PF-127水凝胶联合骨髓间充质干细胞外泌体来源miR-132诱导成骨分化修复牙槽骨缺损

doi:10.3969/j.issn.1006-6233.2024.05.06

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Abstract Objective: To investigate the effect of PF-127 hydrogel combined with bone marrow mesenchymal stem cell-derived exosome-miR-132 on the repair of alveolar bone defects by inducing osteogenic differentiation. Methods: Bone marrow mesenchymal stem cells (BMSCs) were cultured and transfected with anti-miR-132 and anti-miR-NC plasmids. Bone mesenchymal stem cell-derived exosomes (BMSCs-exo) were prepared and identified for expression of marker proteins by Western blot. PF-127 hydrogel and BMSCs-Exo complexes were prepared and BMSCs uptake was detected by PKH67 labeling; osteogenic ability of BMSCs was detected by alizarin red staining; reverse transcription-polymerase chain reaction (RT-PCR) was used to detect miR-132 expression and mRNA expression of alkaline phosphatase (ALP), osteocalcin (OCN), and Runt-related transcription factor 2 (Runx2) in cells. An alveolar bone defect model was established in rats and the rats were randomly divided into three groups: PF-127 hydrogel group, PF-127 hydrogel+BMSCs-exo-anti-miR-NC group, and PF-127 hydrogel+BMSCs-exo-anti-miR-132 group. Micro-computed tomography (micro-CT) was used to evaluate the alveolar bone defect; Western blot was used to detect the expression of ALP, OCN, and Runx2 proteins in alveolar bone tissue. Results: miR-132 expression was significantly lower in the anti-miR-132 group BMSCs than in the anti-miR-NC group (P<0.05), indicating successful transfection of anti-miR-132 into BMSCs. Exosome markers CD9 and CD63 were significantly expressed in the BMSCs-exo-anti-miR-NC and BMSCs-exo-anti-miR-132 groups. Compared with the BMSCs-exo-anti-miR-NC group, miR-132 expression was significantly lower in the BMSCs-exo-anti-miR-132 group (P<0.05). Compared with the PF127 hydrogel group, miR-132 expression was significantly lower in the BMSC-exo-anti-miR-NC group, PF127 hydrogel+BMSC-exo-anti-miR-NC group, and PF127 hydrogel+BMSC-exo-anti-miR-132 groups, and the relative activity of alizarin red staining, Runx2, ALP, and OCN mRNA expression in cells were significantly increased, with the most significant changes in the PF127 hydrogel+BMSC-exo-anti-miR-132 group (P<0.05). Compared with the control group, BV/TV ratio, BMD, Tb.N, Tb.Th, and Runx2, ALP, and OCN protein expression in cells were significantly higher in the PF-127 hydrogel+BMSCs-exo-anti-miR-NC and PF127 hydrogel+BMSCs-exo-anti-miR-132 groups, and Tb.Sp was significantly lower (P<0.05), with the most significant changes in the PF127 hydrogel+BMSCs-exo-anti-miR-132 group. Conclusion: PF-127 hydrogel combined with bone marrow mesenchymal stem cell-derived exosome-miR-132 to interfere with miR-132 expression can promote osteogenic differentiation of bone marrow mesenchymal stem cells, thereby promoting the repair of alveolar bone defects in rats.

Key words： Alveolar bone defect PF-127 hydrogel Bone marrow mesenchymal stem cells Exosomes MiR-132 Osteogenic differentiation

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	QIU Jingyi
	YUAN Jing

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QIU Jingyi,YUAN Jing. PF-127 Hydrogel Combined with Bone Marrow Mesenchymal Stem Cell-Derived Exosome-miR-132 to Induce Osteogenic Differentiation for Repair of Alveolar Bone Defects[J]. HeBei Med, 2024, 30(5): 731-737.

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http://www.hbyxzzs.cn/EN/10.3969/j.issn.1006-6233.2024.05.06 OR http://www.hbyxzzs.cn/EN/Y2024/V30/I5/731

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