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前脂肪细胞来源的外泌体通过降低 miR-223 的表达来促进成骨细胞分化。

Exosomes derived from preadipocytes improve osteogenic differentiation, potentially via reduced miR‑223 expression.

机构信息

Department of Oral Implantology, Stomatology Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310058, P.R. China.

Department of Conservative Dentistry and Endodontics, Stomatology Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310058, P.R. China.

出版信息

Mol Med Rep. 2019 Feb;19(2):951-958. doi: 10.3892/mmr.2018.9760. Epub 2018 Dec 13.

Abstract

Insufficient bone volume remains a key issue when using dental implants. Adipose tissue‑derived stem cells (ADSCs) can accelerate bone healing when combined with dental implants. To improve the application of ADSCs for dental uses, the present study aimed to identify optimal implantation conditions. Mesenchymal stem cell‑derived exosomes can induce naïve stem cells to differentiate through the osteogenic lineage. In the present study, exosomes derived from 3T3L1 preadipocytes (3T3L1‑exo) were purified and characterized. The effects and potential mechanisms of 3T3L1‑exo on 3T3L1 cell ossification were examined by reverse transcription‑quantitative polymerase chain reaction, western blotting, electron microscopy, RNA sequencing and histological analysis. The current study confirmed that 3T3L1‑exo enhanced 3T3L1 preadipocyte osteogenic differentiation, as revealed by upregulation of osteogenic differentiation‑associated genes and increased Alizarin Red staining. Furthermore, the microRNA (miR) expression profiles of 3T3L1‑exo and 3T3L1 preadipocytes were sequenced and compared. The results of a further analysis demonstrated that miR‑223 expression was reduced in 3T3L1 preadipocytes stimulated by 3T3L1‑exo compared with in unstimulated cells. This finding suggested that 3T3L1‑exo promoted 3T3L1 bone formation by decreasing miR‑223 through a competitive mechanism, another miRNA, or another factor. The mechanism by which miR‑223 is decreased warrants further investigation. In conclusion, the application of 3T3L1‑exo may be useful for investigating preadipocyte‑induced bone regeneration.

摘要

当使用牙种植体时,骨量不足仍然是一个关键问题。脂肪组织来源的干细胞(ADSCs)与牙种植体结合使用时可以加速骨愈合。为了提高 ADSC 在牙科中的应用,本研究旨在确定最佳的植入条件。间充质干细胞衍生的外泌体可以通过成骨谱系诱导幼稚干细胞分化。在本研究中,分离并鉴定了来自 3T3L1 前脂肪细胞(3T3L1-exo)的外泌体。通过逆转录-定量聚合酶链反应、western blot、电子显微镜、RNA 测序和组织学分析,研究了 3T3L1-exo 对 3T3L1 细胞成骨的作用及其潜在机制。本研究证实 3T3L1-exo 增强了 3T3L1 前脂肪细胞的成骨分化,这表现为成骨分化相关基因的上调和茜素红染色的增加。此外,对 3T3L1-exo 和 3T3L1 前脂肪细胞的 microRNA(miR)表达谱进行了测序和比较。进一步分析的结果表明,与未刺激细胞相比,受 3T3L1-exo 刺激的 3T3L1 前脂肪细胞中 miR-223 的表达降低。这一发现表明,3T3L1-exo 通过竞争性机制、另一种 miRNA 或另一种因素降低 miR-223 来促进 3T3L1 骨形成。miR-223 降低的机制值得进一步研究。总之,3T3L1-exo 的应用可能有助于研究前脂肪细胞诱导的骨再生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c81/6323233/de50f9824f78/MMR-19-02-0951-g00.jpg

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