Labeling fibrin fibers with microparticle sized beads alter single fibrin fiber lysis, external clot lysis, and produce large fibrin aggregates upon lysis

Abstract

BackgroundFluorescent beads are often used as a tool for visualizing fibrin fibers and can mimic the size of microparticles in the blood. Studies showed microparticles alter the appearance and behavior of whole blood clot systems. ObjectivesHere we investigate the effect of beads on fibrin fiber lysis and extensibility to enhance understanding of this common research technique and as a biomimetic system for fibrin-microparticle interaction. MethodsWe used fluorescence microscopy, atomic force microscopy (AFM), and scanning electron microscopy (SEM) to quantify changes in lysis, extensibility, and clot structure of fibrin fibers and clots in the presence and absence of beads. Results and ConclusionsFibrin clot structure and lysis were altered in the presence of beads. Fibrin clots formed with beads had a higher fiber density, smaller fibers, and smaller pores. The rate of lysis for clots was reduced when beads were present. Lysis of bead-labeled individual fibers showed that beads, at concentrations similar to those reported for microparticles in the blood, cause a subset of fibers to resist lysis. In the absence of beads, all fibers lyse. These results demonstrate that beads alter fiber lysis through both a change in fibrin clot structure as well as changes to individual fiber lysis behavior. Additionally, the lysis of clots with beads produced large fibrin aggregates. This data encourages researchers to use careful consideration when labeling fibrin fibers with fluorescent beads and suggests that particles binding fibrin(ogen) in the bloodstream may be an underappreciated mechanism increasing the risk of thrombosis.