Experimental Study On Blood Coagulation And Wound Healing By Pulsed Plasma Jet

In this thesis,a pulsed non-thermal plasma jet reactor developed by the laboratory was used.Based on the cellular mechanism of platelet function,the mechanism of platelet in promoting blood coagulation was explored.At the same time,the potential application value in wound healing was verified by animal experiments.The device is composed of a pulsed power supply and a jet reactor.Microsecond pulses with frequency between 1-30 kHz and voltage between 0-12 kV can be output by the power supply.The jet length used in this paper is about 5cm,and the distribution of the active particles is tested by spectral analysis.The excitation temperature of jet electron is 3616 K.In terms of blood coagulation,in order to study the mechanism of plasma promoting blood coagulation,experiments of plasma treating whole blood and separating platelets were carried out.Based on the size of membrane structure and the absence of nucleus,it is speculated that the structure is composed of platelets.A series of experiments in vitro were carried out to study the mechanism of plasma stimulating platelet function.The results show that the contents of the plasma are reduced by TEM.No label mass spectrometry analysis of platelet supernatant revealed increased release of many proteins associated with platelet aggregation.Among them,CXC chemokine RTCK1 increased the most,which is closely related to platelet activation.Flow cytometry showed that platelet aggregation increased after plasma treatment.When PKH26 and PKH67 fluorescent dyes were combined with flow cytometry,it was found that the double-colored events in the 120s plasma group were 63.7%,and only 17.0%in the control group.The aggregation increased slightly after half an hour,which proved that the process was not reversible.This indicates that plasma treatment can continuously activate platelets and stimulate their biological function in blood agglutination.In order to verify the promoting effect of plasma on chronic wound healing,streptozotocin was used to build a diabetic rat model,and plasma treatment was used in the experimental group and helium treatment in the control group.After that,the healing was tested.On day 7,14,and 21,the wound tissues were fixed and treated with hematoxylin-eosin staining,Mason trichromatic staining,and immunohistochemical staining to evaluate wound re-epithelialization,collagen deposition,neovascularization,and growth factor expression.The results showed that plasma treatment resulted in faster epithelial tissue formation,collagen deposition,and more complete wound structure.The wound dynamics results showed that there were differences in wound area ratios at day 1,3,6,11,13,15,and 16(P<0.05).There were significant differences in wound area ratios on days 5,14 and 17-21(P<0.01).The wound healing was faster in the plasma treatment group.At the same time,this study also noted that plasma treatment can reduce the formation of scar.Hematoxylin-eosin staining and immunohistochemical staining showed that plasma can affect the TGF-β/Smad2/3 pathway and reduce the amount ofα-SMA,thereby reducing tissue fibrosis and scarring.