Tethering a laminin peptide to a crosslinked collagen scaffold for biofunctionality.
Cell adhesion peptide regulates various cellular functions like proliferation, attachment, and spreading. The cellular response to laminin peptide (PPFLMLLKGSTR), a motif of laminin-5 alpha3 chain, tethered to type I collagen, crosslinked using microbial transglutaminase (mTGase) was investigated. mTGase is an enzyme that initiates crosslinking by reacting with the glutamine and lysine residues on the collagen fibers stabilizing the molecular structure. In this study that tethering of the laminin peptide in a mTGase crosslinked collagen scaffold enhanced cell proliferation and attachment. Laminin peptide tethered crosslinked scaffold showed unaltered cell morphology of 3T3 fibroblasts when compared with collagen and crosslinked scaffold. The triple helical structure of collagen remained unaltered by the addition of laminin peptide. In addition a dose-dependent affinity of the laminin peptide towards collagen was seen. The degree of crosslinking was measured by amino acid analysis, differential scanning calorimeter and fourier transform infrared spectroscopy. Increased crosslinking was observed in mTGase crosslinked group. mTGase crosslinking showed higher shrinkage temperature. There was alteration in the fibrillar architecture due to the crosslinking activity of mTGase. Hence, the use of enzyme-mediated linking shows promise in tethering cell adhesive peptides through biodegradable scaffolds.
Damodaran
Gopinath
G
National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland.
Collighan
Russell
R
Griffin
Martin
M
Pandit
Abhay
A
D016475:3T3 Cells; D000199:Actins / Q000378:metabolism; D000818:Animals; D002417:Cattle; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002942:Circular Dichroism; D003094:Collagen / Q000378:metabolism* / Q000648:ultrastructure; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003599:Cytoskeleton / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D007797:Laminin / Q000378:metabolism*; D051379:Mice; D018625:Microscopy, Atomic Force; D010455:Peptides / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D054457:Tissue Scaffolds; D011503:Transglutaminases / Q000378:metabolism
D000199:Actins; D003432:Cross-Linking Reagents; D007797:Laminin; D010455:Peptides; D003094:Collagen; D011503:Transglutaminases