Artificial Blood

UNIT-II
What is the need of artificial Blood ?

• Blood is key component of surgery and treatment of injury, and its

availability is critical for survival in the presence of severe blood
losses.
To fill gap between availability and need of blood; artificial blood is
used.
• Risks: ranging from about 3% (3 adverse outcomes per 100 units
transfused) for minor reactions.
• Still, it has not been possible to obtain an artificial fluid with the same
properties as blood, primarily due to its cellular nature.
ARTIFICIAL BLOOD
• 3% (3:100 units transfused) minor reaction and 0.001% fatal hemolytic
reaction.
• For medical care delivery:-availability of blood is one unit (0.5 l) per 20
person-year.
• According to WHO availability and use is limited to one unit per 100
person-year.
• Possibility of transmitting infectious disease ex. Hepatitis (0.002%) and
HIV (1:400000)
• Used to mirror the functions of biological blood.
• The main aim of artificial blood is to provide an alternative to blood
transfusions, which is transferring blood from one individual to another.
• Carries oxygen
• It does not contain RBC’s, or WBC’s
• Alternate to blood transfusion
• Eliminating the human to human transfer of blood
Uses
• Acts like red blood cells to transport oxygen
• Can be derived from red blood cells of humans or animals
• Gets its oxygen carrying properties from “perflourocarbons”
• At present only three products are in clinical trials for blood
replacement
• Side effects :hemoglobin-based products gives hypertension
Advantages Disadvantages

• Demand for blood transfusions • Currently more expensive

is rising • May increase chance of heart
• No chance of HIV or other attack
blood transmitted diseases
• Army medical care
• Lasts longer in storage
• May cost less in the future
Exchange of Oxygen

• Understanding of exchange of
oxygen in the microcirculation
are still in research phase.
• In-vivo methods are used to
understand how oxygen is
distributed to the tissues by the
microscopic blood vessels. hamster skinfold model:
• Tissue under study consists of used to quantify blood flow,
skeletal muscle and connective blood and tissue pO2,
leukocyte activation, blood
tissue. Arterial and venous
vessel permeability to
catheters are used to monitor
macromolecules, and blood
systemic data vessel tone throughout the
arterioles, capillaries, and
venules
 Distribution of Transport Properties in the Circulation & plasma expanders

→ Artificial blood or blood substitutes are

plasma expanding fluids that carry
more oxygen in comparison to water or
plasma.
→ Its physical properties are different from
natural blood.
→ The transport properties of blood, such
as hydraulic pressure, partial
pressure of oxygen, oxygen content
and viscosity, are distributed in normal
condition.
→ The distribution is matched with micro
and macro circulation system through
its life.
→ The distribution of blood pressure
influences the cellular composition of
blood vessels, and is regulated by the Distributed nature of transport properties in the
so-called myogenic response, which hamster skinfold microcirculation

causes vessels to constrict, thus Most arteries and veins in the body are
increasing viscous losses, if pressure innervated by sympathetic adrenergic
increases and vice versa. nerves, that cause the observed changes in
vascular function
 Contd..
Examples of transport properties
distribution
→Hydraulic blood pressure, which changes
continuously in the circulation due to
viscosity variation.
→The BP distribution influences the
cellular composition of blood vessels which
is regulated by myogenic response (vessel
constriction occurs when BP increases with
decreased viscosity).
→Blood viscosity and blood flow velocity
in the circulation are also distributed such
like shear stress is uniform throughout the
circulation system.
→This is because of continuous variation of
hematocrit value (% of red cells in your
bloods) of the blood (it is about half the
value in capillaries then in larger blood Distributed nature of transport properties in the
vessels due to the Faraheus–Lindquist effect hamster skinfold microcirculation
).
 Contd..
Examples of transport properties
distribution
→In larger arterioles, blood viscosity is
about 3.5 to 4.0 centipoise (cP),
→while in the smaller arterioles viscosity
falls to about that of plasma (1.1 to 1.2 cP).
→Dependency of shear stress on
viscosity: Shear stress and shear stress
dependent release of endothelial dependent
relaxing factors (EDRF-NO as strong
vasodilator (Endothelium Derived Relaxing
Factor nitric oxide ) and prostaglandins),
depends on viscosity since the circulation is
designed to maintain shear stress constant.
→The molecule of plasma expander do not
depend on Fahraeus–Lindquist effect:-
causing the whole microcirculation with a
uniform viscosity and thus changing shear Distributed nature of transport properties in the
stress distribution. hamster skinfold microcirculation
 The Distribution of Oxygen in the Circulation System

Krogh model: This model shows how gases are exchanged between blood flowing
in a cylindrical conduit, the single capillary, and a surrounding tissue cylinder.

