Cold - induced platelet aggregation in vivo and its inhibition by a nonionic surface active substance

-eAJeSqO -BJ

Jeh)e

1 IQ JO

seJn+eJedwe+ -6e

‘JeheHOq

~103

eq+ seM

‘MOU

pe+eqnDul o+

o+

unouy

ueyM

seti

uehe eq+

eJnsodxe

enp

q3ns

40

Gu!+e$lGe e4e6eJ66e

pue

~U~IJJJ~M

pun04

eJeM

uol+e6eJ6

uol+lppe

Aq

6ullle~s

eJefi

peule+qo sly1

edeqs

‘se6ueqD

6uluu!6eq

AGoloydJow

04

*(6’8’~‘9’~‘~) 40

Alsnolhqo

pettJesqo se

s!q+

o+

?? (ll’ol) -leq+!e

s+ele+eld seDue+sqns

elqeJedwo3

poolq

uol+ewJo)sueJI AJn4ueD

o+

I!$lln

AIsnoeue+uods

+e

eDnpu!

edeys

woJ+

(L’S) ~01

+ele+eid

se6uey3

uneJpq+ln 3slp

•(G’P’E’z’~)

-ea eJe

‘(9)

IeDlJeilds

04.

lecJ!boloqdJow eq.c

e~uls

+ele.+eld

u!

p!d

unouy

IleM

suo!$eJe+lv

uo~+3npoJ+u~

*sule+oJd eueJqwew eq+ $0 se6ueya leuol+ewJo&uo3 o+ enp sl s+ele+eld uo ~103 40 +DeJJe eq+ +ec(+ slseq+odAq eL(+ +Joddns Aew s+ 1nseJ eq+ su !e+oJd eueJqwew +ele+el d ez!l!qe+s o+ pesoddns sl eDue+sqns s/q4 eauls ?? uo!ys .l.ue+ae+Jns -e+ +uepuedep esop e u! (89 j Uj31uoJnld) lelJ+snpu! ue 40 uol+aeru! snoueAeJ+ul uodn Jeeddeslp o+ uees seM a051 40 eJn+eJedwe+ poolq e +e +ueseJd uol+eSeJ66e +ele+eld ?? +unoD e+e6eJ66e u! eseeJ3ep.e Aq peMollo4 SPA a04 JO 0~ o+ u~op 6ul1003 Jeq+JnJ seeJeqM a051 40 eJn+eJedwe$ poolq e 4e peAJesqo sen uo!+e6 -eJ66e 40 wnw lxew v ‘300~ punoJe $0 SeJn+eJedWe+ +e Al -snoeue+uods e+e6eJ66e o+ pun04 eJeM s+ele+eld ?? a05 o+ gc WOJ+ eslnde+s peloo3 se& poolq s]q+ we+sAs 6u!loo3 leeJodJoDeJ+xe ue q6noJy+ edessed Ag *poolq eu!ueD ~u]Mo~+ UJ pelpn+s ueeq seq s+ele+eld uo SeJn+eJedWe+ 4ueJe++ip 40 +=i+e eu+ enb!uve+ 3!J+Deieo+oqd e ~+IM

IP 40

eu6om

A+lsJehlun

-

0005

a

‘6~

‘ASololsAyd

esseJ+S-43o]-+Jeqoa

leDl6oloq+ed

kJ3NNflkl8

33NVlS9nS

CINV

‘tl

3hl13V

SlI CINV Ohlh

NOlllglHNl

NI

pue t13NN38

33VJtlflS

?? 13v,ki~s8v

‘euboloa IewJou

40

e+n+!+sui

‘rl’>l

31NOINON

NOllV33U33V

V A9

1EllZIlVld

Cl33nClNI

-

Cl703

UOUldJad

‘SS%Id

’ aLI1

GL6r

‘zQC-TCC

‘66

‘z’ron

COLD-INDUCED

332 tfons tro

were

made

with

techniques.

vestigate

the

Furthermore,

The

poiyoxyethyiene upon

platelets

platelet present

effect the

PLATELET

of

effect

study

cold of

rich

on

at

low

plasma

temperatures

using

therefore platelets

a noniontzed

- poiyoxypropyiene

Vo1,2,~0.4

AGGREGATION

was

surface

in vl-

conducted

in flowing

condensate was

various

to

whole

active (PiuronicR

In-

blood.

substance F 68'1

examined,

Methods Platelet

aggregation

in flowing

blood

was

measured

wlth

a photo-

8 Ll oscilloscope

Schematic drawing Arrows techntque. the chamber.

