多孔纯钛表面掺锶羟基磷灰石涂层对骨髓间充质干细胞粘附的影响（英文）

发布时间：2018-03-02 16:22

本文选题：细胞粘附　切入点：骨髓间充质干细胞　出处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2017年09期 　论文类型：期刊论文

【摘要】：目的:观察电化学沉积的纳米结构掺锶羟基磷灰石涂层表面对大鼠骨髓间充质干细胞早期粘附行为的影响。创新点:首次观察电化学沉积的掺锶羟基磷灰石涂层表面骨髓间充质干细胞的早期粘附行为,并对相关基因进行了检测。方法:纯钛表面经过喷砂和双重酸处理,形成多孔粗糙结构。用电化学方法在其粗糙表面沉积羟基磷灰石涂层(HA)和掺锶羟基磷灰石涂层(Sr-HA)。用贴壁法将4周大鼠股骨骨髓间充质干细胞分离进行原代培养后将细胞接种到多孔纯钛,HA和Sr-HA表面培养1、6和24小时。用场发射扫描电镜(SEM)观察细胞的形貌特点。用异硫氰酸荧光素(FITC)标记的鬼笔环肽进行免疫荧光染色标记细胞骨架,Hoechst 33258进行细胞核染色,激光共聚焦荧光显微镜进行拍照后使用Image J进行细胞的计数和图形分析。用实时荧光定量逆转录聚合酶链式反应(RT-q PCR)测定24、48和72小时不同实验组中骨髓间充质干细胞中FAK、vinculin、integrin β1和integrin β3的基因表达,并进行统计学分析。结论:SEM观察结果显示,骨髓间充质干细胞在三种表面都能正常黏附、生长和增殖。在Sr-HA组表面,细胞粘附、细胞活力和细胞骨架的铺展都较粗糙表面组和HA组有较显著的提高。RT-q PCR结果显示,在各个时间点Sr-HA组表面骨髓间充质干细胞的FAK、vinculin、integrin β1和integrin β3的基因表达与粗糙组有显著性差异,且在24小时后与HA组亦有显著性差异。综上所述,掺锶羟基磷灰石纳米涂层具有较好的生物相容性,可以促进大鼠骨髓间充质干细胞在纯钛表面的早期粘附。
[Abstract]:Objective: to observe the effect of Sr-doped hydroxyapatite coating on the early adhesion behavior of rat bone marrow mesenchymal stem cells by electrochemical deposition. Early adhesion behavior of surface bone marrow mesenchymal stem cells, Methods: the surface of pure titanium was treated by sand blasting and double acid treatment. The porous rough structure was formed. Hydroxyapatite (HA) and strontium doped hydroxyapatite (Sr-HAN) were deposited on the rough surface by electrochemical method. Bone marrow mesenchymal stem cells (MSCs) were isolated by adherent method for 4 weeks and cultured in primary culture. The cells were cultured on porous pure titanium HA and Sr-HA for 1 h and 24 h. The morphology of the cells was observed by field emission scanning electron microscopy (SEM). The cells were labeled with fluorescein isothiocyanate (FITC) and labeled with phalanophorin. Cytoskeleton Hoechst 33258 was used for nuclear staining. Image J was used for cell counting and graphic analysis after laser confocal fluorescence microscopy was taken. Bone marrow mesenchymal dryness was measured by real-time fluorescence quantitative reverse transcriptase polymerase chain reaction (RT-q PCR) in different experimental groups for 48 and 72 hours. The gene expression of integrin 尾 _ 1 and integrin 尾 _ 3 in FAK vinculin was detected. Conclusion: bone marrow mesenchymal stem cells can adhere to, grow and proliferate normally on all three surfaces. The expression of FAKvinculin integrin 尾 1 and integrin 尾 3 on the surface of bone marrow mesenchymal stem cells in Sr-HA group was significantly higher than that in rough surface group and HA group. The results showed that the gene expression of FAKvinculin 尾 1 and integrin 尾 3 on the surface of bone marrow mesenchymal stem cells in Sr-HA group was significantly different from that in rough group at different time points. In conclusion, strontium hydroxyapatite nano-coating has good biocompatibility and can promote the early adhesion of rat bone marrow mesenchymal stem cells on the surface of pure titanium.
【作者单位】： Department
【基金】：supported by the Zhejiang Provincial Natural Science Foundation of China(No.LQ13H140001)
【分类号】：R783.1

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