成骨样细胞与聚乳酸体外复合培养的实验研究

目的：研究成骨样细胞与生物可降解聚合物基质的关系，用聚乳酸(polylactic acid，PLA)构建细胞——聚合物复合体的适用性。方法：自新生Wistar大鼠颅盖骨分离成骨样细胞，并将其种植到PLA方片上复合培养。结果：复合培养第7天HE染色显示，鳞状细胞出现。第10、13天，细胞数据明显增多，鳞状细胞占主体，细胞排列规则，极向一致。结论：成骨样细胞在PLA上显示良好粘附力和形态并能增殖生长。聚乳酸可以构建细胞——聚合物复合体并有望成为组织工程化骨中细胞外基质替代物。

The Experimental Study of Culture of Osteoblast-like Cell on PLA Substrate

Objective: The purpose of our study is to investigate the behavior of osteoblast-like cell cultred with biodegradable polymer substrate and the feasibility of building cell-polymer construct with polylactic acid (PLA). Methods: PLA was made in the form of films. SEM was used to examine the surface and section feature of the polymer film. Osteablast-like cells were derived from newborn Wistar rat calvaria. Cells were then seeded on the PLA films and grown for 13 days. HE stain and SEM were used to evaluate the morphology of cells. Qualified Gomori's Calcium-Cobalt method was adapted to dye the alkaline phosphatase (ALP) in the cytoplasm of cells. Results: SEM shows the surface of the polymer is rough. On the seventh day, cells became squamous-like. Cells began to contact with each other by their processes, demonstrating a network-like structure on the polymer. On the tenth and thirteenth day, the number became notably larger. Squamous-like cells dominated. The arrangement of cells became regular. Cell showed the same polarizarion. By ALP dyeing, cytoplasm became positively black. SEM observed three types of cell morphology: bipolar, multipolar and polygonal cells. Conclusion: The results presented here suggest that the osteoblast-like cells on PLA substrate demonstrated good sign of adhesion, morphology and growth. This in vitro study has provided a positive step towards in vivo implantation and the success of tissue-engineered growth of bone.