1

FIG, 1 of the measuring chamber and the indicate'dtrection of blood flow

BASF Wyandotte Corp., Wyandotte, Michigan Dusseldorf, Germany by C.H. Erbsibh,

48192,

measuring through

supplied

COLD-INDUCED

vo1.2,~0.4

electrical

method

transparency

of

surrounding ring

red

chamber

glass

capillary ringe

isolated a

the

pump

tains changes

cells,

(B)

from

The

the on

transparent of

is based

polnting can

flow

by

photocells,

a

When the

through

flow

Through

(A)

is

capillary

light,

Thereby,

this

by a sy(ID

235

~1

projected blood

monitor

transmitted

the

a sillconized

capillary

tubelet

of

higher

a measu-

constantly

the

the

the

that

photocells

_@ /d\%.I

by

con-

typical they

pro-

transducer

pump

t

a.femoralis

I-

upon

upstream.

of

pressure

I

of

333

to

flow

be drawn

part

narrowest

of

to

centerline

fixed blood

compared

is made

In the

aggregates

Intensity

technique

aggregates

chamber

two

AGGREGATION

Blood

Is

chamber

(PI.

microscope

platelet

(Flg.1).

capillary

The

(12).

PLATELET

v.femoralis

I

1

II

A

I

Windkessel

cooling

system

thermistor

probe

chamber

FIG. 2 Experimental set up of the extraoorporeal circuit with the cooling system and the measuring chamber. Flow rate through the system was malntalned by the roller pump at a rate of 6 ml/mln. The tublngs used were PVC tublngs or made from sillcone rubber; all glass ware used was slllconited.

COLD-INDUCED

334

duce

characteristic

and

2)

and

counted The 26

The

and

heparlnlted

diately

with

after min.

started

stream

of

S'C,

blood

up

Each

(ampl.1

impulses

injected

Into

in Rtngers

0.1,

0.25,

can

was

recorded

0.5

over

the

be

was

to

the

ml

brachlal to

g/kg a 20

50

and

body

mln

decrease values

a rate

of

system,

min

wlthln twice.

Immewith-

was

the

located

up-

of

perfusion

to

30,25,20,15,10,

for

of

to

37'C. series for F 68

PluronicR

concentrations

a

After-

monitored

solutions

Platelet

at

5 min.

mln

was

These

measuBlood

in a second

a solution

weight.

6 ml/min

(Fig.21,

20

aggregation

final

was

In se-

therefore

vein

30

vein,

give

of

of

to

maintained

or

count

(131,

by a probe

rewarmed

15'C

at

stepwise

was

-

heparin.

femoral

cooled

once

Cronkite

blood

20

pentobarbl-

Platelet

cooling

After

level

mg/kg

found

pumped

the

weighing

to control

of

was

was

repeated

solution

and

of

chamber.

Thereupon

pertod,

was

returning

to

blood

down

cooled

count

continuously

system

was

and

a slllconized

temperature

procedure

Brecher

artery

dogs

with 25 R2 Vetren ,

administration

monltored

cooling

was

of

returned

36'C

5 mongrel

mg/kg

procedure

measuring

of

on

platelet

femoral

and

the

the

a 5 mfn were

after

was

temperature

This

the

a'Wlndkessel',

chamber,

wards

method

cooling

the

temperature

20

heparinltatlon

1 hour

through

and

amplification

standardized

anesthetized

with

the

The

from

ring

after

into

performed

were

experiments

Blood

(1,s.)

were

animals

determined

in 60

which

vo1.2,No.4

AGGREGATION

electronically.

kg.

veral

signals

conversion

experiments

tal

PLATELET

were of

made

0.05,

aggregation

period.

