Multifunctional chitosan-based hydrogels: characterization and evaluation of biocompatibility and biodegradability in vitro

N. Manko, M. Lootsik, V. Antonyuk, I. Ivasechko, N. Skorokhyd, H. Kosiakova, O. Mehed’, T. Horid’ko, N. Hula, O. Klyuchivska, R. Panchuk, N. Pokhodylo, О. Barabash, T. Dumych, R. Stoika
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Abstract

Creation of novel remedies efficient in supporting wound healing remains an actual task in pharmacology. Hydrogels showed high efficiency in wound healing and tissue regeneration due to viscosity, elasticity and fluidity that provide them with functional characteristics similar to that in extracellular matrix. The aim of the study was to create chitosan-based hydrogels functionalized with different components (chondroitin-6-sulfate, hyaluronic acid, N-stearoylethanolamine) and to estimate their biocompatibility and biodegradabili­ty in vitro. For the first time, a lipid substance N-stearoylethanolamine (NSE) known as suppressor of pro-inflammatory cytokines expression was used as hydrogel component (1.95 mg/g). FTIR analysis confirmed the complexation of chitosan molecule with hyaluronate, chondroitin-6-sulfate, NSE. MTT-test and Trypan blue exclusion test were used to study hydrogels cytotoxicity towards human cells of different tissue origin. Biodegradability of hydrogels was evaluated using direct hydrogel contact with cells and cell-independent degradation. It was shown that chondroitin-6-sulfate (<2 mg/ml), hyaluronic acid (<2 mg/ml) and NSE (26 µg/ml) did not demonstrate significant toxic effects towards pseudonormal human cells of the MCF10A, HaCat, HEK293 lines and mouse cells of the Balb/3T3 line. The studied hydrogels were stable in saline solution, while in a complete culture medium containing 10% fetal bovine blood serum they underwent degradation in >24 h. The identified biodegradability of the chitosan-based hydrogels is important for the release of noncovalently immobilized NSE into biological medium. Further studies on laboratory animals with experimental wounds are expected to explore the potential of created hydrogels as anti-inflammatory and wound-healing agents. Keywords: biodegradability, chitosan hydrogels, chondroitin-6-sulfate, FTIR analysis, human pseudonormal cells, hyaluronic acid, N-stearoylethanolamine, toxicity
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基于壳聚糖的多功能水凝胶:体外生物相容性和生物降解性的表征与评估
创造出有效支持伤口愈合的新型疗法仍然是药理学的一项实际任务。水凝胶具有粘度、弹性和流动性,其功能特性与细胞外基质相似,因此在伤口愈合和组织再生方面表现出很高的效率。这项研究的目的是用不同的成分(软骨素-6-硫酸、透明质酸、N-硬脂酰乙醇胺)创建壳聚糖基水凝胶,并在体外评估其生物相容性和生物降解性。水凝胶成分中首次使用了抑制促炎细胞因子表达的脂质物质 N-硬脂酰乙醇胺(NSE)(1.95 毫克/克)。傅立叶变换红外光谱分析证实了壳聚糖分子与透明质酸、6-硫酸软骨素和 NSE 的复合物。MTT 试验和胰蓝排除试验用于研究水凝胶对不同组织来源的人体细胞的细胞毒性。通过水凝胶与细胞的直接接触和细胞独立降解,对水凝胶的生物降解性进行了评估。结果表明,6-硫酸软骨素水凝胶(24 小时)的生物降解性对非共价固定的 NSE 释放到生物介质中非常重要。对实验动物伤口的进一步研究有望探索所制备的水凝胶作为抗炎和伤口愈合剂的潜力。关键词:生物降解性、壳聚糖水凝胶、6-硫酸软骨素、傅立叶变换红外分析、人类假正常细胞、透明质酸、N-硬脂酰乙醇胺、毒性
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