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Masquelet Technique: Effects of Spacer Material and Micro-topography on Factor Expression and Bone Regeneration

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Abstract

We and others have shown that changing surface characteristics of the spacer implanted during the first Masquelet stage alters some aspects of membrane development. Previously we demonstrated that titanium (TI) spacers create membranes that are better barriers to movement of solutes > 70 kDa in size than polymethyl methacrylate (PMMA) induced-membranes, and roughening creates more mechanically compliant membranes. However, it is unclear if these alterations affect the membrane’s biochemical environment or bone regeneration during the second stage. Ten-week-old, male Sprague–Dawley rats underwent an initial surgery to create an externally stabilized 6 mm femoral defect. PMMA or TI spacers with smooth (~ 1 μm) or roughened (~ 8 μm) surfaces were implanted. Four weeks later, rats were either euthanized for membrane harvest or underwent the second Masquelet surgery. TI spacers induced thicker membranes that were similar in structure and biochemical expression. All membranes were bilayered with the inner layer having increased factor expression [bone morphogenetic protein 2 (BMP2), transforming growth factor beta (TGFβ), interleukin 6 (IL6), and vascular endothelial growth factor (VEGF)]. Roughening increased overall IL6 levels. Ten-weeks post-engraftment, PMMA-smooth induced membranes better supported bone regeneration (60% union). The other groups only had 1 or 2 that united (9–22%). There were no significant differences in any micro computed tomography or dynamic histology outcome. In conclusion, this study suggests that the membrane’s important function in the Masquelet technique is not simply as a barrier. There is likely a critical biochemical, cellular, or vascular component as well.

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Abbreviations

PMMA:

Polymethyl methacrylate—traditional spacer material, also known as bone cement

TI:

Titanium—experimental spacer material

PBS:

Phosphate buffered saline—wash solution

DAPI:

4′,6-Diamidino-2-phenylindole—nuclear stain

TGFβ:

Transforming growth factor beta—positive regenerative protein

BMP2:

Bone morphogenetic protein 2—positive regenerative protein—promotes osteogenic differentiation

VEGF:

Vascular endothelial growth factor—positive regenerative protein—promotes angiogenesis

IL6:

Interleukin 6—negative regenerative protein—proinflammatory factor

microCT:

Micro computed tomography

BV/TV :

Bone volume/total volume fraction—fraction of volume of interest filled with bone

TV :

Total volume—total volume of interest

BV :

Bone volume—bone within the total volume of interest

BMD :

Bone mineral density—average mineral density of both bone and space within the volume of interest

TMD :

Tissue mineral density—average mineral density of only bone within the volume of interest

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Acknowledgments

We would like to thank Brendon King and Stephanie Podgorny for their efforts on these projects as part of the STARS Summer Program for High School Students (data collection). This work was supported by the Washington University Musculoskeletal Research Center (NIH P30 AR057235) as well as direct funding from the AO Foundation (AO Start-up Grant S-15-190M) and Saint Louis University (Presidential Research Fund).

Conflict of interest

Dr. J. Tracy Watson has intellectual property rights with and receives royalties from Smith and Nephew, Zimmer Biomet, and Advanced Orthopaedic Solutions. He has intellectual property rights with and is a Consultant for Advanced Orthopaedic Solutions. None of these are direct conflicts of interest to this research. All other authors have no conflicts to declare.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Saint Louis University School of Medicine, 1402 S. Grand Blvd, Schwitalla Hall M176, St. Louis, MO, 63104, USA

    Zacharie Toth, Matt Roi, Emily Evans, J. Tracy Watson, Daemeon Nicolaou & Sarah McBride-Gagyi

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  1. Zacharie Toth
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Correspondence to Sarah McBride-Gagyi.

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Associate Editor Eric M. Darling oversaw the review of this article.

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Toth, Z., Roi, M., Evans, E. et al. Masquelet Technique: Effects of Spacer Material and Micro-topography on Factor Expression and Bone Regeneration. Ann Biomed Eng 47, 174–189 (2019). https://doi.org/10.1007/s10439-018-02137-5

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