Low Temperature Plasma Treatment of Rat Blood is Accompanied by Platelet Aggregation

Abstract

Growing evidence has suggested the feasibility and effectiveness of blood coagulation with low temperature plasma (LTP) at atmospheric pressure. In the present study we examined morphological changes and contents released out from platelet after LTP treatment to investigate the mechanism of LTP accelerate blood coagulation. Changes of isolated rat platelets treated with LTP had been detected by flow cytometry, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Additionally, a scheme that the mixture of different color fluorescent stained platelets treated by LTP and then detected by flow cytometry had proposed to quantitatively assess plasma triggered platelets aggregation. Furthermore, we had used label free quantitative mass spectrometry to identify changes in the supernatant proteome released from LTP treated platelets. On the surface of LTP treatment hastened whole rat blood clots, a layer of semi-transparent film had formed, which had been demonstrated to form from platelet-like membrane structure by ultrastructural analysis with TEM. Flow cytometry combined with fluorescent staining confirmed that LTP treatment promoted platelet aggregation. The observation of plasma treated stained platelets under fluorescence confocal microscopy also confirmed this judgment. We separated the supernatant from the platelets treated with LTP, and then analyzed their differences using mass spectrometry. Compared with control group, Gene ontology (GO) analysis showed that the concentration of 16 protein molecules in the experimental group was increased. LTP treatment can promotes platelet aggregation and the release of platelet activating proteins, which is the potential mechanism of speeding up blood coagulation.