钛表面锶改性后骨髓基质干细胞黏附、迁移、增殖及成骨相关基因的表达  

Effects of strontium-modified titanium surfaces on adhesion, migration and proliferation of bone marrow mesenchymal stem cells and expression of bone formation-related genes

作　　者：刘春栋[1] 申晓青[1] 张艳丽 张小根[1] 吴补领[2,3] Liu Chundong;Shen Xiaoqing;Zhang Yanli;Zhang Xiaogen;Wu Buling(Department of Stomatology,Zhujiang Hospital,Southern Medical University,Guangzhou 510280,Guangdong Province,China;Department of Stomatology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,Guangdong Province,China;College of Stomatology,Southern Medical University,Guangzhou 510515,Guangdong Province,China)

机构地区：[1]南方医科大学珠江医院口腔科,广东省广州市510280 [2]南方医科大学南方医院口腔科,广东省广州市510515 [3]南方医科大学口腔医学院,广东省广州市510515

出　　处：《中国组织工程研究》2020年第7期1009-1015,共7页Chinese Journal of Tissue Engineering Research

基　　金：广东省医学科研基金项目(B2016040)，项目负责人：刘春栋~~

摘　　要：背景:锶是一种既刺激骨形成、又抑制骨吸收的元素,纯钛种植体表面锶改性促进骨质疏松状态下种植体骨结合是当前研究的热点,具有非常重要的意义。目的:探讨改良锶改性纯钛表面对大鼠骨髓基质干细胞黏附、铺展、迁移、增殖及成骨相关基因表达的影响。方法:将TA2纯钛片打磨清洗后,依次放于5mol/L NaOH溶液、100mmol/L醋酸锶溶液内处理,并以600℃或者700℃进行热处理1h,去离子水80℃水化处理(W)24 h,以纯钛为对照,共分为5组,分别为Ti、Sr600、Sr600W、Sr700、Sr700W组。采用全骨髓培养法获取大鼠骨髓基质干细胞,将改性钛片置于24孔培养板中与细胞悬液培养,CCK-8法检测骨髓基质干细胞的黏附和增殖情况。骨髓基质干细胞与钛片共培养24 h后对细胞肌动蛋白进行染色,观察细胞黏附伸展形态;通过细胞划痕实验和荧光染色,观察骨髓基质干细胞在各组钛片表面迁移的能力;qRT-PCR法检测各组细胞Ⅰ型胶原、Runx2、骨保护蛋白、RANKL、骨桥蛋白的mRNA表达。结果与结论:锶改性纯钛片可以促进骨髓基质干细胞的铺展、迁移。在5d和7d时,锶改性钛片表面细胞增殖较纯钛组均明显升高(P<0.001)。在5d时,Sr700组的细胞增殖数目较Sr600组高(P<0.01)。在14 d时,锶改性纯钛片可以促进骨髓基质干细胞Ⅰ型胶原、Runx2、骨桥蛋白的表达。结果表明,改良锶改性纯钛可以促进大鼠骨髓基质干细胞的黏附、铺展、迁移、增殖及成骨相关基因的表达,水化处理对大鼠骨髓基质干细胞成骨向分化具有促进作用。BACKGROUND:Strontium promotes bone formation,and reduces bone resorption.Strontium-modified titanium implant surface has been the focus of research in implant osseointegration under osteoporotic conditions.OBJECTIVE:To investigate the cell adhesion,stretch,and migration of advanced strontium-modified titanium surfaces using bone marrow mesenchymal stem cells isolated from rats.METHODS:Strontium-modified titanium surfaces were produced by sequential treatments with 5 mol/L NaOH,100 mmol/L strontium acetate,and heated at 600 or 700℃ for 1 hour.After heat treatment,half the samples were soaked in deionized water at 80℃ for 24 hours,then washed and dried.The pure titanium served as control group,and there were five groups.The whole bone marrow adherence method was used to separate bone marrow mesenchymal stem cells from the rats.The modified titanium sheets were placed in 24-well plates and cultured in cell suspension.Cell adhesion and cell proliferation were assessed using cell counting kit-8 assay.After bone marrow mesenchymal stem cells were with the titanium sheet for 24 hours,the actin was stained to observe cell adhesion and stretch.The migration of bone marrow mesenchymal stem cells on different titanium surfaces was investigated using cell scratch test and fluorescence staining.The expression levels of collagen type?,Runx2,osteoprotegerin,RANKL and osteopontin were detected by qRT-PCR.RESULTS AND CONCLUSION:Strontium-modified titanium could promote the stretch and migration of bone marrow mesenchymal stem cells.The number of proliferative cells in the Sr700 group was significantly higher than that in the Sr600 group at 5 days(P<0.01).On day 14,strontium-modified titanium promoted the expression of collagen type?,Runx2,and osteopontin.In summary,strontium-modified titanium can promote adhesion,spreading,migration and proliferation of rat bone marrow mesenchymal stem cells.Moreover,hydration treatment can enhance the osteogenic activity of rat bone marrow mesenchymal stem cells.

关 键 词：纯钛 锶改性 骨髓基质干细胞 细胞黏附 细胞增殖 成骨相关基因 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]