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

Du,Wenhua; Su,Lingkai; Zhang,Ning; Wang,Huiming

doi:10.3892/mmr.2018.9760

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

Authors:
- Wenhua Du
- Lingkai Su
- Ning Zhang
- Huiming Wang
View Affiliations

Affiliations: 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, Stomatology Department, Meitan General Hospital, Beijing 100088, P.R. China
Published online on: December 13, 2018 https://doi.org/10.3892/mmr.2018.9760
Pages: 951-958
Copyright: © Du et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

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.

View Figures

View References

1	Bressan E, Botticelli D, Sivolella S, Bengazi F, Guazzo R, Sbricoli L, Ricci S, Ferroni L, Gardin C, Velez JU and Zavan B: Adipose-derived stem cells as a tool for dental implant osseointegration: An experimental study in the dog. Int J Mol Cell Med. 4:197–208. 2015.PubMed/NCBI
2	Padial-Molina M, O'Valle F, Lanis A, Mesa F, Dohan Ehrenfest DM, Wang HL and Galindo-Moreno P: Clinical application of mesenchymal stem cells and novel supportive therapies for oral bone regeneration. Biomed Res Int. 2015:3413272015. View Article : Google Scholar : PubMed/NCBI
3	Théry C, Zitvogel L and Amigorena S: Exosomes: Composition, biogenesis and function. Nat Rev Immunol. 2:569–579. 2002. View Article : Google Scholar : PubMed/NCBI
4	Grayson WL, Bunnell BA, Martin E, Frazier T, Hung BP and Gimble JM: Stromal cells and stem cells in clinical bone regeneration. Nat Rev Endocrinol. 11:140–150. 2015. View Article : Google Scholar : PubMed/NCBI
5	James AW: Review of signaling pathways governing MSC osteogenic and adipogenic differentiation. Scientifica (Cairo). 2013:6847362013.PubMed/NCBI
6	Qin Y, Sun R, Wu C, Wang L and Zhang C: Exosome: A novel approach to stimulate bone regeneration through regulation of osteogenesis and angiogenesis. Int J Mol Sci. 17:E7122016. View Article : Google Scholar : PubMed/NCBI
7	Baglio SR, Rooijers K, Koppers-Lalic D, Verweij FJ, Pérez Lanzón M, Zini N, Naaijkens B, Perut F, Niessen HW, Baldini N and Pegtel DM: Human bone marrow- and adipose-mesenchymal stem cells secrete exosomes enriched in distinctive miRNA and tRNA species. Stem Cell Res Ther. 6:1272015. View Article : Google Scholar : PubMed/NCBI
8	Gernapudi R, Yao Y, Zhang Y, Wolfson B, Roy S, Duru N, Eades G, Yang P and Zhou Q: Targeting exosomes from preadipocytes inhibits preadipocyte to cancer stem cell signaling in early-stage breast cancer. Breast Cancer Res Treat. 150:685–695. 2015. View Article : Google Scholar : PubMed/NCBI
9	Fleury A, Martinez MC and Le Lay S: Extracellular vesicles as therapeutic tools in cardiovascular diseases. Front Immunol. 5:3702014. View Article : Google Scholar : PubMed/NCBI
10	Yu B, Zhang X and Li X: Exosomes derived from mesenchymal stem cells. Int J Mol Sci. 15:4142–4157. 2014. View Article : Google Scholar : PubMed/NCBI
11	Narayanan R, Huang CC and Ravindran S: Hijacking the cellular mail: Exosome mediated differentiation of mesenchymal stem cells. Stem Cells Int. 2016:38086742016. View Article : Google Scholar : PubMed/NCBI
12	Cui Y, Luan J, Li H, Zhou X and Han J: Exosomes derived from mineralizing osteoblasts promote ST2 cell osteogenic differentiation by alteration of microRNA expression. FEBS Lett. 590:185–192. 2016. View Article : Google Scholar : PubMed/NCBI
13	Wang Z, Ding L, Zheng XL, Wang HX and Yan HM: DC-derived exosomes induce osteogenic differentiation of mesenchymal stem cells. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 22:600–604. 2014.(In Chinese). PubMed/NCBI
14	Ekström K, Omar O, Granéli C, Wang X, Vazirisani F and Thomsen P: Monocyte exosomes stimulate the osteogenic gene expression of mesenchymal stem cells. PLoS One. 8:e752272013. View Article : Google Scholar : PubMed/NCBI
15	Yeo RW, Lai RC, Zhang B, Tan SS, Yin Y, Teh BJ and Lim SK: Mesenchymal stem cell: An efficient mass producer of exosomes for drug delivery. Adv Drug Deliv Rev. 65:336–341. 2013. View Article : Google Scholar : PubMed/NCBI
16	Lopatina T, Bruno S, Tetta C, Kalinina N, Porta M and Camussi G: Platelet-derived growth factor regulates the secretion of extracellular vesicles by adipose mesenchymal stem cells and enhances their angiogenic potential. Cell Commun Signal. 12:262014. View Article : Google Scholar : PubMed/NCBI
17	Cai Z, Yang F, Yu L, Yu Z, Jiang L, Wang Q, Yang Y, Wang L, Cao X and Wang J: Activated T cell exosomes promote tumor invasion via Fas signaling pathway. J Immunol. 188:5954–5961. 2012. View Article : Google Scholar : PubMed/NCBI
18	Wan DC, Shi YY, Nacamuli RP, Quarto N, Lyons KM and Longaker MT: Osteogenic differentiation of mouse adipose-derived adult stromal cells requires retinoic acid and bone morphogenetic protein receptor type IB signaling. Proc Natl Acad Sci USA. 103:12335–12340. 2006. View Article : Google Scholar : PubMed/NCBI
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
20	Guan X, Gao Y, Zhou J, Wang J, Zheng F, Guo F, Chang A, Li X and Wang B: miR-223 regulates adipogenic and osteogenic differentiation of mesenchymal stem cells through a C/EBPs/miR-223/FGFR2 regulatory feedback loop. Stem Cells. 33:1589–1600. 2015. View Article : Google Scholar : PubMed/NCBI
21	Marolt D, Knezevic M and Novakovic GV: Bone tissue engineering with human stem cells. Stem Cell Res Ther. 1:102010. View Article : Google Scholar : PubMed/NCBI
22	Lai RC, Yeo RW, Tan SS, Zhang B, Yin Y, Kwan Sze NS, Choo A and Lim SK: Mesenchymal stem cell exosomes: The future MSC-based therapy? Mesench Stem Cell Ther. 39–61. 2012.
23	Yuan X, Wang X, Chen C, Zhou J and Han M: Bone mesenchymal stem cells ameliorate ischemia/reperfusion-induced damage in renal epithelial cells via microRNA-223. Stem Cell Res Ther. 8:1462017. View Article : Google Scholar : PubMed/NCBI
24	Chu M, Wu R, Qin S, Hua W, Shan Z, Rong X, Zeng J, Hong L, Sun Y, Liu Y, et al: Bone marrow-derived MicroRNA-223 works as an endocrine genetic signal in vascular endothelial cells and participates in vascular injury from kawasaki disease. J Am Heart Assoc. 6:e0048782017. View Article : Google Scholar : PubMed/NCBI
25	Berenstein R, Nogai A, Waechter M, Blau O, Kuehnel A, Schmidt-Hieber M, Kunitz A, Pezzutto A, Dörken B and Blau IW: Multiple myeloma cells modify VEGF/IL-6 levels and osteogenic potential of bone marrow stromal cells via Notch/miR-223. Mol Carcinog. 55:1927–1939. 2016. View Article : Google Scholar : PubMed/NCBI