【摘要】：背景:骨髓間充質(zhì)干細胞因其多向分化潛能廣泛應(yīng)用于組織工程領(lǐng)域,miRNA對于促進骨髓間充質(zhì)干細胞成骨分化具有重要作用。目的:探究miR-218和miR-26a對大鼠骨髓間充質(zhì)干細胞成骨分化的影響,為誘導骨髓間充質(zhì)干細胞成骨分化的相關(guān)研究及臨床應(yīng)用提供前期依據(jù)和選材參考。方法:提取Wistar大鼠雙下肢股骨骨髓,分離培養(yǎng)骨髓間充質(zhì)干細胞傳至第3代備用。miR-218、miR-26a和聚乙烯亞胺(PEI)以特定比例混合孵育形成miRNA/PEI復合物,設(shè)立陰性對照組。結(jié)果與結(jié)論:(1)大鼠骨髓間充質(zhì)干細胞生長狀態(tài)良好;(2)MTT法篩選miRNA mimics轉(zhuǎn)染適宜濃度為50 nmol/L;(3)qRT-PCR檢測成骨相關(guān)基因堿性磷酸酶、Ⅰ型膠原的mR NA表達水平增高(P0.05);(4)與空白對照組和陰性對照組相比,堿性磷酸酶染色后胞質(zhì)均產(chǎn)生了較為明顯的著色;(5)茜素紅染色出現(xiàn)較多礦化結(jié)節(jié)。結(jié)果顯示,以PEI為載體構(gòu)建的miR-218/PEI復合物、miR-26a/PEI復合物及miR-218+miR-26a/PEI共轉(zhuǎn)染復合物對骨髓間充質(zhì)干細胞的成骨分化具有一定的促進作用,相同條件下miR-26a成骨能力稍優(yōu)于miR-218。
[Abstract]:Background: bone marrow mesenchymal stem cells (BMSCs) are widely used in tissue engineering for their multidirectional differentiation potential. MiRNA plays an important role in promoting osteogenic differentiation of BMSCs. Objective: to investigate the effects of miR-218 and miR-26a on osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs), and to provide a preliminary basis and reference for clinical application in inducing osteogenic differentiation of BMSCs. Methods: bone marrow of both lower extremities of Wistar rats were isolated and cultured. Bone marrow mesenchymal stem cells were isolated and transferred to the third generation. MiR-218miR-26a and polyethyleneimide (PEI) were incubated in a specific proportion to form miRNA/PEI complex, and negative control group was set up. Results and conclusion: (1) Rat bone marrow mesenchymal stem cells (BMSCs) grew well; (2) the suitable concentration of miRNA mimics transfection was 50 nmol/L; (3) qRT-PCR to detect alkaline phosphatase (ALP), which was associated with osteogenesis. Compared with the control group and the negative control group, the mR NA expression level of type I collagen increased significantly (P0.05); (4), the cytoplasm of alkaline phosphatase staining was significantly stained; (5) alizarin red staining showed more mineralized nodules. The results showed that the miR-218/PEI complex constructed with PEI as the carrier could promote the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and the osteogenic ability of miR-26a was slightly better than that of miR-218. under the same conditions.
【作者單位】： 吉林大學口腔醫(yī)院牙發(fā)育及頜骨重塑吉林省重點實驗室;北京大學口腔醫(yī)學院;
【基金】：國家自然科學基金項目(81320108011) 吉林省省級產(chǎn)業(yè)創(chuàng)新專項資金項目(2016C044-3) 吉林省自然科技基金項目(20170101093JC) 大學生創(chuàng)新創(chuàng)業(yè)訓練計劃(2015781125)~~
【分類號】：R68
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本文編號：2235184