破骨细胞TRPV5通道对体外降解生物珊瑚人工骨的影响  

Effect of osteoclast TRPV5 channel on in vitro degradation of biological coral artificial bone

作　　者：崔红旺 王良盛 温鹏 孟志斌[2] Cui Hongwang;Wang Liangsheng;Wen Peng;Meng Zhibin(Trauma Medical Center,The First Affiliated Hospital of Hainan Medical University,Haikou 570102,Hainan Province,China;Department of Spine Surgery,The First Affiliated Hospital of Hainan Medical University,Haikou 570102,Hainan Province,China)

机构地区：[1]海南医学院第一附属医院创伤医学中心,海南省海口市570102 [2]海南医学院第一附属医院脊柱外科,海南省海口市570102

出　　处：《中国组织工程研究》2023年第21期3337-3342,共6页Chinese Journal of Tissue Engineering Research

基　　金：海南省重点研发计划项目(ZDYF2021SHFZ084),项目负责人:崔红旺。

摘　　要：背景:生物珊瑚人工骨是一种修复长段骨缺损的良好替代材料,但其在体内的降解吸收速率与新生骨的生长速率不够匹配,因而其在临床上的应用受到限制。目的:探讨破骨细胞TRPV5通道对生物珊瑚人工骨降解的影响。方法:取生长状态良好的小鼠单核巨噬细胞株RAW264.7,接种于生物珊瑚人工骨片上,加入破骨细胞诱导分化培养液,观察有破骨细胞出现后,分别加入含0,50,500,5000μmol/L钌红(TRPV5通道抑制剂)的破骨细胞诱导分化培养液。培养一定时间后,激光共聚焦显微镜下观察TRPV5通道蛋白表达,扫描电镜下观察生物珊瑚人工骨片上的吸收陷窝,应用Western Blot检测细胞TRPV5通道蛋白表达。结果与结论:①激光共聚焦显微镜:TRPV5表达于破骨细胞的胞浆和胞膜,随着钌红浓度的增加,破骨细胞上TRPV5表达减少;②扫描电镜:0 nmol/L钌红组骨片上可见大片连续的骨吸收陷窝,其他浓度钌红组骨片上的骨吸收陷窝减少,并散在分布,并且随着钌红浓度的升高,骨陷窝面积逐渐减少;③Western Blot检测:与0 nmol/L钌红组比较,其他浓度钌红组TRPV5通道蛋白表达明显降低(P<0.05或P<0.01);④结果表明:破骨细胞在生物珊瑚人工骨片上生长良好,TRPV5可作为调控生物珊瑚人工骨降解速率的靶点。BACKGROUND:Biological coral artificial bone is a good alternative material for repairing long bone defects.However,the degradation and absorption rate of biological coral artificial bone in the human body does not match the growth rate of new bone,which leads to the limitation of its clinical application.OBJECTIVE:To explore the effect of osteoclast TRPV5 on the degradation of biological coral artificial bone.METHODS:Monocyte macrophage strain RAW264.7 from mouse was inoculated on biological coral artificial bone slices,which were co-cultured with osteoclasts.After the appearance of osteoclasts was observed,the osteoclast-induced differentiation medium containing 0,50,500,5000μmol/L ruthenium red(TRPV5 channel inhibitor)was added separately.After culturing for a certain period of time,the expression of TRPV5 channel protein was observed under a laser confocal microscope.The absorption lacuna on the biological coral artificial bone slice was observed under a scanning electron microscope.The expression of TRPV5 channel protein in cells was detected by western blot assay.RESULTS AND CONCLUSION:(1)Laser scanning confocal microscope:TRPV5 could be expressed on the cytoplasm and membrane of osteoclasts.With the increase of ruthenium red concentration,the expression of TRPV5 on osteoclasts decreased.(2)Scanning electron microscope:Large continuous bone resorption lacuna was seen on the bone slices in the 0 nmol/L ruthenium red group.The bone resorption lacuna on the bone slices in other concentrations of ruthenium red decreased and was scattered,and with the increase of the concentration of ruthenium red,the area of the bone lacuna gradually decreased.(3)Western blot assay:Compared with 0 nmol/L ruthenium red group,the expression of TRPV5 channel protein in other concentrations of ruthenium red groups was significantly decreased(P<0.05 or P<0.01).(4)The results confirm that osteoclasts grow well on biological coral artificial bone,and TRPV5 can be used as a target to regulate the degradation rate of biological coral ar

关 键 词：生物珊瑚人工骨 降解 破骨细胞 TRPV5通道 骨缺损 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R-331