Germination Study of Some Protein-Based Gels Obtained from By-Products from the Leather Industry on Tomato and Pepper Seeds.

This study aimed to evaluate the biostimulant effects of three protein-based gels, GHC 1-B (20% gelatin (GPU-B) obtained by thermal hydrolysis from residual untanned leather and 80% collagen hydrolysates (HCE-B) obtained by alkaline-enzymatic hydrolysis from residual bovine-tanned leather), GHC 2-B (40% keratin hydrolysate (HKU-B) obtained by alkaline-enzymatic hydrolysis from sheep wool + 40% HCE-B + 20% GPU-B), and GHC 3-B (20% GPU-B + 80% hydrolyzed collagen (HPU-B) obtained by thermal and enzymatic hydrolysis from residual untanned leather). A germination study was carried out on pepper and tomato seeds at concentrations of 1%, 3%, and 10%. As a result of the study, it was found that all three protein-based gels showed a stimulatory effect on the tomato seeds at a 1% concentration, where the Gi (germination index) was ˂100%. The GHC 2-B variant had the highest stimulatory effect (Gi-190.23%). Pepper seeds have proven to be more sensitive to the gel's composition. The concentration at which it proved to be non-inhibitory (Gi-88.29%) was 1% in the case of GHC 2-B. It was found that the presence of hydrolyzed keratin in the composition can be a plus compared to the other two protein gels tested due to its composition, which is richer in phytonutrient compounds (e.g., sulfur molecules).

Development of gallic acid-loaded ethylcellulose fibers as a potential wound dressing material.

In this study, novel fibers were designed based on ethylcellulose (EC), loaded with different concentrations of gallic acid (GA) using the electrospinning technique, in order to investigate the potential of these materials as wound dressings. The chemical structure and morphology, along with the antimicrobial and biocompatibility tests of the EC_GA fibers were investigated. To observe the chemical interactions between the components, fourier transform infrared spectroscopy (FTIR) was used. The morphological analyzes were performed using scanning electron microscope (SEM). The uniaxial tensile test machine was used to obtain mechanical performance of the fibers. MTT assay was applied to get the biocompatibility properties of the fibers and antimicrobial test was applied to obtain the antimicrobial activity of the fibers. Based on the obtained results, the highest viability value of 67.4 % was obtained for 10%EC_100GA on the third day of incubation, demonstrating that with the addition of a higher concentration of GA, the cell viability increases. The antimicrobial tests, evaluated against Staphylococcus (S.) aureus, Escherichia (E.) coli, Pseudomonas (Ps.) aeruginosa and Candida (C.) albicans, showed a >90 % microbial reduction capacity correlated with a logarithmic reduction ranging from 0.63 to 1, for 10%EC_100 GA. In vitro release tests of GA from the fibers showed that GA was totally released from 10%EC_100 GA fibers after 2880 min, demonstrating a controlled release profile. These findings demonstrated that EC_GA fibers may be suitable for application in biomedical fields such as wound dressing materials. However, further studies should be performed to increase the biocompatibility properties of the fibers.

Antimicrobial Wound Dressings based on Bacterial Cellulose and Independently Loaded with Nutmeg and Fir Needle Essential Oils.

The aim of the present study was to obtain antimicrobial dressings from bacterial cellulose loaded with nutmeg and of fir needle essential oils. The attractive properties of BC, such as biocompatibility, good physicochemical and mechanical stability, and high water absorption, led to the choice of this material to be used as a support. Essential oils have been added to provide antimicrobial properties to these dressings. The results confirmed the presence of oils in the structure of the bacterial cellulose membrane and the ability of the materials to inhibit the adhesion of Staphylococcus aureus and Escherichia coli. By performing antibacterial tests on membranes loaded with fir needle essential oil, we demonstrated the ability of these membranes to inhibit bacterial adhesion to the substrate. The samples loaded with nutmeg essential oil exhibited the ability to inhibit the adhesion of bacteria to the surface of the materials, with the 5% sample showing a significant decrease. The binding of essential oils to the membrane was confirmed by thermal analysis and infrared characterization.

In Vivo Acute Toxicity and Immunomodulation Assessment of a Novel Nutraceutical in Mice.

