Feasibility Of Heparin Anticoagulation Platelet-rich Plasma In Patients Undergoing Cardiopulmonary Bypass

Part ⅠQuality comparison of platelet-rich plasma preserved by different anticoagulants during cardiopulmonary bypassObjective:In this study,the quality of platelets obtained by autologous blood cell separation through venous drainage of blood cells before and before extracorporeal circulation is evaluated in vitro,and the changes of platelet mass at different times under the normal temperature are observed.Compared with the conventional citrate anticoagulant PRP and the control group EDTA anticoagulation whole blood,apheresis platelet concentrates,and stopping whole blood.To observe the effect of different anticoagulants on the separation and preservation of autologous platelet rich plasma,and to explore the safety and feasibility of the application of heparin anticoagulant plasma in patients with cardiopulmonary bypass.Methods:From January 2017 to August of 2017,20 adult patients who were selected in our hospital were selected as the subjects.The same patients were divided into citric acid anticoagulant group(SC-PRP)and heparin anticoagulant group(Heparin-PRP),and the blood samples collected by EDTA anticoagulant tube were 1 in the control group.The small plate was 2 in the control group,and compared with the whole blood(3)in the control group after cardiopulmonary bypass,20 cases in each group.After inward anaesthesia,20ml was injected into several citric acid anticoagulant tubes and heparin anticoagulant tubes by internal jugular vein catheterization.Each tube contained 2.7 ml whole blood and 0.3ml anticoagulant,mixed,200g centrifugal force,centrifugation for 10 minutes.After statically indwelling,the upper plasma was removed to another centrifuge tube,after two centrifugation,abandoned.The supernatant layer was extracted from plasma lower layer to make platelet rich plasma(PRP).Sysmex blood cell analyzer was used to detect five indexes of platelet count in each group.Blood gas and electrolytes,blood sugar,lactic acid and osmotic pressure were measured by Nova biomedical blood gas analyzer.The concentration,maximum aggregation and maximum aggregation time of two groups of Adenosine diphosphate,ADP,Arachidonic acid,AA,and adrenaline(Adrenaline,ADR)were detected with light density Turbidimetry in two groups of adenosine(Arachidonic acid,AA)and adrenaline(Adrenaline,ADR),and the low permeability was measured by platelet aggregation apparatus.The value of Hypotonic shock response(HSR).The differences of platelet indexes among different groups were compared.Results:For the same sample of the same sample,heparin and citric acid anticoagulant platelet rich plasma can achieve satisfactory separation effect.The concentration of platelets is about 2-3 times the concentration of whole blood platelets,and the concentration of concentrated platelet PLT is higher than PRP.Compared with EDTA anticoagulant,both citrate and heparin anticoagulant could cause reversible platelet aggregation,and PLT,MPV,PDW,PLCR and PCT were lower,compared with EDTA anticoagulants(P<0.05).When the anticoagulant of anticoagulant in citric acid group and heparin group was added to EDTA anticoagulant,PLT,MPV,PDW,PLCR and PCT were all recovered in varying degrees,and there were significant differences(P<0.05).The MPV and PDW of PRP in heparin group PRP and citric acid group were all larger than those in concentrated platelet,and the PDW in heparin group was larger than that in citric acid group(P<0.05).After the end of cardiopulmonary bypass(CPB),the PLT of whole blood decreased significantly(P<0.05)than that before CPB(P<0.05),and PDW,PLCR and PCT decreased significantly after CPB(P<0.05).The pH value of Heparin-PRP was higher than that of SC-PRP(P<0.05),PC02 was lower than that of SC-PRP group(P<0.01),HCO3-was higher than SC-PRP group(P<0.01).Compared with the concentrated platelet group,the PH of Heparin-PRP was higher(P<0.05)and HCO3-was far higher than that of concentrated platelets(P<0.01);the pH value of SC-PRP was higher than that of the concentrated platelets group(P<0.05),PCO2 and HCO3-were greater than those of the concentrated platelets group(P<0.01).The pH value of the heparin PRP group and the group PRP of citrate was higher than that of the whole blood group(P<0.01),and the PCO2 of the heparin group and the PRP group of citrate was lower than that of the whole blood group(P<0.01).There was a significant difference between the two groups of anticoagulants between the PRP and the whole blood between pH and PCO2(P<0.01).HCO3-was also statistically significant(P<0.05).Compared with concentrated platelet group and citric acid group,heparin group Na+was lower,K+,Ca++,Mg++ and Cl-were