Bhardwaj Nandana, Singh Yogendra Pratap, Devi Dipali, Kandimalla Raghuram, Kotoky Jibon, Mandal Biman B
Seri-Biotechnology Unit, Life Science Division, Institute of Advanced Study in Science and Technology, Guwahati-781035, India.
J Mater Chem B. 2016 Jun 7;4(21):3670-3684. doi: 10.1039/c6tb00717a. Epub 2016 May 3.
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 μm) 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.
关节软骨损伤是肌肉骨骼治疗中最棘手的临床问题之一，因为其自我修复和再生能力有限。在本研究中，制备了源自非桑蚕柞蚕的3D多孔丝素蛋白支架，并检测了其支持软骨组织工程的能力。此外，还利用家蚕和蓖麻蚕的丝素蛋白支架进行了对比研究。在此，对制备的支架进行了全面表征，并比较了接种原代猪软骨细胞并在体外培养2周的丝素蛋白支架内软骨组织的形成情况。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 - 150μm），其结构稳定性增强。制备的支架表现出改善的力学性能，并通过后续实验揭示了改善的热性能和降解性能。代谢活性和荧光显微镜研究表明，柞蚕和蓖麻蚕丝素蛋白支架对软骨细胞的附着和增殖支持更好。生化分析表明，柞蚕丝素蛋白支架中硫酸化糖胺聚糖（sGAGs）和II型胶原蛋白的产量显著高于蓖麻蚕和家蚕支架（p < 0.001）。此外，组织化学和免疫组织化学研究表明sGAGs的积累增加和胶原蛋白II的表达增强。此外，大鼠皮下模型中的支架在植入8周后表现出体内生物相容性。综上所述，这些结果证明了柞蚕非桑蚕丝素蛋白支架的积极特性，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。