Achieving and maintaining a well-balanced immune system has righteously become an insightful task for the general population and an even more fundamental goal for those affected by immune-related diseases. Since our immune functions are indispensable in defending the body against pathogens, diseases and other external attacks, while playing a vital role in maintaining health and modulating the immune response, we require an on-point grasp of their shortcoming as a foundation for the development of functional foods and novel nutraceuticals. Seeing that immunoceuticals are considered effective in improving immune functions and reducing the incidence of immunological disorders, the main focus of this study was to assess the immunomodulatory properties and possible acute toxicity of a novel nutraceutical with active substances of natural origin on C57BL/6 mice for 21 days. We evaluated the potential hazards (microbial contamination and heavy metals) of the novel nutraceutical and addressed the acute toxicity according to OECD guidelines of a 2000 mg/kg dose on mice for 21 days. The immunomodulatory effect was assessed at three concentrations (50 mg/kg, 100 mg/kg and 200 mg/kg) by determining body and organ indexes through a leukocyte analysis; flow cytometry immunophenotyping of lymphocytes populations and their subpopulations (T lymphocytes (LyCD3+), cytotoxic suppressor T lymphocytes (CD3+CD8+), helper T lymphocytes (CD3+CD4+), B lymphocytes (CD3-CD19+) and NK cells (CD3-NK1.1.+); and the expression of the CD69 activation marker. The results obtained for the novel nutraceutical referred to as ImunoBoost indicated no acute toxicity, an increased number of lymphocytes and the stimulation of lymphocyte activation and proliferation, demonstrating its immunomodulatory effect. The safe human consumption dose was established at 30 mg/day.

Electrically Triggered Quercetin Release from Polycaprolactone/Bismuth Ferrite Microfibrous Scaffold for Skeletal Muscle Tissue.

Skeletal muscle tissue engineering presents a promising avenue to address the limitations pertaining to the regenerative potential of stem cells in case of injury or damage. The objective of this research was to evaluate the effects of utilizing novel microfibrous scaffolds, containing the compound quercetin (Q), on skeletal muscle regeneration. Morphological test results showed us that the combination of bismuth ferrite (BFO), polycaprolactone (PCL), and Q were bonded and well-ordered with each other, and a uniform microfibrous structure was obtained. Antimicrobial susceptibility testing of PCL/BFO/Q was conducted, and microbial reduction was found to be over 90% in the highest concentration of Q-loaded microfibrous scaffolds with the most inhibitory effect on S. aureus strains. Further, biocompatibility was investigated by performing MTT testing, fluorescence testing, and SEM imaging on mesenchymal stem cells (MSCs) to determine whether they could act as suitable microfibrous scaffolds for skeletal muscle tissue engineering. Incremental changes in the concentration of Q led to increased strength and strain, allowing muscles to withstand stretching during the healing process. In addition, electrically conductive microfibrous scaffolds enhanced the drug release capability by revealing that Q can be released significantly more quickly by applying the appropriate electric field, compared with conventional drug-release techniques. These findings suggest a possible use for PCL/BFO/Q microfibrous scaffolds in skeletal muscle regeneration by demonstrating that the combined action of both guidance biomaterials was more successful than Q itself acting alone.

Evaluation of the Putative Duplicity Effect of Novel Nutraceuticals Using Physico-Chemical and Biological In Vitro Models.

Nutraceuticals are experiencing a high-rise use nowadays, which is incomparable to a few years ago, due to a shift in consumers' peculiarity tendencies regarding the selection of alternatives to Western medicine, potential immunity boosters, or gut-health promoters. Nutraceuticals' compositions and actual effects should be proportional to their sought-after status, as they are perceived to be the middle ground between pharma rigor and naturally occurring actives. Therefore, the health benefits via nutrition, safe use, and reduction of potential harm should be the main focus for manufacturers. In this light, this study assess the nutritional profile (proteins, fats, fibers, caloric value, minerals) of a novel formulated nutraceutical, its physico-chemical properties, FTIR spectra, antioxidant activity, anthocyanins content, and potential hazards (heavy metals and microbiological contaminants), as well as its cytotoxicity, adherence, and invasion of bacteria on HT-29 cells, as well as its evaluation of beneficial effect, potential prebiotic value, and duplicity effect on gut microbiota in correlation with Regulation (EC) No 1924/2006. The results obtained indicate the growth stimulation of Lb. rhamnosus and the inhibitory effects of E.coli, Ent. Faecalis and Lc. lactis. The interaction between active compounds suggested a modulator effect of the intestinal microbiota by reducing the number of bacteria that adhere to epithelial cells or by inhibiting their growth.

Crosslinked Collagenic Scaffold Behavior Evaluation by Physico-Chemical, Mechanical and Biological Assessments in an In Vitro Microenvironment.

