- Contatti Medical
Transcrição
- Contatti Medical
""O TrallsfusiOI/ Medicil/i?, 2005, 15, 319-322 ORIGINAL doi: 1O.1111íj.l365-3148.2005.00594.x . _'_n. -. - é"";:; ARTICLE The effect of continuous-flow automated plateletpheresis on fibrinolytic activity of danar plasma P. Radziwon*t, B. Boczkowska-Radziwon*,A. Lipska*, M. Smoktunowicz* and J. Kloczkot * Regiollal Ce11lrefor Transfusiol/ Medicine, BialJ'stok, Receil'ed 26 Janllary 2005; acceptedfor publicatiol/ and t Deptart11lent of Hael7latology, Medical Uni\'ersity of BialJ'stok, Bialystok, Poland 7 March 2005 SUMMARY.Blood circulating in extracorporeal circuit of lhe apheresis seis has a contact with an artificial surface. The data on lhe influence of plateletpheresis on fibrinolytic activity are very limited and difficult to interpret. The aim of our study was to estimate lhe effect of plateJetpheresis on lhe activation of fibrinolysis. Plateletpheresis was performed in 17 hea1thy blood donors using continuous-flow cell separator COM.TEC (Fresenius, Bad Homburg, Germany). Before and after plateletpheresis, blood samples were taken and markers of fibrinolysis (PAP, t-PA, PAI-I) as well as factor XII activity have been l11easured. We observed statistically significant decrease in t-PA and factor XII activities anel' plateletpheresis. There were no signifiGani changes in concentrations of t-PA, PAI-l and PAr as well as PAI-l activity anel' plateletpheresis. Plateletpheresis perforl11edby COM.TEC cell separator has very little, if any, effect on lhe activation of fibrinolysis. The mechanism of lhe inhibition of t-PA activity needs further investigations. During extracorporeal circulation occurring in haemodialysis, cardiopuhnonary bypass or apheresis bloud colllacl,; wilh lhe artiCicÜl1, on!y in parI biocompatible anel reJativcly large surface of disposablc seis. Plasma proteins, e.g. coagulation factors, adsorb on lhe foreign surface (Vroman et aI., 1971; Forbes, 1981). Protein deposition is followed by lhe adhesion of blood cells: platelets, leucocytes and erythrocytes (Vroman, 1987; Koepke et aI., 1981). The interaction with lhe surface affects enzyme activity (induces factor XII activation) and cell functions (MulvihiII et ai., 1990). There are reports suggesting that fibrinogen, vou Willebrand factor and fibronectin stimulate platelet interaction with artificial surfaces, whereas albumin, ceruloplasmin and transferrin inhibit it (Kim et ai., 1977; Elam & Nygren, 1992). Kobayashi et aI. (l9?3) observed increased coagulaliou activity anel' plateletpheresis performed with continuous-flow blood separator CS-3000 (Baxter Healthcare, Deerfield, IL, USA). In contrary, Slohlawctz c/ oi. (1999) cliel nol detecl significant changcs in blooel coaglllalion inelllcecl by plalcJelpheresis perforl11ed on lhe two other blood separators Key words: COM.TEC, facto r XII, fibrinolysis, PAI-I, PAP, plateletpheresis, t-PA. - AMICUS (Fenwal Division, Baxter Healthcare) and MCS 3p (Haemonetics, Braintree, MA, USA). The data on lhe influence of plate1etpheresis on fibrinolytic activity are very limited and are difficult to interpret (Kobayashi et ai., 1993; Stohlawetz et ai., 1999). There are no reports about lhe effects of plateletpheresis on t-PA concentration and activity and on PAI-l activity. The aim of this study was to estimate lhe effect of plateletpheresis performed on lhe new automated blood separator - COM.TEC (Fresenius, Bad Homburg, Germany) - on lhe main activator and ',J _'_';:'"" .