本文選題：細(xì)胞粘附　切入點(diǎn)：骨髓間充質(zhì)干細(xì)胞　出處：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2017年09期 　論文類型：期刊論文

【摘要】：目的:觀察電化學(xué)沉積的納米結(jié)構(gòu)摻鍶羥基磷灰石涂層表面對(duì)大鼠骨髓間充質(zhì)干細(xì)胞早期粘附行為的影響。創(chuàng)新點(diǎn):首次觀察電化學(xué)沉積的摻鍶羥基磷灰石涂層表面骨髓間充質(zhì)干細(xì)胞的早期粘附行為,并對(duì)相關(guān)基因進(jìn)行了檢測(cè)。方法:純鈦表面經(jīng)過(guò)噴砂和雙重酸處理,形成多孔粗糙結(jié)構(gòu)。用電化學(xué)方法在其粗糙表面沉積羥基磷灰石涂層(HA)和摻鍶羥基磷灰石涂層(Sr-HA)。用貼壁法將4周大鼠股骨骨髓間充質(zhì)干細(xì)胞分離進(jìn)行原代培養(yǎng)后將細(xì)胞接種到多孔純鈦,HA和Sr-HA表面培養(yǎng)1、6和24小時(shí)。用場(chǎng)發(fā)射掃描電鏡(SEM)觀察細(xì)胞的形貌特點(diǎn)。用異硫氰酸熒光素(FITC)標(biāo)記的鬼筆環(huán)肽進(jìn)行免疫熒光染色標(biāo)記細(xì)胞骨架,Hoechst 33258進(jìn)行細(xì)胞核染色,激光共聚焦熒光顯微鏡進(jìn)行拍照后使用Image J進(jìn)行細(xì)胞的計(jì)數(shù)和圖形分析。用實(shí)時(shí)熒光定量逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(RT-q PCR)測(cè)定24、48和72小時(shí)不同實(shí)驗(yàn)組中骨髓間充質(zhì)干細(xì)胞中FAK、vinculin、integrin β1和integrin β3的基因表達(dá),并進(jìn)行統(tǒng)計(jì)學(xué)分析。結(jié)論:SEM觀察結(jié)果顯示,骨髓間充質(zhì)干細(xì)胞在三種表面都能正常黏附、生長(zhǎng)和增殖。在Sr-HA組表面,細(xì)胞粘附、細(xì)胞活力和細(xì)胞骨架的鋪展都較粗糙表面組和HA組有較顯著的提高。RT-q PCR結(jié)果顯示,在各個(gè)時(shí)間點(diǎn)Sr-HA組表面骨髓間充質(zhì)干細(xì)胞的FAK、vinculin、integrin β1和integrin β3的基因表達(dá)與粗糙組有顯著性差異,且在24小時(shí)后與HA組亦有顯著性差異。綜上所述,摻鍶羥基磷灰石納米涂層具有較好的生物相容性,可以促進(jìn)大鼠骨髓間充質(zhì)干細(xì)胞在純鈦表面的早期粘附。
[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.
【作者單位】： Department
【基金】：supported by the Zhejiang Provincial Natural Science Foundation of China(No.LQ13H140001)
【分類號(hào)】：R783.1

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