BLOOD VASCULATURE,

MEGAKARYOCYTES AND
PLATELETS
LESSON 1:
a)Blood Vessels, Structure &
Physiology
1) Arteries and Veins
2)Arterioles and Venules
3)Capillaries

b) Endothelium
OVERVIEW OF HEMOSTASIS AND THROMBOSIS
The maintenance of circulatory hemostasis is achieved
through the process of balancing bleeding (hemorrhage) and
clotting (thrombosis).

4 Major Components of Hemostasis:

1. Vascular System
2. Platelets (Thrombocytes)
3. Blood Coagulation Factors
4. Fibrinolysis and ultimate tissue repair
Processes involved in hemostasis following
injury to a small blood vessel:
1. Blood vessel spasm
2. Formation of a platelet plug
3. Contact among damaged blood vessel, blood platelet, and
coagulation proteins
4. Development of a blood clot around the injury
5. Fibrinolytic removal of excess hemostatic material to
reestablish vascular integrity
Blood Vasculature: Structure and Function
• Types of Blood Vessels:
• Arteries and Veins
• Arterioles and Venules
• Capillaries
Blood Vasculature: Structure and Function
Tissue Zones:
1. Tunica adventitia / Tunica
Externa
2. Tunica media
3. Tunica intima / Tunica interna
VASCULATURE PHYSIOLOGY:
• The Role of Vasoconstriction in Hemostasis
• Vasoconstriction is a reflex in which blood vessels narrow to
increase blood pressure.
• Caused by thromboxane A2 from activated platelets and injured
epithelial cells, nervous system reflexes from pain, and direct
injury to vascular smooth muscle.
• Epinephrine and serotonin promotes vasoconstriction
VASCULATURE PHYSIOLOGY:
• The Role of Endothelium
• Regulates the permeability of the inner vessel wall and provides the
principal stimulus to thrombosis following injury to a blood vessel.
• Involved in the clotting process by producing or storing clotting
components.
• Rich with plasminogen activator, which, if appropriately stimulated, is
released and activates plasminogen, which ensures rapid lysis of fibrin
clots.
• Elaborates prostacyclin,which is synthesized by the endothelium from
prostaglandin precursors and strongly inhibits platelet aggregation and
adhhesion.
THE ENDOTHELIUM
• The Endothelins - are produced in a variety of tissues, where they act
as modulators of vasomotor tone, cell proliferation, and hormone
production.
• 3 Members of the Endothelin Family:
1. Endothelin-1 - is the only family member produced in endothelial cells and
is also produced in vascular smooth muscle cells.
2. Endothelin-2 - is produced predominantly within the kidney and intestine,
with smaller amounts produced in the myocardium, placenta, and uterus.
3. Endothelin-3 - has been found in high concentrations in the brain and may
regulate important functions, such as proliferation and development in
neurons and astrocytes. It also is found throughout the gastrointestinal tract
and in the lung and kidney.
THE ENDOTHELIAL DYSFUNCTION
• The Endothelial Dysfunction - play an important role in the initiation,
progression, and clinical complications of various forms of
inflammatory and degenerative vascular diseases.
• Stimuli of Endothelial Dysfunction:
1. Immunoregulatory Substances (Tumor Necrosis Factor/TNF & Interleukin-
1/IL1)
2. Viral Infection and Transformation
3. Bacterial Toxins
4. Cholesterol / Oxidatively modified lipoproteins
THE ENDOTHELIAL DYSFUNCTION
• Disruption of the endothelium directly activates all four
components of hemostasis.
• After this event, the following events take place:
1. Initially, rapid vasoconstriction for up to 30 minutes reduces blood flow and
promotes contact activation of platelets and coagulation factors.
2. In the second phase, platelets adhere immediately to the exposed
subendothelial connective tissue, particularly collagen.
The aggregated platelets enhance sustained vasoconstriction by releasing
thromboxane A2 and vasoactive amines, including serotonin and epinephrine.
THE ENDOTHELIAL DYSFUNCTION

3. In the third phase, coagulation is initiated through both the intrinsic and
extrinsic systems.
4. Finally, fibrinolysis occurs following the release of tissue plasminogen
activators (t-PAs) from the vascular wall. Fibrinolytic removal of excess
hemostatic material is necessary to reestablish vascular integrity.
MAINTENANCE OF VASCULAR INTEGRITY
• Essentail Factors for Vascular Integrity
1. Circulating functional platelets
2. Adrenocorticosteroids
3. Ascorbic Acid

