基于CRISPR/Cas9技术靶向沉默miR-382-5p表达介导BMSC骨向分化及促进骨修复的机制研究

The treatment for nonunion or bone defect has been a major challenge in clinical practice. There are still many problems in traditional treatment methods. In research on bone tissue engineering, however, how to improve proliferation and osteogenic differentiation of seed cells has recently become the focus of researchers. More and more research had demonstrated that microRNA participated in regulation of bone repair and reconstruction. Our preliminary studies had tentatively demonstrated that miR-382-5p might participate in regulation of proliferation, apoptosis and osteogenic differentiation of BMSCs through regulating TAZ expression. This research intended to further clarify the specific molecular biological mechanism by which miR-382-5p directly regulated TAZ to affect proliferation and osteogenic differentiation of BMSCs, and assess the potential value of stem cell sheets prepared from BMSCs underwent knockout of miR-382-5p with CRISPR/Cas9 in the treatment for fracture through in vivo tests on rats nonunion models.

骨缺损或骨不连的治疗是临床面临的一大难题，传统的治疗手段还存在诸多问题，而骨组织工程中的研究中，如何提高种子细胞的增殖和成骨分化能力近来成为研究者们的关注重点。越来越多的研究表明microRNA可参与骨修复和重建的调控过程。我们前期研究初步证实了miR-382-5p可能通过调控TAZ的表达，参与调控BMSC细胞的增殖和成骨分化过程。本项目拟进一步明确miR-382-5p通过对TAZ的直接调控从而影响BMSC细胞的增殖于成骨分化能力的具体分子生物学机制，并通过大鼠骨不连模型的体内实验，评估基于CRISPR/Cas9技术沉默miR-382-5p的BMSC细胞制备成的干细胞片应用于骨修复治疗的潜在价值。

骨缺损或骨不连的治疗是临床面临的一大难题，传统的治疗手段还存在诸多问题，而骨组织工程中的研究中，如何提高种子细胞的增殖和成骨分化能力近来成为研究者们的关注重点。越来越多的研究表明microRNA可参与骨修复和重建的调控过程。我们通过研究初步证实了miR-382-5p通过调控TAZ的表达，参与调控BMSC细胞的增殖和成骨分化过程。明确了miR-382-5p通过对TAZ的直接调控从而影响BMSC细胞的增殖于成骨分化能力的具体分子生物学机制，并通过大鼠骨不连模型的体内实验，证实了基于CRISPR/Cas9技术沉默miR-382-5p的BMSC细胞制备成的干细胞片应用于骨修复治疗具有临床应用价值