Skip to main content

Advertisement

SpringerLink
Log in

MicroRNA-218 Promotes Osteogenic Differentiation of Mesenchymal Stem Cells and Accelerates Bone Fracture Healing

  • Original Research
  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Abstract

As a regulator of osteogenesis, microRNA-218 (miR-218) is reported to promote osteogenesis of mesenchymal stem cells (MSCs). However, the in vivo osteogenic effect of miR-218 remains elusive. In this study, miR-218 was confirmed to promote osteogenic differentiation of MSCs by stimulating the alkaline phosphatase activity, calcium nodule formation, and osteogenic marker gene expression. For in vivo study, the miR-218-overexpressing BMSCs were locally administrated into the fracture sites in a femur fracture mouse model. Based on the X-rays, micro-computed tomography, mechanical testing, histology, and immunohistochemistry examinations, miR-218 overexpression improved new bone formation and accelerated fracture healing. These findings suggest that miR-218 may be a promising therapeutic target for bone repair in future clinical applications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Marsell R, Einhorn TA (2011) The biology of fracture healing. Injury 42:551–555

    Article  PubMed  PubMed Central  Google Scholar 

  2. Phillips AM (2005) Overview of the fracture healing cascade. Injury 36(Suppl 3):S5–S7

    Article  PubMed  Google Scholar 

  3. Keating JF, Blachut PA, O’Brien PJ, Court-Brown CM (2000) Reamed nailing of Gustilo grade-IIIB tibial fractures. J Bone Joint Surg Br 82:1113–1116

    Article  PubMed  CAS  Google Scholar 

  4. Marsh D (1998) Concepts of fracture union, delayed union, and nonunion. Clin Orthop Relat Res S22–S30

  5. Granero-Molto F, Weis JA, Miga MI, Landis B, Myers TJ, O’Rear L, Longobardi L, Jansen ED, Mortlock DP, Spagnoli A (2009) Regenerative effects of transplanted mesenchymal stem cells in fracture healing. Stem Cells 27:1887–1898

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Lin W, Xu L, Zwingenberger S, Gibon E, Goodman SB, Li G (2017) Mesenchymal stem cells homing to improve bone healing. J Orthop Transl 9:19–27

    Google Scholar 

  7. Ambros V (2004) The functions of animal microRNAs. Nature 431:350–355

    Article  PubMed  CAS  Google Scholar 

  8. Zhang JF, Fu WM, He ML, Xie WD, Lv Q, Wan G, Li G, Wang H, Lu G, Hu X, Jiang S, Li JN, Lin MC, Zhang YO, Kung HF (2011) MiRNA-20a promotes osteogenic differentiation of human mesenchymal stem cells by co-regulating BMP signaling. RNA Biol 8:829–838

    Article  PubMed  CAS  Google Scholar 

  9. Li H, Li T, Fan J, Li T, Fan L, Wang S, Weng X, Han Q, Zhao RC (2015) miR-216a rescues dexamethasone suppression of osteogenesis, promotes osteoblast differentiation and enhances bone formation, by regulating c-Cbl-mediated PI3K/AKT pathway. Cell Death Differ 22:1935–1945

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Trompeter HI, Dreesen J, Hermann E, Iwaniuk KM, Hafner M, Renwick N, Tuschl T, Wernet P (2013) MicroRNAs miR-26a, miR-26b, and miR-29b accelerate osteogenic differentiation of unrestricted somatic stem cells from human cord blood. BMC Genomics 14:111

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Li H, Li T, Wang S, Wei J, Fan J, Li J, Han Q, Liao L, Shao C, Zhao RC (2013) miR-17-5p and miR-106a are involved in the balance between osteogenic and adipogenic differentiation of adipose-derived mesenchymal stem cells. Stem Cell Res 10:313–324

    Article  PubMed  CAS  Google Scholar 

  12. Huang J, Zhao L, Xing L, Chen D (2010) MicroRNA-204 regulates Runx2 protein expression and mesenchymal progenitor cell differentiation. Stem Cells 28:357–364

    PubMed  PubMed Central  Google Scholar 

  13. Sun Y, Xu L, Huang S, Hou Y, Liu Y, Chan KM, Pan XH, Li G (2015) mir-21 overexpressing mesenchymal stem cells accelerate fracture healing in a rat closed femur fracture model. BioMed Res Int 2015:412327

    PubMed  PubMed Central  Google Scholar 

  14. Sun Y, Xu J, Xu L, Zhang J, Chan K, Pan X, Li G (2017) MiR-503 promotes bone formation in distraction osteogenesis through suppressing Smurf1 expression. Sci Rep 7:409

    Article  PubMed  PubMed Central  Google Scholar 

  15. Tu M, Tang J, He H, Cheng P, Chen C (2017) MiR-142-5p promotes bone repair by maintaining osteoblast activity. J Bone Miner Metab 35:255–264

