similar to those of amyloid.

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Polymeric nanoparticles (NPs) are known to facilitate intracellular uptake of drugs to improve their efficacy, with minimum bioreactivity. The goal of this study was to assess cellular uptake and trafficking of PLGA NPs and chitosan (Chi)-covered PLGA NPs in Madin-Darby bovine kidney (MDBK) and human colorectal adenocarcinoma (Colo 205) cells. Both PLGA and Chi-PLGA NPs were not cytotoxic to the studied cells at concentrations up to 2500 g/mL. The positive charge conferred by the chitosan deposition on the PLGA NPs improved NPs uptake by MDBK cells. In this cell line, Chi-PLGA NPs colocalized partially with early endosomes compartment and showed a more consistent perinuclear localization than PLGA NPs. Kinetic uptake of PLGA NPs by Colo 205 was slower than that by MDBK cells, detected only at 24 h, exceeding that of Chi-PLGA NPs. This study offers new insights on NP interaction with target cells supporting the use of NPs as novel nutraceuticals/drug delivery systems in metabolic disorders or cancer therapy. (c) 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 3599-3611, 2015.

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The aim of this work was to characterize the physico-chemical properties of 3-hydroxyflavone (3-HF) in a silver nanoparticles complex (SNPs) using UV-vis and Fluorescence spectroscopy, Atomic Force Microscopy (AFM) and Transmission Electron Microscopy (TEM) analysis. One also evaluated its effect on the cell viability and morphology of L929 mouse fibroblast cells in vitro. The contribution of the carrier protein, Bovine Serum Albumin (BSA) to 3-HF properties has also been investigated. 3-HF in BSA/SNPs systems presented no cytotoxic effect in L929 mouse fibroblast cells at any of the tested concentrations. The results are discussed with relevance to the oxidative stress process.

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The Romanian propolis was extracted in five different media, respectively, in water (pH 6.8), glycine buffer (pH 2.5), acetate buffer (pH 5), phosphate buffer (pH 7.4) and carbonate buffer (pH 9.2). The extracts presented different amounts of flavonoids and phenolic acids, increasing pH leading to higher concentrations of active compounds. Five variants of gold nanoparticles suspensions based on different pH Romanian propolis aqueous extracts were successfully synthesized. The obtained nanoparticles presented dimensions between 20 and 60 nm in dispersion form and around 18 nm in dried form, and different morphologies (spherical, hexagonal, triangular). Fourier transform infrared spectroscopy proved the attachment of organic compounds from propolis extracts to the colloidal gold suspensions and X-ray diffraction certified that the suspensions contain metallic gold. The obtained propolis gold nanoparticles do not exhibit any antibacterial or antifungal activity, but presented different catalytic activities and toxicity on tumour cells.

Abstract

Recent studies provided evidence that mesenchymal stem cells (MSCs) have regenerative potential in cutaneous repair and profound immunomodulatory properties making them a candidate for therapy of neuroimmunologic diseases. Neuromyelitis optica (NMO) is an autoimmune, demyelinating central nervous system disorder characterized by a longitudinally extensive spinal cord lesion. A 46-year-old male diagnosed with NMO had relapses with paraplegia despite treatment and developed two stage IV pressure ulcers (PUs) on his legs. The patient consented for local application of autologous MSCs on PUs. MSCs isolated from the patient's bone marrow aspirate were multiplied in vitro during three passages and embedded in a tridimensional collagen-rich matrix which was applied on the PUs. Eight days after MSCs application the patient showed a progressive healing of PUs and improvement of disability. Two months later the patient was able to walk 20 m with bilateral assistance and one year later he started to walk without assistance. For 76 months the patient had no relapse and no adverse event was reported. The original method of local application of autologous BM-MSCs contributed to healing of PUs. For 6 years the patient was free of relapses and showed an improvement of disability. The association of cutaneous repair, sustained remission of NMO and improvement of disability might be explained by a promotion/optimization of recovery mechanisms in the central nervous system even if alternative hypothesis should be considered. Further studies are needed to assess the safety and efficacy of mesenchymal stem cells in NMO treatment.

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Novel biocompatible and antimicrobial composites based on poly(lactic acid) (PLA), hydrolyzed collagen (HC) and silver nanoparticles (AgNPs) were prepared by melt processing. Tributyl o-acetyl citrate (ATBC) was used as plasticizer, for improving the processability of PLA. The influence of HC and AgNPs on the PLA bionanocomposites was investigated in terms of biocompatibility, antifungal activity and water contact angle measurements. Surface morphology by SEM and the identification of AgNPs by UV-Vis were also presented. The investigated bionanocomposites exhibited the characteristic plasmonic effect of silver nanoparticles. All composites showed a high degree of biocompatibility. Sample containing HC 5 wt.% and AgNPs showed a significant antifungal property, inhibiting fungal adhesion and mature biofilm development. The increased hydrophilicity determined by contact angle analysis for the PLA/HC10/AgNPs composite did no contributed significantly to improving of anti-adherence effect. 10 wt.% HC promoted fungal colonization and mature biofilm development.

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