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HAEMOSTASIS (VIRCHOW’S TRIAD) (VIRCHOW’S TRIAD) Blood vessel (vasoconstriction) Blood vessel (vasoconstriction) Platelets (adhesion and aggregation) C l ti d Fib i l i Coagulation and Fibrinolysis Prof. Dr. Herman Hariman, Ph.D (U.K), SpPK (KH) Prof. Dr. Adi Koesoema Aman, SpPK (KH)
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Platelets (adhesion and aggregation)ocw.usu.ac.id/course/download/1110000096-hematology-and-immunology... · • Hess test (commonly used): pressure isHess test (commonly used): pressure

Mar 31, 2019

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Page 1: Platelets (adhesion and aggregation)ocw.usu.ac.id/course/download/1110000096-hematology-and-immunology... · • Hess test (commonly used): pressure isHess test (commonly used): pressure

HAEMOSTASIS(VIRCHOW’S TRIAD)(VIRCHOW’S TRIAD)

Blood vessel (vasoconstriction)Blood vessel (vasoconstriction)Platelets (adhesion and aggregation)

C l ti d Fib i l iCoagulation and Fibrinolysis

Prof. Dr. Herman Hariman, Ph.D (U.K), SpPK (KH)Prof. Dr. Adi Koesoema Aman, SpPK (KH)

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HAEMORRHAGICHAEMORRHAGIC SCREENING TEST

A Global test that may predict what happened with thepp

Blood vessel (vasoconstriction)( )Platelets (adhesion and aggregation)

Coagulation and FibrinolysisCoagulation and Fibrinolysis

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BLOOD VESSELBLOOD VESSEL

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BLOOD VESSELSMEASUREMENT

TORNIQUET TESTBLEEDING TIMEBLEEDING TIME

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Tourniquet TestC ill f iliCapillary-fragility-test

• Hess test (commonly used): pressure isHess test (commonly used): pressure is put midway between systole and diastole for 5 minutes Reading is done 5 minutesfor 5 minutes. Reading is done 5 minutes after the release of the cuff

• Rumple Leed (X): the same pressure as• Rumple Leed (X): the same pressure as above but for 15 minutes and reading is done straight away after the release of thedone straight away after the release of the cuff

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Pressure midway between systole and diastoleFor 5 minutes (Hess test)o 5 u es ( ess es )

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InterpretationInterpretation

• (+): small red dots difficult to be seen(+): small red dots difficult to be seen• (++): bigger dots and easily seen

( ) t l il b t l it• (+++): not only easily seen but also quite prominent and sometimes confluent

• (++++): true bruising

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Bleeding TimeBleeding Time

• As a global test to measure roughlyAs a global test to measure roughly• Platelet’s function

Pl t l t’ b• Platelet’s number

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Put blood pressure at 40 mmHg

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First blood drop is wipedN t bl d i ti d ff b filt 30Next, blood is tipped off by filter paper every 30 sec

Normal is less than 5 min

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PLATELETPLATELET

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NUMBER (USUALLY SEVERE)

PLATELET

FUNCTION (LESS SEVERE)

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Bleeding TimeBleeding Time

• As a global test to measure roughlyAs a global test to measure roughly• Platelet’s function

Pl t l t’ b• Platelet’s number

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PLATELET NUMBER IS MEASURED FROM FULL BLOOD COUNT

CELL COUNTER

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PLATELET FUNCTION IS TESTED USING PLATELET AGGREGOMETER

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PLATELETSPLATELETS

PLATELET ADHESION PLATELET AGGREGATIONPLATELET AGGREGATION

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Platelet Aggregation● Platelet adhesion is mediated by vWF. This activates platelets

causing release of TxA2

● During activation, a receptor for fibrinogen becomes exposed onthe platelet membrane.

●Activated platelets release by ADP, Epinephrin, serotonin:

Fibrinogen ADP/ATPvWF Serotonin dense core granulesvWF SerotoninFactor V Ca2+

Factor VIIIPl t l t d i d th f t (PDGF) t h liα

-gra

nule

s dense core granules

Platelet derived growth factor (PDGF) ~ promotes healingPlatelet factor IV – prevents formation of active thrombin

inhibitor from heparin and anti-thrombin III.

α

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AGGREGATION OF PLATELETAGGREGATION OF PLATELET

IS MEASURED AFTER ADDING

THE PLATELET AGONISTS

SUH AS

ATPEPINEPHRINSEROTONINRISTOCETIN

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Graph seen in the platelet aggregometer after adding of some agonists

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Basic Principle ofpvon Willebrand’s Factor Test

• FUNCTIONAL TEST• Ristocetin co-factor assay dg Plt aggregometer

• ANTIGEN TEST (immunoassay)• Radioimmunoassay (RIA)• Enzyme-linked immunoassay (ELISA)

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Basic Principle ofBasic Principle ofFibrinogen test

• Among other coagulation factors, Fibrinogen has no enzymatic activity and hence there is no functional test but only detection of protein

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Intrinsic pathway

XIIa

TF

Extrinsic PathwayXIa

ProthrombinVIIa

TF

IXa

XaVa

VIIIaV

VIII

Fibrinogen FibrinThrombin Soft clot

gXIIIa

FibrinHard clot

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FIBRINOGEN ASSAYSFIBRINOGEN ASSAYS

• Radial immunodifussionRadial immunodifussion• Clauss method (pure clotting-based)• Blomback and Blomback (+thrombin –Ca++)Blomback and Blomback (+thrombin Ca++)

soluble and unstabilized. The tyrosine content is measured after adding urea.g

• Ratnoff and Menzies: Fibrinogen is clotted into Fibrin in glass beads, washed and bolied +NaOH, tyrosine measured with Fiolin Calcateu-Phenol reagent

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COAGULATION AND FIBRINOLYSIS

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I t i i th E t i i thIntrinsic pathway Extrinsic pathway

Final Common pathwayFinal Common pathwayX, V,

Prothrombin Thrombin

Fibrinogen Fibrin

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I t i i th E t i i thIntrinsic pathway Extrinsic pathway

Final Common pathwayFinal Common pathwayX, V,

Prothrombin Thrombin

Fibrinogen Fibrin

Prothrombin Time (PT)

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I t i i th E t i i thIntrinsic pathway Extrinsic pathway

Final Common pathwayFinal Common pathwayX, V,

Prothrombin Thrombin

Fibrinogen Fibrin

Activated Partial Thromboplastin Time (APTT)

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I t i i th E t i i thIntrinsic pathway Extrinsic pathway

Final Common pathwayFinal Common pathwayX, V,

Prothrombin Thrombin

Fibrinogen Fibrin

Thrombin Time (T.T)

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BASIC HAEMORRHAGIC SCREENING TEST

MEASURES ROUGHLYMEASURES ROUGHLY• Full Blood Count• Torniquet test• Bleeding timeBleeding time• Prothrombin time

A ti t d ti l th b l ti ti• Activated partial thromboplastin time• Thrombin time

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To give a rough idea of what happened in disturbedhappened in disturbed

HAEMOSTASIS

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THANK YOU