Experimental Studies On The Construction And Application Of Engineering Epiphyseal Plate In Epiphyseal Plate Injury

Part I: The seed cells of epiphyseal plate tissue engineering -- the methods of culture the epiphyseal plate chondrocyte in vitro and its cellular phenotype in rabbits.Objective To establish a kind of method about culture the epiphyseal plate chondrocyte in vitro, so that we can obtain the seed cells for the useage of epiphyseal plate tissue engineering.Methods The cells we used come from proximal epiphyseal plate of tibia of rabbits. Before we culture the cells, we used Trypsin and Collagenese II . Checking up biological characteristics of the cells by applying the ways of microscope and MTT test. Identifying the cells by applying the ways of safrarine"o"staining and PCR test.Results We have observed the conditions of the growth of the epiphyseal plate chondrocyte. The test of safrarine"o"staining shows positive. The test of PCR shows the expreesion of II collagen. The test of MTT shows the biological properties of the cells decrease from the 12th day.Conclusions We have established a method of culture the epiphyseal plate chondrocyte, at the same time we have identified that the cells were the epiphyseal plate chondrocyte.Part II: The construction of engineering epiphyseal plate ----the study on the biocompatibility of epiphyseal plate chondrocyte with scaffold PLA and epiphyseal plate chondrocyte with scaffold SF.Objective To explore the way of construction about engineering epiphyseal plate using epiphyseal plate chondrocyte and scaffold.Methods To construct two engineering epiphyseal plates by culturing the epiphyseal plate chondrocyte on scaffold PLA and scaffold SF. Checking up biological characteristics of the cells by applying the ways of scanning electron- microscope, laser scan confocal microscope and MTT test.Results The epiphyseal plate chondrocyte grew well on scaffold PLA and scaffold SF. The function of the tow scaffold is similar.Conclusions Both scaffold PLA and scaffold SF have good biocompatibility and can be used as ideal scaffold in tissue engineering.Part III: The animal experimental study on the application in epiphyseal plate injury of the two engineering epiphyseal plates.Objective To observe the role of engineering epiphyseal plate in the treatment of epiphyseal plate injury.Methods The two engineering epiphyseal plates were implanted into the model of epiphyseal plate injury. Evaluating the role by histology, BrdU and X-ray. At the same time, comparing engineering epiphyseal plate to scaffold .Results The degree of deformity in engineering epiphyseal plate group was less that scaffold group ,whereas the degree of deformity in scaffold group less than control group .The two engineering epiphyseal plate groups are similar , and the two scaffold groups are also similar.Conclusions It is feasible to treat epiphyseal plate injury by engineering epiphyseal plate, and it is better than by scaffold.Partâ…£: The methods of culture and identification of the human epiphyseal chondrocyte.Objective To establish a kind of method about culture and identification of the human epiphyseal chondrocyte.Methods The cells we used come from epiphysis of polydactyly. Before we culture the cells, we used Trypsin and Collagenase II . Checking up biological characteristics of the cells by applying the ways of MTT test. Identifying the cells by applying the ways of safrarine"o"staining, Alcian blue test and immuno- histochemistry staining for type II collagen.Results We have observed the conditions of the growth of the human epiphyseal chondrocyte. The test of safrarine"o"staining and Alcian blue shows positive. The test of immunohistochemistry staining for type II collagen shows the expression of II collagen. The test of MTT shows the biological properties of the cells decrease from the 15th day.Conclusions We have established a method of culture the human epiphyseal chondrocyte , at the same time we have identified that the cells were the human epiphyseal chondrocyte.