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壳聚糖支架复合绵羊鼻软骨细胞构建组织工程化软骨

Characterization of Chitosan-Based Scaffolds Seeded with Sheep Nasal Chondrocytes for Cartilage Tissue Engineering.

机构信息

Faculty of Chemical Engineering and Technology, University of Zagreb, Marulićev trg 19, p.p.177, 10001, Zagreb, Croatia.

Faculty of Science, University of Zagreb, Horvatovac102a, 10001, Zagreb, Croatia.

出版信息

Ann Biomed Eng. 2021 Jun;49(6):1572-1586. doi: 10.1007/s10439-020-02712-9. Epub 2021 Jan 6.

Abstract

The treatment of cartilage defect remains a challenging issue in clinical practice. Chitosan-based materials have been recognized as a suitable microenvironment for chondrocyte adhesion, proliferation and differentiation forming articular cartilage. The use of nasal chondrocytes to culture articular cartilage on an appropriate scaffold emerged as a promising novel strategy for cartilage regeneration. Beside excellent properties, chitosan lacks in biological activity, such as RGD-sequences. In this work, we have prepared pure and protein-modified chitosan scaffolds of different deacetylation degree and molecular weight as platforms for the culture of sheep nasal chondrocytes. Fibronectin (FN) was chosen as an adhesive protein for the improvement of chitosan bioactivity. Prepared scaffolds were characterised in terms of microstructure, physical and biodegradation properties, while FN interactions with different chitosans were investigated through adsorption-desorption studies. The results indicated faster enzymatic degradation of chitosan scaffolds with lower deacetylation degree, while better FN interactions with material were achieved on chitosan with higher number of amine groups. Histological and immunohistochemical analysis of in vitro engineered cartilage grafts showed presence of hyaline cartilage produced by nasal chondrocytes.

摘要

软骨缺损的治疗仍然是临床实践中的一个挑战。壳聚糖基材料已被认为是适合软骨细胞黏附、增殖和分化形成关节软骨的微环境。利用鼻软骨细胞在适当的支架上培养关节软骨,作为软骨再生的一种有前途的新策略而出现。除了优良的性能外,壳聚糖缺乏生物活性,例如 RGD 序列。在这项工作中,我们制备了具有不同脱乙酰度和分子量的纯壳聚糖和蛋白修饰壳聚糖支架,作为培养绵羊鼻软骨细胞的平台。纤连蛋白(FN)被选为一种黏附蛋白,以提高壳聚糖的生物活性。通过吸附-解吸研究,对制备的支架进行了微观结构、物理和生物降解性能的表征,并研究了 FN 与不同壳聚糖的相互作用。结果表明,脱乙酰度较低的壳聚糖支架的酶降解速度更快,而具有更多胺基的壳聚糖与材料的 FN 相互作用更好。体外工程软骨移植物的组织学和免疫组织化学分析表明,鼻软骨细胞产生了透明软骨。

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