Formation Of Tissue Engineered Cartilage With Silk And Articular Chondrocytes In Vitro

Limited regenerative capacity of cartilage tissue makes treatment of chondral lesions difficult. The current methods to treat catilage lesion may relieve symptoms, but can not regenerate the injured tissue. Tissue engineering techniques is new resolvent of sufferer form cartilage lesion. It include seed cell, the coculture of scaffold and cells, implantation of cell and scaffold compound. It is the prerequisite to harvest seed cells maintaining stable phenotype, high proliferous activity and with broad extensive source to construct engineering cartilage in vitro. and a major challenge to find a appropriate scaffold confronting tissue engineering researchers. In this work we studied the methods of chondrocytes isolation and the protential of silk as scaffold of engineered cartilage. 1 In this study rabbit articular chondrocytes were harvested by methods of tissue block and enzyme digestion by stages. Chondrocytes by tissue block shew short shuttle-like, and maintained such shape after 7 passages, while chondrocytes of enzyme digestion presented different shapes, and became fibre-like little by little. 2 Chondrocytes were isolated with 0.2% collengasen type â…¡confected by serum-free Ham-F12 medium and PBS free-Ca2+,Mg2+. By the method of digestion by stages, the lost of cells decreased, and cells maintained higher viability. the chondrocytes isolated with collengase type confected by serum-free medium were more actative than that of phosphate buffer free-Ca2+,Mg2+. There was significant difference between two groups. 3 Furthermore, we used silk scaffold to construct engineered cartilage. The growth and proliferation of chondrocytes in silk scaffold were compared with that of chondrpcytes in collagen. The silks were digested by trypsine and coated by collagen typeâ…¡. Chondrocytes attached silk scaffold during the process of surge. The growth of cells were observed under microscope. And the compound of chondrocytes and collagen gel were observed in histology slice. The results shew silks presented favourable attachment to chondrocytes and kept cells grow and proliferate. There were much extracellular surrounding chondrocytes. In initial culture, chondrocytes in collagen gel grew better than that in silk scaffold. However collagen scaffolds degraded and lost their structure, silk constructs retained their structure throughout the culture after two weeks of incubation. Therefore above of that we can conclude that silk can serve asscaffold for cartilage tissue engineering.