聚乳酸和海藻酸钙体外复合培养人成骨细胞的对比研究

Comparative Study of Polylactic Acid vs Calcium Alginate Cultured with Osteoblasts from Human Periosteum in vitro

目的对比聚乳酸、海藻酸钙与人成骨细胞的生物相容性,为骨组织工程中生物材料的选择提供依据。方法人骨膜成骨细胞体外培养成功后,无菌条件下与聚乳酸、海藻酸钙复合培养,依加入的成骨细胞浓度不同将海藻酸钙组分为3个亚组,于培养2、4、6、8d行组织形态学观察、骨钙素含量、细胞增殖、碱性磷酸酶活性、细胞蛋白含量测定以及细胞周期分析,并行碱性磷酸酶染色和钙结节四环素染色。结果①成骨细胞能在聚乳酸、海藻酸钙上正常黏附、生长、繁殖,其功能不受影响,且海藻酸钙组的以上指标均高于对照组和聚乳酸组,差异有显著性意义(P<0.05);②海藻酸钙成骨细胞高浓度组与低浓度组同时点相比,以上指标均优于低浓度组(P<0.05);结论①聚乳酸、海藻酸钙对成骨细胞的生长和功能无干扰作用,具有良好的生物相容性,且海藻酸钙的生物相容性优于聚乳酸。②在海藻酸钙中加入高浓度成骨细胞其效果更佳。

Objective To study the biocompatibility of polylactic acid and calcium alginate cultured with osteoblasts from human periosteum in vitro in bone tissue engeneering.Methods The successfully cultured osteoblasts in vitro (2×10^(4)/ml) were divided into three groups: control group, polylactic acid group and calcium alginate group (three subgroups according to the density of the added osteoblasts: calcium alginate group 1, 2×10^(4)/ml osteoblasts; calcium alginate group 2, 5×10^(4)/ml osteoblasts; calcium alginate group 3, 10×10^(4)/ml osteoblasts). On the day 2, 4, 6, 8, the morphology, cell proliferation, osteocalcin content, alkaline phosphatase activity, protein content and cell cycle were observed. The ALP staining and the tetracycline staining were done.Results The osteoblasts could adhere, grow and proliferate, and their function was not affected. Between control group and polylactic acid group as well as calcium alginate group 1, there was significant difference in cell proliferation, osteocalcin content and alkaline phosphatase activity (P<0.05, P<0.01). The number of osteoblasts in S stage in calcium alginate group 1 was much more than in control group and polylactic acid group, so was the osteogenic nod number.Conclusion Both polylactic acid and calcium alginate have good biocompatibility with osteoblast from human periosteum. Further more, calcium alginate can stimulate cell proliferation. They can be used as a biomaterial for bone tissue engineering. Addition of high concentration of osteoblasts into the three calcium alginate could achieved best effects.