Potential of silk fibroin/chondrocyte constructs of muga silkworm Antheraea assamensis for cartilage tissue engineering
Potential of silk fibroin/chondrocyte constructs of muga silkworm Antheraea assamensis for cartilage tissue engineering
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DOI：
10.1039/c6tb00717a
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发表时间：
2016-06-07
影响因子：
7
通讯作者：
Mandal, Biman B.
中科院分区：
文献类型：
--
作者：
Bhardwaj, Nandana;Singh, Yogendra Pratap;Mandal, Biman B.
关节软骨损伤由于其有限的自我修复和再生能力而成为肌肉骨骼治疗学中最令人困惑的临床问题之一。在这项研究中，3D多孔丝素蛋白支架来源于非桑树muga蚕assamensis和检查其支持软骨组织工程的能力。此外，家蚕和蓖麻蚕丝素蛋白支架用于比较研究。在本文中，制造的支架被彻底表征，并比较了与原代猪软骨细胞接种并在体外培养2周的丝素蛋白支架内的软骨组织形成。表面形态和结构构象研究揭示了高度互连的多孔结构（孔径80-150 μ m），其结构内具有增强的稳定性。所制造的支架表现出改善的机械性能，并随后进行了连续的实验，以揭示改善的热性能和降解性能。A.丝素蛋白支架assamensis和P. ricini支持更好的软骨细胞附着和增殖，如代谢活性和荧光显微镜研究所示。生化分析表明，A. assamensis丝素蛋白支架，然后是蓖麻青霉和B。Mori支架组（P < 0.001）。此外，组织化学和免疫组织化学研究表明，增强积累的sGAG和胶原II的表达。此外，在大鼠皮下模型中的支架植入8周后显示出体内生物相容性。总之，这些结果证明了来自A. assamensis，并表明其适合作为基于软骨细胞的软骨修复的有前途的支架。
Articular cartilage damage represents one of the most perplexing clinical problems of musculoskeletal therapeutics due to its limited self-repair and regenerative capabilities. In this study, 3D porous silk fibroin scaffolds derived from non-mulberry muga silkworm Antheraea assamensis were fabricated and examined for their ability to support cartilage tissue engineering. Additionally, Bombyx mori and Philosamia ricini silk fibroin scaffolds were utilized for comparative studies. Herein, the fabricated scaffolds were thoroughly characterized and compared for cartilaginous tissue formation within the silk fibroin scaffolds seeded with primary porcine chondrocytes and cultured in vitro for 2 weeks. Surface morphology and structural conformation studies revealed the highly interconnected porous structure (pore size 80-150 mu m) with enhanced stability within their structure. The fabricated scaffolds demonstrated improved mechanical properties and were followed-up with sequential experiments to reveal improved thermal and degradation properties. Silk fibroin scaffolds of A. assamensis and P. ricini supported better chondrocyte attachment and proliferation as indicated by metabolic activities and fluorescence microscopic studies. Biochemical analysis demonstrated significantly higher production of sulphated glycosaminoglycans (sGAGs) and type II collagen in A. assamensis silk fibroin scaffolds followed by P. ricini and B. mori scaffolds (p < 0.001). Furthermore, histochemistry and immunohistochemical studies indicated enhanced accumulation of sGAGs and expression of collagen II. Moreover, the scaffolds in a subcutaneous model of rat demonstrated in vivo biocompatibility after 8 weeks of implantation. Taken together, these results demonstrate the positive attributes from the non-mulberry silk fibroin scaffold of A. assamensis and suggest its suitability as a promising scaffold for chondrocyte based cartilage repair.