含硅涂层镁合金对骨髓干细胞生物活性的影响  

Effect of silicon-coated magnesium alloy on the biological activity of bone marrow stem cells in vitro

作　　者：杨小明 谢俊杰 李梅[2] 林潇[3] 谭丽丽[4] 杨柯[4] Yang Xiaoming;Xie Junjie;Li Mei;Lin Xiao;Tan Lili;Yang Ke(Department of Orthopaedics,Quanzhou First Hospital affiliated Fujian Medical University,Quanzhou 362000,China;Guangdong Provincial People′s Hospital,Guangzhou 510030,China;Institute of Orthopedics,Soochow University,Suzhou 215026,China;Institute of Metal Research,Chinese Academy of Sciences,Shenyang 110016,China)

机构地区：[1]福建医科大学附属泉州第一医院骨科,362000 [2]广东省人民医院骨科实验室,广州510030 [3]苏州大学骨科研究所,215026 [4]中国科学院金属研究所,沈阳110016

出　　处：《中华实验外科杂志》2021年第11期2100-2103,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察含硅涂层镁合金体外对小鼠骨髓间充质干细胞生物活性的影响。方法实验根据材料和培养液的不同分为硅涂层镁合金组(A组)、裸镁合金组(B组)、医用钛合金组(C组)和空白对照组(D组)。倒置相差显微镜观察细胞形态,采用细胞计数试剂盒(CCK-8)检测细胞增殖活力,扫描电镜观察细胞在材料表面的黏附情况,成骨分化能力通过碱性磷酸酶(ALP)、矿化结节及成骨相关基因检测评价。组间比较采用单因素方差分析,两两比较采用LSD-t检验。结果扫描电镜下见硅涂层镁合金表面呈粗糙、多孔形态;材料浸提液培养后各组细胞轮廓清晰、形态良好。在培养的第5天,各组的吸光度(A)值分别为A组(0.380)、B组(0.383)、C组(0.384)和D组(0.398),细胞毒性检测差异无统计学意义(F=1.051,P>0.05);扫描电镜观察见A组材料表面细胞生长粘附形态良好。细胞培养10 d,茜素红染色结果提示A组细胞的矿化程度最高,A值为0.307,明显高于B、C、D组的0.237、0.105和0.114(t=5.538、16.032、15.330,P<0.01)。成骨相关基因表达量检测结果提示A组的ALP、骨钙蛋白和Ⅰ型胶原基因的相对表达量分别为4.015、6.425和6.679,明显高于B组(2.150、3.516、5.264,t=32.297、21.965、14.167,P<0.01),C组(1.043、1.025、1.063,t=51.461、40.774、56.222,P<0.01)和D组(1.162、1.004、1.005,t=49.400、40.933、56.799,P<0.01)。结论含硅涂层镁合金材料在体外具有良好的生物活性,有利于细胞的生长、黏附、增殖和分化。Objective To observe the effect of silicon coated magnesium alloy on the bioactivity of mouse bone marrow stem cells in vitro.Methods The experiments were divided into 4 groups according to different materials and culture medium:silicon-containing coated alloy group(group A),uncoated alloy group(group B),titanium alloy group(group C),and negative control group(group D).The cell morphology was observed by a microscopy.The cell viability was detected by cell counting kit-8(CCK-8),and cell attachment morphology on the surface of different samples was observed by scanning electron microscope(SEM).To evaluate the osteogenic differentiation ability of cell,the activity of alkaline phosphatase(ALP),mineralized nodules and osteogenic related genes was detected.One-Way ANOVA and LSD-t test were used for the comparison between multiple groups.Results The surface characterization of silicon coated alloy showed a rough and porous layer by SEM.After cultured with the extract,cells grew well and presented good shape in all groups by a microscopy.On the 5th day of culture,the absorbance(A)values of each group were as follows:group A(0.380),group B(0.383),group C(0.384)and group D(0.398),there was no significant difference(F=1.051,P>0.05).Cells in groups A presented a good cell extension under SEM.The results of Alizarin Red staining showed that the mineralized degree was the highest in group A(A values=0.307),which was significantly higher than that in group B,C and D(0.237,0.105,0.114,t=5.538,16.032,15.330,P<0.01).The expression values of ALP,osteocalcin and collagen typeⅠgenes in group A were 4.015,6.425 and 6.679,respectively,which were significantly higher than group B(2.150,3.516,5.264,t=32.297,21.965,14.167,P<0.01),group C(1.043,1.025,1.063,t=51.461,40.774,56.222,P<0.01)and group D(1.162,1.004,1.005,t=49.400,40.933,56.799,P<0.01).Conclusion The silicon-containing coated magnesium alloy showed a good biological activity in vitro,which is conducive to cell growth,adhesion,proliferation and differentiation.

关 键 词：硅涂层 镁合金 生物活性 

分 类 号：R318.08[医药卫生—生物医学工程]