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Nifedipine and phenytoin induce matrix synthesis, but not proliferation, in intact human gingival connective tissue ex vivo

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Journal of Cell Communication and Signaling Aims and scope

Abstract

Drug-induced gingival enlargement (DIGE) is a fibrotic condition that can be caused by the antihypertensive drug nifedipine and the anti-seizure drug phenytoin, but the molecular etiology of this type of fibrosis is not well understood and the role of confounding factors such as inflammation remains to be fully investigated. The aim of this study was to develop an ex vivo gingival explant system to allow investigation of the effects of nifedipine and phenytoin alone on human gingival tissue. Comparisons were made to the histology of human DIGE tissue retrieved from individuals with DIGE. Increased collagen, fibronectin, and proliferating fibroblasts were evident, but myofibroblasts were not detected in DIGE samples caused by nifedipine and phenytoin. In healthy gingiva cultured in nifedipine or phenytoin-containing media, the number of cells positive for p-SMAD2/3 increased, concomitant with increased CCN2 and periostin immunoreactivity compared to untreated explants. Collagen content assessed through hydroxyproline assays was significantly higher in tissues cultured with either drug compared to control tissues, which was confirmed histologically. Matrix fibronectin levels were also qualitatively greater in tissues treated with either drug. No significant differences in proliferating cells were observed between any of the conditions. Our study demonstrates that nifedipine and phenytoin activate canonical transforming growth factor-beta signaling, CCN2 and periostin expression, as well as increase collagen density, but do not influence cell proliferation or induce myofibroblast differentiation. We conclude that in the absence of confounding variables, nifedipine and phenytoin alter matrix homeostasis in gingival tissue explants ex vivo, and drug administration is a significant factor influencing ECM accumulation in gingival enlargement.

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Acknowledgments

The authors wish to extend special thanks to Dr. Sandhu (Western University, London, ON, Canada) who supplied all the gingival tissues used in the study and Sarah Michelson (Western University, London, ON, Canada) for quantifying PCNA-positive and total number of cells in the tissues from patient samples. This work was funded by the Natural Sciences and Engineering Research Council of Canada (Grant Number: 355615–2009) and the Canadian Foundation for Innovation Leaders Opportunity Fund (Grant no: 18742) to DWH. SSK is a recipient of a Canadian Institutes of Health Research Doctoral Award scholarship. DWH is a recipient of the Ontario Ministry of Research and Innovation Early Researcher Award.

Competing interests

The authors declare no conflict of interests.

Author contributions

SSK performed the research, explant culture, immunohistochemistry, hydroxyproline assay, data analysis, prepared the figures, and wrote the manuscript. DWH designed the research study, as well as wrote and revised the manuscript.

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Authors and Affiliations

  1. Department of Anatomy & Cell Biology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada

    Shawna S. Kim, Sarah Michelsons & Douglas W. Hamilton

  2. Graduate Program of Biomedical Engineering, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada

    Kendal Creber & Douglas W. Hamilton

  3. Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada

    Michael J. Rieder

  4. Division of Oral Biology, Schulich School of Medicine and Dentistry, The University of Western Ontario, Dental Sciences Building, London, ON, N6A 5C1, Canada

    Douglas W. Hamilton

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  1. Shawna S. Kim
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  2. Sarah Michelsons
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  4. Michael J. Rieder
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Correspondence to Douglas W. Hamilton.

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Kim, S.S., Michelsons, S., Creber, K. et al. Nifedipine and phenytoin induce matrix synthesis, but not proliferation, in intact human gingival connective tissue ex vivo. J. Cell Commun. Signal. 9, 361–375 (2015). https://doi.org/10.1007/s12079-015-0303-9

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