University of Illinois, Urbana, 61802, USA.
Acta Biomater. 2010 Oct;6(10):3988-97. doi: 10.1016/j.actbio.2010.05.016. Epub 2010 May 23.
The biocompatibility of chitosan and its similarity to glycosaminoglycans (GAG) make it attractive for cartilage tissue engineering. We have previously reported improved chondrogenesis but limited cell adhesion on chitosan scaffolds. Our objectives were to produce chitosan scaffolds coated with different densities of type II collagen and to evaluate the effect of this coating on mesenchymal stem cell (MSC) adhesion and chondrogenesis. Chitosan fibrous scaffolds were obtained by a wet spinning method and coated with type II collagen at two different densities. A polyglycolic acid mesh served as a reference group. The scaffolds were characterized by Fourier-transform infrared spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and type II collagen content. Constructs were analyzed after MSCs seeding via live/dead assay, weight and DNA evaluations, SEM, and TEM. Constructs were cultured in chondrogenic medium for 21 days prior to quantitative analysis (weight, DNA, and GAG), SEM, TEM, histology, immunohistochemistry, and quantitative real time polymerase chain reaction. The cell attachment and distribution after seeding correlated with the density of type II collagen. The cell number, the matrix production, and the expression of genes specific for chondrogenesis were improved after culture in collagen coated chitosan constructs. These findings encourage the use of type II collagen for coating chitosan scaffolds to improve MSCs adhesion and chondrogenesis, and confirm the importance of biomimetic scaffolds for tissue engineering.
壳聚糖的生物相容性及其与糖胺聚糖（GAG）的相似性使其成为软骨组织工程的理想选择。我们之前已经报道了壳聚糖支架上改善的软骨生成，但细胞黏附有限。我们的目的是制备不同密度 II 型胶原蛋白涂覆的壳聚糖支架，并评估这种涂层对间充质干细胞（MSC）黏附和软骨生成的影响。壳聚糖纤维支架通过湿法纺丝法获得，并在两种不同密度下涂覆 II 型胶原蛋白。聚乙二醇酸网作为参考组。通过傅里叶变换红外光谱、扫描电子显微镜（SEM）、透射电子显微镜（TEM）和 II 型胶原蛋白含量对支架进行了表征。通过活/死检测、重量和 DNA 评估、SEM 和 TEM 分析 MSC 接种后的构建体。构建体在软骨形成培养基中培养 21 天，然后进行定量分析（重量、DNA 和 GAG）、SEM、TEM、组织学、免疫组织化学和定量实时聚合酶链反应。接种后的细胞附着和分布与 II 型胶原蛋白的密度相关。在涂覆有胶原蛋白的壳聚糖支架中培养后，细胞数量、基质生成和软骨生成特异性基因的表达均得到改善。这些发现鼓励使用 II 型胶原蛋白涂覆壳聚糖支架以改善 MSC 黏附和软骨生成，并证实仿生支架对于组织工程的重要性。
