Lecture on Hemostasis by Dr. Roomi

BLOOD PHYSIOLOGYHEMOSTASIS

ByDr. Mudassar Ali Roomi (MBBS, M. Phil.)

Assistant Professor Physiology

Hemostasis-video

Hemostasis • Hemostasis: The term hemostasis means prevention of blood

loss after a vessel is severed or ruptured.• Events in Hemostasis: hemostasis is achieved by several

mechanisms: – (1) vascular constriction (vasoconstriction)– (2) formation of a platelet plug (weak clot)– (3) formation of a blood clot as a result of blood coagulation (strong

clot)– (4) eventual growth of fibrous tissue (organization) into the blood

clot to close the hole in the vessel permanently.– (5) Lysis of blood clot (fibrinolysis)– (6) Healing of vessel wall

PRIMARY AND SECONDARY HEMOSTASIS

• Primary hemostasis includes the platelet and vascular response to vessel injury.

• Secondary hemostasis includes the coagulation factor response to such injury.

Vascular Constriction

• Immediately after a blood vessel has been cut or ruptured, the trauma to the vessel wall itself causes the smooth muscle in the wall to contract; this instantaneously reduces the flow of blood from the ruptured vessel.

• The contraction results from – (1) local myogenic spasm, – (2) local autacoid factors from the traumatized tissues and blood platelets (e.g.

thromboxane A2)– (3) nervous reflexes. The nervous reflexes are initiated by pain nerve impulses

or other sensory impulses that originate from the traumatized vessel or nearby tissues.

– However, even more vasoconstriction• The more severely a vessel is traumatized, the greater the degree of

vascular spasm. • The spasm can last for many minutes or even hours, during which time

the processes of platelet plugging and blood coagulation can take place.

Formation of the Platelet Plug

• If the cut in the blood vessel is very small—indeed, many very small vascular holes do develop throughout the body each day—the cut is often sealed by a platelet plug, rather than by a blood clot

• The time which is required to make the platelet plug is call as bleeding time. Its normal value is 1-6 minutes. MCQ

Physical and Chemical Characteristics of Platelets

• Platelets (also called thrombocytes) are minute discs 1 to 4 micrometers in diameter.

• They are formed in the bone marrow from megakaryocytes, which are extremely large cells of the hematopoietic series in the marrow.

• The normal concentration of platelets in the blood is between 150,000 and 300,000 per mm3.

• Platelets have many functional characteristics of whole cells, even though they do not have nuclei and cannot reproduce.

Platelets

Comparative sizes of blood cells Megakaryocyte is the largest cell in the bone marrow

Physical and Chemical Characteristics of Platelets

• In their cytoplasm are such active factors as:– (1) actin and myosin molecules, which are contractile proteins similar– to those found in muscle cells, and still another contractile protein,

thrombosthenin, that can cause the platelets to contract; – (2) residuals of both the endoplasmic reticulum and the Golgi apparatus that

synthesize various enzymes and especially store large quantities of calcium ions; – (3) mitochondria and enzyme systems that are capable of forming adenosine

triphosphate (ATP) and adenosine diphosphate (ADP); – (4) enzyme systems that synthesize prostaglandins, which are local hormones

that cause many vascular and other local tissue reactions; – (5) an important protein called fibrin-stabilizing factor, which is important for

clot retraction– (6) a growth factor (PDGF) that causes vascular endothelial cells, vascular

smooth muscle cells, and fibroblasts to multiply and grow, thus causing cellular growth that eventually helps repair damaged vascular walls.

Physical and Chemical Characteristics of Platelets

• The cell membrane of the platelets is also important. On its surface is a coat of glycoproteins that repulses adherence to normal endothelium and yet causes adherence to injured areas of the vessel wall, especially to injured endothelial cells and even more so to any exposed collagen from deep within the vessel wall.

• In addition, the platelet membrane contains large amounts of phospholipids that activate multiple stages in the blood-clotting process.

