转化生长因子β1对多孔钽/MG63成骨样细胞复合物细胞增殖及分泌功能的影响

摘要

背景：前期研究已得出国产多孔钽无毒性，具有良好的生物相容性，具有促成骨作用。目的：观察转化生长因子β1对国产多孔钽/MG63成骨样细胞复合物中成骨细胞生长增殖、细胞周期及分泌功能的影响，方法：取生长状态良好的第3代 MG63成骨样细胞悬液，按1×109 L-1的细胞浓度接种于多孔钽材料上，分别以含0，0.5，5，10µg/L 转化生长因子β1的培养基培养。CCK-8法检测培养1-13 d 成骨细胞增殖水平；培养5，9 d，扫描电镜及透射电镜观察培养细胞形态及超微结构，检测细胞分泌Ⅰ型胶原水平。结果与结论：①0.5，5，10µg/L 转化生长因子β1均可促进多孔钽上成骨细胞的增殖，其中以5µg/L转化生长因子β1的促增殖作用最明显；②在5µg/L 转化生长因子β1作用下，成骨细胞在多孔钽支架表面及微孔内形成叠层黏附生长，向材料表面伸出较多的伪足，细胞分泌的基质基本覆盖多孔钽表面，胞浆内可见大量粗面内质网、游离核糖体及致密分布的线粒体，高尔基复合体较发达，并且胞质内出现较多基质小泡；③5µg/L 转化生长因子β1组培养5，9 d 的Ⅰ型胶原水平高于0.5及10µg/L 转化生长因子β1组(P <0.05)；④结果表明：转化生长因子β1可促进国产多孔钽表面 MG63成骨样细胞的增殖，其中以5µg/L 转化生长因子β1对成骨细胞的促增殖及Ⅰ型胶原分泌作用最明显。%BACKGROUND: Previous studies have demonstrated that the Chinese porous tantalum made in China has non-toxicity and good biocompatibility, which can promote osteogenesis. OBJECTIVE: To investigate the effects of transforming growth factor β1 on proliferation, cel cycle and secretion of osteoblasts on porous tantalum/MG63 osteoblast-like cel composites. METHODS: Passage 3 MG63 osteoblast-like cel suspension (1×109/L) was seeded onto the porous tantalum, then the cel composites were inoculated in the medium with 0, 0.5, 5 and 10 µg/L transforming growth factor β1, respectively. The proliferation of osteoblasts was detected by cel counting kit-8 assay at 1-13 days after inoculation; the cel morphology and ultrastructure observed by scanning electron microscope and transmission electron microscopy; and level of col agen type I detected by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSON: 0.5, 5, 10 µg/L transforming growth factor β1 could promote the osteoblast proliferation, and cel proliferation in the 5 µg/L transforming growth factor β1 group was higher than that in the other groups; in the 5 µg/L transforming growth factor β1 group, laminated osteoblasts adhered on the surface and grew into inner of porous tantalum, which extended more pseudopodia toward the scaffold; osteoblasts-secreted matrix could cover the scaffold and numerous rough endoplasmic reticulum, free ribosomes, dense mitochondria, Golgi apparatus as wel as matrix vesicles could be found in the cytoplasm. In addition, the level of col agen type I in the 5 µg/L transforming growth factor β1 group was significantly higher than that in the other groups (P < 0.05). These results indicate that transforming growth factor β1 can promote proliferation, and col agen type I secretion of osteoblasts on porous tantalum/MG63 osteoblast-like cel composites, and the optimum mass concentration of transforming growth factor β1 is 5 µg/L.