[Experimental research on repair of rabbit articular cartilage deffects with composite of autologous cell-carriers].
摘要
OBJECTIVE: To study the effect of the repair of rabbit articular cartilage defects by the composite of chondrogenic induction of autologous MSCs and autologous "two-phase" bone matrix gelatin (BMG). METHODS: Twenty-four healthy adult New Zealand rabbits weighing 2 to 3 kg were divided into group A, B and C with 8 in each. Autologous MSCs derived from group A were cultured in vitro and observed under inverted phase contrast microscope when enough cells through trypsinization transferring in vitro were obtained. Then the growth curves of 1, 3 and 5 passage culture of MSCs were drawn. The 3rd passage MSCs were induced into chondrogenic differentiation by adding TGF-beta1 (10 ng/mL), IGF-1 (10 ng/mL) and vitamin C (50 ng/mL) in vitro. At 8 days after induction, the features of chondrocytes were observed under inverted phase contrast microscope,and immunohistochemical staining and Mallory staining were made. Getting out part of the ilium of group A and B, according to the method of Urist, the "two-phase" BMG was acquired. Chondrogenic induction of autologous MSCs was inoculated into the corresponding BMG to set up a composite of cell-carrier, and then it was observed through scanning electric microscope after 3 days of culture. The model of articular cartilage defects of rabbits was made: in group A,autologous cell-carriers were implanted; in group B, there only existed autologous BMG; in group C,there was nothing. At 8, 12 weeks after operation, the gross, HE staining and immunohistochemical staining were made,and grading scales were evaluated according to Wakitani histological grading method. RESULTS: Features of MSCs were as follows: the shape of primary cells was shot-spindled and of passage cells was long. As to the growth curves of 1, 3 and 5 passage culture of MSCs,passage cells grew slowly for 3 days after being passaged and went into log-growth during the 3rd and the 7th days and into plateau later, but the 3rd passage cells grew best. Observation of MSCs after