壳聚糖修饰后的纳米羟基磷灰石作为BMP-2输送载体转染细胞的研究

目的：研究羟基磷灰石（hydroxyapatite,HA）纳米粒载体经壳聚糖（chitosan,CS）修饰后介导骨形态发生蛋白2（Bone morphogenetic protein2,BMP2）基因转染成骨细胞后的表达及表达产物对成骨细胞的影响。方法：用壳聚糖对羟基磷灰石纳米粒进行表面修饰;复苏成骨细胞株并进行传代培养,采用HA-CS纳米粒载体转染技术,将BMP-2目的基因导入成骨细胞,RT-PCR法鉴定BMP-2的整合和表达;检测细胞中碱性磷酸酶（alkaline phoaphatase,ALP）及骨钙素（osteocalcin,OCN）活性。结果：RT-PCR结果显示BMP-2基因在成骨细胞中稳定表达,而且表达产物增加了成骨细胞中碱性磷酸酶及骨钙素活性。结论：HA-CS纳米粒作为输送载体可以安全地将目的基因BMP-2导入成骨细胞中并稳定表达,同时增加了成骨细胞的生物学活性。

Surface modification of Nano-hydroxyapatite as carrier of BMP-2 to cell transfection

Objective:To investigate the expression of bone morphagenetic protein 2(BMP-2) gene mediated by hydroxyapatite(HA) nanoparticles with chitosan(CS) into osteoblast and the impact of BMP-2 on osteoblast.Methods:The surface of HA nanoparticles was modified by CS,and the osteoblast was recovered and cultured.The BMP-2 gene was transfected into osteoblast mediated by HA-CS nanoparticles,and RT-PCR was used to identify the expression of BMP-2.The activity of osteoblast was studied by alkaline phoaphatase(ALP) and osteocalcin(OCN).Results:RT-PCR demonstrated the BMP-2 gene was steadily expressed in the osteoblast and enhanced the bone ALP group activity and increased the content of OCN after transfection.Conclusion:HA-CS nanoparticle as a carrier can safely transfect target gene BMP-2 into osteoblast and express steadily by itself and increase the osteoblast biological activity.