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
Effects of strontium-doped calcium phosphate loaded with BMP-2 on proliferation and function of osteoblasts
  
DOI:10.3969/j.issn.1006.7108.2018.07.009
中文关键词:  骨形成蛋白-2    成骨细胞  增殖  功能  磷酸钙
英文关键词:Bone morphogenetic protein-2  Strontium  Osteoblast  Proliferation  Function  Calcium phosphate
基金项目:安徽省教育厅高校自然科学研究重点项目(KJ2017A266);安徽省自然科学基金(1708085QH209)
作者单位
陶周善1 周皖舒2* 江云云1 吴兴净1 王林1 杨民1 谢加兵1 徐祝军1 1.皖南医学院第一附属医院弋矶山医院创伤骨科安徽 芜湖 241000 2.皖南医学院第二附属医院老年病科安徽 芜湖 241000 
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中文摘要:
      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7d,和Con组比较,CPC 组和BSCPC组的成骨细胞数目明显增加(P<0.05),且以BSCPC组成骨细胞数目最多(P<0.05);共培养14d及21d,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(P<0.05),且以BSCPC组细胞钙化能力最强及ALP活性最高(P<0.05);共培养7d,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(P<0.05),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(P<0.05)。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。
英文摘要:
      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.
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