Medical School of Chinese PLA, Chinese PLA General Hospital, No. 28 Fuxing Road, Haidian District, Beijing, 100853, China.
Department of Orthopedics, National Clinical Research Center for Orthopedics, Sports Medicine and Rehabilitation, Chinese PLA General Hospital, Beijing, 100853, China.
J Orthop Surg Res. 2023 Jul 29;18(1):546. doi: 10.1186/s13018-023-04021-y.
Osteoporosis remains a significant clinical challenge worldwide. Recent studies have shown that exosomes stimulate bone regeneration. Thus, it is worthwhile to explore whether exosomes could be a useful therapeutic strategy for osteoporosis. The purpose of this study was to investigate the effects of exosomes derived from human umbilical cord mesenchymal stem cells (hucMSCs) on osteoblast proliferation and differentiation.
Exosomes were isolated from hucMSCs. Bioinformatics analysis was performed to identify the differentially expressed lncRNAs in myeloma-derived mesenchymal stem cells. Plasmids encoding LINC00520 or short hairpin RNA of LINC00520 were transfected into hucMSCs and then exosomes were isolated. After human osteoblasts hFOB1.19 were exposed to the obtained exosomes, cell survival, cell cycle, apoptosis and calcium deposits of hFOB1.19 cell were detected by MTT, 7-aminoactinomycin D, Annexin V-FITC/propidium iodide and Alizarin red staining, respectively.
In hFOB1.19 cells, 10 × 10/mL hucMSC-derived exosomes inhibited cell proliferation, arrested cell cycle, and promoted apoptosis, while hucMSCs or 1 × 10/mL exosomes promoted cell proliferation, accelerated cell cycle, and promoted calcium deposits and the expression of OCN, RUNX2, collagen I and ALP. In hFOB1.19 cells, exosomes from hucMSCs with LINC00520 knockdown reduced the survival and calcium deposits, arrested the cell cycle, and enhanced the apoptosis, while exosomes from hucMSCs overexpressing LINC00520 enhance the proliferation and calcium deposits and accelerated the cell cycle.
LINC00520 functions as a modulator of calcium deposits, and exosomes derived from hucMSCs overexpressing LINC00520 might be a novel therapeutic approach for osteoporosis.
骨质疏松症仍然是全球范围内一个重大的临床挑战。最近的研究表明,外泌体能刺激骨再生。因此,探索外泌体是否可以成为骨质疏松症的一种有用的治疗策略是值得的。本研究旨在探讨人脐带间充质干细胞(hucMSCs)来源的外泌体对成骨细胞增殖和分化的影响。
从 hucMSCs 中分离出外泌体。进行生物信息学分析,以鉴定骨髓瘤来源的间充质干细胞中差异表达的 lncRNA。将编码 LINC00520 的质粒或 LINC00520 的短发夹 RNA 转染到 hucMSCs 中,然后分离出外泌体。将获得的外泌体暴露于人成骨细胞 hFOB1.19 后,通过 MTT、7-氨基放线菌素 D、Annexin V-FITC/碘化丙啶和茜素红染色分别检测 hFOB1.19 细胞的细胞存活、细胞周期、细胞凋亡和钙沉积。
在 hFOB1.19 细胞中,10×10/mL hucMSC 来源的外泌体抑制细胞增殖,使细胞周期停滞,并促进细胞凋亡,而 hucMSCs 或 1×10/mL 外泌体则促进细胞增殖,加速细胞周期,并促进钙沉积和 OCN、RUNX2、胶原 I 和 ALP 的表达。在 hFOB1.19 细胞中,LINC00520 敲低的 hucMSCs 来源的外泌体减少了细胞的存活和钙沉积,使细胞周期停滞,并增强了细胞凋亡,而 LINC00520 过表达的 hucMSCs 来源的外泌体则增强了细胞的增殖和钙沉积,并加速了细胞周期。
LINC00520 作为钙沉积的调节剂发挥作用,并且过表达 LINC00520 的 hucMSCs 来源的外泌体可能成为骨质疏松症的一种新的治疗方法。