    Article  PubMed  CAS  Google Scholar 

  16. Lee WY, Li N, Lin S, Wang B, Lan HY, Li G (2016) miRNA-29b improves bone healing in mouse fracture model. Mol Cell Endocrinol 430:97–107

    Article  PubMed  CAS  Google Scholar 

  17. Amin ND, Bai G, Klug JR, Bonanomi D, Pankratz MT, Gifford WD, Hinckley CA, Sternfeld MJ, Driscoll SP, Dominguez B, Lee KF, Jin X, Pfaff SL (2015) Loss of motoneuron-specific microRNA-218 causes systemic neuromuscular failure, vol 350. Science, New York, pp 1525–1529

    Google Scholar 

  18. Hassan MQ, Maeda Y, Taipaleenmaki H, Zhang W, Jafferji M, Gordon JA, Li Z, Croce CM, van Wijnen AJ, Stein JL, Stein GS, Lian JB (2012) miR-218 directs a Wnt signaling circuit to promote differentiation of osteoblasts and osteomimicry of metastatic cancer cells. J Biol Chem 287:42084–42092

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Zhang WB, Zhong WJ, Wang L (2014) A signal-amplification circuit between miR-218 and Wnt/beta-catenin signal promotes human adipose tissue-derived stem cells osteogenic differentiation. Bone 58:59–66

    Article  PubMed  CAS  Google Scholar 

  20. Zhang Y, Xie RL, Croce CM, Stein JL, Lian JB, van Wijnen AJ, Stein GS (2011) A program of microRNAs controls osteogenic lineage progression by targeting transcription factor Runx2. Proc Natl Acad Sci USA 108:9863–9868

    Article  PubMed  Google Scholar 

  21. Xu L, Meng F, Ni M, Lee Y, Li G (2013) N-cadherin regulates osteogenesis and migration of bone marrow-derived mesenchymal stem cells. Mol Biol Rep 40:2533–2539

    Article  PubMed  CAS  Google Scholar 

  22. Huang S, Xu L, Sun Y, Wu T, Wang K, Li G (2015) An improved protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. J. Orthop. Transl. 3:26–33

    Google Scholar 

  23. Liang WC, Wang Y, Xiao LJ, Wang YB, Fu WM, Wang WM, Jiang HQ, Qi W, Wan DC, Zhang JF, Waye MM (2014) Identification of miRNAs that specifically target tumor suppressive KLF6-FL rather than oncogenic KLF6-SV1 isoform. RNA Biol. 11:845–854

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  24. Guo J, Zhang JF, Wang WM, Cheung FW, Lu YF, Ng CF, Kung HF, Liu WK (2014) MicroRNA-218 inhibits melanogenesis by directly suppressing microphthalmia-associated transcription factor expression. RNA Biol 11:732–741

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Xu D, Xu L, Zhou C, Lee WY, Wu T, Cui L, Li G (2014) Salvianolic acid B promotes osteogenesis of human mesenchymal stem cells through activating ERK signaling pathway. Int J biochem Cell Biol 51:1–9

    Article  PubMed  CAS  Google Scholar 

  26. Rui YF, Lui PP, Wong YM, Tan Q, Chan KM (2013) Altered fate of tendon-derived stem cells isolated from a failed tendon-healing animal model of tendinopathy. Stem Cells Dev 22:1076–1085

    Article  PubMed  CAS  Google Scholar 

  27. Huang S, Xu L, Sun Y, Zhang Y, Li G (2015) The fate of systemically administrated allogeneic mesenchymal stem cells in mouse femoral fracture healing. Stem Cell Res Ther 6:206

    Article  PubMed  PubMed Central  Google Scholar 

  28. He YX, Zhang G, Pan XH, Liu Z, Zheng LZ, Chan CW, Lee KM, Cao YP, Li G, Wei L, Hung LK, Leung KS, Qin L (2011) Impaired bone healing pattern in mice with ovariectomy-induced osteoporosis: a drill-hole defect model. Bone 48:1388–1400

    Article  PubMed  Google Scholar 

  29. Gomez-Barrena E, Rosset P, Lozano D, Stanovici J, Ermthaller C, Gerbhard F (2015) Bone fracture healing: cell therapy in delayed unions and nonunions. Bone 70:93–101

    Article  PubMed  Google Scholar 

  30. Waki T, Lee SY, Niikura T, Iwakura T, Dogaki Y, Okumachi E, Oe K, Kuroda R, Kurosaka M (2016) Profiling microRNA expression during fracture healing. BMC Musculoskelet Disord 17:83

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Gay I, Cavender A, Peto D, Sun Z, Speer A, Cao H, Amendt BA (2014) Differentiation of human dental stem cells reveals a role for microRNA-218. J Periodontal Res 49:110–120

    Article  PubMed  CAS  Google Scholar 

  32. Orimo H (2010) The mechanism of mineralization and the role of alkaline phosphatase in health and disease. J Nippon Med Sch 77:4–12

