Research Of Cultivating Tissue-Engineered Cartilage In Rotating Bioreactor

Objective:To evaluate the advantage of cultivating tissue-engineered cartilage in a rotating bioreactor(HFB-40)Methods:The articular cartilage taken from New Zealand rabbits under sterile condition was sheared to the size of about 1mm×1mm×1mm. Chondrocytes were isolated from the articular cartilage by digesting it with 0.25% trypsin and 0.2% type II collagenase in sequence .After proliferation in vitro, chondrocytes of the second generation were seeded into PLGA scaffolds at the density of 4×10~7 /ml. Each PLGA template seeded with chondrocytes was cultivated in the bioreactor(A) or flasks(B) for 4 weeks. The chondrocyte-PLGA constructs were harvested. The content of GAG ,DNA and the percentage of type II collagen dyeing area were detected.Results: The chondrocyte-PLGA constructs grew much better in the bioreactor than in the flasks. HE and TB stain results showed significant differences between group A and B.The type II collagen dyeing area of group A and B were respectively (94.71±0.46) % and (63.87±1.39) % . And the GAG conent of group A and B were respectively 459.167±4.682 and 355.282±5.110 .The content of DNA werel4.25±0.147 and 14.25±0.147. There are significant dfferences of the type II collagen dyeing area, the conent of GAG and DNA between group A and B (P<0.01).The chondrocyte-PLGA constructs of group A which was much thicker and smoother, produced much more type II collagen and GAG than group B.Conclusion:It is advantageous for the tissue-engineered cartilage to be cultivated in the bioreactor which can provide more suitable culture environment for chondrocyte-PLGA constructs.