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Mechanical microenvironments and protein expression associated with formation of different skeletal tissues during bone healing

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Abstract

Uncovering the mechanisms of the sensitivity of bone healing to mechanical factors is critical for understanding the basic biology and mechanobiology of the skeleton, as well as for enhancing clinical treatment of bone injuries. This study refined an experimental method of measuring the strain microenvironment at the site of a bone injury during bone healing. This method used a rat model in which a well-controlled bending motion was applied to an osteotomy to induce the formation of pseudarthrosis that is composed of a range of skeletal tissues, including woven bone, cartilage, fibrocartilage, fibrous tissue, and clot tissue. The goal of this study was to identify both the features of the strain microenvironment associated with formation of these different tissues and the expression of proteins frequently implicated in sensing and transducing mechanical cues. By pairing the strain measurements with histological analyses that identified the regions in which each tissue type formed, we found that formation of the different tissue types occurs in distinct strain microenvironments and that the type of tissue formed is correlated most strongly to the local magnitudes of extensional and shear strains. Weaker correlations were found for dilatation. Immunohistochemical analyses of focal adhesion kinase and rho family proteins RhoA and CDC42 revealed differences within the cartilaginous tissues in the calluses from the pseudarthrosis model as compared to fracture calluses undergoing normal endochondral bone repair. These findings suggest the involvement of these proteins in the way by which mechanical stimuli modulate the process of cartilage formation during bone healing.

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Acknowledgments

The authors would like to acknowledge Paul Barbone and the Boston University Immunohistochemistry Core Facility for their technical support. Funding was provided by the National Institutes of Health (NIH) AR53353 (EFM), the National Science Foundation Civil, Mechanical and Manufacturing Innovation Division (NSF CMMI) 1266243 (EFM), and the Boston University Clinical and Translational Science Institute (BU CTSI) Grant #UL1RR025771.

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Authors and Affiliations

  1. Department of Mechanical Engineering, Boston University, Boston, MA, USA

    Gregory J. Miller & Elise F. Morgan

  2. Department of Orthopaedic Surgery, Boston University School of Medicine, Boston, MA, USA

    Louis C. Gerstenfeld & Elise F. Morgan

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  1. Gregory J. Miller
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  2. Louis C. Gerstenfeld
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Correspondence to Elise F. Morgan.

Electronic supplementary material

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10237_2015_670_MOESM1_ESM.pdf

Supplementary Material provides additional details and figures regarding the surgical and mechanical stimulation protocols, specimen preparation and image capture for strain measurements, the accuracy of the measurements of the displacement and strain, the sampling and variability of the strain fields and corresponding histology slides, as well as additional relative frequency and logistic regression plots for Edil, max and Edil, min (Pdf 1463 KB)

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Miller, G.J., Gerstenfeld, L.C. & Morgan, E.F. Mechanical microenvironments and protein expression associated with formation of different skeletal tissues during bone healing. Biomech Model Mechanobiol 14, 1239–1253 (2015). https://doi.org/10.1007/s10237-015-0670-4

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  • DOI: https://doi.org/10.1007/s10237-015-0670-4

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