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The present study evaluated three green extraction methods, accelerated solvent extraction (ASE), ultrasound-assisted extraction (UAE), and laser irradiation extraction (LE), for the polyphenolic compounds and vitamin C extraction of Cornus mas L. and Crataegus monogyna fruit extracts. The polyphenols and vitamin C of extracts were quantified using HPLC-DAD, and the total phenolic content, flavonoid content, antioxidant activity (DPPH and reducing power), and antidiabetic activity were also studied. The antidiabetic activity was examined by the inhibition of alpha-amylase and alpha-glucosidase, and in vitro on a beta TC cell line (beta-TC-6). The results showed significant differentiation in the extraction yield between the methods used, with the ASE and LE presenting the highest values. The C. mas fruit extract obtained by ASE exhibited the best antioxidant activity, reaching an IC50 value of 31.82 +/- 0.10 mu g/mL in the DPPH assay and 33.95 +/- 0.20 mu g/mL in the reducing power assay. The C. mas fruit extracts obtained by ASE and LE also have the highest inhibitory activity on enzymes associated with metabolic disorders: alpha-amylase (IC50 = 0.44 +/- 0.02 mu g/mL for the extract obtained by ASE, and 0.11 +/- 0.01 mu g/mL for the extract obtained by LE at combined wavelengths of 1270 + 1550 nm) and alpha-glucosidase (IC50 of 77.1 +/- 3.1 mu g/mL for the extract obtained by ASE, and 98.2 +/- 4.7 mu g/mL for the extract obtained by LE at combined wavelengths of 1270 + 1550 nm). The evaluation of in vitro antidiabetic activity demonstrated that the treatment with C. mas and C. monogyna fruit extracts obtained using ASE stimulated the insulin secretion of beta-TC-6 cells, both under normal conditions and hyperglycemic conditions, as well. All results suggest that C. mas and C. monogyna fruit extracts are good sources of bioactive molecules with antioxidant and antidiabetic activity.

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Purpose: In this study, wound dressings were designed using zinc-modified marine collagen porous scaffold as host for wild bilberry (WB) leaves extract immobilized in functionalized mesoporous silica nanoparticles (MSN). These new composites were developed as an alternative to conventional wound dressings. In addition to the antibacterial activity of classic antibiotics, a polyphenolic extract could act as an antioxidant and/or an anti-inflammatory agent as well. Methods: Wild bilberry leaves extract was prepared by ultrasound-assisted extraction in ethanol and its properties were evaluated by UV-Vis spectroscopy (radical scavenging activity, total amount of polyphenols, flavonoids, anthocyanins, and condensed tannins). The extract components were identified by HPLC, and the antidiabetic properties of the extract were evaluated via alpha-glucosidase inhibitory activity. Spherical MSN were modified with propionic acid or proline moieties by post-synthesis method and used as carriers for the WB leaves extract. The textural and structural features of functionalized MSN were assessed by nitrogen adsorption/desorption isotherms, small-angle XRD, SEM, TEM, and FTIR spectroscopy. The composite porous scaffolds were prepared by freeze drying of the zinc-modified collagen suspension containing WB extract loaded silica nanoparticles. Results: The properties of the new composites demonstrated enhanced properties in terms of thermal stability of the zinc-collagen scaffold, without altering the protein conformation, and stimulation of NCTC fibroblasts mobility. The results of the scratch assay showed contributions of both zinc ions from collagen and the polyphenolic extract incorporated in functionalized silica in the wound healing process. The extract encapsulated in functionalized MSN proved enhanced biological activities compared to the extract alone: better inhibition of P. aeruginosa and S. aureus strains, higher biocompatibility on HaCaT keratinocytes, and anti-inflammatory potential demonstrated by reduced IL-1 beta and TNF-alpha levels. Conclusion: The experimental data shows that the novel composites can be used for the development of effective wound dressings.

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The present study evaluated the antioxidant and antidiabetic properties of Medicago sativa and Solidago virgaurea extracts enriched in polyphenolic compounds. The extracts were obtained by accelerated solvent extraction (ASE) and laser irradiation. Then, microfiltration was used for purification, followed by nanofiltration used to concentrate the two extracts. The obtained extracts were analyzed to determine their antioxidant activity using DPPH radical scavenging and reducing power methods. The antidiabetic properties have been investigated in vitro on a murine insulinoma cell line (beta-TC-6) by the inhibition of alpha-amylase and alpha-glucosidase. M. sativa obtained by laser irradiation and concentrated by nanofiltration showed the highest DPPH center dot scavenging (EC50 = 105.2 +/- 1.1 mu g/mL) and reducing power activities (EC50 = 40.98 +/- 0.2 mu g/mL). M. sativa extracts had higher inhibition on alpha-amylase (IC50 = 23.9 +/- 1.2 mu g/mL for concentrated extract obtained after ASE, and 26.8 +/- 1.1), while S. virgaurea had the highest alpha-glucosidase inhibition (9.3 +/- 0.9 mu g/mL for concentrated extract obtained after ASE, and 8.6 +/- 0.7 mu g/mL for concentrated extract obtained after laser extraction). The obtained results after evaluating in vitro the antidiabetic activity showed that the treatment with M. sativa and S. virgaurea polyphenolic-rich extracts stimulated the insulin secretion of beta-TC-6 cells, both under normal conditions and under hyperglycemic conditions as well. This paper argues that M. sativa and S. virgaurea polyphenolic-rich extracts could be excellent natural sources with promising antidiabetic potential.

