Bacterial cellulose (BC) is a natural polymer with properties suitable for tissue engineering and possible applications in scaffold production. However, current procedures have limitations in obtaining BC pellicles with the desired structural, physical, and mechanical properties. Thus, this study analyzed the optimal culture conditions of BC membranes and two types of processing: draining and oven-drying. The aim was to obtain BC membranes with properties suitable for a wound dressing material. Two studies were carried out. In the preliminary study, the medium (100 mL) was inoculated with varying volumes (1, 2, 3, 4, and 5 mL) and incubated statically for different periods (3, 6, 9, 12, and 18 days), using a full factorial experimental design. Thickness, uniformity, weight, and yield were evaluated. In the optimization study, a Box–Behnken design was used. Two independent variables were used: inoculum volume (X1: 1, 3, and 5 mL) and fermentation period (X2: 6, 12, and 18 d) to determine the target response variables: thickness, swelling ratio, drug release, fiber diameter, tensile strength, and Young’s modulus for both dry and moist BC membranes. The mathematical modelling of the effect of the two independent variables was performed by response surface methodology (RSM). The obtained models were validated with new experimental values and confirmed for all tested properties, except Young’s modulus of oven-dried BC. Thus, the optimal properties in terms of a scaffold material of the moist BC were obtained with an inoculum volume of 5% (v/v) and 16 d of fermentation. While, for the oven-dried membranes, optimal properties were obtained with a 4% (v/v) and 14 d of fermentation.
Anthocyanins are extensively studied for their health-related properties, including antibacterial activity against urinary tract infections (UTI). Among common fruits, blueberries, with their remarkable antioxidant capacity, are one of the richest sources. Anthocyanin-rich extracts were obtained from four varieties: Snowchaser, Star, Stella Blue and Cristina Blue, grown in the hot climate of Southern Spain. Their total anthocyanins contents (TAC) were determined spectrophotometrically, and the anthocyanin profile by ultra high performance liquid chromatography—tandem mass spectrometer (UHPLC-MS/MS). Their antioxidant activity was assessed by oxygen radical absorbance capacity (ORAC) assay, while antibacterial activity against strains isolated from UTI patients was assessed in vitro, helping to select the varieties with the highest bioactive potential. Star showed the highest TAC and antioxidant activity (1663 ± 159 mg of cyanidin-3-O-glucoside (cy-3-O-glu) equivalents/100 g fresh weight (FW), 6345 ± 601 μmol Trolox equivalents (TE)/100 g FW, respectively), followed by Cristina Blue, Stella Blue and Snowchaser. As far as we know, this is the first time that cyanidin-3-rutinoside has been identified in blueberries. The extracts inhibited all the tested strains, MICs ranging from 0.4 mg/mL (for Stella Blue extract against UTI P. aeruginosa) to 9.5 mg/mL (for all extracts against UTI K. pneumoniae ssp. pneumoniae). This is the first study that assessed in vitro the antibacterial activity of blueberries against Klebsiella pneumoniae, Providencia stuartii and Micrococcus spp. strains isolated from UTI.
The probiotic properties and the viability of lactic acid bacteria of fermented dairy products can be improved by addition of bioactive compounds originating from natural sources (e.g. goji berries). This study aimed to evaluate how goji berries and honey affect the sensorial quality of yoghurt, the chemical properties, the viability of lactic acid bacteria (LAB) and the concurrent microflora development. Two types of yoghurts (yoghurt with goji berries and yoghurt with honey and goji berries) were developed. The addition of honey affected the entire yoghurt microflora including LAB, manifesting bactericidal effect. The addition of goji berries maintained the viability of LAB at probiotic levels (10 6 -10 7 log CFU/ml) during 21 days of storage; compared to classic yoghurt, LAB viability decreased during storage at 10 3 log CFU/ml. Goji berries also improved sensory acceptance of consumers. The results obtained in this study collect information that enables the use of goji berries as enhancer of probiotic levels in yoghurt, while honey can provide bacteriostatic/bactericidal effect for contaminants.
The probiotic properties and the viability of lactic acid bacteria of fermented dairy products can be improved by addition of bioactive compounds originating from natural sources (e.g. goji berries). This study aimed to evaluate how goji berries and honey affect the sensorial quality of yoghurt, the chemical properties, the viability of lactic acid bacteria (LAB) and the concurrent microflora development. Two types of yoghurts (yoghurt with goji berries and yoghurt with honey and goji berries) were developed. The addition of honey affected the entire yoghurt microflora including LAB, manifesting bactericidal effect. The addition of goji berries maintained the viability of LAB at probiotic levels (106-107 log CFU/ml) during 21 days of storage; compared to classic yoghurt, LAB viability decreased during storage at 103 log CFU/ml. Goji berries also improved sensory acceptance of consumers. The results obtained in this study collect information that enables the use of goji berries as enhancer of probiotic levels in yoghurt, while honey can provide bacteriostatic/bactericidal effect for contaminants.
Tissue engineering promotes tissue regeneration through biomaterials that have excellent properties and have the potential to replace tissues. Many studies show that bacterial cellulose (BC) might ensure tissue regeneration and substitution, being used for the bioengineering of hard, cartilaginous and soft tissues. Bacterial cellulose is extensively used as wound dressing material and results show that BC is a promising tissue scaffold (bone, cardiovascular, urinary tissue). It can be combined with polymeric and non-polymeric compounds to acquire antimicrobial, cell-adhesion and proliferation properties. To ensure proper tissue regeneration, the material has to be: biocompatible, with minimum tissue reaction and biodegradability; bio-absorbable, to promote tissue development, cellular interaction and grow; resistant to support the weight of the newly formed tissue. Its versatile structure, physical and biochemical properties can be adjusted by adapting the bacteria culturing conditions. The main biomedical applications seem to be as hard (bone, dental), fibrocartilaginous (meniscal) and soft tissue (skin, cardiovascular, urinary) substituents. This paper reviews the current state of knowledge, challenges and future applications of BC and its biomedical potential in veterinary medicine. It was focused on the main uses in regeneration and scaffold tissue replacement and, although BC showed promising results, there is a lack of successful results of BC use in clinical practice. Most studies were performed only at experimental level and further research is needed for BC to enter clinical veterinary practice.
Cylindrospermopsin (CYN) and microcystins (MC) are cyanotoxins that can occur simultaneously in contaminated water and food. CYN/MC-LR mixtures previously investigated in vitro showed an induction of micronucleus (MN) formation only in the presence of the metabolic fraction S9. When this is the case, the European Food Safety Authority recommends a follow up to in vivo testing. Thus, rats were orally exposed to 7.5 + 75, 23.7 + 237, and 75 + 750 μg CYN/MC-LR/kg body weight (b.w.). The MN test in bone marrow was performed, and the standard and modified comet assays were carried out to measure DNA strand breaks or oxidative DNA damage in stomach, liver, and blood cells. The results revealed an increase in MN formation in bone marrow, at all the assayed doses. However, no DNA strand breaks nor oxidative DNA damage were induced, as shown in the comet assays. The histopathological study indicated alterations only in the highest dose group. Liver was the target organ showing fatty degeneration and necrotic hepatocytes in centrilobular areas, as well as a light mononuclear inflammatory periportal infiltrate. Additionally, the stomach had flaking epithelium and mild necrosis of epithelial cells. Therefore, the combined exposure to cyanotoxins may induce genotoxic and histopathological damage in vivo.
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