BASIC PRINCIPLES OF HEMOSTASIS PLATELET ACTIVATION AND PLUG

FORMATION INVOLVES
Thrombopoeitin  Adhesion
 Hormone responsible for the production  Shape change
of platelets  Secretion
 Aggregation
Thrombopoiesis 
 Direct production of platelets from the PHYSIOLOGY
cytoplasm of megakaryocyte via
endomitosis Shape : Biconvex disk like diameter about 2-
4 um average cubage 8 um3
Endomitosis
 There is a nuclear division without Complicated Structure: under the electronic
cytoplasm microscope, there are a-granule dense body
, lysin peroxide enzyme , opening tubular
Platelets system , dense tubular system canalicular,
 Primary defense against bleeding etc.
 Circulate in resting state
 Have minimal interaction with other Dense Body: Contains ADP, ATP ,
blood components or the vessel wall 5-HT,Ca2+, epinephrine , etc.
 Morphology of resting platelet is smooth
, discoid COMPONENTS
 Actin and Myosin molecules
 When stimulated by endothelial damage  Residuals of both the Endoplasmic
, platelets become activated and then : Reticulum and Golgi apparatus
- Become round and sticky  Mitochondria and enzymes that are
- Build a hemostatic plug capable of forming ATP
- Provide reaction surface proteins  Fibrin-stabilizing factor
that make fibrin  A coat of glycoproteins that repulses
- Aid in wound healing adherence to normal endothelium
 minute discs , 1 to 4 um in diameter  Half Life of 8 to 12 days
 Formed in the bone marrow from
megakaryocyte N:C
 NORMAL CONCENTRATION : 150,000- Stages of Platelets Size Ratio
400,000 per uL 20-50 10:1
 1 megakaryocyte = develop 2,000 -4,000 Megakaryoblast um
platelets 20-60 4:1 -7:1
 1/3 in spleen ; 2/3 in circulation Promegakaryocyte um
Granular 30-90 2:11 –
 Life Span : 7-10 days
Megakaryocyte um 1:1
Vitamin K 40-120 Less
Mature um than 1:1
 Vitamin important for coagulation
Platelets / 1-4 um
Thrombocyte (2.5)
 PHYSIOLOGICAL CHARACTERISTICS OF  Provide surface membrane proteins
PLATELETS to attach to other platelets, bind
collagen , and sub-endothelium.
a. Thrombocyte adhesion
 Its membrane glycoprotein (GP,IX PLATELET STRUCTURE
and GPIIa), collagen
(underendothelial structure), vWF Chromosome
(plasma component),fibrinogen are  granular and centrally located
involved in adhesion Hyalomere
b. Thrombocyte aggregation  surrounds the chromosome , non-
 Induced by physiological factors such granular and clear to light blue
as ADP, thromboxane A2 (TXA2),  Platelets are composed of 60% CHON,
epinephrine, 5-HT, histamine, 8% CHO and various minerals , water and
collagen,thrombin,prostacyclin, etc. nucleotide
and by pathological factors like
bacteria, virus, immune complex, SYSTEM COMPONENTS
drugs, etc.
c. Thrombocyte release 1. Peripheral Zone
 ADP, ATP, 5-HT, Ca2+ released from 2. Sol-Gel Zone
dense body , and β-platelet 3. Organelle Zone
globin,PF4,vWF, fibrinogen,PFV, 4. Membranous Zone
PDGF,thrombin sensitive protein
from α-granule, and acid protein A. Peripheral Zone
hydrolyzed enzyme , tissue  Responsible for platelet adhesion and
hydrolyzed enzyme from lysosome aggregation
d. Thrombocyte contraction
 Loose platelet thrombus could turn Glycocalyx
into compact platelet thrombus by -amorphous extension coat
Ca2+ release and cytoskeleton GpIb – receptor for vWB factor
movement (filament/canaliculus) GpIIb/IIIa – receptor for fibrinogen
within platelet. GpVa- receptor for thrombin

PLATELET FUNCTION Thrombus

 Presence of clot in the vessels
 Provide negatively charged surface
for factor X and prothrombin Embolus
activation  Presence of clot in circulation
 Release substances that mediate
vasoconstriction, platelet B. Sol-Gel Zone
aggregation, coagulation and  For contraction and support
vascular repair
 a) Microfilaments
- actin and myosin (actomyosin) Hemorrhage
[clot retraction]  Opposite of hemostasis, first stage of
b) Microtubules wound healing
- tubulin
- responsible for maintaining the ROLES OF PLATELET IN HEMOSTASIS
shape of platelets
1. Activated platelets supply lecithoid
C. Organelle Zone (phospholipid) surface for blood
A. Alpha Granules clotting factor and involve in
- Amplifies reaction to injury, activating factor X and prothrombin
normally 20-200 platelet 2. Surface of platelet membrane
- Platelet factor IV combine with many blood clotting
- PDGF factor, such as fibrinogen, FV,
- Fibrinogen FXI,FXIII to speed up coagulation
- Factor V 3. Activated platelets release α-granule
- Von Willebrand Factor which contains fibrinogen to
- B-thrombopoeitin intensity fibrin forming and blood
- Fibronectin coagulation
- Platelet adhesion 4. Activated platelets contract clot with
B. Dense Granules (C2PA2S) its contractive protein to solidify
- Calcium blood coagulation.
- Catecholamines
- Pyrophosphate FORMATION OF PRIMARY
- ADP ( Stimulates platelet HEMOSTATIC PLUG
aggregation)
- 5- hydroxytrypthamine
- Serotonin (for vasoconstriction)

