hVEGF基因修饰的SD大鼠BMSCs与胶原膜BME-10X复合物的成骨能力研究

目的：观察人血管内皮生长因子（hVEGF165）基因的骨髓基质细胞（BMSCs）与胶原膜BME‐10X复合的相容性及该复合物移植在体内的成骨情况，为其能否修复牙周骨质缺损提供依据。方法在体外将hVEGF165基因转染的SD大鼠BM SCs种植于胶原膜BM E‐10X复合膜上，使用激光共聚焦扫描显微镜、荧光显微镜、扫描电镜及组织学方法进行观察，并将该复合物植入裸鼠皮下，于4、8周处死裸鼠取材制成组织切片，H‐E染色后观察异位体内成骨和血管生成的情况。结果复合物培养24 h后见目的细胞在胶原膜中贴附、伸展，各层BM E‐10X胶原膜上均有数量不等的细胞附着。转染细胞在胶原膜表面复层生长，并进入内部间隙中。复合物植入裸鼠皮下后，可见新生胶原纤维、类骨质样结构和新生血管的形成。结论 hVEGF165基因转染 BMSCs 在胶原膜BM E‐10X上生长良好，细胞‐胶原复合物在裸鼠体内具有异位成骨及促进新生血管形成能力。

The Complex Constructed by hVEGF165 Genes Transfected BMSCs andCollagen Membrane(BME-10X)Enhanced Osteogenesis and Angiogenesis

Objective To investigate the possibility of forming new bones in rats by incorporating the hVEGF165 transfected BMSCs with collagen membrane (BME‐10X) to form a tissue‐engineered cell and matrix complex ,and transplanting it into Sprague Dawley rats . Methods The hVEGF165 transfected BMSCs were cultured in BME‐10X collagen matrix for 3 days . Histological examination was performed with light microscopy and H‐E staining . Cell attachment was analyzed by fluorescence microscope ,con‐focal laser scanning microscope ,and scanning electron microscope . T he gene‐modified tissue‐engineered complex ,which was constructed by transfected BMSCs and BME‐10X ,was implanted into subcutaneous sites in nude mice . After 4 and 8 weeks ,the animals were sacrificed and the tissue‐engineered complex was evaluated by histology . Results When the transfected cells were seeded in BME‐10X ,it was noted that cell adhered and stretched well after 24 h of culture . Investigation under tomographic scanning with confocal laser scanning confocal microscope showed that cell and material integrated well . The study in vivo showed that when BMSCs were transplanted in the subcutaneous sites of nude mice ,the transfected BMSC complex formed more contents of ossification and angiogenesis than those in the un‐transfected group . Conclusion In this study the BME‐10X showed a good biocompatibility with hVEGF165 transfect‐ed BMSCs . BMSCs transfected with VEGF165 gene showed enhanced osteogenesis and angiogenesis both in v itro and in v ivo .