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Fundamental insight into the effect of carbodiimide crosslinking on cellular recognition of collagen-based scaffolds

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Bax, DV 
Davidenko, N 
Gullberg, D 
Hamaia, SW 
Farndale, RW 


Research on the development of collagen constructs is extremely important in the field of tissue engineering. Collagen scaffolds for numerous tissue engineering applications are frequently crosslinked with 1-ethyl-3-(3-dimethylaminopropyl-carbodiimide hydrochloride (EDC) in the presence of N-hydroxy-succinimide (NHS). Despite producing scaffolds with good biocompatibility and low cellular toxicity the influence of EDC/NHS crosslinking on the cell interactive properties of collagen has been overlooked. Here we have extensively studied the interaction of model cell lines with collagen I-based materials after crosslinking with different ratios of EDC in relation to the number of carboxylic acid residues on collagen. Divalent cation-dependent cell adhesion, via integrins α1β1, α2β1, α10β1 and α11β1, were sensitive to EDC crosslinking. With increasing EDC concentration, this was replaced with cation-independent adhesion. These results were replicated using purified recombinant I domains derived from integrin α1 and α2 subunits. Integrin α2β1-mediated cell spreading, apoptosis and proliferation were all heavily influenced by EDC crosslinking of collagen. Data from this rigorous study provides an exciting new insight that EDC/NHS crosslinking is utilising the same carboxylic side chain chemistry that is vital for native-like integrin-mediated cell interactions. Due to the ubiquitous usage of EDC/NHS crosslinked collagen for biomaterials fabrication this data is essential to have a full understanding in order to ensure optimized collagen-based material performance.



cell adhesion, collagen, EDC crosslinking

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Acta Biomaterialia

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European Research Council (320598)
British Heart Foundation (SP/15/7/31561)
British Heart Foundation (RG/15/4/31268)
British Heart Foundation (None)
British Heart Foundation (None)
This work was supported by the British Heart Foundation (Grant NH/11/1/28922, RG/15/4/31268, SP/15/7/31561 and RG/09/003/27122) and the ERC Advanced Grant 320598 3D-E. D. V. Bax is funded by the Peoples Programme of the EU 7th Framework Programme (RAE no: PIIF-GA-2013-624904) and was supported by an EPSRC IKC Proof of Concept Award.
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