Results Fig.

3 shows

tion

during

II give

the

the

the

dardized

cooling

output

impulses

signals

2 VetrenR activity

a representative

from

of from

example

procedure. the the

photocell

Is a heparin-llke per mllllgram

two

The

the original

In this

photocells,

impulse II.

of

#lg. channel

shaper

which

first

signals

preparatlon

with

145

registra-

channel III

I and the

stan-

is triggered clearly

IU of

by

dtscer-

heparin

COLD-INDUCED

vo1.2,~0.4

nlble ded

in when

their the

real

circuit

at

blood

a

amplitude

PLATELET

from

the

temperature

of

the

was

to

2O'C.

lowered

temperature

of

around

erythrocyte

blood A

335

AGGREGATION

noise

perfusing maximum

15'C

whereas

of

the

were

extracorpo-

signals upon

recor-

occured

further

36OC

3ooc

FIG. 3 Representative example of the signal pattern obtained upon At each temperature level stepwlse cooling of the blood. channel I and II gfve output of photocell 1 and 2; channel signals, Bottom: time marks In sec. III: standardized

336

COLD-INDUCED

cooling

down

to

10 and

PLATELET

5'C

the

AGGREGATION

number

of

vo1.2,~0.4

signals

decreased

11 measurements

(Flg.4).

mar-

kedly, This

result

ring

stepwise

ted

at

vious

each from

temperatures around

15'C

lowered

to

could

be confirmed

cooling level

of

this.fig, of and

procedure

platelet

temperature

over

a remarkable

around.

20°C,

decreased

10 and

In

aggregates

a 5 min

aggregation

Aggregation

again

when

blood

coun-

period,

As

ob-

started

at

blood

reached

the

were

Du-

Its

maximum

temperature

5'C,

240

200 f

.: * 160 z a

37-35

31-29

26-24

21-19

16-14

11-9

6-4

temperature,OC

i‘lG. 4 Effect of different temperatures (abscissa) on platelets In flowtng canine blood during stepwise cooling, Ordinate gives aggregate count per 5 mln (averages and sd).

Fig.

5 gives

rements

averages

in which

canine

and

standard

blood

after

deviations cooling

from down

to

11 measu4'C

was

was

vo1.2,No.4

COLD-INDUCED

PLATELET

337

AGGREGATION

temperature,OC I

I

I

1

2

3

I

0

I

I

I

5

6

7

I

4

I

I

I

I

8 min

9

10

11

1f

I

I

12

13

14 70

FIG. 5 Effect of different temperatures on platelets In flow ing canine blood during continuous rewarming of the cooling system (mean values and sd). Aggregate count per 30 set is p lotted against temperature and time.

rewarmed

continuously,

aggregates

counted

perature, gation

As

A

Fig. R nlc

count

not

maximum

following was

obvlous

could

24'C.

per

be of

In

this

30

set

from

fig., 1s

thls

observed

at

aggregation

experiments,

therefore,

plotted

fig.,

too,

against a

therefore,

found a

number

time

below again

at

temperature

of

7

of

and

slgnlficant

temperatures was

the

tem-

aggreand

16'C.

above In

approx.

the 15'C

chosen. 6

shows

F 68 per

the

upon 2

mln

effect

of

cold-induced Is

plotted

different platelet against

concentrations aggregation, time.

