Heparin coating reduces cell activation and mediator release in an in vitro venovenous bypass model for liver transplantation

Abstract

We used an in vitro model for venovenous bypass in a prospective, randomized study to analyze the effect on leukocytes cell activation after coating the total blood contact surface with covalently bound heparin. In ten experiments heparin-coated circuits were used, and in ten other experiments noncoated circuits were used. Monocyte cytokine production and neutrophil myeloperoxidase release were analyzed. Monocytes were isolated using anti-CD14 paramagnetic beads, and oligo(dT)₂₅ beads were used to isolate mRNA before subsequent reverse transcription and semiquantitative amplification of various cytokines in order to determine time-related changes in expression during bypass. After 2 h, mRNAs for IL-1β and IL-6 were highly upregulated in noncoated compared to heparin-coated circuits. Little or no change was seen in the expression of other cytokines. IL-1β and IL-6 were measured in plasma after 12 h and reflected the upregulated mRNAs in noncoated circuits. A significantly reduced release of myeloperoxidase was observed in coated versus noncoated circuits. This indicates that heparin-coated surfaces reduce cellular activation and the release of inflammatory mediators.