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Ovariectomized Rats with Established Osteopenia have Diminished Mesenchymal Stem Cells in the Bone Marrow and Impaired Homing, Osteoinduction and Bone Regeneration at the Fracture Site

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Abstract

We investigated deleterious changes that take place in mesenchymal stem cells (MSC) and its fracture healing competence in ovariectomy (Ovx)-induced osteopenia. MSC from bone marrow (BM) of ovary intact (control) and Ovx rats was isolated. 99mTc-HMPAO (Technitium hexamethylpropylene amine oxime) labeled MSC was systemically transplanted to rats and fracture tropism assessed by SPECT/CT. PKH26 labeled MSC (PKH26-MSC) was bound in scaffold and applied to fracture site (drill-hole in femur metaphysis). Osteoinduction was quantified by calcein binding and microcomputed tomography. Estrogen receptor (ER) antagonist, fulvestrant was used to determine ER dependence of osteo-induction by MSC. BM-MSC number was strikingly reduced and doubling time increased in Ovx rats compared to control. SPECT/CT showed reduced localization of 99mTc-HMPAO labeled MSC to the fracture site, 3 h post-transplantation in Ovx rats as compared with controls. Post-transplantation, Ovx MSC labeled with PKH26 (Ovx PKH26-MSC) localized less to fracture site than control PKH26-MSC. Transplantation of either control or Ovx MSC enhanced calcein binding and bone volume at the callus of control rats over placebo group however Ovx MSC had lower efficacy than control MSC. Fulvestrant blocked osteoinduction by control MSC. When scaffold bound MSC was applied to fracture, osteoinduction by Ovx PKH26-MSC was less than control PKH26-MSC. In Ovx rats, control MSC/E2 treatment but not Ovx MSC showed osteoinduction. Regenerated bone was irregularly deposited in Ovx MSC group. In conclusion, Ovx is associated with diminished BM-MSC number and its growth, and Ovx MSC displays impaired engraftment to fracture and osteoinduction besides disordered bone regeneration.

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Acknowledgments

The authors are thankful for the technical assistance provided by Kavita Singh at the Confocal Microscopy Facility, SAIF division, CSIR-CDRI.

Conflict of Interest

Although in no way related to this study, NC has received research support from GlaxoSmithKline Consumer Health Care, Gurgaon, India and served as an advisory board member of Alkem Laboratories Ltd, India. All other authors have no conflict of interest.

Funding

Authors acknowledge funding from Council of Scientific and Industrial Research (CSIR) to [N.C.] (BSC0201, ASTHI); Research fellowship grants from the Department of Biotechnology [DT], Indian Council of Medical Research [MPK] and CSIR (S.P.C).

Supporting grants

Council of Scientific and Industrial Research, Indian Council of Medical Research and Department of Biotechnology, Government of India.

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Authors and Affiliations

  1. Division of Endocrinology and Centre for Research in Anabolic Skeletal Targets in Health and Illness (ASTHI), CSIR-Central Drug Research Institute, B.S. 10/1, Sector-10, Jankipuram Extension, Lucknow, India

    Deepshikha Tewari, Mohd Parvez Khan, Shyamsundar P. China, Mahesh C. Tewari, Geet K. Nagar, Omeje E. Ogechukwu & Naibedya Chattopadhyay

  2. Departments of Chemical Engineering, CRNTS and Biosciences and Bioengineering, Indian Institute of Technology-Bombay, Powai, Mumbai, 400076, India

    Nitin Sagar, Atul K. Singh & Jayesh R. Bellare

  3. Department of Nuclear Medicine, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Raebareli Road, Lucknow, 226014, UP, India

    Subhash C. Kheruka, Sukanta Barai & Sanjay Gambhir

  4. Sophisticated and Analytical Instrumentations Facility, CSIR- Central Drug Research Institute, B.S. 10/1, Sector-10, Jankipuram Extension, Lucknow, India

    Achchhe L. Vishwakarma

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  1. Deepshikha Tewari
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Correspondence to Naibedya Chattopadhyay.

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Tewari, D., Khan, M.P., Sagar, N. et al. Ovariectomized Rats with Established Osteopenia have Diminished Mesenchymal Stem Cells in the Bone Marrow and Impaired Homing, Osteoinduction and Bone Regeneration at the Fracture Site. Stem Cell Rev and Rep 11, 309–321 (2015). https://doi.org/10.1007/s12015-014-9573-5

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