Construction of injectable tissue engineered cartilage in vitro / 第三军医大学学报

ABSTRACT

Objective To observe the biological behavior of chondrocytes in chitosan based hydrogel in vitro and to evaluate the possibility of chitosan and glycerophosphate mixed gel using as injectable scaffolds in tissue engineering.Methods A novel injectable thermosetting gel was made of chitosan and glycerophosphate mixture.Chondrocytes isolated by enzymic digestion from the articular cartilage of the young rabbits aged 3-4 weeks were cultured in monolayer system.After primary culture and subculturing,the cells were seeded in chitosan based gel in concentration of 5?10~7/ml in vitro.Characteristics of chondrocytes during 4-week culture were observed with invert microscope.MTT was used to detect the seed growth after one-week culture.In 4th week,after haematoxylin eosine and toluidine blue staining,the artifical cartilage was performed type Ⅱ collagen immunohistochemistry and observed by scanning electron microscope.Results Chondrocytes retained their round morphology,multiplied,and produced matrix in chitosan and glycerophosphate mixture gels.The results of immunohistochemistry were strongly positive.Conclusion That most chondrocytes appeared to occupy large areas of rough chitosan based gels,guarantees a suitable environment where chondrocytes secrete matrix macromolecules and preserve their round morphology in vitro.Chitosan and glycerophosphate mixture gel may be a vehicle for injectable tissue-engineered cartilage.