title: Characterization of a microbial transglutaminase cross-linked type II collagen scaffold.
abstract: This study investigated the effect on the mechanical and physicochemical properties of type II collagen scaffolds after cross-linking with microbial transglutaminase (mTGase). It is intended to develop a collagen-based scaffold to be used for the treatment of degenerated intervertebral discs. By measuring the amount of epsilon-(gamma-glutamyl)lysine isodipeptide formed after cross-linking, it was determined that the optimal enzyme concentration was 0.005% (w/v). From the production of covalent bonds induced by mTGase cross-linking, the degradation resistance of type II collagen scaffolds can be enhanced. Rheological analysis revealed an almost sixfold increase in storage modulus (G') with 0.005% (w/v) mTGase cross-linked scaffolds (1.31 +/- 0.03 kPa) compared to controls (0.21 +/- 0.01 kPa). There was a significant reduction in the level of cell-mediated contraction of scaffolds with increased mTGase concentrations. Cell proliferation assays showed that mTGase crosslinked scaffolds exhibited similar cytocompatibility properties in comparison to non-cross-linked scaffolds. In summary, cross-linking type II collagen with mTGase imparted more desirable properties, making it more applicable for use as a scaffold in tissue engineering applications.
authors.affiliation: National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland.
authors.forename: Damien M
authors.initials: DM
authors.lastname: O Halloran
authors.forename: Russell J
authors.initials: RJ
authors.lastname: Collighan
authors.forename: Martin
authors.initials: M
authors.lastname: Griffin
authors.forename: Abhay S
authors.initials: AS
authors.lastname: Pandit
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001426:Bacterial Proteins / Q000502:physiology* / Q000633:toxicity; D001672:Biocompatible Materials; D049109:Cell Proliferation; D002645:Chickens; D024043:Collagen Type II / Q000737:chemistry / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000378:metabolism*; D004151:Dipeptides / Q000737:chemistry / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D013302:Streptomyces / Q000201:enzymology; D023822:Tissue Engineering; D011503:Transglutaminases / Q000502:physiology* / Q000633:toxicity
chemical_list: D001426:Bacterial Proteins; D001672:Biocompatible Materials; D024043:Collagen Type II; D003432:Cross-Linking Reagents; D004151:Dipeptides; C001299:epsilon-(gamma-glutamyl)-lysine; D011503:Transglutaminases