金属离子对成骨细胞凋亡、细胞周期分布及ALP分泌的影响  

EFFECT OF METAL IONS Co^(2+) AND Cr^(3+) ON OSTEOBLAST APOPTOSIS,CELL CYCLE DISTRIBUTION,AND SECRETION OF ALKALINE PHOSPHATASE

作　　者：戴闽[1] 周通华[1] 熊皞[1] 邹文楠[2] 詹平[1] 付文峰[1] 

机构地区：[1]南昌大学第一附属医院骨二科,南昌330006 [2]南昌大学建筑工程学院工程力学实验中心

出　　处：《中国修复重建外科杂志》2011年第1期56-60,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(30760255)~~

摘　　要：目的金属磨损产物对人工关节的无菌性松动有影响,通过观察Co2+、Cr3+对小鼠成骨细胞凋亡、细胞周期以及在离子刺激下对成骨细胞分泌ALP的影响,为防治人工关节置换术后假体周围骨溶解提供实验依据。方法体外培养小鼠颅骨成骨细胞MC3T3-E1至第3～5代,调整细胞密度至5×105个/mL,根据干预方法不同将细胞分为两组。实验组:成骨细胞于含10%FBS的α-MEM培养基中培养,待细胞完全贴壁后,加入CoCl2与CrCl3混合溶液;对照组:成骨细胞于含10%FBS的α-MEM培养基中培养。于培养12、24、48h后,采用流式细胞仪检测细胞凋亡率及周期分布,ELISA法检测培养上清液中ALP含量。结果培养12、24、48h,实验组凋亡率分别为13.90%±0.52%、14.80%±0.41%、13.40%±0.26%,对照组分别为8.56%±0.31%、8.19%±0.24%、2.15%±0.11%,差异均有统计学意义(P<0.05)。各时间点实验组细胞G2M期分布比例明显较对照组降低,G0G1期分布比例较对照组增多,差异均有统计学意义(P<0.05)。培养12、24、48h,实验组ALP含量检测吸光度(A)值分别为0.955±0.052、0.624±0.041、0.498±0.026,对照组分别为1.664±0.041、1.986±0.024、2.192±0.041,两组比较差异均有统计学意义(P<0.05)。结论 Co2+、Cr3+对成骨细胞的细胞周期分布有明显影响,抑制其增殖,使细胞大部分处于G0G1期,同时增加其凋亡率,抑制成骨细胞释放ALP。Objective Metal wear products cause the aseptic loosening of joint prosthesis.To investigate the effect of metal ions Co2＋ and Cr3＋ on the osteoblast apoptosis,cell cycle distribution,and secretion of alkaline phosphatase（ALP）,and to search a method to prevent and treat aseptic loosening.Methods The mouse calvarial osteoblasts（MC3T3-E1） were cultured in vitro to 3-5 generations（5 × 105 cells/mL） and divided into 2 groups：the experimental group and the control group.The osteoblasts were cultured in α-MEM medium containing 10%FBS（the control group）,and the mixed solution of CoCl2 and CrCl3 was added after the osteoblasts cultured in α-MEM medium containing 10%FBS attached completely（the experimental group）.At 12,24,and 48 hours after culture,the osteoblast apoptosis and the cell cycle distribution were assessed by flow cytometry;and ELISA method was applied to detect ALP content in serum supernatant.Results At 12,24,and 48 hours after culture,the apoptosis rates in the experimental group（13.90%± 0.52%,14.80%± 0.41%,and 13.40%± 0.26%） were significantly higher than those in the control group（8.56%± 0.31%,8.19%± 0.24%,and 2.15%± 0.11%）,（P 0.05）;G2M（dividing phase） distribution ratio significantly decreased and G0G1（dormancy stage） distribution ratio significantly increased when compared with those in the control group（P 0.05）;and the absorbency（A） values of ALP were 0.955± 0.052,0.624± 0.041,and 0.498± 0.026 in the exprimental group,and were 1.664± 0.041,1.986± 0.024,and 2.192± 0.041 in the control group,showing significant differences between 2 groups（P 0.05）.Conclusion Metal ions Co2＋ and Cr3＋ have a marked effect on osteoblasts cell cycle distribution,which can make most of the cells to be in dormancy stage（G0G1）,up-regulate the apoptosis rate and inhibit the releasion of ALP from osteoblasts.

关 键 词：假体无菌性松动 金属离子 成骨细胞 细胞周期分布 细胞凋亡 ALP 

分 类 号：R687.4[医药卫生—骨科学]