    Article  PubMed  CAS  Google Scholar 

  33. Gordon JA, Tye CE, Sampaio AV, Underhill TM, Hunter GK, Goldberg HA (2007) Bone sialoprotein expression enhances osteoblast differentiation and matrix mineralization in vitro. Bone 41:462–473

    Article  PubMed  CAS  Google Scholar 

  34. Orimo H, Shimada T (2008) The role of tissue-nonspecific alkaline phosphatase in the phosphate-induced activation of alkaline phosphatase and mineralization in SaOS-2 human osteoblast-like cells. Mol Cell Biochem 315:51–60

    Article  PubMed  CAS  Google Scholar 

  35. Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR, de Crombrugghe B (2002) The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell 108:17–29

    Article  PubMed  CAS  Google Scholar 

  36. Tu Q, Valverde P, Chen J (2006) Osterix enhances proliferation and osteogenic potential of bone marrow stromal cells. Biochem Biophys Res Commun 341:1257–1265

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  37. Zhao Z, Zhao M, Xiao G, Franceschi RT (2005) Gene transfer of the Runx2 transcription factor enhances osteogenic activity of bone marrow stromal cells in vitro and in vivo. Mol Ther 12:247–253

    Article  PubMed  CAS  Google Scholar 

  38. Cao Y, Zhou Z, de Crombrugghe B, Nakashima K, Guan H, Duan X, Jia SF, Kleinerman ES (2005) Osterix, a transcription factor for osteoblast differentiation, mediates antitumor activity in murine osteosarcoma. Cancer Res 65:1124–1128

    Article  PubMed  CAS  Google Scholar 

  39. Sowa AK, Kaiser FJ, Eckhold J, Kessler T, Aherrahrou R, Wrobel S, Kaczmarek PM, Doehring L, Schunkert H, Erdmann J, Aherrahrou Z (2013) Functional interaction of osteogenic transcription factors Runx2 and Vdr in transcriptional regulation of Opn during soft tissue calcification. Am J Pathol 183:60–68

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The work was partially supported by grants from Research Grants Council of the Hong Kong SAR, China (Project No. 14119115, 14160917, N_CityU102/15, T13-402/17-N); National Natural Science Foundation of China (81772404, 81430049, and 81772322); Hong Kong Innovation Technology Commission Funds (ITS/UIM-305). This study was also supported in part by SMART program, Lui Che Woo Institute of Innovative Medicine.

Author information

Author notes

  1. Liu Shi and Lu Feng have equal contribution to this article.

Authors and Affiliations

  1. Department of Orthopaedics & Traumatology, Li Ka Shing Institute of Health Sciences and Lui Che Woo Institute of Innovative Medicine, Faculty of Medicine, Prince of Wales Hospital, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, People’s Republic of China

    Liu Shi, Lu Feng, Yang Liu, Wei-ping Lin & Gang Li

  2. Department of Orthopaedics, Zibo Central Hospital, 54 West Gongqingtuan Road, Zibo, Shandong, People’s Republic of China

    Ji-qiang Duan

  3. Key Laboratory of Orthopaedics and Traumatology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, The First Clinical Medical College, Guangzhou University of Chinese Medicine, Guangzhou, People’s Republic of China

    Jin-fang Zhang

  4. The CUHK-ACC Space Medicine Centre on Health Maintenance of Musculoskeletal System, The Chinese University of Hong Kong Shenzhen Research Institute, Shenzhen, People’s Republic of China

    Gang Li

  5. Li Ka Shing Institute of Health Institute, Prince of Wales Hospital, The Chinese University of Hong Kong, Room 904, 9/F, Shatin, Hong Kong SAR, People’s Republic of China

    Gang Li

Authors
  1. Liu Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lu Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ji-qiang Duan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wei-ping Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jin-fang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Gang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

JZ and GL designed and supervised all the experiments; LS, LF, and YL conducted experiments, analyzed the data, and prepared the manuscript. JD and WL analyzed the IHC staining blindly. All authors reviewed and approved the manuscript.

Corresponding authors

Correspondence to Jin-fang Zhang or Gang Li.

Ethics declarations

Conflict of interest

Liu Shi, Lu Feng, Yang Liu, Ji-qiang Duan, Wei-ping Lin, Jin-fang Zhang, and Gang Li declare that none of them have any conflict of interest.

Human and Animal Rights and Informed Consent

The animal experiemts were carried out with full ethical approval of The Chinese University of Hong Kong animal ethical committee  accorign to the laws and regulations for animal experiemts of Hong Kong.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 370 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shi, L., Feng, L., Liu, Y. et al. MicroRNA-218 Promotes Osteogenic Differentiation of Mesenchymal Stem Cells and Accelerates Bone Fracture Healing. Calcif Tissue Int 103, 227–236 (2018). https://doi.org/10.1007/s00223-018-0410-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00223-018-0410-8

Keywords

Search

Navigation