Different kinetics of the respiratory burst response in granulocytes, induced by serum from blood coagulated in contact with polymer materials

Tubes of different polymer materials were filled with blood collected by ve nous puncture. The blood was allowed to clot for 10 min, and the serum was collected. Complement activation was demonstrated through assessment of the C3-level by radial immunodiffusion. Phospholipid fingerprints were made af ter lipid extraction of serum and separation by thin-layer chromatography. The granulocyte fraction of venous blood was separated on a Percoll gradien t and the cells were either loaded with a calcium probe, or incubated with luminol. These cells were used as a biological test for inflammatory mediat ors. Serum from blood coagulated in contact with different materials was ad ded to the test cells. The intracellular calcium level was recorded by Calc ium Green-1(R) fluorescence and the respiratory burst of the test cells was recorded by luminol-amplified chemiluminescence. Serum from blood coagulat ed in contact with glass tubes, methylised glass tubes and teflon (PTFE) tu bes induced a transient increase of the cellular calcium level, indicating a G protein-coupled activation of the test cells. Serum from blood coagulat ed in contact with glass tubes, methylised glass tubes, and PTFE tubes prim ed the test cells for a subsequent f-MLP response. Serum from blood coagula ted in contact with polyurethane and polypropylene induced a direct biphasi c respiratory burst response in the test cells and serum from blood coagula ted in contact with methylised glass induced a direct monophasic respirator y burst response in the test cells. Complement activation was demonstrated after blood contact with hydrophobic glass and PTFE. Different fingerprints of phospholipid content were found in sera after blood contact with differ ent materials. The data show that different inflammatory mediators are rele ased during blood coagulation in contact with different materials. The meth od may be valuable as a screening test for blood compatibility of materials . (C) 1999 Elsevier Science Ltd. All rights reserved.