静磁场对纯钛表面成骨细胞和巨噬细胞的影响  

Effect of Static Magnetic Field on Osteoblasts and Macrophages on Pure Titanium

作　　者：蒋沐岐 孙宇轩 孙亚生 聂志祥 张贞 胡丽 JIANG Muqi;SUN Yuxuan;SUN Yasheng;NIE Zhixiang;ZHANG Zhen;HU Li(Department of Stomatology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China;Wuhan National High Magnetic Field Center,Huazhong University of Science and Technology,Wuhan 430074,China)

机构地区：[1]华中科技大学同济医学院附属协和医院口腔医学中心,湖北武汉430022 [2]华中科技大学国家脉冲强磁场科学中心,湖北武汉430074

出　　处：《口腔医学研究》2024年第6期512-518,共7页Journal of Oral Science Research

基　　金：武汉强磁场学科交叉基金(编号:WHMFC202208);华中科技大学研究生创新基金(编号:YCJJ202201052)。

摘　　要：目的:本研究旨在探讨静磁场对纯钛表面成骨细胞及巨噬细胞的影响及机制。方法:在0 mT和100 mT静磁场作用下,细胞分别被接种到孔板表面和大颗粒喷砂酸蚀处理钛片表面。CCK-8法检测静磁场对成骨细胞及巨噬细胞活力的影响;细胞骨架荧光染色检测静磁场下成骨细胞及巨噬细胞的形态改变;免疫荧光染色检测静磁场下成骨细胞的骨特异性转录因子2(runt-related transcription factor 2,Runx2)蛋白表达;实时荧光定量聚合酶链反应(real-time fluorescence quantitative PCR,RT-qPCR)检测成骨细胞中成骨相关基因及巨噬细胞中免疫相关基因表达;流式细胞术检测巨噬细胞极化相关蛋白CD86、CD206表达水平。结果:100 mT静磁场可以有效促进钛片表面成骨细胞增殖,促进成骨相关基因碱性磷酸酶(alkaline phosphatase,ALP)、Ⅰ型胶原蛋白(collagen typeⅠ,COL-1)、Runx2、骨保护蛋白(osteoprotegerin,OPG)表达,增强成骨相关蛋白Runx2合成;100 mT静磁场会促进巨噬细胞增殖,且能促进巨噬细胞第3天时向M1型极化,在第7天时会抑制炎症相关基因肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)表达。结论:静磁场具有促进大颗粒喷砂酸蚀处理钛片表面成骨细胞增殖、矿化能力,以及促进钛表面巨噬细胞从前期促炎作用向后期抗炎作用的转变能力。Objective:To explore the effect and mechanism of static magnetic field on osteoblasts and macrophages on pure titanium surface.Methods:Under the static magnetic fields of 0 mT and 100 mT,cells were cultured on the surface of tissue culture well plates and sandblasting and acid etching(SLA-treated)titanium sheets,respectively.CCK-8 method was used to detect the effect of static magnetic field on the viability of osteoblasts and macrophages.Cytoskeleton fluorescence staining was used to detect the morphological changes of osteoblasts and macrophages under static magnetic field.Immunofluorescence staining was used to detect the expression of runt-related transcription factor 2(Runx2),protein in osteoblasts under static magnetic field.The expression of osteogenesis-related genes in osteoblasts and immune-related genes in macrophages was detected by quantitative real-time fluorescence quantitative PCR(RT-qPCR).The expression levels of polarization-related proteins CD86 and CD206 in macrophages were detected by flow cytometry.Results:The static magnetic field of 100 mT could effectively promote the proliferation of osteoblasts on the titanium sheets,promote the expression of osteogenesis-related genes alkaline phosphatase(ALP),collagen typeⅠ(COL-1),Runx2,and osteoprotegerin(OPG),and enhance the synthesis of osteogenesis-related protein Runx2.The static magnetic field of 100mT could promote the proliferation of macrophages,promote the polarization of macrophages to M1 type on the 3rd day,and inhibit the expression of inflammation-related genes tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)on the 7th day.Conclusion:Static magnetic field has the ability to promote the proliferation and mineralization of osteoblasts on the SLA-treated titanium sheets,and to promote the transformation of macrophages from pro-inflammatory to anti-inflammatory on the titanium sheets.

关 键 词：静磁场 钛 成骨 免疫 

分 类 号：R783.1[医药卫生—口腔医学]