• It has a half life in the blood of 8 to 12 days.• it is eliminated from the circulation mainly by the tissue

macrophage system (reticulo-endothelial system). Most of the platelets are removed by macrophages in the spleen and liver.

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Aggregation of platelets into a

thrombus

Platelets

Endothelial cells

Platelets adhering to subendothelial space

Platelet thrombus

Normal platelets in flowing blood

Platelets adhering to damaged endothelium

and undergoing activation

Platelet Adhesion and Activation

Subendothelial space

Shape change during activation

• Activated platelets change in shape to become more star shaped, and pseudopods form on their surface. Thus they assume a stellate shape

• Fig: Scanning electron micrograph of blood cells. From left to right: human erythrocyte, activated thrombocyte (platelet), leukocyte .

Granule secretion by platelets

• Activated platelets excrete the contents of their granules into surrounding blood.

• There are three types of granules:– dense (or delta) granules: (containing ADP or ATP, calcium,

and serotonin)– lambda granules – similar to lysosomes and contain several

hydrolytic enzymes.– Alpha granules (containing P-selectin, platelet factor

4, transforming growth factor-β, platelet-derived growth factor, fibronectin, B thromboglobulin, vWF, fibrinogen, and coagulation factors V and XIII).

Blood coagulation

• more than 50 substances are involved in coagulation.

1. procoagulants = promote/favor coagulation e.g. all the clotting factots

2. anticoagulants = inhibit/disfavor coagulation e.g heparin, warfain, protein C, protein S, antithrombin III.

General Mechanism.three essential steps: • (1) formation of prothrombin activator:

In response to rupture of the vessel or damage to the blood itself, a complex cascade of chemical reactions occurs in the blood involving more than a dozen blood coagulation factors. The net result is formation of a complex of activated substances collectively called prothrombin activator.

• (2) The prothrombin activator catalyzes conversion of prothrombin into thrombin.

• (3) The thrombin acts as an enzyme to convert fibrinogen into fibrin fibers that enmesh platelets, blood cells, and plasma to form the clot.

Extrinsic pathway of blood clotting

• Prothrombin activator is generally considered to be formed in two ways: – (1) by the extrinsic pathway that begins with

trauma to the vascular wall and surrounding tissues

– (2) by the intrinsic pathway that begins in the blood itself.

Extrinsic pathway of blood clotting

• 1. Release of tissue factor III from Traumatized /injured tissue

• 2. Activation of Factor X—role of Factor VII and tissue factor.

• 3. Effect of activated Factor X (Xa) to form prothrombin activator—role of Factor V.

Note: there is positive feedback effect of thrombin, acting through Factor V, to accelerate the entire process once it begins.

Intrinsic pathway of blood clotting

Intrinsic pathway of blood clotting

• 1. Blood trauma or exposure to collagen or glass tube causes (1) activation of Factor XII

• 2. Activation of Factor XI.• 3. Activation of Factor IX by

activated Factor XI.• 4. Activation of Factor X—role

of Factor VIII.• 5. Action of activated Factor X

to form prothrombin activator—role of Factor V

Difference between

Extrinsic pathway 1. Triggered by injury to tissue

2. Quick or urgent system3. Factors III, VII are present4. Factors VIII, XI, XI, XII are

absent5. extrinsic pathway is checked

by PT test.6. This pathway is affected by

Warfarin anticoagulant

Intrinsic pathway1. Stimulated by Blood trauma or

exposure to collagen or glass tube2. Slow or delayed system3. Factors III, VII are absent4. Factors VIII, XI, XI, XII are

present5. intrinsic pathway is checked by

APTT test.6. This pathway is affected by

heparin anticoagulant

clot retraction

• it expresses serum fluid from blood clot• platelets are necessary for clot retraction as

they release factor XIII (fibrin stabilizing factor) which is required for clot retraction.

