Immobilization of heparin to EDC/NHS-crosslinked collagen. Characterization and in vitro evaluation

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Wissink, M.J.B. and Beernink, R. and Pieper, J.S. and Poot, A.A. and Engbers, G.H.M. and Beugeling, T. and Aken, W.G. van and Feijen, J. (2001) Immobilization of heparin to EDC/NHS-crosslinked collagen. Characterization and in vitro evaluation. Biomaterials, 22 (2). pp. 151-163. ISSN 0142-9612

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Abstract:In the present study, heparin immobilization to a non-cytotoxic crosslinked collagen substrate for endothelial cell seeding was investigated. Crosslinking of collagen using N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) resulted in a material containing 14 free primary amino groups per 1000 amino acid residues (E/N14C). At a fixed molar ratio NHS : EDC of 0.6, the amount of heparin covalently immobilized to E/N14C increased with increasing molar ratios of EDC to heparin carboxylic acid groups (Hep-COOH), to a maximum of approximately 5–5.5 wt% at a ratio of 2. Upon incubation in cell culture medium of endothelial cells, 4 to 7% of the immobilized heparin was released during 11 days.
Immobilization of increasing amounts of heparin to E/N14C progressively reduced activation of contact activation proteases. Optimal anticoagulant activity, as measured by thrombin inhibition, was obtained after heparin immobilization using a ratio of EDC to Hep-COOH of 0.2–0.4 (14–20 mg heparin immobilized per gram of collagen). Platelets deposited to (heparinized) E/N14C showed only minor spreading and aggregation, although heparin immobilization slightly increased the number of adherent platelets. The results of this study suggest that heparin immobilization to EDC/NHS-crosslinked collagen may improve the in vivo blood compatibility of this material.
Item Type:Article
Copyright:© 2001 Elsevier
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Science and Technology (TNW)
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Link to this item:http://purl.utwente.nl/publications/74443
Official URL:http://dx.doi.org/10.1016/S0142-9612(00)00164-2
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