改良以聚羟基乙酸为支架同种异体组织工程化塑形软骨的构建

Improvement upon construction of tissue-engineered allogeneic cartilage molded with polyglycolic acid as the scaffold]

OBJECTIVE: To improve the method for constructing allogeneic molded cartilage by means of tissue engineering techniques. METHODS: The chondrocytes from the rib and articular cartilage of infant rabbits were harvested by type II collagenase digestion, followed by in vitro cell culture for 3 to 4 passages. The chondrocytes were then prepared into cell suspension and seeded onto C -and O -shaped pre-molded polyglycolic acid (PGA) scaffolds form chondrocyte-PGA composites, which were subsequently cultured in vitro for 7 to 10 d before implanted subcutaneously into adult rabbits. Improvement was made upon conventional shaping and implantation procedures. Morphological observation and cartilage regeneration assessment were conducted at different time points following the implantation, in comparison with the observation by conventional shaping and implantation methods. RESULTS: During in vitro cell culture, the rate of viable chondrocytes in the final cell suspension was (92+/-2)% after well-controlled prolongation of digestion trypsin, similar to the viable cell rate (93+/-2) % by traditional procedures (P>0.05). Gross observation found milk-white, newly generated cartilage which had good flexibility 4 weeks after implantation, and after 8 weeks and later, the cartilage took on the color of porcelain-white. Histological examination showed a few inflammatory cells around the newly generated immature cartilage 4 weeks after implantation, and the inflammation abated when the newly generated cartilage acquired similar histological properties to that of the original cartilage 8 weeks postoperatively and later. After 16 weeks, no blood vessel or capillaries were visible within the new cartilage. CONCLUSION: The chondrocyte viability is not affected when the cells are treated with well-controlled prolonged digestion with trypsin during in vitro cell culture. Improved PGA scaffolds shaping and the implantation procedure facilitate the regeneration of the cartilage after the implantation of the composites.