significantly higher(P<0.01).Blood glucose and lactic acid in heparin group and citric acid group were all in normal range,significantly lower than those in platelet concentrating group,with significant difference(P<0.01).The osmotic pressure of heparin group was lower than that of citric acid group and platelet concentrating group(P<0.01).Compared with the platelet concentrates,the recovery rate of hypotonic shock reaction of the two groups was higher than that of the PRP group(P<0.01).There was no significant difference between heparin PRP group and citric acid PRP group(P>0.05).The 60s,180s and 300s aggregation rates and maximum aggregation rates of AA and ADP in heparin group were better than those in the platelet concentrating group(P<0.05).The aggregation rate of ADP and ADR in the citric acid group,the 300s aggregation rate and the maximum aggregation rate of AA,ADP and ADR in the citric acid group were all better than those in the platelet concentrating group(P<0.05),respectively.The 300s aggregation rate and the maximum aggregation rate of AA,ADP and ADR in the citric acid group were better than those in the concentrated platelet group(P<0.05).Compared with SC-PRP,Heparin-PRP has higher 60s aggregation rate,60s,180s,300s aggregation rate and maximum aggregation rate of AA than SC-PRP(P<0.05).Conclusion:Compared with EDTA anticoagulation,citrate and heparin anticoagulation both cause reversible aggregation of platelets,but this aggregation is reversible after EDTA entry.In heparin group,PDW,MPV and PLCR of PRP were slightly larger than those of PRP and platelet concentrates of citric acid group.The PLT,PDW,PLCR and PCT of whole blood before CPB were better than that of whole blood.In heparin group,the preservation effect of most ions was closer to physiological level,lactic acid and blood glucose were slightly higher than those of other groups,but they were all within normal range.The pH of heparin PRP was higher than that of citric acid group PRP and platelet concentrating group,which was caused by the difference between PCO2 and HCO3-.The aggregation of AA and ADP in the heparin PRP group was better than that of the group PRP,and the aggregation of ADR in group PRP was better.The aggregation of AA,ADP and ADR in the two groups was superior to that of the concentrated platelets group.The recovery rate of hypotonic shock reaction of two groups of PRP was higher than that of the concentrated platelet group,and there was no significant difference between heparin PRP group and citric acid PRP group.Heparin can be used as a platelet anticoagulant during cardiac surgery to preserve PRP.Part ⅡThe quality changes of platelet rich plasma preserved by two groups of anticoagulants at different timeObjective:The first part compares the preservative effects of various anticoagulants on PRP.In this stage,the platelet rich plasma with heparin anticoagulant and citric acid anticoagulant will be further evaluated in the condition of prolonged temperature preservation and the change of platelet quality,and platelet morphologic indexes at different time points under electron microscope are added to explore the platelet rich plasma in heparin anticoagulation.How long is it safe and effective for the patient?Methods:From September 2017 toFebruary 2018,20 adult patients were selected as the research object in our hospital,and the same patients were divided into citric acid anticoagulant group and heparin anticoagulant group at the same time.After inward anaesthesia,30ml was injected into several citric acid anticoagulant tubes and heparin anticoagulant tubes by internal jugular vein catheterization.Each tube contained 2.7 ml whole blood and 0.3ml anticoagulant,mixed,200g centrifugal force,centrifugation for 10 minutes.After statically indwelling,the upper plasma was removed to another centrifuge tube,after two centrifugation,abandoned.The upper layer and the lower layer of plasma were made into platelet rich plasma(PRP).After being stored in centrifuge tubes,the corresponding indexes were detected at the time points of 2 hours,4 hours,8 hours and 24 hours at the time of sampling,including five items of platelet count in two groups,and the two groups of blood gas and electrolytes,blood glucose,lactate,osmotic pressure and other indicators;two groups of hypotonic shock test;two groups in ADP,AA and epinephrine induced maximum aggregation.The shape and size of platelets were observed by electron microscopy at the time and 6 hours after sampling.The differences of platelet indexes between the two groups were compared.Results:The PRP,PDW,MPV and PLCR of the two groups were significantly different at different times(P<0.05).Compared with this group Oh,only the PDW and MPV of the 4H are