Each

curve

of

Pluro-

Aggregate gives

mean

338

COLD-INDUCED

PLATELET

AGGREGATION

Vol.Z,No.4

FIG. 6 Effect of different concentrations of PluronlcR F 68 on coldinduced (15OC) platelet aggregation (averages and sdl In flowing canine blood, Number of platelet aggregates counted per 2 mln Is plotted against time In mtn. Arrows indicate time of InJgcl-ion of PluronicR F 68.

values F 68

and In the

al

vein

up

to

of

the

effect ge

standard

had

0.1

concentration no

g/kg

jectlon

and

0.25

of

the

decrease

Its

aggregation

0.05

g/kg

g/kg). R

F 68

In aggregate lnhlbtttng

5 measurements.

g/kg

InJected

Increase

resulted

F 68

aggregation

In addltlon, in this count effect

already

obvlous

was

the

reduction maxlmum

highest

2 min

concentration and

prolonged

the

brachl-

concentration

its

In the

highest

was

Into

fncreaslng

showed

R

Pluronlc

ln the

In an

PluronlcR

platelet

10.5

Pluronic

from

effect,

counted,

cold-Induced

admtnlstered

of

stgntflcant

aggregates on

deviations

dosa-

after

in-

used

duratlon

slgnlflcantly.

of

COLD-INDUCED

VO~.Z,NO.~

PLATELET

339

AGGREGATION

Discussion When

the

lowered

temperature to

20°C

be

strongest

decreases reduced with takes of

to

the

whole

account

blood

Inducing

that

through

the

cooling

to

plasma,,

authors

emphasize

down

4-6'C

plasma

cooled

warmed

and

In some the

of

that

amount

of

sence

of

which

inhibit

posed

that

formational that The

of

of

for

active

Is a nonionized

this

agent

in fatty

they

rich re-

Alexander

aggregation, cold

on

and

specific

With

not

by the too,

platelets

membrane

require their

a

in that

occur

in the

same

compounds

H owever,

but

The

demonstrate

is not

serotonin

into

significantly

could

did

infused

blood

other,

inhibited

ADP

they

sup-

med iated

results

prote ins

ab-

by

in a con-

simi lar to

ADP. substance

weight

detergent

used

as

of

approx.

Is used

emulsions

(151,and

are

system,

decreased chilled

platelets

was

cooling

in the

present

polyoxyethylene-polyoxypropylene

a molecular

tions

of

cellular

change

surface

genators

and

15'C

to each

and

chilled

effect

at

adhere

ADP-induced

postulated

with

to

fibrinogen

release

rich

In platelet

when

the

in flowing

Kattlove the

the

through

observed

Ca++

of

chamber

platelet

ets

only

flowing

measuring

of

Is

If one

by

(1 ml/m In) was

ACD

passage

platelets

experlments

aggregatlon

the

its

aggregates

indicating

vitro

experiments

before

and

plate

aggregate

(11)

platelets

stirring

that

blood

in agreement

Alexander

system

by

gradually

flowing are

proves

simultaneously.

addltlonal

number

hlgh

to

stlrred

blood

obtained

the

and

is

appears

of cold

15'C

of

blood

aggregation

results

agitation

is comparable These

of

and

canine

effect

around

These

Kattlove

the

that

of

5'C.

of

flowing

platelet

temperature

10 or

observations

into

remarkable

temperatures

when

further

heparlnized

aggregation

at

again

the

the

a first This

spontaneously, to

of

condensate In medical

as 'an emulsifier

(141,

protective

cardiopulmonary bypass (16). R F 68 proved to dlmlnl-sh nlc

8,350.

as

defoaming

agent

sludglng

and

and

applica-

stabilization

substance

against

In experimental

investigation

in oxy-

hemolysis cold

injury

infarction

during Pluro-

resulting

342

COLD-INDUCED

PLATELET

AGGREGATION

vo1.2,~o.b

19.

DOWBEN, R.M. and KOEHLER, W.R. The interaction of a nonpropertles of the Ionic detergent with protein, I. Physical protein detergent cpmplex, Arch, Blochem. Blophys. 93, 496, 1961.

20,

JIRGENSONS, proteins: dispersion

B, Effect of detergents on the conformatfon of I. An abnormal Increase of the optical rotatory constant, Arch. Btochem, Blophys, 94, 59, 1961,