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Amino Acids

, Volume 46, Issue 7, pp 1751–1761 | Cite as

Cross-linking of collagen I by tissue transglutaminase provides a promising biomaterial for promoting bone healing

  • Dario Fortunati
  • David Yi San Chau
  • Zhuo Wang
  • Russell John Collighan
  • Martin GriffinEmail author
Original Article

Abstract

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.

Keywords

Tissue transglutaminase Cross-linking Collagen I HOB Integrins 

Abbreviations

TGs

Transglutaminases

TG2

Tissue transglutaminase

HOB

Human osteoblasts

Collagen I

COL I

FAK

Focal adhesion kinase

PKCα

Protein kinase α

hBMP7

Human bone morphogenetic protein 7

bALP

Bone alkaline phosphatase

ECM

Extra cellular matrix

MMPs

Matrix metalloproteinases

AFM

Atomic force microscopy

RGD

GRGDTP

RAD

GRADSP

GAPDH

Glyceraldehyde 3-phosphate dehydrogenase

OPN

Osteopontin

VEGF

Vascular endothelial growth factor

RT

Reverse transcription

TIMP

Tissue inhibitors of MMPs

VCAN

Versican

VTN

Vitronectin

HA

Hydroxyapaptite

Si-CaP

Silicate-substituted calcium phosphate

PE

Polyethylene

Notes

Acknowledgments

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.

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Copyright information

© Springer-Verlag Wien 2014

Authors and Affiliations

  • Dario Fortunati
    • 1
  • David Yi San Chau
    • 1
  • Zhuo Wang
    • 1
  • Russell John Collighan
    • 1
  • Martin Griffin
    • 1
    Email author
  1. 1.School of Life and Health SciencesAston UniversityBirminghamUK

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