June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Tissue-engineered human cornea as a model to study wound healing: how is that process altering matrix metalloproteinases expression?
Author Affiliations & Notes
  • Camille Couture
    Department of Ophtalmology, CHU of Quebec Research Center, Laval University, Quebec, QC, Canada
    Department of Surgery, Laval University, Quebec, QC, Canada
  • Patrick Carrier
    Department of Ophtalmology, CHU of Quebec Research Center, Laval University, Quebec, QC, Canada
  • Julien Patenaude
    Department of Ophtalmology, CHU of Quebec Research Center, Laval University, Quebec, QC, Canada
    Department of Surgery, Laval University, Quebec, QC, Canada
  • Karine Zaniolo
    Department of Ophtalmology, CHU of Quebec Research Center, Laval University, Quebec, QC, Canada
  • Lucie Germain
    Department of Ophtalmology, CHU of Quebec Research Center, Laval University, Quebec, QC, Canada
  • Sylvain Guérin
    Department of Ophtalmology, CHU of Quebec Research Center, Laval University, Quebec, QC, Canada
  • Footnotes
    Commercial Relationships Camille Couture, None; Patrick Carrier, None; Julien Patenaude, None; Karine Zaniolo, None; Lucie Germain, None; Sylvain Guérin, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 736. doi:https://doi.org/
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      Camille Couture, Patrick Carrier, Julien Patenaude, Karine Zaniolo, Lucie Germain, Sylvain Guérin; Tissue-engineered human cornea as a model to study wound healing: how is that process altering matrix metalloproteinases expression?. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):736. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: The reepithelialization of the corneal surface, occurring mainly after an injury to the eye, requires an important extracellular matrix (ECM) remodeling which is suspected to be regulated by the matrix metalloproteinases (MMPs) -9 and -2. The goal of this study was to analyze the gene expression and enzymatic activity of MMP-9 and MMP-2 during wound healing using tissue-engineered human corneas as a model.

Methods: The self-assembly approach was used to produce human tissue-engineered corneas (hTECs) as previously described (Germain et al., Pathobiol. 1999). hTECs (N=11) were wounded with a 8-mm diameter biopsy punch and deposited on another reconstructed human corneal stroma to allow wound closure on a natural ECM. Undamaged hTECs (N=3) were used as controls. After 4 to 6 days of culture, hTECs were separated in 3 sections: the central (designed as the wound), the internal and the external rings, using trepans of 8-mm, 13-mm and 19-mm diameters, respectively. Total RNA was extracted from the cells isolated from each of the three sections to perform microarrays and qPCR analysis. Culture media was collected each day during wound healing to monitor MMP-9 and MMP-2 activities by gel zymography.

Results: Expression of both the MMP-2 and MMP-9 genes was found to increase in the central ring compared with their level in the external ring, as revealed by microarray and qPCR analyses. Consistent with these results, the MMP-2 and MMP-9 enzymatic activities increased from day 0 to day 4 during the wounding process. Most of all, microarray analyses identified other deregulated target genes whose respective protein products may contribute to the efficiency of the wound healing process.

Conclusions: This study will improve our understanding of the cellular and molecular mechanisms that modulate human corneal wound healing by exploiting a new, innovative 3D reconstructed tissue much closer to the native cornea.

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