-' .I' - ,.. . ...,~ . '\~.:;;,~ lhe main inhibitor of fibrinolysis - t-PA and PAI-l as well as on factor XII activity. Correspondence: Piotr Radziwon, MD, PhD, Regional Centre for Transfusion Medicine, Sklodowskiej rolando Te!.: +48 85 7447002; fax: +48 85 7447133; e-mail: [email protected] ~ 2005 BlackweIl Publishing Ltd 23, PL - 15-950Bialystok, MA TERIALS AND METHODS Seventeen hea1thy donors, aged 21-40 years, who had not taken any drugs for at least 2 weeks prior to blood sampling, underwent standard thrombapheresis procedure. The automated continuous-flow cell separator 319 .' ; 4 ...".0 320 P. Radzi1l'01let a!. " -C .:> esis (11= 17) Parameter Mean :I: SD Blood flow (mL min-l) Blood volume processed (mL) Donation time (min) ACD consull1ption (mL) PLT yieId (x 1011) 46.60 :I: 12 2601.5 :I: 314.4 58.9 :I: 14.3 338:1: 42 3.2 :I: 0.3 fr PAI-] Table 1. The main parametersof performed thrombapher- ,': , , ", The concentration and activity of PAI-I were measured using COALIZA PAIol and COATEST PAI from Chromogenix-Instrumentation Laboratory SpA, Italy, respectively. o: d!~. ~úr1 .'<-J '~{ PAP Concentration of PAP was measured using Imuclone PAP Elisa kit (American Diagnostica, Greenwich, CT, USA),. COM.TEC (Fresenius) with single needle apheresis sei (S5L) were used. Table I shows lhe main parameters of performed procedures. Whole blood was anticoagulated with ACD-A at average blood : ACD-A fatia of 9 : I. The investigations were perfonl1ed with approval of lhe Committee on lhe Ethics of Research in Hul11an Experimentation at Medical Academy in Bialystok, Poland and under lhe guidelines of lhe Helsinki DecIaration for human research. Factor XII actil'ity , Coagulation factor XII deficient plasma (human) (Dade Behring, Marburg, Gerl11any) and activated partial thromboplastin assay has been applied for lhe meaSUrel11entof fartar XII activity. The results were expressed in percentage of lhe narro. ' Statistics The data are presented as mean value :f: standard deviation (SD). For statistical evaluation of lhe results Wilcoxon's test was used. Changes in l11easured parameters were corrected for haemodilution. Blood collectioll and p/'epa/'atioll o/ platelet poor plasma (P PP) Venous blood was drawn without stasis from donors before and 5 min after thrombapheresis, into syringes containing either sodium citrate (0.32% final concentration) for lhe measurement of topA, PAIol and PAP concentrations or 0.5 M citrate buffer, pH 4.3 (StabilyteTM Biopool, Bray, Ireland) for lhe measuremcnt ol' toPA and PAI. 1 activities, Platclet poor plasma was prepared by centrifugatian af whole blood at 2000 x g for 10 min RESUL TS We observed significant decrease of toPA ano factor XII activity after thrombapheresis, whereas its concentration remained unchanged. There were no significant changes in lhe concentratians of PAP ano PAIol as well as PAIol activity. The data of lhe measured parameters are presented in Table 2. toPA DISCUSSION The concentration and activity of toPA were measured using commerciaIly available kits (COALIZA toPA and COATEST toPA, respectively) from Chromogenix-Instrumentation Laboratory SpA, .. -,_It~IY'd ~~~~~tJ High-molecular-weight kininogen, prekaIlikrein, facter XII and factor XI recognized as lhe 'contact system' require contact with artificial, negatively charged surfaces for zymogene activation in vitro. : -:w : ';:~-;'F"'rabl~' 2. T~e data of measuredparameters of fibrinolysis(mean1['80,':'; = 17) Parameter Before After PAP concentration (ng mL -I) toPA concentration (ng rol-I) toPA activity (IU mL -I) PAI-1 concentration (ng mL-I) PAI-1 activity (AV mL -I) Factor XII activity (%) 21.15 :I: 11.9 3.70:1:11.2 5.36:1: 3.3 46.18:1: 30.50 6.88 :I: 4.58 99.20:1: 16.45 21.00:1: 11.7 3.18:1:0.8 1.17:1: 0.6 42.82:1: 29.21 6.21 :I: 4.31 78.69 :I: 14.92 j;,. .