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Effects of daily restraint with and without injections on skeletal properties in C57BL/6NHsd mice

Lab Animal volume 46, pages 299–301 (2017)Cite this article

Animal model experiments have an essential place in the infrastructure of biomedical science. The literature is replete with papers studying various physiological and organ systems in which animals are manipulated via administration of novel compounds. These studies typically involve groups of animals that are injected with placebo compounds. As there are studies that demonstrate that restraint and injection can affect behavior and corticosteroid levels in rodents1,2, the basis of such placebo injections is to control for any potential effects caused by handling and injecting the experimental animals. But these stressors may not adversely affect all studies equally. While placebo injections make sense for studies that are focused on outcomes which may be directly or indirectly affected by stress hormones, the value of placebo injections is less clear for other studies. If placebo groups are not necessary for some studies, this would reduce both the number of animals used in research and the need to handle/inject a significant number of animals.

Bone is a dynamic organ that undergoes continual renewal throughout life3. The breakdown of this process leads to conditions such as osteoporosis, where bones lose mass and mechanical properties, ultimately leading to fracture. The mouse has become a highly utilized research model in skeletal biology due to the ease of genetic manipulation to answer mechanistic questions. Due to the relatively slow changes that occur in bone, most interventional studies involve treatment durations that last weeks or months. Studies of bone physiology often include a control group (no manipulation) in addition to a placebo group (administration of vehicle), though changes of bone are relatively slow and likely not influenced by the stressors associated with restraint and injection. The goal of the current study was to examine the effects of saline injections in mice, specifically on skeletal properties, and to determine if a reduction of animal use can be affected through the elimination of a placebo group that does not provide meaningful comparison data. Our working hypothesis was that daily handling and injection would not significantly alter bone mass or mechanical properties compared to non-intervention controls.

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Figure 1: Effect of daily animal handling and injection on body mass (a), thymus mass/body mass ratio (b) and neutrophil/lymphocyte ratio (c).
Figure 2: Effect of daily animal handling and injection on trabecular bone volume (a) and cortical bone area (b).
Figure 3: Effect of daily animal handling and injection on ultimate load of the tibia and femur.

References

  1. Turner, P. V., Brabb, T., Pekow, C. & Vasbinder, M. A. J. Am. Assoc. Lab. Anim. Sci. 50, 600–613 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Balcombe, J. P., Barnard, N. D. & Sandusky, C. Contemp. Top. Lab. Anim. Sci. 43, 42–51 (2004).

    CAS  PubMed  Google Scholar 

  3. Robling, A., Castillo, A. & Turner, C. H. Annu. Rev. Biomed. Eng. 8, 455 (2006).

    Article  CAS  Google Scholar 

  4. Kramer, K. et al. Altern. Lab. Anim. 32 Suppl. 1A, 177–181 (2004).

    CAS  PubMed  Google Scholar 

  5. Hankenson, F. C. et al. J. Am. Assoc. Lab. Anim. Sci. 52, 277–285 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Horn, M. J., Hudson, S. V., Bostrom, L. A. & Cooper, D. M. J. Am. Assoc. Lab. Anim. Sci. 51, 781–788 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Trammell, R. A., Cox, L. & Toth, L. A. Comp. Med. 62, 172–178 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Augustsson, H., Lindberg, L. & Höglund, A. U. Lab. Anim. 36, 271–281 (2002).

    Article  CAS  Google Scholar 

  9. Trammell, R. A. & Toth, L. A. Comp. Med. 61, 492–498 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Bollinger, A. P and Everds, N. The Laboratory Mouse. (Hedrich, H.J., ed.) Academic Press, Cambridge, MA. (2004).

    Google Scholar 

  11. Davis, A. K., Maney, D. L. & Maerz, J. C. Funct. Ecol. 22, 760–772 (2008).

    Article  Google Scholar 

  12. Bouxsein, M. L. et al. J. Bone Miner. Res. 20, 1085–1092 (2005).

    Article  Google Scholar 

  13. Berman, A. G., Clauser, C. A., Wunderlin, C., Hammond, M. A. & Wallace, J. M. PLoS ONE 10, e0130504–16 (2015).

    Article  Google Scholar 

  14. O'Brien, C. A. et al. Endocrinology 145, 1835–1841 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported in part by NIH grants AR62002 (M.R.A.) and AR067221 (J.M.W.). The microCT utilized in this experiment was purchased through a NIH S10 grant (OD 016208). The authors would like to thank Karen Boschen for performing the blinded behavioral assessment.

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Authors and Affiliations

  1. Laboratory Animal Resource Center, Indiana University School of Medicine, Indianapolis, IN

    Rachel A Larsen, Jessica L Peveler, Joshuah B Klutzke & Debra L Hickman

  2. Department of Cellular & Integrative Physiology, Indiana University School of Medicine, Indianapolis, IN

    Debra L Hickman

  3. Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, IN

    Mohammad W Aref, Drew M Brown & Matthew R Allen

  4. Department of Biomedical Engineering, Indiana University-Purdue University of Indianapolis, Indianapolis, IN

    Joseph M Wallace & Matthew R Allen

  5. Roudebush Veterans Administration Medical Center, Indianapolis, IN

    Matthew R Allen

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  1. Rachel A Larsen
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Correspondence to Matthew R Allen.

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Larsen, R., Peveler, J., Klutzke, J. et al. Effects of daily restraint with and without injections on skeletal properties in C57BL/6NHsd mice. Lab Anim 46, 299–301 (2017). https://doi.org/10.1038/laban.1295

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