higher than the Oh value(P<0.05).The 2H and 24h of PDW in Heparin-PRP have significant differences compared with 0h,and there are significant differences in each time.The pH value of Heparin-PRP in 0h,2h and 4H is higher than that in SC-PRP(P<0.05).Na+,K+,Ca++,Mg++ ions,lactate and osmotic pressure of SC-PRP were significantly different from Heparin-PRP at all times(P<0.05).There was a significant difference between the two groups of blood glucose in Oh and 2H(P<0.05).Compared with the Oh in this group,the pH values of the two groups increased first and then decreased,and the pH values of SC-PRP were significantly different from those of Oh at all times(P<0.05).The Ca++ of Heparin-PRP decreased in 2H,4H and 8h(P<0.05),the blood glucose decreased,and there was a significant difference between 8h and 24h(P<0.05);the lactic acid increased gradually with time(P<0.05),but within 8 hours,the biochemical indexes of Heparin-PRP were in the normal range.The blood glucose of SC-PRP showed a decreasing trend in 4h,8h and 24h(P<0.05),and lactic acid increased gradually with time(P<0.05).The osmotic pressure of SC-PRP in 8h is lower than that of Oh(P<0.05).The maximum aggregation rate of AA and ADP of SC-PRP was lower than Heparin-PRP(P<0.05)at most times;ADR aggregation rate was higher in Oh and 2H than in heparin group(P<0.05).From the beginning of 4h,the maximum aggregation rate of AA,ADP and ADR in two groups of platelets decreased with time,and there was a significant difference compared with that of Oh in this group(P<0.05),but the value within 8 hours was still within the normal range.The HSR results of the two groups of hypotonic shock were in the normal range,and there was no statistical difference between the two groups at all times(P>0.05);there was a downward trend in the group with time,but there was no significant difference between the groups in 8h,and there was a significant difference between the two groups and Oh(P<0.05).Scanning electron microscopy showed that the platelet volume of Heparin-PRP was slightly larger than that of SC-PRP and platelet concentrating group(P<0.05),and there was no significant difference between groups after 6h.There was no significant difference in the scores of Oh and 6h between the two groups.It was higher than that of the concentrated platelets group.The two groups increased after 6 hours,and the citrate group had longer and more pseudo feet.The maximum number of pseudo foot in the concentrated platelets group was the longest.The morphology of the three groups of platelets was different in transmission electron microscope.Platelet concentrates platelets have more platelet and more pseudo feet.Platelets and heparin platelets of citric acid platelets have more disc like platelets and smaller and shorter pseudo feet.The volume of heparin platelets under transmission electron microscope was not significantly different from that of the other two groups(P>0.05).Conclusion:PDW,MPV and PLCR of Heparin-PRP were slightly higher than SC-PRP at all times,and PDW,MPV and PLCR tended to increase as time went on.The pH value of Heparin-PRP was higher than that of citric acid group,and the electrolytes were all in physiological range.The lactate values of each time were slightly higher than those of SC-PRP.The electrolyte content of SC-PRP is different from that of the normal value.As time went on,the calcium and blood sugar values of Heparin-PRP and SC-PRP decreased gradually,and lactic acid increased gradually,but they were all within normal range in the first 8 hours.The maximum aggregation rate of Heparin-PRP in AA and ADP is slightly better than that of SC-PRP,while the maximum aggregation rate of ADR is lower than SC-PRP.There was no significant difference in the HSR results between the two groups at all times.The group showed a decreasing trend with time,but there was no significant difference in the 8h group.The results of scanning electron microscopy showed that the platelet volume of Heparin-PRP was slightly greater than that of SC-PRP.There was no significant difference between the two groups of platelets in Oh.After 6h,the two groups of pseudo feet were all increased,and the most of the pseudo foot in the concentrated platelets group was the longest.The results of transmission electron microscopy were similar to that of the concentric group.The number of round feet was longer in SC-PRP and Heparin-PRP groups,but less in the number of pseudo feet.The diameters of the three groups were not statistically different from those of SEM.With the extension of time,Heparin-PRP and SC-PRP had their own advantages and disadvantages.Most of heparin’s indexes were within normal range within 8 hours.Heparin-PRP is relatively safe within 8 hours.