负载BMP-2掺锶磷酸钙复合材料对成骨细胞增殖及功能的影响

目的探索负载BMP-2掺锶磷酸钙复合材料对成骨细胞增殖及功能的影响。方法获取SD大鼠成骨细胞,随机分为空白对照组(Con组)、磷酸钙组(CPC组)和复合材料组(BSCPC组);培养基中分别添加安慰剂、磷酸钙和负载BMP-2掺锶磷酸钙共培养一段时间,通过CCK-8法检测成骨细胞的增殖情况,碱性磷酸酶(alkalinephosphatase,ALP)及茜素红染色观察细胞的功能状态,蛋白电泳观察骨保护素(osteoprotegerin,OPG)、核因子κB受体活化因子(receptor activator of nuclear factor-KB,RANK)及核因子κB受体活化因子配体(ligand of receptor activator of nuclear factor-κB,RANKL)蛋白的表达情况。结果共培养1、3、5和7 d,和Con组比较,CPC组和BSCPC组的成骨细胞数目明显增加(P0.05),且以BSCPC组成骨细胞数目最多(P0.05);共培养14 d及21 d,和Con组比较,CPC组和BSCPC组的成骨细胞的矿化能力及ALP活性明显增加(P0.05),且以BSCPC组细胞钙化能力最强及ALP活性最高(P0.05);共培养7 d,和Con组比较,CPC组和BSCPC组的成骨细胞的OPG表达明显增加,而RANK及RANKL蛋白表达明显降低(P0.05),且以BSCPC组的成骨细胞蛋白OPG、RANK及RANKL蛋白表达量改善最为显著(P0.05)。结论负载BMP-2掺锶磷酸钙复合材料促进成骨细胞增殖分化和改善细胞活性和功能。

Effects of strontium-doped calcium phosphate loaded with BMP-2 on proliferation and function of osteoblasts

Objective To investigate the effect of strontium-doped calcium phosphate loaded with BMP-2 on the proliferation and function of osteoblasts. Methods The osteoblasts of SD rats were obtained and randomly divided into blank control group (Con group), calcium phosphate group (CPC group), and composite group (BSCPC group). Placebo, calcium phosphate, and strontium-doped calcium phosphate loaded with BMP-2 were added to culture medium for co-culture for a period of time. The proliferation of osteoblasts was detected using CCK-8 assay. Alkaline phosphatase (ALP) and Alizarin red staining were used to observe the functional status of the cells. Protein electrophoresis was used to observe the expression of osteoprotegerin (OPG), RANK, and RANKL. Results Compared with Con group, the number of osteoblasts in CPC group and BSCPC group increased significantly (P<0.05), and the number of osteoblasts was the highest in BSCPC group (P<0.05). Compared with Con group, the mineralization and ALP activity of osteoblasts in CPC group and BSCPC group increased significantly (P<0.05), and the cells in BSCPC group had the highest calcification ability and the highest ALP activity (P<0.05). Compared with Con group, the OPG expression of osteoblasts in CPC group and BSCPC group increased significantly, while the expression of RANK and RANKL protein decreased significantly (P<0.05). The expression levels of OPG, RANK and RANKL protein improved most significantly in BSCPC group (P<0.05). Conclusion Strontium-doped calcium phosphate composites loaded with BMP-2 promotes osteoblast proliferation and differentiation and improves cell activity and function.