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Subchondral mesenchymal stem cells from osteoarthritic knees display high osteogenic differentiation capacity through microRNA-29a regulation of HDAC4

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Abstract

Subchondral bone deterioration and osteophyte formation attributable to excessive mineralization are prominent features of end-stage knee osteoarthritis (OA). The cellular events underlying subchondral integrity diminishment remained elusive. This study was undertaken to characterize subchondral mesenchymal stem cells (SMSCs) isolated from patients with end-stage knee OA who required total knee arthroplasty. The SMSCs expressed surface antigens CD29, CD44, CD73, CD90, CD105, and CD166 and lacked CD31, CD45, and MHCII expression. The cell cultures exhibited higher proliferation and greater osteogenesis and chondrogenesis potencies, whereas their population-doubling time and adipogenic lineage commitment were lower than those of bone marrow MSCs (BMMSCs). They also displayed higher expressions of embryonic stem cell marker OCT3/4 and osteogenic factors Wnt3a, β-catenin, and microRNA-29a (miR-29a), concomitant with lower expressions of joint-deleterious factors HDAC4, TGF-β1, IL-1β, TNF-α, and MMP3, in comparison with those of BMMSCs. Knockdown of miR-29a lowered Wnt3a expression and osteogenic differentiation of the SMSCs through elevating HDAC4 translation, which directly regulated the 3′-untranslated region of HDAC4. Likewise, transgenic mice that overexpressed miR-29a in osteoblasts exhibited a high bone mass in the subchondral region. SMSCs in the transgenic mice showed a higher osteogenic differentiation and lower HDAC4 signaling than those in wild-type mice. Taken together, high osteogenesis potency existed in the SMSCs in the osteoarthritic knee. The miR-29a modulation of HDAC4 and Wnt3a signaling was attributable to the increase in osteogenesis. This study shed an emerging light on the characteristics of SMSCs and highlighted the contribution of SMSCs in the exacerbation of subchondral integrity in end-stage knee OA.

Key messages

  • Subchondral MSCs (SMSCs) from OA knee expressed embryonic stem cell marker Oct3/4.

  • The SMSCs showed high proliferation and osteogenic and chondrogenic potencies.

  • miR-29a regulated osteogenesis of the SMSCs through modulation of HDAC4 and Wnt3a.

  • A high osteogenic potency of the SMSCs existed in mice overexpressing miR-29a in bone.

  • Aberrant osteogenesis in SMSCs provides a new insight to subchondral damage in OA.

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Acknowledgements

This study was partially supported by grants [MOST104-2314-B-182A-006-MY3] from the Ministry of Science & Technology, [NHRI-EX106-10436SI] from the National Health Research Institute, and [CMRPG8B0873; CMRPG8E0651-3; and CLRPG8B00421-3] from Chang Gung Memorial Hospital, Taiwan. We are grateful to Dr. Pei-Chin Chuang and Mr. Shun-Hung Tseng for providing the flow cytometry system and the Center for Laboratory Animals, Kaohisung Chang Gung Memorial Hospital, Taiwan, for the use of their facilities.

Grants [NHRI-EX106-10436SI] from the National Health Research Institute, [MOST103-2314-B-182A-053] from the Ministry of Science & Technology, and [CLRPG8B0043, CMRPG8E1321-3, and CMRPG8E0651-3] from Chang Gung Memorial Hospital, Taiwan

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Authors and Affiliations

  1. Department of Medical Research, Kaohisung Chang Gung Memorial Hospital, 123, Ta-Pei Road, Niao-Sung District, Kaohsiung, 83303, Taiwan

    Wei-Shiung Lian, Yu-Shan Chen, Yi-Chih Sun, Shing-Long Wu, Huei-Jing Ke & Feng-Sheng Wang

  2. Core Laboratory for Phenomics and Diagnostics, Department of Pediatrics, Chang Gung University College of Medicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

    Wei-Shiung Lian, Yu-Shan Chen, Yi-Chih Sun, Shing-Long Wu, Huei-Jing Ke & Feng-Sheng Wang

  3. Department of Orthopedic Surgery, Chang Gung University College of Medicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

    Ren-Wen Wu, Mel S. Lee & Jih-Yang Ko

  4. Graduate Institute of Clinical Medical Sciences, Chang Gung University College of Medicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan

    Jih-Yang Ko & Feng-Sheng Wang

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  1. Wei-Shiung Lian
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  2. Ren-Wen Wu
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  3. Mel S. Lee
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Correspondence to Jih-Yang Ko or Feng-Sheng Wang.

Ethics declarations

Experimental protocols were approved by the IRB of Chang Gung Memorial Hospital (no. 104-5248B and no. 106-2251C). Informed consent was obtained from all patients with end-stage knee OA who required total knee replacement. Experiments involving laboratory animals were approved by the IACUC of Kaohsiung Chang Gung Memorial Hospital (IACUC no. 2014120401).

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The authors declare that they have no conflict of interest.

Additional information

Wei-Shiung Lian and Ren-Wen Wu contributed equally to this study.

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Lian, WS., Wu, RW., Lee, M.S. et al. Subchondral mesenchymal stem cells from osteoarthritic knees display high osteogenic differentiation capacity through microRNA-29a regulation of HDAC4. J Mol Med 95, 1327–1340 (2017). https://doi.org/10.1007/s00109-017-1583-8

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  • DOI: https://doi.org/10.1007/s00109-017-1583-8

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