Anca Oancea

Several strategies promote phyllosphere colonization by soil-born Trichoderma plant-beneficial strains. One of these strategies is foliar spraying with suspensions containing large amounts of chlamydospores—spores with thick cell wall structures that make them highly resistant to harsh environmental conditions. Trichoderma biomass was produced by cultivation on a cornmeal medium and compared with the biomass produced on potato dextrose broth by microscopic and thermogravimetric analyses. The analyses revealed increased chlamydospore content and thermostability in the fungal biomass produced on the corn meal medium. The Trichoderma suspension rich in chlamydospores was sprayed on bitter gourd (Momordica charantia) leaves at two inoculant concentrations, 10⁶ and 10⁸ ufc/mL. The effect of these treatments on the plant physiological parameters, leaf photosynthetic pigments, polyphenol and flavonoid contents, antioxidant activities of the leaves and fruits, and yield was compared to the control (plants sprayed with water) and to the experimental treatment involving spraying with 10⁸ ufc/mL of propagules produced in potato dextrose broth. The effect of chlamydospore-rich suspensions on plant physiological parameters was more pronounced and long-lasting compared with the other treatments. The treatment with chlamydospore-rich suspension enhanced the accumulation of polyphenols and flavonoids in the leaves (by 17% and 50%, respectively) and fruits (by 18% and 31%, respectively) and increased the antioxidant activity. The Trichoderma treatment increased the yield by +25.33–53.07%. The application of the foliar treatment with Trichoderma suspensions did not modify the cytocompatibility of the extracts from the fruits determined on the L929 cells.

Several strategies promote phyllosphere colonization by soil-born Trichoderma plant-beneficial strains. One of these is foliar spraying with suspensions containing large amounts of chlamydospores – spores with a thick cell wall structure that makes them highly resistant to harsh environmental conditions. Trichoderma biomass was produced by cultivation on cornmeal medium and compared by microscopic and thermogravimetric analysis with the biomass produced in potato dextrose broth. Both analyses revealed higher chlamydospores content and thicker mycelium in the fungal biomass produced on cornmeal medium. The Trichoderma suspension rich in chlamydospores was sprayed on bitter gourd (Momordica charantia) leaves at two concentration levels, 106 and 108 ufc/ml. The effect of these treatments on plant physiological parameters, leaf photosynthetic pigments, polyphenol and flavonoid contents, antioxidant activity of leaves and fruits and yield was compared to the control (plants sprayed with water) and to the experimental treatment sprayed with 108 ufc propagules produced in potato dextrose broth. The effect of chlamydospore-rich suspensions on plant physiological parameters was more pronounced and long-lasting compared with the other treatments. The treatment with chlamydospore-rich suspension enhanced the accumulation of polyphenols and flavonoids in the leaves and fruits and increased the antioxidant activity. The application of the foliar treatment with Trichoderma suspensions did not modify the cytocompatibility of the extracts from fruits determined on L929 cells.

Biocompatible drug-delivery systems for soft tissue applications are of high interest for the medical and pharmaceutical fields. The subject of this research is the development of hydrogels loaded with bioactive compounds (inulin, thyme essential oil, hydro-glycero-alcoholic extract of Vitis vinifera, Opuntia ficus-indica powder, lactic acid, citric acid) in order to support the vaginal microbiota homeostasis. The nanofibrillar phyto-hydrogel systems developed using the biocompatible polymers chitosan (CS), never-dried bacterial nanocellulose (NDBNC), and Poloxamer 407 (PX) incorporated the water-soluble bioactive components in the NDBNC hydrophilic fraction and the hydrophobic components in the hydrophobic core of the PX fraction. Two NDBNC-PX hydrogels and one NDBNC-PX-CS hydrogel were structurally and physical-chemically characterized using Fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), and rheology. The hydrogels were also evaluated in terms of thermo-responsive properties, mucoadhesion, biocompatibility, and prebiotic and antimicrobial effects. The mucin binding efficiency of hydrogel base systems was determined by the periodic acid/Schiff base (PAS) assay. Biocompatibility of hydrogel systems was determined by the MTT test using mouse fibroblasts. The prebiotic activity was determined using the probiotic strains Limosilactobacillus reuteri and Lactiplantibacillus plantarum subsp. plantarum. Antimicrobial activity was also assessed using relevant microbial strains, respectively, E. coli and C. albicans. TEM evidenced PX micelles of around 20 nm on NDBNC nanofibrils. The FTIR and XRD analyses revealed that the binary hydrogels are dominated by PX signals, and that the ternary hydrogel is dominated by CS, with additional particular fingerprints for the biocompounds and the hydrogel interaction with mucin. Rheology evidenced the gel transition temperatures of 18–22 °C for the binary hydrogels with thixotropic behavior and, respectively, no gel transition, with rheopectic behavior for the ternary hydrogel. The adhesion energies of the binary and ternary hydrogels were evaluated to be around 1.2 J/m² and 9.1 J/m², respectively. The hydrogels exhibited a high degree of biocompatibility, with the potential to support cell proliferation and also to promote the growth of lactobacilli. The hydrogel systems also presented significant antimicrobial and antibiofilm activity.

