Cross-linking of collagen I by tissue transglutaminase provides a promising biomaterial for promoting bone healing
- 1.1k Downloads
Transglutaminases (TGs) stabilize proteins by the formation of ε(γ-glutamyl)lysine cross-links. Here, we demonstrate that the cross-linking of collagen I (COL I) by tissue transglutaminase (TG2) causes an alteration in the morphology and rheological properties of the collagen fibers. Human osteoblasts (HOB) attach, spread, proliferate, differentiate and mineralize more rapidly on this cross-linked matrix compared to native collagen. When seeded on cross-linked COL I, HOB are more resistant to the loss of cell spreading by incubation with RGD containing peptides and with α1, α2 and β1 integrin blocking antibodies. Following adhesion on cross-linked collagen, HOB show increased phosphorylation of the focal adhesion kinase, and increased expression of β1 and β3 integrins. Addition of human bone morphogenetic protein to HOB seeded on TG2 cross-linked COL I enhanced the expression of the differentiation marker bone alkaline phosphatase when compared to cross-linked collagen alone. In summary, the use of TG2-modified COL I provides a promising new scaffold for promoting bone healing.
KeywordsTissue transglutaminase Cross-linking Collagen I HOB Integrins
- Collagen I
Focal adhesion kinase
Protein kinase α
Human bone morphogenetic protein 7
Bone alkaline phosphatase
Extra cellular matrix
Atomic force microscopy
Glyceraldehyde 3-phosphate dehydrogenase
Vascular endothelial growth factor
Tissue inhibitors of MMPs
Silicate-substituted calcium phosphate
This work was financially supported by the European Community programme TRACKS where D. F. was a recipient of a Marie Curie postdoctoral Fellowship with contract MRTN-CT-2006-036032 and by the Grant EPSRC (GR/S21755/02). We would like to thank Professor John Mitchell and Dr Matthew Boyd (University of Nottingham) for their kind help with the COL I gel rheology. We would like to thank Mr. A. Parchure for the technical assistance with the β1 integrin detection.
Conflict of interest
No competing financial interest exists.
- Chan KL, Khankhel AH, Thompson RL, Coisman BJ, Wong KH, Truslow JG, Tien J (2013) Crosslinking of collagen scaffolds promotes blood and lymphatic vascular stability. J Biomed Mater Res A. doi: 10.1002/jbm.a.34990
- Piercy-Kotb SA, Mousa A, Al-Jallad HF, Myneni VD, Chicatun F, Nazhat SN, Kaartinen MT (2012) Factor XIIIA transglutaminase expression and secretion by osteoblasts is regulated by extracellular matrix collagen and the MAP kinase signaling pathway. J Cell Physiol 227:2936–2946PubMedCrossRefGoogle Scholar
- Wang Z, Collighan RJ, Gross SR, Danen EH, Orend G, Telci D, Griffin M (2010) RGD-independent cell adhesion via a tissue transglutaminase-fibronectin matrix promotes fibronectin fibril deposition and requires syndecan-4/2 and α5β1 integrin co-signaling. J Biol Chem 285:40212–40229PubMedCentralPubMedCrossRefGoogle Scholar