→ It is assumed that most of the oxygen is exchanged at tissue level.

→ Assumption explains existence of large blood/tissue oxygen gradients in a tissue
capillaries.
→ However, capillary/tissue O2 gradients are maximal in the lung (50 mmHg/μm) and
minimal in the tissues (0.5 mmHg/μm).
→ Most tissue capillaries appear to be in near oxygen concentration equilibrium with
the tissue; thus, large oxygen gradients are not present, suggesting that capillaries
may not be the primary mechanism for tissue oxygenation.

Phosphorescence quenching:- Po2 measurement is done by this method optically in the

microvessels and the surrounding tissue.
When Phosphorescence quenching method is used in conjunction with hamster skinfold
model, it allows for in-vivo analysis of the assumptions made in the Krogh model.
Results from Phosphorescence quenching method along with
hamster model and previous findings from other laboratories

→Most of the oxygen is delivered by the arterioles and that little

oxygen is contributed by the capillaries.
→Capillary blood pO2 is only slightly higher (about 5 mmHg)
than tissue pO2.
→Arterio/venous capillary pO2 differences are very small because
tissue pO2 is essentially uniform, and capillaries are close to
pO2 equilibrium with the tissue.
→The only tissue domain where pO2 displays large gradients is
the immediate neighborhood of the microvessels (vessels with
diameter 80 μm and smaller), a tissue compartment whose main
constituent is the microvascular wall.
→A major portion of blood oxygen exits the circulation via the
arterioles.
 Oxygen Gradients in the Arteriolar Wall

→Hypothesis is made that arteriolar wall consists of high

metabolism tissue (endothelium & smooth muscles) and
therefore a large oxygen sink occurs at this location.
→From the above assumption, it is believed that sharp gradients
are present in arterioles, but not in capillaries and venules.
→Oxygen exiting from arterioles is driven by sharp oxygen
gradients at the arteriolar wall, which is consistent with the
hypothesis.

Note:- By above hypothesis and findings design of artificial blood is

conceptualized.
 Conceptualization for the design of artificial blood
→ Endothelium and smooth muscle serve as a metabolic barrier to the passage
of oxygen from blood to tissue, which in part protects the tissue from the
high oxygen content (pO2) of blood.
→ One of the goals of basal tissue perfusion is to supply oxygen to the
endothelium and smooth muscle.
→ Oxygenation of working tissue results from three events:
• Lowering of the vessel wall metabolism
• Increased perfusion with oxygenated blood
• Deployment of a biochemical process that protects the tissue from high pO 2
levels
→ Due to basal membrane conditions, tissue capillaries partially supply oxygen
to the tissue.
→ The physical and biological properties of blood affect the oxygen
consumption of the vessel walls.
 Mathematical Modeling of Blood/Molecular Oxygen
Carrying Plasma Expander
→ Mathematical modeling is based upon previous assumptions.
→ The mathematical model is based on oxygen mass balance in following
vessel segments (Major artery ---->>>Capillaries).
→ The objective of mathematical model is to calculate the oxygen tension of
blood in the capillaries, which should correspond to tissue pO2. The
expression is given as

• Equation express how the total loss of oxygen from the arterial and arteriolar
network (Kn)is related to transport properties of the blood vessels and blood.
• Total oxygen exit is the summation of losses from (n )individual vascular segments
(i).
• This equation gives the functional relationship between transport parameters that
determine capillary blood pO2 for given changes in the physical properties of blood.
Mathematical Modeling of Blood/Molecular Contd..
Oxygen Carrying Plasma Expander

• μ: blood viscosity
• F(Htc, C): Concentration of hemoglobin (red blood cells + molecular)
• mt:Maximum amount of oxygen that can be dissolved in blood under normal
conditions
• D: Diffusion constant for oxygen in tissue
• α: Solubility of oxygen in tissue
• go :Oxygen consumption by the vessel wall
• Li :Length of each vessel segment
• di is the diameter of each segment
• ni: is the slope of the oxygen dissociation curve for hemoglobin.
Mathematical Modeling of Blood/Molecular Contd..
Oxygen Carrying Plasma Expander

• The equation shows two different group terms.