Wound healing-associated difficulties continue to drive biotechnological creativeness into complex grounds. The sophisticated architecture of skin wound sites and the intricate processes involved in the response to the use of regenerative devices play a critical role in successful skin regeneration approaches and their possible outcomes. Due to a plethora of complications involved in wound healing processes as well as the coordination of various cellular mechanisms, biomimetic approaches seems to be the most promising starting ground. This study evaluates the behavior of a crosslinked, porous collagen scaffold obtained by lyophilization and dehydrothermal reticulation (DHT). We address the key physio-chemical and mechanical factors, such as swelling, density and porosity, mechano-dynamic properties, SEM and TG-DSC, as well as important biological outcomes regarding scaffold biocompatibility and cellular metabolic activity, cytokine expression in inflammation, apoptosis and necrosis, as well as hemocompatibility and biodegradation. The mechanical and visco-elastic behavior are correlated, with the samples found to present similar thermal behavior and increased rigidity after DHT treatment. High biocompatibility rates were obtained, with no inflammatory stimulation and a reduction in necrotic cells. Higher percentages of cellular early apoptosis were observed. The hemocompatibility rate was under 2%, coagulation effects expressed after 4 min, and the DHT scaffold was more resistant to the biodegradation of collagenase compared with the untreated sample.

Microbial Contamination and Survival Rate on Different Types of Banknotes.

In the COVID-19 pandemic context, numerous concerns have been raised regarding the hygienic status of certain objects we interact with on a daily basis, and especially cash money and their potential to harbor and transmit pathogenic bacteria. Therefore, in the present study, we analyzed different currency bills represented by British pounds (5 £, 10 £ and 20 £), Romanian lei (1 leu, 5 lei and 10 lei), U.S. dollars (1 $, 5 $ and 10 $) and Euros (5 €, 10 € and 20 €) in order to evaluate the bacterial survival rate and bacterial adherence. We used five reference microorganisms by American Type Culture Collection (ATCC, Manassas, VA, USA): Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 8739, Enterococcus sp. ATCC 19952, Salmonella enterica subsp. enterica serovar Typhi ATCC 6539, and Listeria monocytogenes ATCC 7644. Microorganisms were selected in accordance with the criteria of prevalence, pathogenicity, opportunism, and incidence. However, Maldi-TOF analysis from samples taken from the banknotes revealed only a few of the common pathogens that are traditionally thought to be found on banknotes. Some of the most important factors for the survival of pathogenic agents on surfaces are the presence of organic matter, temperature and humidity. Our data showed that Salmonella enterica survived 72 h on every banknote tested, while L. monocytogenes tended to improve persistence in humid conditions. Survival rate is also influenced by the substrate composition, being lower for polymer-based banknotes especially for Salmonella enterica, Listeria monocytogenes and Enterococcus sp. The adherence of bacterial strains was lower for polymer-based banknotes British pounds and Romanian Leu, in contrast to the cotton-based U.S dollars and Euro banknotes. The risk of bacterial contamination from the banknote bills is high as indicated by both a strong survival capacity and low adherence of tested bacteria with differences between the two types of materials used for the tested banknotes.

Electrically Triggered Drug Delivery from Novel Electrospun Poly(Lactic Acid)/Graphene Oxide/Quercetin Fibrous Scaffolds for Wound Dressing Applications.

The novel controlled and localized delivery of drug molecules to target tissues using an external electric stimulus makes electro-responsive drug delivery systems both feasible and desirable, as well as entailing a reduction in the side effects. Novel micro-scaffold matrices were designed based on poly(lactic acid) (PLA) and graphene oxide (GO) via electrospinning. Quercetin (Q), a natural flavonoid, was loaded into the fiber matrices in order to investigate the potential as a model drug for wound dressing applications. The physico-chemical properties, electrical triggering capacity, antimicrobial assay and biocompatibility were also investigated. The newly fabricated PLA/GO/Q scaffolds showed uniform and smooth surface morphologies, without any beads, and with diameters ranging from 1107 nm (10%PLA/0.1GO/Q) to 1243 nm (10%PLA). The in vitro release tests of Q from the scaffolds showed that Q can be released much faster (up to 8640 times) when an appropriate electric field is applied compared to traditional drug-release approaches. For instance, 10 s of electric stimulation is enough to ensure the full delivery of the loaded Q from the 10%PLA/1%GO/Q microfiber scaffold at both 10 Hz and at 50 Hz. The antimicrobial tests showed the inhibition of bacterial film growth. Certainly, these materials could be loaded with more potent agents for anti-cancer, anti-infection, and anti-osteoporotic therapies. The L929 fibroblast cells cultured on these scaffolds were distributed homogeneously on the scaffolds, and the highest viability value of 82.3% was obtained for the 10%PLA/0.5%GO/Q microfiber scaffold. Moreover, the addition of Q in the PLA/GO matrix stimulated the production of IL-6 at 24 h, which could be linked to an acute inflammatory response in the exposed fibroblast cells, as a potential effect of wound healing. As a general conclusion, these results demonstrate the possibility of developing graphene oxide-based supports for the electrically triggered delivery of biological active agents, with the delivery rate being externally controlled in order to ensure personalized release.