A lack of these factors produces fragility of the vessels, which

makes them prone to disruption.
MAINTENANCE OF VASCULAR INTEGRITY
The integrity of arterioles and venules depends on
vasoconstriction, the formation of a plug of fused platelets
over the injury, and the formation of a fibrin clot.
Vasoconstriction is of ultimate importance in damaged
arteries. Veins, which contain 70% of the blood volume, may
rupture with a slight increase in hydrostatic pressure.
LESSON 2:
a) Megakaryopoiesis
Hematopoietic Growth
Factors and Interleukins affecting
Megakaryocytes and Platelets
THE MEGAKARYOCYTIC CELL SERIES
• Megakaryocytopoiesis proceeds initially through a phase
characterized by mitotic division of a progenitor cell,
followed by a wave of nuclear endoreduplication.
• Endoreduplication is the process in which chromosomal
material (DNA) and the other events of mitosis occur without
subsequent division of the cytoplasmic membrane into
identical daughter cells.
THE MEGAKARYOCYTIC CELL SERIES
• Thrombopoietin - the hormone thought to stimulate the
production and maturation of megakaryocytes, which in
turn produce platelets, has recently been purified and
cloned.
• Thrombopoietin activity results from several different
cytokines: erythropoietin, IL-3, and granulocyte-
macrophage colony-stimulating factor (GM-CSF). These
substances have been shown to be able to increase
megakaryocyte size, maturational stage, and ploidy.
MEGAKARYOBLAST
SIZE: 10-24 mm
NUCLEUS: Round
Nucleoli: 2-6
Chromatin: Homogeneous, loosely
organized
CYTOPLASM: Basophilic
Granules: Absent by Wright stain
N/C RATIO: 3:1
REFERENCE INTERVAL:
Bone Marrow: 20% of megakaryocyte
precursors in bone marrow
Peripheral Blood: 0%
PROMEGAKARYOCYTE
SIZE: 15-40 mm
NUCLEUS: Indented
Nucleoli: Variable
Chromatin: Condensed
CYTOPLASM: Basophilic
Granules: Present
N/C RATIO: 1:2
REFERENCE INTERVAL:
Bone Marrow: 25% of megakaryocyte
precursors in bone marrow
Peripheral Blood: 0%
MEGAKARYOCYTE
• Megakaryocytes are the largest
bone marrow cells, ranging up to
160 mm in size. The nuclear-
cytoplasmic (N:C) ratio can be as
high as 1:12.
• Nucleoli are no longer visible.
• Distinctive feature of the
megakaryocyte is that it is
multilobular, not multinucleated.
• The fully mature lobes of the
megakaryocyte shed platelets from
the cytoplasm on completion of
maturation.
PLATELETS
• Platelets have an average
diameter of 2 to 4 mm, with
younger platelets being larger
than older ones.
• In contrast to megakaryocytes,
platelets have no nucleus.
• The cytoplasm is light blue, with
evenly dispersed, fine red-purple
granules.
PLATELETS (continuation...)
• An inactive or unstimulated platelet circulates as a thin, smooth-
surfaced disc.
• Platelets circulate at the center of the flowing bloodstream through
endothelium-lined blood vessels without interacting with other
platelets or with the vessel wall.
• Stronger stimulation causes platelets to become sticky without losing
their discoid shape; however, changes in shape to an irregular
sphere with spiny pseudopods will occur with additional stimulation.
• This alteration in cellular shape is triggered by an increase in the level of
cytoplasmic calcium.
LESSON 3:
a) Cellular ultrastucture of a
mature Platelet
b) Platelet kinetics, function, life
span, normal values
CELLULAR ULTRASTRUCTURE OF A MATURE
PLATELET
• THE GLYCOCALYX
• or a FLUFFY COAT - these surrounds the cellular membrane of the
platelet externally.
• unique among the cellular components of the blood.
• composed of plasma proteins and carbohydrate molecules that are
related to the coagulation, complement, and fibrinolytic systems.
CELLULAR ULTRASTRUCTURE OF A MATURE
PLATELET
• MICROFILAMENTS & MICROTUBULES
• located directly beneath the cell membrane of platelet
• provide structure of the platelet to maintain its discoid shape
• maintains the posin of the organelles
• secondary system of microfilaments is functional in internal
organization and secretion of blood coagulation products, such
as fi brinogen.
CELLULAR ULTRASTRUCTURE OF A MATURE
PLATELET
• GRANULES
• Alpha Granules - most abundant
• contains:
• heparin-neutralizing platelet factor 4 (PF 4)
• beta-thromboglobulin
• platelet-derived growth factor
• platelet fibrinogen
• fibronectin
• von Willebrand factor (vWF)
• thrombospondin
CELLULAR ULTRASTRUCTURE OF A MATURE
PLATELET
• GRANULES
• Dense or Delta Granules
• contains:
• serotonin
• adenosine diphosphate (ADP)
• adenosine triphosphate (ATP)
• calcium.
• Lysosomes - store hydrolase enzymes
CELLULAR ULTRASTRUCTURE OF A MATURE
PLATELET
Other Cytoplasmic Constituents
• Contractile proteins, including actomyosin
(thrombosthenin), myosin, and filamin,
• Glycogen
• Enzymes of the glycolytic and hexose pathways
PLATELET KINETICS, LIFESPAN & NORMAL
VALUES
• An average megakaryocyte produces about 1,000 to 2,000
platelets.
• Marrow transit time / maturation period of megakaryocyte - 5
days
• Platelets initially enter the spleen & remains for 2 days.
• Platelets are in either the circulating blood or the active splenic
pool
• Approximately 2/3 of the total number of platelets are in systemic circulation
• The remaining 1/3 exists as pool of platelets in the spleen that free exchange with
the general circulation
PLATELET KINETICS, LIFESPAN & NORMAL
VALUES
• A normal person has an average of 250 × 109/L (range, 150 ×
109/L to 450 × 109/L) platelets in the systemic circulation.
• Platelet turnover or effective thrombopoiesis averages 35 ×
109/L ± 4.3 × 109/L/day.
• The life span of a mature platelet is 9.0 days ± 1 day.
• At the end of their life span, platelets are phagocytized by the
liver and spleen and other tissues of the mononuclear phagocytic
system.