• Clot contraction is accelerated by thrombin and Ca++

Causes of Excessive Bleeding

A. vitamin K deficiency: B. hemophilia

C. thrombocytopenia

A. vitamin K deficiency

– Factor II, VII, IX, X are vitamin K-dependant factors. Vitamin K does not synthesize these factors rather it activates them.

– In the absence of vitamin K, subsequent insufficiency of these coagulation factors in the blood can lead to serious bleeding tendencies.

– Vitamin K (a fat soluble vitamin) is continually synthesized in the intestinal tract by bacteria, so that vitamin K deficiency seldom occurs in the normal person as a result of vitamin K absence from the diet (except in neonates before they establish their intestinal bacterial flora).

– vitamin K deficiency often occurs as a result of poor absorption of fats from the gastrointestinal tract.

– Vit. K deficiency increases the clotting time– Warfarin (coumarin) is oral anti-coagulant and it acts by antagonizing the

synthesis of vitamin K dependant factors.

B. hemophilia• it is a bleeding disease that occurs almost exclusively in males (boys). It is X-linked

recessive disease.• In 85 per cent of cases, it is caused by an abnormality or deficiency of Factor VIII; this type

of hemophilia is called hemophilia A or classic hemophilia. About 1 of every 10,000 males in the United States has classic hemophilia.

• In the other 15 per cent of hemophilia patients, the bleeding tendency is caused by deficiency of Factor IX (Chrismas disease).

• Both of these factors are transmitted genetically by way of the female chromosome X. Therefore, almost never will a woman have hemophilia because at least one of her two X chromosomes will have the appropriate genes. If one of her X chromosomes is deficient, she will be a hemophilia carrier, transmitting the disease to half of her male offspring and transmitting the carrier state to half of her female offspring.

• Bleeding can occur after minor trauma• bleeding can often last for days after extraction of a tooth. Hematoma can form in knee

joint.• Therapy that is truly effective is injection of purified Factor VIII. The cost of Factor VIII is

high• Factor VIII is present in intrinsic pathway (checked by APTT test).

Inheritance of hemophilia

C. thrombocytopenia• It means the presence of very low numbers of platelets in the circulating

blood. People with thrombocytopenia have a tendency to bleed, as do hemophiliacs, except that the bleeding is usually from many small venules or capillaries, rather than from larger vessels as in hemophilia.

• As a result, small punctate hemorrhages occur throughout all the body tissues. The skin of such a person displays many small, purplish blotches, giving the disease the name thrombocytopenic

• purpura. • As stated earlier, platelets are especially important for repair of minute

breaks in capillaries and other small vessels.• Ordinarily, bleeding will not occur until the number of platelets in the

blood falls below 50,000/ml, rather than the normal 150,000 to 300,000. Levels as low as 10,000/ml are frequently lethal.

C. Thrombocytopenia (continued)• Most people with thrombocytopenia have the disease known

as idiopathic thrombocytopenia, which means thrombocytopenia of unknown cause. In most

• of these people, specific antibodies have formed and react against the platelets themselves to destroy them.

• Treatment: – Relief from bleeding for 1 to 4 days can often be effected in a

patient with thrombocytopenia by giving fresh whole blood transfusions that contain large numbers of platelets.

– splenectomy is often helpful, sometimes effecting almost complete cure because the spleen normally removes large numbers of platelets from the blood.

VII. Thromboembolic Conditions

A. thrombus = abnormal clot that develops in the blood vessel

B. embolus = free flowing clotC. causes of thromboembolic conditions (Virchow’s

triad) 1. roughened/injured endothelial surface of a vessel 2. slow flowing blood (stasis of flow) 3. hypercoagulability

Femoral Venous Thrombosis/deep vein thrombosis (DVT) and

Massive Pulmonary Embolism• the immobility of patients confined to bed plus the practice of propping

the knees with pillows often causes intravascular clotting because of blood stasis in one or more of the leg veins for hours at a time. Then the clot grows, mainly in the direction of the slowly moving venous blood, sometimes growing the entire length of the leg veins and occasionally even up into the common iliac vein and inferior vena cava.

