In this study, lemon balm (Melissa officinalis L.) and dill (Anethum graveolens L.) essential oils (EOs) were encapsulated into collagen hydrolysates extracted from bovine tendons and rabbit skins, both mixed with chitosan (CS) by using the coaxial electrospinning technique for potential wound dressing applications. The morphology and chemical composition of the electrospun nanofibers were investigated using scanning electron microscopy (SEM) and attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR). The antimicrobial activity of the dill EO and lemon EO, as well as the electrospun samples loaded with essential oils was determined by disk diffusion assay against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Enterococcus faecalis ATCC 29212, and Salmonella typhimurium ATCC 14028 bacterial strains; Candida albicans ATCC 10231 and Candida glabrata ATCC 90028 yeast strains; and Aspergillus brasiliensis ATCC 9642 fungal strain. In vivo biocompatibility testing of the collagen hydrolysate-chitosan/essential oil electrospun nanofibers was based on the determination of the hematological, biochemical, and immunological profile and the evaluation of the influence produced on the oxidative stress in white Swiss mice. The synergetic effect of dill and lemon balm EOs can improve the antimicrobial activity of collagen hydrolysate-chitosan nanofibers against the most important bacterial strains. The in vivo test results suggested a good biocompatibility of electrospun samples based on collagen hydrolysate extracted from bovine tendons or rabbit skin mixed with chitosan and containing dill and/or lemon balm essential oils as encapsulated bioactive compounds.

This research aimed to obtain biocompatible and antimicrobial nanofibres based on concentrated collagen hydrolysate loaded with thyme or oregano essential oils as a natural alternative to synthesis products. The essential oils were successfully incorporated using electrospinning process into collagen resulting nanofibres with diameter from 471 nm to 580 nm and porous structure. The presence of essential oils in collagen nanofibre mats was confirmed by Attenuated Total Reflectance -Fourier Transform Infrared Spectroscopy (ATR-FTIR), Ultraviolet–visible spectroscopy (UV–VIS) and antimicrobial activity. Scanning Electron Microscopy with Energy Dispersive Spectroscopy analyses allowed evaluating the morphology and constituent elements of the nanofibre networks. Microbiological tests performed against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans showed that the presence of essential oils supplemented the new collagen nanofibres with antimicrobial properties. The biocompatibility of collagen and collagen with essential oils was assessed by in vitro cultivation with NCTC clone 929 of fibroblastic cells and cell viability measurement. The results showed that the collagen and thyme or oregano oil composites have no cytotoxicity up to concentrations of 1000 μg·mL⁻¹ and 500 μg mL⁻¹, respectively. Optimization of electrospinning parameters has led to the obtaining of new collagen electrospun nanofibre mats loaded with essential oils with potential use for wound dressings, tissue engineering or protective clothing.

The aim of this paper was to select keratin hydrolysate with bioactive properties by using the enzymatic hydrolysis of wool. Different proteolytic enzymes such as Protamex, Esperase, and Valkerase were used to break keratin molecules in light of bioactive additive preparation. The enzymatic keratin hydrolysates were assessed in terms of the physico-chemical characteristics related to the content of dry substance, total nitrogen, keratin, ash, cysteic sulphur, and cysteine. The influence of enzymatic hydrolysis on molecular weight and amino acid composition was determined by gel permeation chromatography (GPC) and gas chromatography-mass spectrometry (GC-MS) analyses. Antimicrobial activity of keratin hydrolysates was analysed against Fusarium spp., a pathogenic fungus that can decrease the quality of plants. The bioactivity of enzymatic hydrolysates was tested on maize plants and allowed us to select the keratin hydrolysates processed with the Esperase and Valkerase enzymes. The ratio of organised structures of hydrolysate peptides was analysed by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) deconvolution of the amide I band and may explain the difference in their bioactive behaviour. The most important modifications in the ATR spectra of maize leaves in correlation with the experimentally proven performance on maize development by plant length and chlorophyll index quantification were detailed. The potential of enzymatic hydrolysis to design additives with different bioactivity was shown in the case of plant growth stimulation.

In this study, the effect of gamma irradiation (10 kGy) on proteins extracted from animal hide, scales, and wool was evidenced by calorimetric (μDSC) and spectroscopic (IR, circular dichroism, and EPR) methods. Keratin was obtained from sheep wool, collagen and bovine gelatin from bovine hide, and fish gelatin from fish scales. The μDSC experiments evidenced that gamma irradiation influences the thermal stability of these proteins differently. The thermal stability of keratin decreases, while a resistance to thermal denaturation was noticed for collagen and gelatins after gamma irradiation. The analysis of the IR spectra demonstrated that gamma irradiation determines changes in the vibrational modes of the amide groups that are associated with protein denaturation, most meaningfully in the case of keratin. As evidenced by circular dichroism for all proteins considered, exposure to gamma radiation produces changes in the secondary structure that are more significant than those produced by UV irradiation. Riboflavin has different effects on the secondary structure of the investigated proteins, a stabilizing effect for keratin and fish gelatin and a destabilizing effect for bovine gelatin, observed in both irradiated and non-irradiated samples. The EPR spectroscopy evidences the presence, in the gamma-irradiated samples, of free radicals centered on oxygen, and the increase in their EPR signals over time due to the presence of riboflavin.

Keratin biomaterials with high molecular weights were intensively investigated but few are marketed due to complex methods of extraction and preparation and limited understanding of their influence on cells behavior. In this context the aim of this research was to elucidate decisive molecular factors for skin homeostasis restoration induced by two low molecular weight keratin hydrolysates extracted and conditioned through a simple and green method. Two keratin hydrolysates with molecular weights of 3758 and 12,400 Da were physico-chemically characterized and their structure was assessed by circular dichroism (CD) and FTIR spectroscopy in view of bioactive potential identification. Other investigations were focused on several molecular factors: α1, α2 and β1 integrin mediated signals, cell cycle progression in pro-inflammatory conditions (TNFα/LPS stimulated keratinocytes and fibroblasts) and ICAM-1/VCAM-1 inhibition in human vascular endothelial cells. Flow cytometry techniques demonstrated a distinctive pattern of efficacy: keratin hydrolysates over-expressed α1 and α2 subunits, responsible for tight bounds between fibroblasts and collagen or laminin 1; both actives stimulated the epidermal turn-over and inhibited VCAM over-expression in pro-inflammatory conditions associated with bacterial infections. Our results offer mechanistic insights in wound healing signaling factors modulated by the two low molecular weight keratin hydrolysates which still preserve bioactive secondary structure.