Gelatin or a mixture of albumin and gelatin has been proposed for the coating of vascular grafts according to their surface thrombogenicity and biocompatibility, and the possibility of biodegradation. Heparin treatment of hemocompatible surfaces improved the patency of prostheses. In this study, different amounts of heparin were immobilized on these protein gels using a water-soluble carbodiimide [1-ethyl-3-(3-dimethylaminopropyl) carbodiimide]. The results showed a coupling of heparin with gelatin and/or albumin at the surface of the gels, stable for as long as 1 month. From 0.20 to 3.60 microg x cm(-2), heparin could be immobilized. The antiproliferative activity of immobilized heparin was controlled toward bovine smooth-muscle cells grown on these gels. Cell growth inhibition was dose dependent, but the percentages of inhibition were lower at day 8 than at day 4 at any heparin concentration used under experimental conditions. Referring to heparin in solution, immobilized heparin displayed an antiproliferative activity that improved the potential interest for coating.

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Studies were carried out to compare the behaviour of human umbilical vein endothelial cells (HUVEC) and L-929 fibroblastic cells towards proteins crosslinked by glutaraldehyde (GTA) or carbodiimide (CDI) proposed for coating of vascular prostheses. CDI crosslinking of bovine serum albumin used alone, or mixed with gelatin, allowed higher rates of cell growth and DNA synthesis than GTA crosslinking independent of cells. Assessment of the plating efficiency revealed a similar behaviour of both cells towards membranes and reference plastic surface in terms of percentages of bound cells. HUVEC proliferation onto CDI crosslinked gelatin and/or albumin membranes did not differ significantly whereas the growth of L-929 was enhanced onto gelatin albumin membranes in comparison with both gelatin membranes and the reference surface. The analysis of DNA synthesis corroborated the results of the growth curves and elicited a delay of the growth phases in HUVEC cultured onto CDI crosslinked membranes, unlike the L-929 fibroblast.

We examined the regulation of procoagulant activity and the production of fibrinolytic components by human vascular endothelium grown on coating membranes of gelatin, pure or mixed with albumin, crosslinked by carbodiimide ((G)C, (AG)C) in comparison with plastic culture dishes. Confluent monolayers were stimulated by human recombinant interleukin (IL-1 beta) and responses in terms of tissue factors like procoagulant activity, tissue plasminogen activator (tPA) and plasminogen activator inhibitor (PAI-1) were followed for up to 72 h. Procoagulant activity of cell extracts displayed similar patterns whatever the substratum tested. Quantitative immunological assays revealed a 2-fold increase in tPA antigen released from monolayers grown on (G)C and on (AG)C compared to cells grown on plastic. Exposure of monolayers to IL-1 beta reduced the secretion of tPA antigen which still reached higher values on coated than on uncoated substratum. We found that the quasi-totality of tPA formed stable complexes with PAI-1, thereby suppressing measurable fibrinolytic activity. IL-1 beta stimulated the release of PAI-1 antigen quantified by immunoassay and the kinetics of secretion were comparable on both coated and uncoated substratum.