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Altered Bone Mechanics, Architecture and Composition in the Skeleton of TIMP-3-Deficient Mice

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Abstract

Tissue inhibitor of metalloproteinases-3 (TIMP-3) maintains a healthy extracellular matrix by regulating matrix metalloproteinases (MMP), disintegrin–metalloproteinases (ADAM), and disintegrin–metalloproteinases with ThromboSpondin-like motifs (ADAMTS) activity. Currently, there is a need for a comprehensive understanding of the effects of TIMP-3 on the bone quality and integrity. In this study, we examined the mechanical, morphological, and compositional properties of TIMP-3 knock out (Timp-3 −/−) mouse bone. We hypothesize that the lack of TIMP-3 plays an important role in maintaining the overall bone integrity. Mechanical properties of humeri, lumbar vertebrae, and femurs from Timp-3 −/− mice were determined using 3-point bending, compression, and notched 3-point bending, respectively. Morphological properties of the humeral cortical and trabecular bone and the caudal vertebrae cortical bone were evaluated using micro-computed tomography, while the composition of the femoral cortical and trabecular bone was examined using Fourier transform infrared spectroscopic imaging. Our results revealed that the integrity of the Timp-3 −/− bone is compromised due to changes in its composition, structure, and mechanics. Reductions in the yield and ultimate load and stress capacity, and loss in bone fracture toughness were attributed to reduced density and thickness, and increased porosity of cortical bone. Thin trabeculae were dense, highly connected, and closely packed in Timp-3 −/− bone. Furthermore, altered cortical and trabecular bone mineralization and increased compositional heterogeneity were found in Timp-3 −/− bone, all being indicative of high bone remodeling. In conclusion, this study suggests that the lack of TIMP-3 is detrimental to bone development and maintenance.

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Acknowledgements

The authors are grateful for the provision of Timp-3 −/− mice by Dr. Rama Khokha (Toronto, Canada). This study was funded by the Florida Institute of Technology and partly supported by Arthritis Research UK (ARUK) grant 20039 awarded to GB and by grants awarded to AAP by ARUK (20581) and Biotechnology and Biological Sciences Research Council (BB/J003727/1).

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Authors and Affiliations

  1. Department of Biomedical Engineering, Florida Institute of Technology, Melbourne, FL, USA

    Brendyn Miller & Alessandra Carriero

  2. Hospital for Special Surgery, New York, NY, USA

    Lyudmila Spevak, Lyudmila Lukashova & Adele Boskey

  3. Department of Comparative Biomedical Sciences, Royal Veterinary College, London, UK

    Behzad Javaheri & Andrew A. Pitsillides

  4. Institute of Aging and Chronic Disease, University of Liverpool, Liverpool, UK

    George Bou-Gharios

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  1. Brendyn Miller
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  2. Lyudmila Spevak
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  3. Lyudmila Lukashova
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  4. Behzad Javaheri
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  5. Andrew A. Pitsillides
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  6. Adele Boskey
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  7. George Bou-Gharios
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  8. Alessandra Carriero
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Correspondence to Alessandra Carriero.

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Conflict of interest

The authors Brendyn Miller, Lyudmila Spevak, Lyudmila Lukashova, Behzad Javaheri, Andrew A. Pitsillides, Adele Boskey, George Bou-Gharios, and Alessandra Carriero declare that they have no conflict of interest.

Human and Animal Rights and Informed Consent

Mice were maintained under standard laboratory conditions and experiments were conducted in compliance with the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines for reporting. Briefly, mice were housed up to 4 per cage in polypropylene cages with wood chip and paper bedding and provided standard rodent maintenance diet and water ad libitum throughout the study. All procedures complied with the UK Animals (Scientific Procedures) Act 1986 and were reviewed and approved by UK Home Office and local ethics committee.

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Miller, B., Spevak, L., Lukashova, L. et al. Altered Bone Mechanics, Architecture and Composition in the Skeleton of TIMP-3-Deficient Mice. Calcif Tissue Int 100, 631–640 (2017). https://doi.org/10.1007/s00223-017-0248-5

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  • DOI: https://doi.org/10.1007/s00223-017-0248-5

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