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Calcified Tissue International

, Volume 97, Issue 5, pp 476–486 | Cite as

Bone Marrow Stress Decreases Osteogenic Progenitors

  • Adeline H. Ng
  • Gurpreet S. Baht
  • Benjamin A. Alman
  • Marc D. GrynpasEmail author
Original Research

Abstract

Age-related bone loss may be a result of declining levels of stem cells in the bone marrow. Using the Col2.3Δtk (DTK) transgenic mouse, osteoblast depletion was used as a source of marrow stress in order to investigate the effects of aging on osteogenic progenitors which reside in the marrow space. Five-month-old DTK mice were treated with one or two cycles of ganciclovir to conditionally ablate differentiated osteoblasts, whereas controls were saline-treated. Treatment cycles were two weeks in length followed by four weeks of recovery. All animals were sacrificed at 8 months of age; bone marrow stromal cells (BMSCs) were harvested for cell culture and whole bones were excised for bone quality assessment. Colony-forming unit (CFU) assays were conducted to investigate the osteogenic potential of BMSC in vitro, and RNA was extracted to assess the expression of osteoblastic genes. Bone quality assessments included bone histomorphometry, TRAP staining, microcomputed tomography, and biomechanical testing. Osteoblast depletion decreased CFU-F (fibroblast), CFU-ALP (alkaline phosphatase), and CFU-VK (von Kossa) counts and BMSC osteogenic capacity in cell culture. Ex vivo, there were no differences in bone mineral density of vertebrae or femurs between treatment groups. Histology showed a decrease in bone volume and bone connectivity with repeated osteoblast depletion; however, this was accompanied by an increase in bone formation rate. There were no notable differences in osteoclast parameters or observed bone marrow adiposity. We have developed a model that uses bone marrow stress to mimic age-related decrease in osteogenic progenitors. Our data suggest that the number of healthy BMSCs and their osteogenic potential decline with repeated osteoblast depletion. However, activity of the remaining osteoblasts increases to compensate for this loss in progenitor osteogenic potential.

Keywords

Aging Osteogenic progenitors Mouse model Bone quality Osteoblast 

Notes

Acknowledgments

This work was supported by a Grant from the Canadian Institutes for Health Research to Marc Grynpas and Benjamin Alman. Adeline Ng was supported by scholarships from the University of Toronto, Lunenfeld-Tanenbaum Research Institute, Ontario Graduate Scholarship fund, and Canadian Institutes for Health Research. The authors wish to thank PuviindranNadesan for his technical assistance.

Compliance with Ethical Standards

Conflict of Interest

Adeline H. Ng, Gurpreet S. Baht, Benjamin A. Alman, and Marc D. Grynpas state that they have no conflicts of interest.

Human and Animal Rights and Informed Consent

All animal work was approved by the Animal Care Committee at the Toronto Center for Phenogenomics.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Adeline H. Ng
    • 1
    • 2
  • Gurpreet S. Baht
    • 3
  • Benjamin A. Alman
    • 3
    • 4
  • Marc D. Grynpas
    • 1
    • 2
    • 5
    Email author
  1. 1.Department of Laboratory Medicine & PathobiologyUniversity of TorontoTorontoCanada
  2. 2.Lunenfeld-Tanenbaum Research Institute, Mount Sinai HospitalTorontoCanada
  3. 3.Program in Developmental and Stem Cell BiologyHospital for Sick ChildrenTorontoCanada
  4. 4.Department of Orthopaedic SurgeryDuke UniversityDurhamUSA
  5. 5.TorontoCanada

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