Processed eggshell membrane powder regulates cellular functions and increase MMP-activity important in early wound healing processes

PLoS One. 2018 Aug 6;13(8):e0201975. doi: 10.1371/journal.pone.0201975. eCollection 2018.

Abstract

Avian eggshell membrane (ESM) is a natural biomaterial that has been used as an alternative natural bandage to cure wounds, and is available in large quantities from egg industries. We have previously demonstrated that processed eggshell membrane powder (PEP), aiming to be used in a low cost wound healing product, possesses anti-inflammatory properties. In this study, we further investigated effects of PEP on MMP activities in vitro (a dermal fibroblast cell culture system) and in vivo (a mouse skin wound healing model). Three days incubation with PEP in cell culture led to rearrangement of the actin-cytoskeleton and vinculin in focal adhesions and increased syndecan-4 shedding. In addition, we observed increased matrix metalloproteinase type 2 (MMP-2) enzyme activation, without effects on protein levels of MMP-2 or its regulators (membrane type 1 (MT1)-MMP and tissue inhibitor of matrix metalloproteinase type 2 (TIMP-2). Longer incubation (10 days) led to increased protein levels of MMP-2 and its regulators. We also observed an increased alpha-smooth muscle actin (α-SMA) production, suggesting an effect of PEP on myofibroblast differentiation. In vivo, using the mouse skin wound healing model, PEP treatment (3 days) increased MMP activity at the wound edges, along with increased MMP-2 and MMP-9 protein levels, and increased keratinocyte cell proliferation. Altogether, our data suggest PEP stimulates MMP activity, and with a positive effect on early cellular events during wound healing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers
  • Cell Differentiation
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Dermis / cytology
  • Disease Models, Animal
  • Egg Shell / chemistry*
  • Enzyme Activation / drug effects
  • Fibroblasts / cytology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Gene Expression
  • Humans
  • Matrix Metalloproteinases / genetics
  • Matrix Metalloproteinases / metabolism*
  • Mice
  • Powders / pharmacology*
  • Stress, Physiological
  • Wound Healing / drug effects*
  • Wound Healing / genetics

Substances

  • Biomarkers
  • Powders
  • Matrix Metalloproteinases

Grants and funding

This work was supported by grants from Norwegian Research Council (NFR 235545) (HPS). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Authors TTV, SBR, MEP are employed by Nofima AS, which is a public Research Institute in Norway. Nofima provided support in the form of salaries for these authors but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. HPS is employed by Biovotec AS. Biovotec AS contributed with salary for HPS and provided the PEP material used but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of this author are articulated in the ‘author contributions’ section.