The Biological Effects of Novel Nutraceuticals with Curcuminoids and Other Plant-Derived Immunomodulators and Pre-Probiotics.

An effective and well-balanced immune system is pivotal for maintaining health. Diet and nutrition can affect the functioning of numerous immune parameters, with direct repercussions on homeostasis. Since our immune functions are indispensable in defending the body against pathogens and thus play a vital role in maintaining health, modulating immune response may well serve as the basis for the development of plant-based functional foods and novel nutraceuticals. This concept is currently utilized in attempts to prevent or mitigate inflammatory reactions via the development of targeted food products or active ingredients since an extended number of phytoconstituents (such as curcuminoids) are associated with beneficial effects on immunity. Immunomodulatory plant-based dietary supplements are considered effective in improving immune functions and reducing the incidence of immunological disorders or imbalances. Therefore, the main focus of this study was to evidence the beneficial biological effects such as antioxidant and antimicrobial, as well as nutritional status, biocompatibility and cell proliferation capacity and immunomodulation of two novel nutraceuticals. The first nutraceutic was based on curcuminoids and other actives from Trigonella foenum- graecum (seeds), Chelidonium majus L. (aerial parts), Taraxacum officinale L. (roots), vitamins (C, D3, A, E) and minerals (zinc) whereas the second one was made of probiotics such as Lactobacillus acidophilus and Bifidobacterium animalis subsp. Lactis combined with actives from Helianthus tuberosus (tubers) and Psyllium/Plantago ovata (husk) as herbal prebiotics.

Chemical Composition, Antipathogenic and Cytotoxic Activity of the Essential Oil Extracted from Amorpha fruticosa Fruits.

The purpose of this paper was to characterize and investigate the antimicrobial potential of Amorpha fruticosa fruits essential oil (EO). The EO was extracted by hydrodistillation, analyzed by GC-MS, and then evaluated for its interaction with microbial and mammalian cells. The antimicrobial activity was assessed against bacterial and fungal strains, in a planktonic and adherent growth state, using qualitative and quantitative assays. The main components identified in A. fruticosa fruits EO were δ-cadinene, γ-muurolene, and α-muurolene. The Gram-positive strains proved to be more susceptible than Gram-negative bacteria and fungal strains. The EO exhibited good antibiofilm activity, inhibiting the microbial adherence to the inert (96-well plates and Foley catheter section) and cellular substrata. The flow cytometry analysis revealed as one of the possible mechanisms of antimicrobial action the alteration of cell membrane hydrophobicity. The cytotoxicity on the L929 cell line occurred at concentrations higher than 0.3 mg/mL. Taken together, our results demonstrate that A. fruticosa fruits EO contains active compounds with selective inhibitory effect on different microbial strains in planktonic and biofilm growth state, explained at least partially by the interference with microbial membranes due to their hydrophobic character.

Eco-Friendly Method for Tailoring Biocompatible and Antimicrobial Surfaces of Poly-L-Lactic Acid.

In this study, a facile, eco-friendly route, in two steps, for obtaining of poly-L-lactic acid/chitosan-silver nanoparticles scaffolds under quiescent conditions was presented. The method consists of plasma treatment and then wet chemical treatment of poly-L-lactic acid (PLLA) films in a chitosan based-silver nanoparticles solution (Cs/AgNp). The changes of the physical and chemical surface proprieties were studied using scanning electron microscopy (SEM), small angle X-Ray scattering (SAXS), Fourier transform infrared spectroscopy (FTIR) and profilometry methods. A certain combination of plasma treatment and chitosan-based silver nanoparticles solution increased the biocompatibility of PLLA films in combination with cell line seeding as well as the antimicrobial activity for gram-positive and gram-negative bacteria. The sample that demonstrated from Energy Dispersive Spectroscopy (EDAX) to have the highest amount of nitrogen and the smallest amount of Ag, proved to have the highest value for cell viability, demonstrating better biocompatibility and very good antimicrobial proprieties.