Department of Biotechnology, Indian Institute of Technology, Kharagpur-721302, West Bengal, India.
Biomaterials. 2012 Apr;33(10):2848-57. doi: 10.1016/j.biomaterials.2011.12.028. Epub 2012 Jan 17.
Adult bone marrow derived mesenchymal stem cells are undifferentiated, multipotential cells and have the potential to differentiate into multiple lineages like bone, cartilage or fat. In this study, polyelectrolyte complex silk fibroin/chitosan blended porous scaffolds were fabricated and examined for its ability to support in vitro chondrogenesis of mesenchymal stem cells. Silk fibroin matrices provide suitable substrate for cell attachment and proliferation while chitosan are promising biomaterial for cartilage repair due to it's structurally resemblance with glycosaminoglycans. We compared the formation of cartilaginous tissue in the silk fibroin/chitosan blended scaffolds with rat mesenchymal stem cells and cultured in vitro for 3 weeks. Additionally, pure silk fibroin scaffolds of non-mulberry silkworm, Antheraea mylitta and mulberry silkworm, Bombyx mori were also utilized for comparative studies. The constructs were analyzed for cell attachment, proliferation, differentiation, histological and immunohistochemical evaluations. Silk fibroin/chitosan blended scaffolds supported the cell attachment and proliferation as indicated by SEM observation, Confocal microscopy and metabolic activities. Alcian Blue and Safranin O histochemistry and expression of collagen II indicated the maintenance of chondrogenic phenotype in the constructs after 3 weeks of culture. Glycosaminoglycans and collagen accumulated in all the scaffolds and was highest in silk fibroin/chitosan blended scaffolds and pure silk fibroin scaffolds of A. mylitta. Chondrogenic differentiation of MSCs in the silk fibroin/chitosan and pure silk fibroin scaffolds was evident by real-time PCR analysis for cartilage-specific ECM gene markers. The results represent silk fibroin/chitosan blended 3D scaffolds as suitable scaffold for mesenchymal stem cells-based cartilage repair.
成体骨髓间充质干细胞是未分化的多能细胞，具有向骨、软骨或脂肪等多种谱系分化的潜能。在本研究中，制备了聚电解质复合物丝素蛋白/壳聚糖共混多孔支架，并研究了其支持间充质干细胞体外软骨生成的能力。丝素蛋白基质为细胞附着和增殖提供了合适的基底，而壳聚糖由于与糖胺聚糖结构相似，是一种有前途的软骨修复生物材料。我们比较了丝素蛋白/壳聚糖共混支架中大鼠间充质干细胞的软骨组织形成情况，并在体外培养 3 周。此外，还利用非桑蚕，野蚕 Antheraea mylitta 和桑蚕 Bombyx mori 的纯丝素蛋白支架进行了对比研究。对构建体进行了细胞附着、增殖、分化、组织学和免疫组织化学评估。扫描电子显微镜观察、共聚焦显微镜和代谢活性表明，丝素蛋白/壳聚糖共混支架支持细胞附着和增殖。3 周培养后，阿利新蓝和番红 O 组织化学和 II 型胶原的表达表明构建体中维持了软骨表型。所有支架中均有糖胺聚糖和胶原蛋白积累，丝素蛋白/壳聚糖共混支架和 A. mylitta 的纯丝素蛋白支架中含量最高。间充质干细胞在丝素蛋白/壳聚糖和纯丝素蛋白支架中的软骨分化通过软骨特异性细胞外基质基因标志物的实时 PCR 分析得到证实。结果表明，丝素蛋白/壳聚糖共混 3D 支架是一种适合间充质干细胞软骨修复的支架。