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The present study aimed to conduct a comparative investigation of the biological properties of phenolic and polysaccharide extracts obtained using an ultrasound-assisted technique from Aloe vera gel and their effects on each stage of the wound healing process in in vitro experimental models. HPLC analysis showed that the phenolic extract contained aloin, ferulic, and caffeic acid, as well as quercetin dihydrate, as major compounds. Capillary zone electrophoresis indicated the prevalence of mannose and glucose in the polysaccharide extract. Cell culture testing revealed the anti-inflammatory properties of the phenolic extract at a concentration of 0.25 mg/mL through significant inhibition of pro-inflammatory cytokines—up to 28% TNF-α and 11% IL-8 secretion—in inflamed THP-1-derived macrophages, while a pro-inflammatory effect was observed at 0.5 mg/mL. The phenolic extract induced 18% stimulation of L929 fibroblast proliferation at a concentration of 0.5 mg/mL, enhanced the cell migration rate by 20%, and increased collagen type I synthesis by 18%. Moreover, the phenolic extract exhibited superior antioxidant properties by scavenging free DPPH (IC50 of 2.50 mg/mL) and ABTS (16.47 mM TE/g) radicals, and 46% inhibition of intracellular reactive oxygen species (ROS) production was achieved. The polysaccharide extract demonstrated a greater increase in collagen synthesis up to 25%, as well as antibacterial activity against Staphylococcus aureus with a bacteriostatic effect at 25 mg/mL and a bactericidal one at 50 mg/mL. All these findings indicate that the phenolic extract might be more beneficial in formulations intended for the initial phases of wound healing, such as inflammation and proliferation, while the polysaccharide extract could be more suitable for use during the remodeling stage. Moreover, they might be combined with other biomaterials, acting as efficient dressings with anti-inflammatory, antioxidant, and antibacterial properties for rapid recovery of chronic wounds. © 2024 by the authors.

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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.

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This study aimed to investigate, for the first time, the chemical composition and antioxidant activity of fluid extracts obtained from three Romanian cultivars of haskap berries (Lonicera caerulea L.) var. Loni, bitter cherries (Prunus avium var. sylvestris Ser.) var. Silva, and pomace from red grapes (Vitis vinifera L.) var. Mamaia, and their capacity to modulate in vitro steatosis, in view of developing novel anti-obesity products. Total phenolic, flavonoid, anthocyanin, and ascorbic acid content of fluid extracts was spectrophotometrically assessed and their free radical scavenging capacity was evaluated using Trolox Equivalent Antioxidant Capacity (TEAC) and free 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical inhibition assays. The Pearson coefficients showed a moderate correlation between the antioxidant activity of fluid extracts and their phenolic content, but a strong correlation between anthocyanin and ascorbic acid content. HPLC analysis identified and quantified the main phenolic compounds of chlorogenic and syringic acid, catechin, and glycosylated kaempferol, apigenin, and quercetin, in variable proportions. An in vitro experimental model of steatosis was developed in HepG2 hepatocytes treated with a mixture of free fatty acids. Cell culture analyses showed that cytocompatible concentrations of fluid extracts could significantly reduce the lipid accumulation and inhibit the reactive oxygen species, malondialdehyde, and nitric oxide secretion in stressed hepatocytes. In conclusion, these results put an emphasis on the chemical compounds' high antioxidant and liver protection capacity of unstudied fluid extracts obtained from Romanian cultivars of bitter cherries var. Silva and pomace of red grapes var. Mamaia, similar to the fluid extract of haskap berries var. Loni, in particular, the positive modulation of fat deposition next to oxidative stress and the lipid peroxidation process triggered by fatty acids in HepG2 hepatocytes. Consequently, this study indicated that these fluid extracts could be further exploited as hepatoprotective agents in liver steatosis, which provides a basis for the further development of novel extract mixtures with synergistic activity as anti-obesity products.

