PF-127 Hydrogel Combined with Bone Marrow Mesenchymal Stem Cell-Derived Exosome-miR-132 to Induce Osteogenic Differentiation for Repair of Alveolar Bone Defects
Objective:To investigate the effect of PF-127 hydrogel combined with bone marrow mesen-chymal stem cell-derived exosome-miR-132 on the repair of alveolar bone defects by inducing osteogenic dif-ferentiation.Methods:Bone marrow mesenchymal stem cells(BMSCs)were cultured and transfected with an-ti-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 BM-SCs 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 estab-lished 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-com-puted 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 ex-pressed in the BMSCs-exo-anti-miR-NC and BMSCs-exo-anti-miR-132 groups.Compared with the BM-SCs-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 hy-drogel+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.
Alveolar bone defectPF-127 hydrogelBone marrow mesenchymal stem cellsExosomesMiR-132Osteogenic differentiation
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