Thrombocytes

Leture 7+8

Course code: Zool-01704

Course Name: Haematology
Department: Zoology
Presented By: Mr. Nabeel Tahir
Platelets (Thrombocytes)
• Platelets are colorless, non-nucleated bodies.
• They are not whole cells, but are fragments of extraordinarily large
bone marrow-bound megakaryocytes.
• Like RBCs and WBCs (except lymphocytes), platelets come from
myeloid stem cells in bone marrow
• Differentiate into giant cells called Megakaryocytes, which break off
membrane bound packets of cytoplasm to form platelets
• The hormone THROMBOPOIETIN (produced by liver) increases the
number of megakaryocytes in the bone marrow, stimulating them to
produce more platelets.
Thrombocyte Development
M e g a k a ry o b la st
F Irs t Id e n tifia b le T h ro m b o c y te P re c u rs o r

Pro m e g a k a ry o c y te

M e g a k a ry o c y te
v e ry la rg e c e ll, la rg e s t in b o n e m a rro w

T h ro m b o c y te
p la te le t s b re a k o f f o f c y to p la s m o f m e g a k a ry o c y te
In Bone Marrow
Platelet Kinetics
• Normal circulating platelet count
– 150K to 400K (150,000/ul to 400,000/ul )
– 90 to 300 K in people of Mediterranean descent

• 1/3 of platelets are sequestered in the spleen

• lifespan of a platelet is 7 to 10 days
• 15 to 45 K platelets are produced daily to maintain steady
state
Platelet structure
1- Surface coat: Mucopolysaccharide (glycosaminoglycans)

2- Plasma membrane: is formed of 3 layers and invaginates inside

to form open membrane system and contain phospholipids. This
membrane is the basis structure of platelet factor 3 (PF3)

3- Beneath the membrane there are:

a- Microfilaments b- Microtubules
Platelet structure
4- Granules : 3 types of granules
- Dense granules contain (ADP) serotonin and calcium.
- Specific (Alpha) granules contain platelet growth factor, beta
thromboglobulin, fibrinogen, clotting factors as V, XIII, platelet factor 4
(PF4) which is heparin antagonist factor.
- Dense tubular system contains calcium and may be site of synthesis of
prostaglandin and thromboxane A2

5-mitochondria, glycogen & lysosomes which contain hydrolytic

enzymes
Platelet structure
Von Willebrand factor (vWF)
• An adhesive protein formed by the vascular endothelial cells and
secreted in the plasma.

• It is formed by megakaryocytes and stored in the platelets.

• Blood clotting factor VIII circulated in plasma bound to vWF

Hemostasis
It is the stoppage of bleeding in case of small vascular
injury.
It depends on:
– vascular phase
– platelet phase
– coagulation phase
The Vascular Phase
• A cut triggers vascular spasm: smooth muscles in the vessel
contract to limit blood loss
• Immediate, 30-minute contraction
• Mechanism

1. Nervous reflexes initiated by pain from traumatized blood

vessel.

2. Local myogenic contraction of smooth muscles due to direct

trauma, the more the traumatized the vessel the greater the
degree of spasm

3. Humoral: through chemical substances released from the site

of injury (serotonin,TxA2, adrenaline,….)
The Platelet Phase
• Upon damage to blood vessel endothelium (which exposes subendothelial
collagen)
1) Platelet adhesion: With the help of von Willebrand factor (VWF) adhere to collagen through platelets
membrane glycoprotein (Ib) and initial release reactions (ADP, TxA2, serotonin)
2) Platelet activation: stimulated by ADP and thromboxane A2. Activated platelets swell, develop
pseudopodia, sticky and discharge their granules (ADP, serotonin, TxA2, catecholamines, vWF, fibrinogen,
…..)
3) Platelet aggregation: Release serotonin and ADP, attract more and more platelets forming a mass of
aggregated platelets (fibrinogen & Gp IIb/IIIa) (platelet cohesion).
4) Formation of a platelet plug large enough to close a small rent in the BV wall. Platelet plug initially is
loose then become firm by contraction of the platelet contractile proteins and by fibrin threads
Platelet Plug Size is Restriction to Injury Site
• Prostacyclin:
• released by intact endothelial cells, inhibits platelet aggregation to the site of
injury only
• Inhibitory compounds:
• released by other white blood cells
• Circulating plasma enzymes:
• break down ADP
• Negative (inhibitory) feedback:
• at high concentration, serotonin blocks ADP action
• Development of blood clot:
• isolates area by sealing it off
Platelet Plug Formation
1) Aspirin inhibit TxA2 synthesis, so inhibit platelet activation and
aggregation
2) Activated platelet expose the platelet membrane phospholipids (PF3)
which is essential for blood coagulation.
3) Normally there is thousands of minute rupture in small BV
(capillaries), that closed by platelet plug formation (purpura)
Platelet Plug Formation

