In vitro screening procedure for characterization of thrombogenic properties of plasma treated surfaces

Estimation of thrombogenic surface properties is an important aspect of hemocompatibility studies. To improve our understanding of interaction between blood and biomaterial surfaces, there is a need to employ standardized methods that are both effective and efficient. This contribution details a systematic approach for the in vitro analysis of plasma modified polymer surfaces and human blood platelet interaction, following the recently introduced ISO 10933-4 guidelines. A holistic multistep process is presented that considers all aspects of testing procedure, including blood collection, platelet function testing, and incubation parameters, right through to a comparison and evaluation of the different methods and analysis available. In terms of detection and analysis, confocal light microscopy is shown to offer many advantages over the widely used scanning electron microscopy technique; this includes simpler, less-invasive sample preparation, and less time-consuming analysis procedure. On the other hand, as an alternative to microscopy techniques, toxicology sulforhodamine B based assay (TOX assay) was also evaluated. It has been shown that the assay could be used for rapid estimation of relative concentration of blood platelets on the surface of plasma treated materials, especially when samples do not allow the implementation of microscopy techniques.