Wang Yi-Xuan, Peng Zhu-Li, Sun Zhi-Wen, Pan Yan-Jun, Ai Hong, Mai Zhi-Hui
Department of Stomatology, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
Department of Orthodontics, Affiliated Stomatology Hospital of Guangzhou Medical University, Guangzhou, China.
Front Bioeng Biotechnol. 2023 Apr 5;11:1127908. doi: 10.3389/fbioe.2023.1127908. eCollection 2023.
This study aimed to determine whether miR-20 promoted osteogenic differentiation in bone marrow-derived mesenchymal stem/stromal cells (BMSCs) and accelerated bone formation in the maxillary sinus bone defect model in rabbits. BMSCs were transfected with miR-20a or anti-miR-20a for 12 h, followed by detection of RUNX2, Sp7 mRNA, bone morphogenetic protein 2 (BMP2), and RUNX2 protein expression. Alkaline phosphatase (ALP) activity and Alizarin Red S staining were used to detect calcified nodule deposition. In the rabbit maxillary sinus bone defect model, miR-20a loaded with AAV and BMP2 protein were mixed with Bio-Oss bone powder for filling the bone defect. At 4 weeks and 8 weeks, bone density was detected by cone beam computed tomography (CBCT), and new bone, osteoblasts, and collagen type 1 were evaluated by hematoxylin and eosin (HE) staining and immunohistochemical (IHC) staining. Overexpression of miR-20a enhanced the mRNA and protein levels of BMP2, RUNX2, and SP7, the activity of ALP, and the levels of matrix mineralization, whereas the levels and activity of the aforementioned factors were decreased by anti-miR-20a treatment of BMSCs. Furthermore, miR-20a significantly increased the bone density, the number of osteoblasts, and the secretion of collagen type 1 in bone defects compared with Bio-Oss bone powder in the rabbit maxillary sinus bone defect model. Overall, miR-20a can induce osteogenic differentiation in BMSCs and accelerate bone formation of maxillary sinus defects in rabbits.
本研究旨在确定miR-20是否能促进骨髓间充质干细胞(BMSCs)的成骨分化,并加速兔上颌窦骨缺损模型中的骨形成。将miR-20a或抗miR-20a转染BMSCs 12小时,随后检测RUNX2、Sp7 mRNA、骨形态发生蛋白2(BMP2)和RUNX2蛋白表达。采用碱性磷酸酶(ALP)活性检测和茜素红S染色检测钙化结节沉积。在兔上颌窦骨缺损模型中,将负载miR-20a的腺相关病毒(AAV)和BMP2蛋白与Bio-Oss骨粉混合用于填充骨缺损。在4周和8周时,通过锥形束计算机断层扫描(CBCT)检测骨密度,并通过苏木精-伊红(HE)染色和免疫组织化学(IHC)染色评估新骨、成骨细胞和Ⅰ型胶原。miR-20a的过表达增强了BMP2、RUNX2和SP7的mRNA和蛋白水平、ALP活性以及基质矿化水平,而用抗miR-20a处理BMSCs则降低了上述因子的水平和活性。此外,在兔上颌窦骨缺损模型中,与Bio-Oss骨粉相比,miR-20a显著增加了骨缺损处的骨密度、成骨细胞数量和Ⅰ型胶原分泌。总体而言,miR-20a可诱导BMSCs的成骨分化并加速兔上颌窦缺损处的骨形成。