• Then, about 1 time out of every 10, a large part of the clot disengages from its attachments to the vessel wall and flows freely with the venous blood through the right side of the heart and into the pulmonary arteries to cause massive blockage of the pulmonary arteries, called massive pulmonary embolism.

• If the clot is large enough to occlude both of the pulmonary arteries at the same time, immediate death ensues. If only one pulmonary artery is blocked, death may not occur, or the embolism may lead to death a few hours to several days later because of further growth of the clot within the pulmonary vessels. But, again, t-PA therapy can be a lifesaver.

deep vein thrombosis (DVT) andMassive Pulmonary Embolism

Disseminated Intravascular Coagulation (DIC)

• In this case the clotting mechanism becomes activated in widespread areas of the circulation

• This often results from the presence of large amounts of traumatized or dying tissue in the body that releases great quantities of tissue factor into the blood.

• This occurs especially in patients with widespread septicemia (Infections) which releases endotoxins that activate the clotting mechanisms.

• the clots are small but numerous, and they plug a large share of the small peripheral blood vessels.

• Plugging of small peripheral vessels greatly diminishes delivery of oxygen and other nutrients to the tissues—a situation that leads to or exacerbates circulatory shock. It is partly for this reason that septicemic shock is lethal in 85 per cent or more of patients.

• Bleeding can eventually result due to consumption of platelets and clotting factors.

Anticoagulants

• A. heparin (Injectible)– It causes the blood-clotting time to increase from a normal of about 6 minutes

to 30 or more minutes. Furthermore, this change in clotting time occurs instantaneously, thereby immediately preventing or slowing further development of a thromboembolic condition. The action of heparin lasts about 1.5 to 4 hours.

– It inactivates factor VIII, IX, X, XI, XII.– It decreases the activity of the factors mainly present in intrinsic pathway

(prolo)• B. coumarins (warfarin)-oral anticoagulant

1. it has liver depressant effect 2. it lowers the levels of prothrombin (II), Factors VII, IX, and X 3. it competes with vit. K

4. Normal coagulation usually returns 1 to 3 days after discontinuing coumarin therapy.

Blood Coagulation Tests

A. bleeding time B. clotting timeC. prothrombin time (PT), D. Activated partial thromboplastin time (APTT

or aPTT).Significance: these tests are usually performed

before surgery. If prolonged, there is increased risk of bleeding.

Bleeding time

• When a sharp-pointed lancet is used to pierce the tip of the finger or lobe of the ear, bleeding ordinarily lasts for 1 to 6 minutes (bleeding time).

• It is determined by Duke’s method• Bleeding time is prolonged by:

1. lack of platelets (thrombocytopenia)2. Von Willebrand’s disease3. Use of aspirin4. Vitamin C deficiency5. Dengue fever

Clotting time• Clotting time: It is the time taken by blood to clot in clan

glass tube.• Blood is collected in clean glass capillary tube and after an

initial wait of 3 minutes its end is broken every 30 seconds until the blood has clotted. By this method, the normal clotting time is 6 to 10 minutes.

• Clotting time is prolonged in:1. Hemophilia2. Liver diseases3. Use of anti-coagulants e.g. (warfarin, heparin)4. Von Willebrand’s disease

Prothrombin time (PT)

• METHOD TO DETERMINE PROTHROMBIN TIME: 1. Blood is immediately oxalated so that none of the

prothrombin can change into thrombin. 2. Then, a large excess of calcium ion and tissue factor is

quickly mixed with the oxalated blood. The excess calcium nullifies the effect of the oxalate, and the tissue factor activates the prothrombin-tothrombin reaction by means of the extrinsic clotting pathway. The time required for coagulation to take place is known as the prothrombin time.

3. PT is prolonged if someone is using warfarin. • The normal prothrombin time is about 12 seconds