Improved physical and biochemical features of a collagen membrane by conjugating with soluble egg shell membrane protein

Biosci Biotechnol Biochem. 2006 Apr;70(4):865-73. doi: 10.1271/bbb.70.865.

Abstract

Soluble egg shell membrane protein (SEP) prepared from the egg shell membrane by performic acid oxidation and pepsin digestion was applied to prepare a membrane conjugated with pepsin-solubilized collagen (PSC) by cross-linking with water-soluble carbodiimide. SEP enabled a conjugated membrane to be constructed with a well-developed network structure having thicker collagen fibrils and higher stability than the PSC membrane alone, as indicated by reduced solubility and collagenase-digestion, increased denaturation temperature, and superior flexibility. The SEP-PSC membrane (SEP:PSC of 1:1, w/w) had lower sweatiness than the PSC membrane. Fibroblasts and keratinocytes adhered to the SEP-PSC-coated wells (SEP:PSC of 1:10, w/w) as a fundamental model of an artificial skin similar to that to the PSC-coated wells due to similar expression level of several cell-adhesion proteins. An SEP-PSC coating accelerated the cell growth of fibroblasts, whereas the cell growth of keratinocytes did not change, or decreased slightly due to more rapid differentiation indicated by a more substantial expression of differentiation marker proteins than a PSC-coating without SEP.

MeSH terms

  • Animals
  • Cell Adhesion
  • Cell Proliferation
  • Cells, Cultured
  • Chickens
  • Collagen / chemistry*
  • Collagen / metabolism
  • Collagen / ultrastructure
  • Egg Proteins / chemistry*
  • Egg Proteins / metabolism*
  • Egg Shell / metabolism*
  • Fibroblasts
  • Humans
  • Membranes / metabolism
  • Microscopy, Electron, Scanning
  • Pepsin A / metabolism
  • Solubility
  • Water

Substances

  • Egg Proteins
  • Water
  • Collagen
  • Pepsin A