临床口腔医学杂志2025，Vol.41Issue(5)：264-269,6.DOI:10.3969/j.issn.1003-1634.2025.05.003
miR-378过表达羊骨髓间充质干细胞复合马鹿角粉3D支架体内成血管的实验研究
In vivo angiogenesis of miR-378 overexpressed sheep bone marrow mesenchymal stem cells combined with red deer antlers powder 3D scaffold
摘要
Abstract
Objective:To investigate the effect of overexpressing miR-378 gene-modified sheep bone marrow mesen-chymal stem cells(BMMSCs)combined with red deer antlers powder 3D scaffolds on vascularization in sheep mandibular de-fects.Methods:Transfected cells and red deer born powder 3D printed scaffold complex were prepared,and the cytotoxicity was detected by CCK-8 method.The adhesion of cells on the scaffold surface was observed by scanning electron microscope and fluorescence microscope.A bilateral mandibular defect model of Altai large-tailed sheep was established,and 8 sheep were randomly divided into 2 groups(n=4):according to the principle of self-control,the first group was implanted with a miR-378-scaffold(BMMSCs composite scaffold transfected with overexpressing miR-378 lentivirus)on one side of the mandi-ble,and the other group was implanted with a BMMSCs scaffold.In the second group,miR-free scaffold was implanted on the other side of the mandible(BMMSCs composite scaffold transfected with negative lentivirus)and BMMSCs scaffold was im-planted in the other mandible.Each group was repeated once.The samples were taken at 4 weeks and 8 weeks after opera-tion,and the angiogenesis of the defect area was observed by qRT-PCR,HE and Masson staining.Results:CCK-8 detection showed that the cell proliferation absorbance values of 1,3,5,7 days in the extraction group were higher than those in the non-extraction group.Scanning electron microscopy showed that the PVA/SF/Red deer antler powder composite scaffold had good three-dimensional void structure.After 5 days of co-culture with the scaffold,the cells in miR-378 group had larger spreading area on the scaffold surface than those in the miR-free group,and more cell filamentous foot process extended into the scaffold.Fluorescence microscopy showed that miR-378 cells had stronger fluorescence expression on scaffolds and more extensive cell coverage than miR-free cells.mRNA expression levels of platelet-derived growth factor(PDGF),hypoxia-indu-cing factor 1α(HIF-1α),vascular endothelial growth factor(VEGF)and angiopoietin-1(Ang-1)in miR-378 group were higher than those in the other two groups.HE and Masson staining results showed that miR-378 group had more angiogenesis at the defect center than the other two groups at 4 and 8 weeks after surgery.Conclusion:The overexpression of miR-378 modified sheep BMMSCs combined with red deer antler powder 3D scaffold can effectively promote the generation of new blood vessels and contribute to the vascularization of tissue engineered bone.关键词
成血管/miR-378/羊骨髓间充质干细胞/马鹿角粉/组织工程Key words
Angiogenesis/miR-378/Sheep bone marrow mesenchymal stem cells/Red deer bone powder/Tissue en-gineering分类
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朱甜甜,吾西坤·吾然,夏衣旦·艾合买提江,韩祥祯,周琦琪,何惠宇,李征..miR-378过表达羊骨髓间充质干细胞复合马鹿角粉3D支架体内成血管的实验研究[J].临床口腔医学杂志,2025,41(5):264-269,6.基金项目
新疆维吾尔自治区科技支疆计划项目(编号:2022E02043) （编号:2022E02043）
