title: The cellular response to transglutaminase-cross-linked collagen.
abstract: Collagen, type I, is a highly abundant natural protein material which has been cross-linked by a variety of methods including chemical agents, physical heating and UV irradiation with the aim of enhancing its physical characteristics such as mechanical strength, thermal stability, resistance to proteolytic breakdown, thus increasing its overall biocompatibility. However, in view of the toxicity of residual cross-linking agents, or impracticability at large scales, it would be more useful if the collagen could be cross-linked by a milder, efficient and more practical means by using enzymes as biological catalysts. We demonstrate that on treating native collagen type I (from bovine skin) with both tissue transglutaminase (TG2; tTG) and microbial transglutaminase (mTG; Streptoverticillium mobaraense) leads to an enhancement in cell attachment, spreading and proliferation of human osteoblasts (HOB) and human foreskin dermal fibroblasts (HFDF) when compared to culture on native collagen. The transglutaminase-treated collagen substrates also showed a greater resistance to cell-mediated endogenous protease degradation than the native collagen. In addition, the HOB cells were shown to differentiate at a faster rate than on native collagen when assessed by measurement of alkaline phosphatase activity and osteopontin expression.
authors.affiliation: School of Biomedical and Natural Sciences, The Nottingham Trent University, Clifton, Nottingham, NG11 8NS, UK.
authors.forename: David Y S
authors.initials: DY
authors.lastname: Chau
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Collighan
authors.forename: Elisabetta A M
authors.initials: EA
authors.lastname: Verderio
authors.forename: Victoria L
authors.initials: VL
authors.lastname: Addy
authors.forename: Martin
authors.initials: M
authors.lastname: Griffin
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D015415:Biomarkers / Q000737:chemistry; D002384:Catalysis; D002417:Cattle; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry*; D024042:Collagen Type I / Q000737:chemistry; D017364:Collagenases / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004151:Dipeptides / Q000737:chemistry; D005347:Fibroblasts / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000737:chemistry; D005780:Gelatin / Q000737:chemistry; D006358:Hot Temperature; D006801:Humans; D010006:Osteoblasts / Q000378:metabolism; D053495:Osteopontin; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D012795:Sialoglycoproteins / Q000737:chemistry; D013997:Time Factors; D011503:Transglutaminases / Q000737:chemistry* / Q000235:genetics*; D014466:Ultraviolet Rays; D014945:Wound Healing
chemical_list: D001672:Biocompatible Materials; D015415:Biomarkers; D024042:Collagen Type I; D003432:Cross-Linking Reagents; D004151:Dipeptides; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; D053495:Osteopontin; C001299:epsilon-(gamma-glutamyl)-lysine; D005780:Gelatin; D003094:Collagen; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D000469:Alkaline Phosphatase; D017364:Collagenases; D019204:GTP-Binding Proteins