Biological safety and therapeutic efficacy of a biopolymer-based multicomponent hydrogel mimicking the extracellular matrix

Background. The development of adhesive wound dressings with regenerative properties for restoring the integrity of damaged skin and mucous membranes is currently a priority area in regenerative medicine and tissue engineering. Objective: to develop a homogeneous, multicomponent collagen-based biopolymer adhesive hydrogel (MCBAH) and to evaluate its safety and efficacy. Materials and methods. Sterile samples of MCBAH were analyzed. The content of volatile organic compounds (VOCs) was determined using a Chromatec-Crystal 5000 gas chromatograph (Chromatec SKB CJSC, Russia). Osmolarity was measured using the Osmometer K-7400 liquid analyzer (Knauer, Germany), and pH was determined with the pH-150MI pH meter (Izmeritel’naya Tekhnika LLC, Russia). Biological safety was assessed in accordance with GOST ISO 10993 and included evaluation of cytotoxicity, pyrogenicity, material-mediated effects, general toxicity (acute and subchronic), as well as sensitizing and irritant effects. In vitro cytotoxicity was assessed using cultures of NIH/3T3 fibroblasts (ATCC® CRL-1658™). The efficacy of MCBAH was evaluated in a rat thermal burn model. Histological examinations were performed on tissues from the heart, lungs, thymus, liver, spleen, kidneys, adrenal glands, gonads, and cerebral cortex. Results. VOCs concentrations did not exceed permissible limits. Osmolarity was 334 ± 20 mOsm/kg, and pH was 7.18 ± 0.11. In vitro studies demonstrated the absence of cytotoxicity. Acute toxicity testing revealed no toxic effects in laboratory animals. Similarly, subchronic toxicity studies showed that exposure to MCBAH did not cause intoxication or mortality. No delayed-type hypersensitivity reactions were observed, indicating the absence of a sensitizing effect. Intradermal administration of MCBAH and its application to the mucous membranes of the hamster cheek pouch, rabbit vagina, and rabbit rectum produced no detectable response, with an irritation index of 0. Following intravenous administration of extracts from MCBAH samples to rabbits, increase in body temperature in all three animals did not exceed 0.1 °C, indicating that the biomimetic hydrogel does not induce a pyrogenic response. Conclusion. In a rat thermal burn model, application of an adhesive biomimetic hydrogel mimicking the extracellular matrix (ECM) immediately after burn induction and daily until complete wound closure significantly accelerated wound healing in the experimental group by 15.3 ± 7.7% (34.9 days) compared with the control group (40.2 days, p < 0.01). Treatment also promoted the formation of a fully developed epithelial layer and accelerated restoration of the original histological structure of the damaged skin. These findings support further investigation of the efficacy of this hydrogel in experimental models of damaged mucous membranes.

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