| The exper imentl study of constructionTissue-engineered cartilage with polylatic acidObjective: The aim of our study is to construct tissue-engineered cartilage with chondrocytes and polylatic acid(PLA)scaffold. Method: 1,TO isolate chondrocytes from rabbitjoint cartilage and culture by modified method. 2,To constructscaffold with polylatic acid (PLA) and sodium chloride (Nacl)mcirone (φ<300 μ), and observe the ultrastructure of the scaffoldwith scanning electron microscope. 3,To construct tissue-engineered cartilage with chondrocytes and polylatic acid (PLA)scaffold. First; Nine pieces of PLA scaffolds discs were dividedinto three groups (group A. B and C). Chondrocytes were seededonto group A and B, and then cultured in F-l2. Group C was alsocultured as the control group but without chondrocytes. After beingcultured for 4 days, group A was evaluated histologically. Second;Six nude mice were divided into two groups randomly (groupexperiment and control). The three discs of group B were implantedinto the subcutaneous of the three nude mice of experimental grouprespectively. Group C was also implanted into three nude mice ofthe control group. Finallyf The tissue-engineered cartilage wereobserved by HE, VG, AB-PAS stain and type Ⅱ collagenbiochemical test 2,4 and 6 weeks after operation respectively.Results: 1. Chondrocyte isolated and cultured by the modifiedmethod do not change it's characteristic of phenotype and shape.2. PLA scaffolds were fulfilled with connected micropores(diameter between 100─300 μm). 3. Scanning electronmicroscope showed that chondrocytes could growth on the wall ofthe micropores 4 days after vitro culture. 4. After cultured in nudemice, it showed that there were many chondrocytes not onlygrowthed in PLA scaffolds, but also secreted type II col1agen fibrinand glycosaminoglycan (GAG). Control group showed that therewere no chondrocytes, type II collogen or glycosaminoglycan(GAG) in PLA scaffolds. Conclusion: Our study demonstratesthat the modified method was more effcient, chondrocytes couldgrowth and regenerate in PLA scaffolds, PLA scaffolds can be usedto form tissue-engineered cartilage. |
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