• Platelets adhesion is potentiated by vWF which is

plasma protein
The Coagulation Phase
• The third mechanism for hemostasis. (begin 20 sec - 2 m)
(within 3- 6 m blood clot is formed)
• A series of reactions in which blood is transformed from a
liquid to a gel
• The main three steps of this series of reactions are:
1) Formation of Prothrombin activator complex: through intrinsic and
extrinsic pathways
2) Conversion of Prothrombin into thrombin
3) Thrombin catalyzes the conversion of fibrinogen into a fibrin
threads
Clotting Factors
• Proteins or ions in plasma required for normal clotting
• The clotting factors are formed in
The liver: Some clotting factors require vit. K in their
synthesis from the liver (factor II,VII,XI and X)
Megakaryocytes: Formation of fibrinogen, Factor V, XIII and
vWF.
Macrophages: Synthesis of factors V, VII, IX and X.
Endothelial cells: Form vWF
Prothrombin activator complex
Formation of prothrombin activator occur by 2 pathways:
I) Extrinsic pathway: initiated by substance outside the blood (tissue
thromboplastin factor III) released due to trauma (tissue damage).
2) Intrinsic pathway: initiated by substances all present in the
plasma (factor XII) & Collagen of injured blood vessel (inside the
body) .
There is cooperation between the 2 pathways in the process
of blood coagulation.
 Coagulation Phase 1:
Two Pathways to Prothrombin Activator
• May be initiated by either the intrinsic or extrinsic pathway
• Triggered by tissue-damaging events
• Each pathway cascades toward factor X
• Once factor X has been activated, it complexes with Ca2+ ions,
PF3, and factor V to form prothrombin activator
A) Extrinsic pathway

Tissue thromboplastin (factor III) released from damaged tissue

and vascular wall (consist of lipoprotein & phospholipids)
1) Factor III activate factor VII to VIIa & form a complex with it.
2) Factor III/VIIa complex in the presence of Ca2+ activate factor X into Xa
3) Xa with factor V, PF3 (platelet phospholipid) and Ca2+ form the
prothrombin activator enzyme complex

4) Prothrombin activator act on prothrombin converting it thrombin

which then act on fibrinogen converting it to fibrin monomer
I (Fibrinogen) II (prothrombin)

III (thromboplastin)
IV (Calcium)
tissue factor

V (Labile factor) VII (stable factor)

VIII (anti-haemophilia globulin)

IX (christmas factor)
X (Stuart prower factor)

XI (plasma thromboplastin antecadent)

XII (Hageman factor)

XIII (fibrin stabilizing factor)

Platelet factor-3 (phospholipid)

High molecular weight kininogen and
vWF
prekallikrein.
B) Intrinsic pathway
Exposure of the blood to damaged endothelium & rough subendothelial collagen
or contact with water wettable surface (glass) initiate the activation of the
intrinsic pathway (slow & terminates in 3 – 6 m) occur in the following steps:
1) Contact activation of factor XII into XIIa
2) XIIa activate factor XI into active XIa
3) XIa activate factor IX into active IXa
4) IXa in the presence of factor VIII and Ca2+ and PF3 activate factor X into Xa
5) Xa with factor V, PF3 (platelet phospholipid) and Ca2+ form the prothrombin activator
enzyme complex

Prothrombin activator act on prothrombin converting it to thrombin which then

act on fibrinogen converting it to fibrin monomer
I (Fibrinogen) II (prothrombin)

III (thromboplastin)
IV (Calcium)
tissue factor

V (Labile factor) VII (stable factor)

VIII (anti-haemophilia globulin)

IX (christmas factor)
X (Stuart prower factor)

XI (plasma thromboplastin antecadent)

XII (Hageman factor)

XIII (fibrin stabilizing factor)

Platelet factor-3 (phospholipid)

vWF High molecular weight kininogen and prekallikrein.

Blood Clot formation
Coagulation Phase 2 & 3:
Common Pathways to the Fibrin Mesh
• Thrombin catalyzes the conversion of fibrinogen into fibrin monomer
that polymerized into fibrin threads. Insoluble fibrin threads
entrapping blood cells, platelets and plasma forming loose blood clot
• Fibrin causes plasma to become a gel-like trap
• Fibrin in the presence of calcium ions & active factor XIII (factor XIIIa)
that:
• Cross-links fibrin
• Strengthens and stabilizes the clot
Role of Ca+2 in blood clotting

Ca+2 required for acceleration of most of blood clotting reactions

1. Absence of Ca2+ prevent blood clotting by the 2 pathways.
2. Citrate and oxalate salts (Ca2+ precipitating agents) can be used as
in vitro anticoagulants.
Clotting Area is Restricted
1. Anticoagulants (plasma proteins):
• antithrombin-III
• Fibrin itself binds thrombin and prevents it from exerting
positive feedback
2. Heparin from endothelium
3. Prostacyclin from endothelium
4. Protein C (activated by thrombomodulin) activates
plasmin
Clot Retraction and Repair
• Clot retraction –
• After clot has formed, platelets contract and pull torn area together,
squeezing out serum
• Stabilizes injury site, facilitates repair
• Takes 30–60 minutes
Repair of BV wall
• Platelet-derived growth factor (PDGF) stimulates rebuilding of blood vessel
wall,
• Fibroblasts form a connective tissue patch
• Stimulated by vascular endothelial growth factor (VEGF), endothelial cells
multiply and restore the endothelial lining
Fibrinolysis
• Slow process of dissolving clot
• thrombin and tissue plasminogen activator (t-PA): activate
plasminogen
• Note that this is the same thrombin that helped activate the fibrin in the first
place
• Plasminogen produces plasmin, which digests fibrin strands