D.Membranous System
a. Dense Tubular System
- Site of arachidonic acid metabolize
- Control center for platelet
activation
- Site of prostaglandins synthesis and
calcium sequestration
- Derived from smooth ER of
megakaryocyte
b. Open Canalicular System
- For the release of granules

Hemostasis
 Cessation of bleeding/ stoppage of
bleeding

Adhesion
 Damage to endothelium exposed blood
to the sub-endothelial tissue matrix with
adhesive molecules.
 Platelt receptor GPIb binds to sub-
endothelium collagen fibers through Von
Willebrand’s Factor (vWF)
 Platelet adherence stops the initial
bleeding
 Dense granules contain large
amounts of ADP
 ADP binds to the platelet membrane
triggering the synthesis and release
of DCA2
 The release of large amounts of ADP
combined with TXA, amplifies the
initial aggregation of platelets into a
large platelet mass.

Aggregation Platelets
 Platelet-to-platelet interaction
Fibrinolysis Fibrinolysis
 Begins 10-20 seconds after vascular
injury and platelet adhesion
 Requires dense granule release from
the adhering platelets
 Requires Ca2+ and ATP BODY
 Requires fibrinogen and fibrinogen
receptors GPIIb and IIIa
MECHANISM:
- ADP released from platelet
cytoplasm upon adherence induces Thrombosis Bleeding
exposure of fibrinogen receptors
GPIIb and IIIa  Excessive  Excessive
- Fibrinogen binds to the exposed coagulation Fibrinolysis
GPIIb and IIIa  Inadequate  Inadequate
- Extracellular Ca2+ dependent fibrinolysis coagulation
fibrinogen bridges from between
adjacent platelets, thereby
promoting platelet aggregation STEPS OF HEMOSTASIS
- This is primary or reversible
aggregation 1. Vascular Spasm (Vasoconstriction)
- Secondary aggregation begins with 2. Platelet Plug formation
the release of dense granules 3. Clot formation (Blood Clotting)
- Secondary aggregation is considered 4. Clot Retraction
irreversible

COMPONENTS OF HEMOSTASIS
Secretion
 Secondary aggregation begins with A. Extravascular
platelet secretion of dense granules - Tissue surrounding the vessel
 - Provides back pressure that
promotes swelling and trapping of B. Zymogen
plasma -Enzyme precursor

FACTORS:
1. The bulk of amount of
surrounding tissue
2. The type of tissue
-skeletal tissue is more
dense than muscular
tissue
3. The tone of the tissue
B. Vascular
- Blood vessel
C. Intravascular
- Coagulation proteins (biochemical,
procoagulant)
C. Cofactor
-Components that acts in the
stabilization and enhancement of
zymogen activation to the active
enzyme form
D. Calcium
E. Serine Proteases
-Active Form
Mitochondria – for energy production
Components of Coagulation Factors

COAGULATION FACTORS 1. Fibrinogen Group (I,V,VIII,XIII)

I Fibrinogen - Calcium dependent
II Prothrombin - Vitamin K dependent
III Tissue Factor - Completely consumed during
IV Calcium Ions coagulation
2. Prothrombrin Group (II , VII, IX, X)
V Proaccelerin
[1972]
- Calcium and Vitamin K dependent
VII Stable Factor - Adsorbed by BaSO4
VIII Anti-hemopholic - Not consumed during coagulation
factor 3. Contact Group ( XII , XI, Prekallikrein
IX Christmas Factor ,MUK)
X Stuart Factor - High molecular weight kininogen
XI Plasma - Kinin system
thromboplastin - Calcium and Vitamin K dependent
antecedent - Not consumed during coagulation
XII Hagemann Factor - Involved in the contact
XIII Fibrin Stabilizing - Not adsorbed by BaSO4
Factor

Coagulation Factors are classified as: Physiological Hemostasis

A. Substrate Definition: The process from vessel bleeding

-Substance on which the enzyme act to automatic hemostasia

Bleeding time : 1- 3 minutes, longer when
platelet decreases
- Time from vessel bleeding to
automatic hemostasia
Primary hemostasis
- Involved with platelets and blood
vessels
Secondary Hemostasis
- involved with coagulation factors
Process of Hemostasis
1. Blood vessel contraction or
convulsion (induced by neuroreflex:
5 –hydroxytryptamine (5-HT);
thromboxane A2 (TXA2); endothelin
(ET).
2. Platelet thrombosis forming (made
by platelet adhesion , aggregation,
release and contraction).
3. Fibrin, clot forming and maintenance
( made by blood coagulation
activation)
HEMOSTATIC SYSTEM
Platelet-Cyclooxygenase Pathway
Arachidonic acid
cycloxygenase
Prostaglandin endoperoxide
Thromboxane synthetase
Thromboxane A2
VASOCONSTRICTION DERIVED
FROM PLATELELETS AND
ENDOTHELIUM
BLOOD COAGULATION
BLOOD CLOTTING FACTOR
 Process of blood flow from flowing
liquid to gel or gelatin
 SERUM: Light yellow fluid after blood
coagulation
 TAKE NOTE: There is no fibrinogen in
SERUM
Blood Coagulation
 A series of complicated
biochemical reactions with
various enzymes
Blood Clotting Factor
 Material which are directly
involved in blood coagulation .
There are 12 factors named
Roman numerals except Ca2+,
phospholipid, other factors being
protein , and except FIII (TF) ,
others are in fresh plasma
synthesized by liver with Vitamin K
TWO TYPES OF BLOOD CLOTTING ENZYMES
 Inactive Type
 Activated Type
(FII,FVII,FIX,FX,FXI,FXII,FXIII)