复合压电薄膜生物相容性及电输出性能与兔骨髓间充质干细胞的成骨分化  

Biocompatibility and electrical output performance of composite piezoelectric film and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells

作　　者：王桢 许顺恩 汤耿[4] 罗四维 滕建祥 谢孟利 何佳林 叶川 Wang Zhen;Xu Shunen;Tang Geng;Luo Siwei;Teng Jianxiang;Xie Mengli;He Jialin;Ye Chuan(Department of Orthopedics,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Research Center of Tissue Engineering and Stem Cell Technique,Guizhou Medical University,Guiyang 550004,Guizhou Province,China;College of Mechanical and Electrical Engineering,Guizhou Normal University,Guiyang 550025,Guizhou Province,China)

机构地区：[1]贵州医科大学附属医院骨科,贵州省贵阳市550004 [2]贵州医科大学,贵州省贵阳市550004 [3]组织工程与干细胞实验中心,贵州省贵阳市550004 [4]贵州师范大学机械与电气工程学院,贵州省贵阳市550025

出　　处：《中国组织工程研究》2024年第13期1969-1975,共7页Chinese Journal of Tissue Engineering Research

基　　金：贵州省科技厅资助项目(黔科合平台人才[2020]6013;黔科合支撑项目[2020]4Y137),项目负责人:叶川~~。

摘　　要：背景:可控的压电效应能促进组织的再生与修复,压电材料在负重组织工程领域应用广泛。目的:制备一种可促进骨组织再生的压电薄膜材料,探究其结构表征、电输出性能、生物相容性及其电输出对兔骨髓间充质干细胞成骨分化的影响。方法:以聚3-羟基丁酸酯4-羟基丁酸酯(poly-3-hydroxybutyrateco/4-hydroxybutyrate,P34HB)为原料,加入质量比分别为0%,5%,10%,15%,20%的锡钛酸钡钙(Ba0.94Ca0.06Sn0.08Ti0.92O3,BCST)粉末,以二氯甲烷为溶剂,通过真空干燥方法制得150-200μm厚度BCST/P34HB压电薄膜材料,经油浴极化后,检测其表面形貌、物相组成、压电系数和开路电压,以及BCST/P34HB在110 Hz、0.25 N力幅度下的电输出对兔骨髓间充质干细胞增殖和成骨分化的影响。结果与结论:①扫描电镜、X射线衍射、水接触角、压电系数检测及电输出性能测试显示,当BCST质量比增加至20%时,BCST/P34HB压电薄膜有着良好的压电性能(d33=5.9 pC/N)及电输出性能(180 mV),更接近500 mV的合适电刺激成骨范围;②活死染色显示,与兔骨髓间充质干细胞共培养第1天,15%组与20%组显示出较少的红色荧光;当培养到第5天时,各组绿色荧光数量较第1天明显增多,10%组、15%组和20%组未见红色荧光,仅0%组和5%组有少量的红色荧光;③阿尔玛蓝染色显示,与兔骨髓间充质干细胞共培养第1,3,5天,随着时间的增加,各组细胞都呈现出增长的趋势,当培养到第5天时,20%组的细胞量明显高于0%组(P<0.05);④成骨诱导第10天,碱性磷酸酶染色结果显示20%组的染色阳性率明显高于0%组(P=0.0001);成骨诱导第21天,茜素红染色和钙结节定量分析显示出了与碱性磷酸酶染色相似的趋势,与0%组相比,15%组和20%组均显示出明显的差异(P<0.01,P<0.0001);⑤结果表明,20%BCST/P34HB薄膜具有良好的压电性能、电输出性能、生物相容性及促骨髓间充质干细胞成骨分化能力。BACKGROUND:Adjustable piezoelectric effect can promote tissue regeneration and repair.Piezoelectric materials are widely used in weight-bearing tissue engineering.OBJECTIVE:To prepare a piezoelectric film material that can promote bone regeneration,and to explore its structural characterization,electrical output performance,biocompatibility,and effect of electrical output on osteogenic differentiation of rabbit bone marrow mesenchymal stem cells.METHODS:Using poly-3-hydroxybutyrateco/4-hydroxybutyrate(P34HB)as raw material,barium calcium stannate titanate powder(Ba0.94Ca0.06Sn0.08Ti0.92O3,BCST)was added according to mass ratios of 0%,5%,10%,15%,and 20%.Dichloromethane was added to solve P34HB,and the thickness of 150-200μm BCST/P34HB piezoelectric film was prepared by vacuum drying method.After polarization in the oil bath,the surface morphology,crystal phase composition,piezoelectric coefficient and open circuit voltage were tested.The effect of BCST/P34HB electrical output at 110 Hz and 0.25 N force on the proliferation and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells was tested.RESULTS AND CONCLUSION:(1)Scanning electron microscopy,X-ray diffraction,water contact angle,piezoelectric coefficient and electrical output performance tests showed that when the mass ratio of BCST increased to 20%,the BCST/P34HB piezoelectric film had good piezoelectric properties(d33=5.9 pC/N)and electrical output performance(180 mV),which was closer to the suitable range of 500 mV for electrical stimulation.(2)Live and dead staining showed that on the first day of co-culture,15%group and 20%group showed less red fluorescence.On the 5th day of culture,the number of green fluorescence in each group was significantly higher than that on the first day,and the red fluorescence was not observed in the 10%,15%and 20%groups,and only a small amount of red fluorescence was observed in the 0%and 5%groups.(3)On the 1st,3rd and 5th days of co-culture with rabbit bone marrow mesenchymal stem cells,Almar blue staining e

关 键 词：骨组织工程 P34HB 压电材料 薄膜材料 成骨分化 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R314