梯度纳米金属纯钛材料调控hAMSCs成骨分化机制不清。前期发现梯度纳米金属纯钛材料处理hAMSCs后，MBNL1与NFYA表达升高且正相关；RIP实验发现MBNL1结合NEAT1；软件预测到miR-22-3p和miR-152-3p分别以相同的MRE与NEAT1、NFYA结合；MBNL1启动子区存在与NFYA的结合位点。推测梯度纳米金属纯钛材料通过MBNL1/NEAT1/miR-22-3p(miR-152-3p)/NFYA反馈环路调控hAMSCs成骨分化。项目拟研究梯度纳米金属纯钛材料作用下MBNL1结合NEAT1，促进其稳定性的机制；NEAT1结合miR-22-3p和miR-152-3p，影响二者对NFYA的负性调控的机制；明确NFYA促进上游MBNL1转录，形成正反馈环路，促进hAMSCs成骨分化作用的机制，为梯度纳米金属纯钛作为新型种植体材料的应用提供理论与实验依据。

The mechanism of gradient nanostructured pure Ti on osteogenic differentiation of hAMSCs is unclear. The expression of MBNL1 and NFYA was increased and positively correlated with each other after culture of hAMSCs with the gradient nanostructured pure Ti. It was confirmed that MBNL1 binds to NEAT1 by RNA immunoprecipitation assay, while miR-22-3p and miR-152-3p is predicted to bind with NEAT1, NFYA through the same MRE, and a putative NFYA binding site is contained in MBNL1 promoter sequence. Given these findings, we would like to propose a novel hypothesis that gradient nanostructured pure Ti could regulate the osteogenic differentiation of hAMSCs via MBNL1/NEAT1/miR-22-3p(miR-152-3p)/NFYA feedback loop. The project aims to study the binding mechanism of MBNL1 with NEAT1 to improve its stability, inquire the negative regulation of NFYA by NEAT1 competitively binding with miR-22-3p and miR-152-3p, expound the transcription role of NFYA on MBNL1, which forms a positive feedback loop to promote osteogenic differentiation of hAMSC cultured on gradient nanostructured pure Ti. Therefore, it will provide theoretical and experimental basis for the application of gradient nanostructured pure Ti as a new implant material.