以壳聚糖-胶原共混膜为三维支架同种异体工程化软骨的构建

目的探讨以壳聚糖-胶原共混膜为三维支架材料的同种异体软骨细胞构建组织工程化软骨的能力。方法将分离、培养、扩传兔软骨细胞，接种在壳聚糖-胶原共混膜上，倒置显微镜下观察细胞在共混膜上的生长情况。体外培养7d后，将细胞-材料复合物种植在新西兰兔皮下，6周取材，对获得的同种异体工程化软骨进行组织学评价。结果兔软骨细胞接种于壳聚糖-胶原共混膜上4h后有贴壁现象出现，细胞呈梭形。培养48h后，软骨细胞分裂增殖越来越多并向周围延伸，培养第7天取材，HE染色示细胞生长良好，呈梭形。体内培养6周取材，HE染色、Masson染色为均一的成熟软骨组织，且共混膜已降解。结论以壳聚糖-胶原共混膜为支架材料同种异体软骨细胞在有免疫力的动物体内可形成工程化软骨。

Fabrication of tissue engineered cartilage using chitosan-collagen scaffold seeded with allogenic chondrocytes

Objective To study the establishing of tissue engineering cartilage with chitosan- collagen scaffold as threedimensional scaffold seeded with allogenic chondrocytes. Methods Rabbit chondrocytes were isolated and cultured in vitro and seeded on chitosan-collagen scaffold. Chondrocytes growth in the scaffold was observed by microscope. Cultured in vitro for 7 days,the scaffold-chondrocyte complexes were transplanted into subcutaneous tissue of adult rabbits. After 6 weeks,gross observation and histology were used to evaluate the results. Results The chondrocytes were seeded into chitosan-collagen scaffold and adhered to the scaffold after 4 hours. After 48 hours ,the proliferation of the chondrocytes inceased gradually and the chondrocytes extended around. After 7 days,the HE stain showed the chondrocytes grew well. After 6 weeks,the HE stain and Masson stain showed the specimen was mature cartilage tissue and the scaffold was disappeared. Conclusion It demonstrates that the engineering cartilage could be successfully established in animals possessed immune function by seeding allogenic chondrocytes onto chitosan-collagen scaffold.