Abstract
BackgroundAltered fibrin clot architecture is increasingly associated with cardiovascular diseases; yet, little is known about how fibrin networks are affected by small molecules that alter fibrinogen structure. Based on previous evidence that S-nitrosoglutathione (GSNO) alters fibrinogen secondary structure and fibrin polymerization kinetics, we hypothesized that GSNO would alter fibrin microstructure.Methodology/Principal FindingsAccordingly, we treated human platelet-poor plasma with GSNO (0.01–3.75 mM) and imaged thrombin induced fibrin networks using multiphoton microscopy. Using custom designed computer software, we analyzed fibrin microstructure for changes in structural features including fiber density, diameter, branch point density, crossing fibers and void area. We report for the first time that GSNO dose-dependently decreased fibrin density until complete network inhibition was achieved. At low dose GSNO, fiber diameter increased 25%, maintaining clot void volume at approximately 70%. However, at high dose GSNO, abnormal irregularly shaped fibrin clusters with high fluorescence intensity cores were detected and clot void volume increased dramatically. Notwithstanding fibrin clusters, the clot remained stable, as fiber branching was insensitive to GSNO and there was no evidence of fiber motion within the network. Moreover, at the highest GSNO dose tested, we observed for the first time, that GSNO induced formation of fibrin agglomerates.Conclusions/SignificanceTaken together, low dose GSNO modulated fibrin microstructure generating coarse fibrin networks with thicker fibers; however, higher doses of GSNO induced abnormal fibrin structures and fibrin agglomerates. Since GSNO maintained clot void volume, while altering fiber diameter it suggests that GSNO may modulate the remodeling or inhibition of fibrin networks over an optimal concentration range.
Highlights
Changes in fibrin clot architecture are increasingly recognized as an important risk factor for cardiovascular disease and thrombotic complications [1,2,3,4,5]
When human platelet-poor plasma was incubated with increasing concentrations of GSNO, we found fiber density remained relatively unchanged over an order of magnitude (0.01–0.1 mM GSNO), followed by a sharp 53.2% decrease in fiber density as GSNO increased to 2.5 mM, and subsequently to zero, as fibrin formation was completely inhibited, Figure 2D
We found that GSNO decreased fibrin fiber density and fibrin branch point density while increasing fiber diameter, resulting in ‘‘coarse’’ fibrin networks, known to be more susceptible to fibrinolysis [36] and have weaker mechanical properties [8]
Summary
Changes in fibrin clot architecture are increasingly recognized as an important risk factor for cardiovascular disease and thrombotic complications [1,2,3,4,5]. The composition of a network’s structural elements, including fiber density, diameter and branch point density [8] are determined by the clotting conditions, including fibrinogen, thrombin and calcium concentrations, FXIIIa activity, pH, ionic strength, temperature, whether fibrin is formed in plasma and by changes in fibrinogen structure [8,10,11,12,13,14,15,16]. Altered fibrin clot architecture is increasingly associated with cardiovascular diseases; yet, little is known about how fibrin networks are affected by small molecules that alter fibrinogen structure. Based on previous evidence that Snitrosoglutathione (GSNO) alters fibrinogen secondary structure and fibrin polymerization kinetics, we hypothesized that GSNO would alter fibrin microstructure
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
