转染特异性报导基因成骨细胞生物学性状及其与骨支架材料生物相容性的研究

The biological characteristics of osteoblasts transfected with specific reporter gene and their biocom-patibility with bone scaffold materials

目的研究转染特异性报导基因成骨细胞的生物学性状，并观察该细胞在骨支架材料上的生长特性和黏附性。方法常规复苏培养转染12×SBE—OC—Luc报导基因的前成骨细胞株OCTl细胞，通过倒置相差显微镜、HE染色、I型胶原免疫组化法染色、碱性磷酸酶（ALP）偶氮偶联法染色、矿化结节茜素红法染色及四环素法染色观察该转基因成骨细胞的生物学性状；并将其与纳米磷酸钙复合骨支架进行体外复合培养，通过扫描电镜观察该转基因成骨细胞在骨支架材料上的生长特性和黏附性。结果转染12×SBE—OC—Luc报导基因的前成骨细胞株OCTl细胞经培养后能贴壁生长，形态和成纤维细胞相似，能分泌胶原基质和ALP，经I型胶原免疫组化法染色及ALP偶氮偶联法染色呈强阳性；并能够形成矿化结节，茜素红法染色及四环素法染色呈阳性；该转基因成骨细胞在骨支架材料上具有良好的生长特性和黏附性，并能正常分化、增殖和成熟，分泌大量胶原基质和钙结节。结论转染12×SBE—OC—Luc报导基因后成骨细胞生物学性状未发生改变，其与骨支架材料亦具有良好的生物相容性。

Objective To study the biological characteristics of osteoblasts transfected with 12 ×SBE-OC-Luc reporter gene and their growth and conglutination on bone scaffold materials. Methods Pre-osteoblast cell strain-OCT-1 cells of transfected 12 × SBE-OC-Luc reporter gene were resuscitated and cultured. HE staining, type I collagen immunohistochemical staining, alkaline phosphatase staining, calcified nodus alizarin-red staining and cyclomycin staining were used to observe the biological characteristics of the transgene osteoblast. The transgene osteoblasts and NM calcium phosphate combined bone scaffolds were cocultured in vitro. The growth and conglutination properties of the transgene osteoblasts on bone scaffold materials were investigated by scan electron microscopy. Results OCT-1 cells （a prc-osteoblast cell line） of trans- fected 12 × SBE-OC-Luc reporter gene grew and sticked onto walls after culture. Their morphous was similar to that of the fibroblasts. They had capacities of secreting collagen and forming calcified nodi. The type I collagen immunohistochemieal staining and alkaline phosphatase staining showed obviously positive. Alizarin-red staining and eyelomycin staining showed positive. The transgene osteoblasts grew and adhered well onto the bone scaffold materials. They could differentiate, proliferate and mature normally, secreting collagen and forming calcified nodi. Conclusion After being tmnsfected with 12 × SBE-OC-Luc reporter gene, the osteoblasts will not change in biological characters, and have good biocompatibility with bone scaffold materials.