Osteochondral regeneration is a major challenge due to the different composition of cartilage and subchondral bone and different biochemical, biomechanical and biological properties. For this reason, a biomimetic scaffold is necessary to provide different biological signals needed to allow for osteochondral regeneration [¹]. This study aims to design novel biodegradable cross-linked composite hydrogels based on gelatin and polysaccharidic components (chondroitin-4-sulphate and hyaluronic acid), mixed with akermanite and enriched with small bioactive molecule (icariin). Akermanite was used as a better alternative to conventional ceramics due to its bone-like apatite formation ability and good bioactivity [²]. Icariin (Ica) flavonoid (from traditional Chinese medicine Epimedium herb) was used as substitute for growth factors to enhance cell proliferation and chondrogenic and osteogenic differentiation [3]. Variants of biodegradable cross-linked composite hydrogel based on gelatin, polysaccharidic components (chondroitin-4-sulphate and hyaluronic acid), in two ratios of 2:0.8:0.2 and 2:0.08:0.02 (w/w/w), were developed and mixed with akermanite, at a ratio of 2:1 (w/w). Subsequently, both composite hydrogel variants were cross-linked with (N,N-(3-dimethylaminopropyl)-N-ethyl carbodiimide (EDC) and enriched with small bioactive molecule (icariin). The obtained cross-linked composite hydrogel variants enriched with Ica were characterized related to enzymatic biodegradation (type I collagenase), swelling capacity, degree of cross-linking (TNBS assay), and morphology (SEM). Their cytocompatibility was evaluated by analyses of cell viability and cellular cycle (flow cytometry), cell proliferation (Neutral Red assay), and cell adhesion to composite hydrogels (SEM) using NCTC clone L929 cell line. The final results show that both cross-linked composite hydrogel variants enriched with Ica presented optimal physicochemical and structural properties to be used as a scaffold for osteochondral healing. Our data did not reveal any toxicity of composite hydrogels in the NCTC cell line within the tested range of concentrations (10–50 mg/mL). Additionally, cells were capable of spreading and proliferating on the surface of composite hydrogels. The designed biodegradable cross-linked composites enriched with Ica are recommended for further studies as natural temporary scaffolds, which can allow both cartilage and subchondral regeneration with implications for the management of osteochondral healing.

The skin, which represents about 16% of the total body mass, acts as a protective barrier against external microbial factors [¹]. Therefore, damaged tissues, especially burns, require rapid local coverage to avoid infections and to ensure the protective barrier function of the skin [²]. The aim of this study was to design and characterize new hydrogel formulations based on natural and synthetic polymers and that were biodegradable and cytocompatible to serve as temporary dressings with regenerative properties for skin wound healing. The proposed experimental variants of the hydrogels are based on mixtures of gelatin (Gel), sodium alginate (Alg), polyvinyl alcohol (PVA), and methylcellulose (MC1500) in different weight ratios: Gel-Alg (1:0.75, g/g), Gel-Alg-PVA (1:0.27:0.18, g/g/g) and Gel-Alg-MC1500 (1:0.26:0.35, g/g/g). Physicochemical and biochemical characterizations were performed to determine the swelling degree, biodegradation in physiological conditions (pH 7.4, 37 °C) and in the presence of collagenase (mimicking the inflamed wounded milieu), viscosity, and syneresis, while their ultrastructure was investigated by SEM analysis [³]. The L929 murine fibroblast culture was used to assess the in vitro cytocompatibility of the hydrogels after 24 h and 48 h of cultivation using quantitative MTT and LDH assays [⁴]. Cell morphology was observed in treated cultures by light microscopy after Giemsa staining. The physicochemical and biochemical analyses indicated that the novel polymeric hydrogels variants had a good swelling capacity due to the presence of Alg, had an adjustable viscosity, and controlled biodegradation over time in both physiological and inflamed conditions. Two mixture variants were outlined: Gel-Alg-PVA with reduced porosity and low biodegradability over time and Gel-Alg-MC1500 with increased porosity and higher biodegradation over time, even in the physiological environment. The SEM morphology observations showed that the hydrogels had a dense and microporous structure, with pores of irregular shapes and sizes, which could ensure skin protection against external microbial agents while also maintaining the required degree of humidity and oxygen exchange with the external environment. In vitro quantitative tests indicated a high degree of cytocompatibility for all of the tested hydrogels, with cell viability percentages higher than 90%. The cell morphology observations revealed that in the presence of hydrogel samples, the L929 murine fibroblasts maintained their normal phenotype, and the cell density was similar to that of the negative control (untreated cells). Overall, our findings indicated that the hydrogels containing synthetic polymers (Gel-Alg-PVA, Gel-Alg-MC1500) had adequate physicochemical, biochemical, and biological properties that should be further tested to determine their role as biomaterials for skin tissue engineering applications.

The aim of this work was to obtain a mixture of polyphenols and polysaccharides, extracted from forest fruit pomace (a side stream of forest fruit juice production) and chia seeds with a synergic antioxidant activity that will be further encapsulated and used in a fortified protein bar

The Romanian economy is dominated by linear value-chains

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 µg/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.

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-α), 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.

Freshwater fish skin and bones are usually by-products of the fish processing industry.