Experimental study of the cartilage formation of immortalized chondrocytes in vivo / 第三军医大学学报

ABSTRACT

Objective To explore the feasibility of the construction and formation of cartilage with immortalized chondrocytes by tissue engineering technology in vitro . Methods Human telomerase reverse transcriptase(hTERT) gene was introduced into rabbit mandibular condylar chondrocytes by eukaryotic vector. After screening with G418, the positive clones were amplified for culture. Normal or immortalized chondrocyte loaded cytoskeleton ? tricalcium phosphate (? TCP) complexes were incubated in vitro for 1～2 d and then implanted into subcutaneous tissue of nude mouse. The complexes of immortalized chondrocyte ? TCP and chondrocyte ? TCP or ? TCP alone were established as the experimental group and control groups respectively. The specimens were harvested within 3 and 6 months after surgical procedure for histological and immunohistochemical observation. Results In experimental groups and control group 1, the complexes packed with cartilage like tissue were found, but there was only a little fiber like tissue formed in control group 2. Immunohistochemistry revealed strong positive staining with safranine O, toluidine blue and collagen type II and obvious formation of cartilage. There was significant difference between the experimental group and the control groups( P