Study On The Application Of Polyvinyl Alcohol/Regenerated Wild Antheraea Pernyi Silk Fibroin Scaffold For Repairing SD Rat Achille Tendon Defect

Nowadays, tendon defect is a common clinical disease. The traditional method of treatment of tendon injury is usually using transplantation of autologous tendon, allogeneic tendon or artificial muscle. Because of lacking of graft source, immune rejection, the repair effect is usually not ideal. Using tendon tissue engineering technology can achieve completly repair.In our previous studies, we preparated regenerated wild Antheraea Pernyi silk fibroin protein(RWSF). This study on the basic of that, cell proliferation toxicity, DNA molecular damage, cell cycle assay, cytoskeletal protein development, ability of collagen expression, the acute toxicity and the biocompatibility will be discussed. Furthermore, in order to evaluation about the feasibility of PVA/RWSF scaffold in the clinical application.The results is as folliwing:1. Assessment on biological safety of PVA/RWSF tendon tissue engineering material. Cell proliferation toxicity, molecular damage and acute toxicitystudy on its were explored. The results of MTT assay showed that the toxicity grade of PVA/RWSF scaffold come with in range of0-1, it comply meet the requirements of tissue engineering materials. The results of gel electrophoresis of single cell showed that no DNA damage with tendon cells cultured in extracts of PVA/RWSF scaffold suggesting that PVA/RWSF had no significant genetic toxicity. Kunming mice were intraperitoneally injected with extracts. The mice from experimental group showed no toxic reaction or weight reduction. Together that, PVA/RWSF scaffold was good biocompatibility, and it can be used in vivo studies.2. Compatibility of PVA/RWSF scaffold was investigated.(1) The biological function of PVA/RWSF was studied. Analysised cell cycle by flow cytometry and evaluated the proliferation index. Experiment results showed that the cell growing on PVA/RWSF were normal diploid cells, suggesting that the scaffold had no impact on the cell cycle. Applicating FITC-phalloidin to detected PVA/RWSF support expression of cytoskeleton protein F-actin. The results showed that the morphology of tendon cell that growing on the PVA/RWSF scaffold was highly spreading, suggesting that the capacity of tendon cell was not affected by PVA/RWSF. The relative expression amount of Col I and Col III, showed that the ability of secretion heterogeneity extracellular matrix of the tendon cells was not affected by PVA/RWSF scaffold.(2) The biocompatibility of PVA/RWSF scaffold was evaluated in vivo. The scaffold was implanted into SD rats in dorsal muscle tissue, gross observation of experimental animals and pathological section were analaysis. The results suggesting PVA/RWSF scaffolds was good biocompatibility and degradation appropriately.(3) The feasibility of PVA/RWSF scaffold in clinical application. SD rat Achille defects model was constructed, with tubular scaffold was used to bridging broken end of Achilles tendon. The results proved that the practicability of the method and the repeatability of implantation operation. Histological evaluation showed that the rate of scaffold degradation and the rate of newborn tendon tissue generation reached a dynamic balance, and the scaffold promoted the secretion of collagen fiber maturity, to a certain extent. It suggesting the feasibility of clinical application of PVA/RWSF scaffold.