title: Improved physical and biochemical features of a collagen membrane by conjugating with soluble egg shell membrane protein.
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.
authors.affiliation: Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan. k-taka@cc.tuat.ac.jp
authors.forename: Tamao
authors.initials: T
authors.lastname: Ino
authors.forename: Makoto
authors.initials: M
authors.lastname: Hattori
authors.forename: Tadashi
authors.initials: T
authors.lastname: Yoshida
authors.forename: Shunji
authors.initials: S
authors.lastname: Hattori
authors.forename: Keiji
authors.initials: K
authors.lastname: Yoshimura
authors.forename: Koji
authors.initials: K
authors.lastname: Takahashi
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D049109:Cell Proliferation; D002478:Cells, Cultured; D002645:Chickens; D003094:Collagen / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D004527:Egg Proteins / Q000737:chemistry* / Q000378:metabolism*; D004528:Egg Shell / Q000378:metabolism*; D005347:Fibroblasts; D006801:Humans; D008566:Membranes / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D010434:Pepsin A / Q000378:metabolism; D012995:Solubility; D014867:Water
chemical_list: D004527:Egg Proteins; D014867:Water; D003094:Collagen; D010434:Pepsin A