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Objectives: The aim of the present study was to design, produce and characterize composite substrates consisting of different formulations of poly(epsilon-caprolactone) (PCL), as a polymer matrix, and silver-containing mesoporous bioactive glasses (Ag-MBGs) with improved properties for bone tissue engineering.Methods: Ag-MBGs were synthesized by an evaporation-induced self-assembly process. Different polymer-to-particles weight ratios were considered (90/10, 80/20, 70/30 wt%). PCL/Ag-MBGs composites substrates were manufactured by melting and molding technique. The effect of Ag-MBGs embedded in the polymer matrix was investigated by morphological (field emission scanning electron microscopy (FE-SEM), SEM and contact angle measurement), structural/functional (small punch and tensile tests), antimicrobial, and in vitro biological analyses.Results: The obtained results highlighted that the inclusion of 10% by weight of Ag-MBGs improved the punching performances as well as the tensile Young's modulus (from 350.3 +/- 32.0 MPa for PCL to 473.5 +/- 41.0 MPa), without negatively altering the tensile strength of the neat PCL. Indeed, small punch test findings indicated that, over a threshold concentration (10% by weight), the Ag-MBGs acted as weak points, rather than reinforcement, because the mechanical properties of the composite substrates decreased. The bacterial growth monitoring showed a clear antimicrobial effect against both Gram-negative and Gram-positive, confirmed by reduced cell viability registered after 24 h (2 x105 CFU/mL for P. aeruginosa and 2.3 x105 CFU/mL for S. aureus). The results were confirmed in terms of adhesion and adherent growth, reduced at day 3 on PCL samples with 10% of Ag/ MBGs. Furthermore, this formulation induced a significant inhibition zone (21 mm for P. aeruginosa, 23 mm for S. aureus). In vitro biological assays confirmed that all formulations of PCL/Ag-MBGs supported periodontal ligament stem cells' viability and differentiation over time. Particularly, substrates with Ag-MBGs at a concentration of 10% and 20% by weight of Ag-MBGs provided higher values of the percentage of Alamar Blue reduction meanwhile, the highest Ag-MBGs concentration induced a higher expression of alkaline phosphatase activity.Significance: Ag-MBGs proved to be suitable candidates as filler at a specific threshold concentration (10% by weight), considering a compromise among physicochemical, antimicrobial, and pro-regenerative features. These findings provide useful data for the design and development of improved biomaterials with optimized properties, suggesting a potential application in maxillofacial bone and/or periodontal tissue repair.

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Hedera helix L. contains phytochemicals with good biological properties which are beneficial to human health and can be used to protect plants against different diseases. The aim of this research was to find the most suitable extraction method and the most favorable parameters for the extraction of different bioactive compounds from ivy leaves. Different extraction methods, namely microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), and conventional heating extraction (CHE), were used. The most suitable method for the extraction of saponins is MAE with an extraction efficiency of 58%, while for carbohydrates and polyphenols, the best results were achieved via UAE with an extraction efficiency of 61.7% and 63.5%, respectively. The antioxidant activity (AA) of the extracts was also determined. The highest AA was obtained via UAE (368.98 +/- 9.01 mu mol TR/gDM). Better results were achieved at 50( degrees)C for 10 min of extraction, using 80% ethanol in water as solvent. In order to evaluate their in vitro cytotoxicity, the extracts richest in bioactive compounds were tested on NCTC fibroblasts. Their influence on the DNA content of RAW 264.7 murine macrophages was also tested. Until 200 mu g/mL, the extracts obtained via UAE and MAE were cytocompatible with NCTC fibroblasts at 48 h of treatment. Summarizing the above, both MAE and UAE can be employed as green and efficient methods for producing extracts rich in bioactive compounds, exhibiting strong antioxidant properties and good noncytotoxic activity.

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A major clinical challenge today is the large number of bone defects caused by diseases or trauma. The development of three-dimensional (3D) scaffolds with adequate properties is crucial for successful bone repair. In this study, we prepared biomimetic mesoporous bioactive glass (MBG)-based scaffolds with and without ceria addition (up to 3 mol %) to explore the biological structure and chemical composition of the marine sponge Spongia Agaricina (SA) as a sacrificial template. Micro-CT examination revealed that all scaffolds exhibited a highly porous structure with pore diameters primarily ranging from 143.5 mu m to 213.5 mu m, facilitating bone ingrowth. Additionally, smaller pores (< 75 mu m), which are known to enhance osteogenesis, were observed. The undoped scaffold displayed the highest open porosity value of 90.83%. Cytotoxicity assessments demonstrated that all scaffolds were noncytotoxic and nongenotoxic toward osteoblast cells. Moreover, scaffolds with higher CeO2 content promoted osteogenic differentiation of dental pulp stem cells, stimulating calcium and osteocalcin secretion. The scaffolds also exhibited antimicrobial and antibiofilm effects against Staphylococcus aureus (S. aureus) as well as drug delivery ability. Our research findings indicated that the combination of MBG, natural biological structure, and the addition of Ce exhibited a synergistic effect on the structure and biological properties of scaffolds for applications in bone tissue engineering.

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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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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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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: 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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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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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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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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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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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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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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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 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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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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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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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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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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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.

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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.

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