Gentamicin supplemented polyvinylidenfluoride mesh materials enhance tissue integration due to a transcriptionally reduced MMP-2 protein expression

BMC Surg. 2012 Jan 13:12:1. doi: 10.1186/1471-2482-12-1.

Abstract

Background: A beneficial effect of gentamicin supplemented mesh material on tissue integration is known. To further elucidate the interaction of collagen and MMP-2 in chronic foreign body reaction and to determine the significance of the MMP-2-specific regulatory element (RE-1) that is known to mediate 80% of the MMP-2 promoter activity, the spatial and temporal transcriptional regulation of the MMP-2 gene was analyzed at the cellular level.

Methods: A PVDF mesh material was surface modified by plasma-induced graft polymerization of acrylic acid (PVDF+PAAc). Three different gentamicin concentrations were bound to the provided active sites of the grafted mesh surfaces (2, 5 and 8 μg/mg). 75 male transgenic MMP-2/LacZ mice harbouring the LacZ reporter gene under control of MMP-2 regulatory sequence -1241/+423, excluding the RE-1 were randomized to five groups. Bilateral of the abdominal midline one of the five different meshes was implanted subcutaneously in each animal. MMP-2 gene transcription (anti-ß-galactosidase staining) and MMP-2 protein expression (anti-MMP-2 staining) were analyzed semiquantitatively by immunohistochemistry 7, 21 and 90 days after mesh implantation. The collagen type I/III ratio was analyzed by cross polarization microscopy to determine the quality of mesh integration.

Results: The perifilamentary ß-galactosidase expression as well as the collagen type I/III ratio increased up to the 90th day for all mesh modifications, whereas no significant changes could be observed for MMP-2 protein expression between days 21 and 90. Both the 5 and 8 μg/mg gentamicin group showed significantly reduced levels of ß-galactosidase expression and MMP-2 positive stained cells when compared to the PVDF group on day 7, 21 and 90 respectively (5 μg/mg: p < 0.05 each; 8 μg/mg: p < 0.05 each). Though the type I/III collagen ratio increased over time for all mesh modifications significant differences to the PVDF mesh were only detected for the 8 μg/mg group at all 3 time points (p < 0.05 each).

Conclusions: Our current data indicate that lack of RE-1 is correlated with increased mesh induced MMP-2-gene expression for coated as well as for non-coated mesh materials. Gentamicin coating reduced MMP-2 transcription and protein expression. For the 8 μg/mg group this effect is associated with an increased type I/III collagen ratio. These findings suggest that gentamicin is beneficial for tissue integration after mesh implantation, which possibly is mediated via RE-1.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Collagen Type I / metabolism
  • Collagen Type III / metabolism
  • Foreign-Body Reaction / metabolism
  • Gentamicins / administration & dosage
  • Gentamicins / pharmacology*
  • Lac Operon
  • Male
  • Matrix Metalloproteinase 2 / genetics
  • Matrix Metalloproteinase 2 / metabolism*
  • Membranes, Artificial
  • Mice
  • Mice, Transgenic
  • Polyvinyls*
  • Protein Synthesis Inhibitors / administration & dosage
  • Protein Synthesis Inhibitors / pharmacology*
  • Random Allocation
  • Repressor Proteins / metabolism
  • Surgical Mesh*
  • Transcription, Genetic / drug effects*
  • Wound Healing / drug effects*
  • beta-Galactosidase / metabolism

Substances

  • Collagen Type I
  • Collagen Type III
  • Gentamicins
  • Membranes, Artificial
  • Polyvinyls
  • Protein Synthesis Inhibitors
  • RE1-silencing transcription factor
  • Repressor Proteins
  • polyvinylidene fluoride
  • beta-Galactosidase
  • Matrix Metalloproteinase 2