PF-127 Hydrogel Combined with Bone Marrow Mesenchymal Stem Cell-Derived Exosome-miR-132 to Induce Osteogenic Differentiation for Repair of Alveolar Bone Defects
QIU Jingyi, YUAN Jing
Beijing Shijitan Hospital, Capital Medical University, Beijing Haidian District 100039, China
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.
[1] Gerami MH,Khorram R,Rasoolzadegan S,et al.Emerging role of mesenchymal stem/stromal cells (MSCs) and MSCs-derived exosomes in bone- and joint-associated musculoskeletal disorders:a new frontier[J].Eur Med Res,2023,28(1):86. [2] 赵长铭,黄萌,王振宁,等.miR-129-5p调控MC3T3-E1细胞成骨分化的体外研究[J].解放军医学院学报,2021,42(5):565-570. [3] 黎裕明,雷爱凤,赵晓晓,等.microRNA-132对人脂肪源性间充质干细胞成骨分化的影响[J].广西医科大学学报,2019,36(2):187-191. [4] Dos Santos DDL,Besegato JF,de Melo PBG,et al.Effect of curcumin-encapsulated Pluronic® F-127 over duo-species biofilm of Streptococcus mutans and Candida albicans[J].Lasers Med Sci,2022,37(3):1775-1786. [5] Piotrowski M,Maras P,Kadtubowski S,et al.Study of the optimal composition and storage conditions of the fricke-XO-pluronic F-127 radiochromic dosimeter[J].Materials (Basel),2022,15(3):984. [6] He L,Zhou Q,Zhang H,et al.PF127 hydrogel-based delivery of exosomal CTNNB1 from mesenchymal stem cells induces osteogenic differentiation during the repair of alveolar bone defects[J].Nanomaterials (Basel),2023,13(6):1083. [7] Wang N,Liu X,Tang Z,et al.Increased BMSC exosomal miR-140-3p alleviates bone degradation and promotes bone restoration by targeting Plxnb1 in diabetic rats[J].Nanobiotechnology,2022,20(1):97. [8] Zou J,Yang W,Cui W,et al.Therapeutic potential and mechanisms of mesenchymal stem cell-derived exosomes as bioactive materials in tendon-bone healing[J].Nanobiotechnology,2023,21(1):14. [9] Liu L,Guo S,Shi W,et al.Bone marrow mesenchymal stem cell-derived small extracellular vesicles promote periodontal regeneration[J].Tissue Eng Part A,202,27(13-14):962-976. [10] Xue ZL,Meng YL,Ge JH.Upregulation of miR-132 attenuates osteoblast differentiation of UC-MSCs[J].Eur Rev Med Pharmacol Sci,2018,22(6):1580-1587. [11] 刘屿,曾莲,王卫红,等.人骨髓间充质干细胞外泌体来源的miR-335-5p促进人牙周膜干细胞的成骨分化:基于下调DKK1表达[J].南方医科大学学报,2023,43(3):420-427. [12] 郭杰坤,方向,彭小明,等.LncRNA-CWF19L1调控miR-132-3p抑制人骨髓间充质干细胞成骨分化的作用研究[J].现代医院,2021,21(5):798-802,806. [13] Chen N,Ren R,Wei X,et al.Thermoresponsive hydrogel-based local delivery of simvastatin for the treatment of periodontitis[J].Mol Pharm,2021,18(5):1992-2003. [14] Zhou Y,Zhang XL,Lu ST,et al.Human adipose-derived mesenchymal stem cells-derived exosomes encapsulated in pluronic F127 hydrogel promote wound healing and regeneration[J].Stem Cell Res Ther,2022,13(1):407.
2024, Vol. 30 
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