- Contatti Medical

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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
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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.
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