~ 'J Significance NS NS P < 0.05 NS NS P < 0.05 @ 2005 Blackwell Publishing Lld, TrallsfusíOIlMedícille, 15,319-322 ..'1 Plateletpheresis and jibrinolysis These surfaces are thought to be a substitute for i, fi possible physiological receptors on lhe biological surface. The 'contact proteins' have been ascribed to have a role in lhe initiation of açule W inflammatory responses following tissue injury as rf activating ~: we~1 ~s in t?e activation of plasma fibrinolysis f'~ (Nlewlarowsh & Prou-Wartelle, 1959; Colman et aI., 1975; Colman & Schmaier, 1997). Factor ~, XlIa and kallikrein cleave plasminogen directly, but I; Goldsmith less efficiently than1978; t-PAMandle or u-PA& (Colman, 1969; .:' et aI., Kaplan, 1979). ,., Bradykinin selectively induces t-PA release from '.~ endothelial ceIls (Smith et aI., 1985). However, not " only lhe electronegativity of lhe surface but algo other physicochemical parameters could influence lhe activation of lhe contact phase system of plasma (Renaux et aI., 1999). The 'damage' to lhe blood is algo directly related to lhe duration of extracorporeal circulation. Extracorporeal circulatory systems applied in ceIl separators differ depending on lhe principIe of lhe apheresis method (e.g. constant or intermittent flow) and on lhe material of lhe disposable set. The pIateletpheresis performed on blood separator CS 3000 induced . significant activation of coagulation, decrease of Clrantiplasmin concentratiol1 and had no effect on plasmin-plasmin inhibitor complex (Kobayashi et aI., 1993). Unfortunately, in that study, t-PA and PAI-I had not been measured. Another published report revealed no effeet 01'extracorporeal circulation of blood separators: AMICUS and MCS 3p on lhe activation of coagulation, on lhe concentration of plasmin-plasmin inhibitor complex as weIl as on lhe PAI-I concentration (Stohlawetz et aI., 1999). However, these two studies are not comparable. They used different cell separators and there were differences in lhe whole blood : anticoagulant (ACO-A) fatias, blood volume processed anel lhe time of extracorporeal circulation. In lhe study of Stohlawetz et aI. (1999), markers of coagulation were measured in blood taken from lhe inflowing and outflowing blood of cell separator after processing a certain flow volume. Kobayashi et ai. (1993) in their stuciy meas\lred lhe effect of thrombapheresis in blood drawn from coIlection line of danar immediately after lhe start and just before lhe end of apheresis. In our study, we investigated lhe effect of plateletpheresis performed with continuous-flow blood separator COM.TEC on t-PA and PAI-l concentration and activity and plasmin-antiplasmin com pIex in platelet donors. We detected decreased factor XII activity, but did not detect significant changes in PAr, PAI-I and t-PA concentrations as well as PAI-l activity after plateletpheresis. The I (j) 2005 Blackwell Publishing ~ Ltd, Transfllsion Medicine, 15, 319-322 321 mechanism of lhe observed decreased activity of t-PA is difficult to explain. In our opinion, it may be due to lhe inhibitory effect of citrate infusion on t-PA activity/release. Nevertheless, thrombapheresis performed with COM.TEC presents a save procedure for lhe danar in terms of activation of fibrinolysis. ACKNOWLEOGMENT This study was supported by Medical University of Bialystok - Grant no. 3-52839L. REFERENCES Colman, R.W. (1969) Activation ofplasminogen by human plasma kallikrein. 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