【摘要】：背景:鹽酸四環(huán)素不僅具有促進(jìn)骨髓間充質(zhì)干細(xì)胞增殖的作用,還具有極強(qiáng)的骨親活性。研究鹽酸四環(huán)素誘導(dǎo)骨髓間充質(zhì)干細(xì)胞向成骨細(xì)胞分化,可為獲得骨組織工程細(xì)胞提供新方法。目的:觀察體外鹽酸四環(huán)素對(duì)大鼠骨髓間充質(zhì)干細(xì)胞向成骨細(xì)胞分化的作用。方法:骨髓全血細(xì)胞貼壁法分離培養(yǎng)大鼠骨髓間充質(zhì)干細(xì)胞,通過細(xì)胞形態(tài)和骨髓間充質(zhì)干細(xì)胞標(biāo)志物CD34、CD45、CD90、CD29流式檢測(cè)對(duì)骨髓間充質(zhì)干細(xì)胞進(jìn)行鑒定;取第3代骨髓間充質(zhì)干細(xì)胞與鹽酸四環(huán)素共培養(yǎng),以含體積分?jǐn)?shù)10%FBS的L-DMEM的完全培養(yǎng)基培養(yǎng)做對(duì)照組。通過檢測(cè)堿性磷酸酶活性、Ⅰ型膠原和骨鈣素免疫組織化學(xué)染色、茜素紅礦化鈣結(jié)節(jié)組織化學(xué)染色、Ⅰ型膠原蛋白和骨鈣素的RT-PCR的基因表達(dá),研究鹽酸四環(huán)素對(duì)骨髓間充質(zhì)干細(xì)胞的成骨誘導(dǎo)分化活性。結(jié)果與結(jié)論:①鹽酸四環(huán)素能夠持續(xù)促進(jìn)骨髓間充質(zhì)干細(xì)胞的堿性磷酸酶活性增加,與對(duì)照組比較具有顯著性差異;②鹽酸四環(huán)素誘導(dǎo)培養(yǎng)骨髓間充質(zhì)干細(xì)胞21 d,Ⅰ型膠原蛋白和骨鈣素染色表達(dá)陽性,茜素紅染色出現(xiàn)紅色鈣結(jié)節(jié),RT-PCR檢測(cè)Ⅰ型膠原蛋白和骨鈣素基因表達(dá);對(duì)照組程陰性表達(dá);③結(jié)果表明,鹽酸四環(huán)素具有明顯促進(jìn)骨髓間充質(zhì)干細(xì)胞成骨誘導(dǎo)分化的作用。
[Abstract]:BACKGROUND: Tetracycline hydrochloride not only has the effect of promoting the proliferation of mesenchymal stem cells in the bone marrow, but also has extremely strong bone affinity. To study the differentiation of bone marrow mesenchymal stem cells into osteoblasts by tetracycline hydrochloride, a new method can be provided for obtaining bone tissue engineering cells. Objective: To observe the effect of tetracycline hydrochloride on the differentiation of bone marrow mesenchymal stem cells into osteoblasts. Methods: The bone marrow mesenchymal stem cells were isolated and cultured by bone marrow whole blood cell wall-wall method, and the bone marrow mesenchymal stem cells were identified by the cell morphology and the bone marrow mesenchymal stem cell markers CD34, CD45, CD90 and CD29. The 3-generation bone marrow mesenchymal stem cells were co-cultured with tetracycline hydrochloride, and the control group was cultured with a total medium of L-DMEM with a volume fraction of 10% FBS. By detecting the activity of alkaline phosphatase, the immunohistochemical staining of type I collagen and bone calcium, the histochemical staining of I-type collagen and bone calcium, the expression of the gene of the RT-PCR of type I collagen and bone calcium, the bone-induced differentiation activity of tetracycline on the mesenchymal stem cells of the bone marrow was studied. Results and Conclusion: Tetracycline hydrochloride can continuously promote the increase of the alkaline phosphatase activity of the mesenchymal stem cells in the bone marrow, and there is a significant difference between the control group and the control group, and the positive expression of the bone marrow mesenchymal stem cells 21d, the type I collagen and the bone calcium in the tetracycline-induced culture of the tetracycline hydrochloride is positive, The red calcium nodules and RT-PCR were used to detect the expression of the type 鈪，

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