• One group is common to all vessel segments and includes blood viscosity,
hematocrit or blood oxygen carrying capacity, and vessel wall metabolism.
• The second group is a summation where each term is specific to each vascular
segment.
 Blood Substitutes and Hemodilution objectives
• In general, a very low hematocrits can survive (proportion, by volume, that
consists of red blood cells), corresponding to losses of the red blood cell
mass of the order of 70%.
• A 30% shortage in blood volume can lead to irreversible shock if not
rapidly corrected.
• Maintenance of normal volume of blood in the body (normovolemia) is the
objective of most forms of blood substitution or replacement by the dilution
of the original blood constituents.
• Hemodilution produces systemic and microvascular phenomena that lie
beneath all forms of blood replacement and provides a physiological
reference for comparison for blood substitutes.
• Hemodilution fluid is classified in 3 groups:-
1. crystalloid solutions
2. colloidal solutions
3. oxygen carrying solutions.
Above materials change the transport properties of blood which effectively
changes tissue oxygenation.
Hemodilution is analyzed in terms of systemic effects , coupled with the
altered composition of blood which influence the transport properties of the
microcirculation.
 Hematocrit and Blood Viscosity
• Blood viscosity is determined by the hematocrit in the larger vessels while
in microcirculation due to weaker value viscosity is not completely depend
on hematocrit, which is given by equation:-

• For microvascular blood viscosity is given by

• Where; μ is the blood viscosity in centipoise

as, bs are parameters that are shear rate and vessel size dependent
am, bm are parameters that are independent of shear rate but are vessel
size dependent
H is hematocrit value
When hematocrit is reduced then viscosity is reduced rapidly which affects BP in larger
vessels
In microcirculation BP is not very much affected.
 Regulation of Capillary Perfusion During Extreme
Hemodilution in Hamster Skinfold Microcirculation by
High Viscosity Plasma Expander
• Functional capillary density (FCD):- (It is the number of capillaries that
possess red blood cell transit in tissue)
• Maintaining FCD in shock is a critical parameter in determining the
outcome in terms of survival vs. nonsurvival, independent of tissue pO2.

-4 h hemorrhagic shock to 40
mmHg and
resuscitated(revived) with their
own blood.
-Timepoint:
Time after induction of
hemorrhage.
B30: 30 min after resuscitation.
B24: 24 h after resuscitation.
∗: Significantly different from
control,
p > .05. +: Significantly
different between survivors and
nonsurvivors, p > .05.
 Crystalloid and Colloidal Solutions as Volume Expanders

-Crystalloids are among the most widely used fluids for volume
replacement.
Ex:- Ringer’s lactate, is administered in volumes that are as much as three
times the blood loss
-The advantage of crystalloid solutions is that large volumes can be given
over a short period of time with low danger of increasing pulmonary
wedge pressure.
-Excess volumes are rapidly cleared from the circulation by diuresis
(condition in which the kidneys filter too much bodily fluid).
-Blood volume replacement with Ringer’s lactate lowers blood viscosity.
-Albumin is used as a plasma expander for emergency volume restitution,
-Due to high-cost the synthetic colloids dextran and hydroxyethyl starch
are used more frequently.
-These materials may cause anaphylactic(severe allergic reaction) reactions
and tend to alter platelet function.
 Artificial Oxygen Carriers

 It is done by use of modified hemoglobins which bind oxygen

chemically and reversibly, and fluorocarbons which have a high
capacity for dissolving oxygen.
 Hemoglobin separated from the red cell membrane (stroma-free
hemoglobin) which carries oxygen with a high affinity.
 Fluorocarbons carry a limited amount of oxygen under normal
atmospheric conditions, but are biologically inert, can be stored at room
temperature, and are excreted as gas through the lungs.
BLOOD SUBSTITUENTS

Blood substituents can serve as :-

•Plasma volume expander
•Replicate the oxygen carrying function of natural blood
Plasma Expanders :- These are compounds, which are either entirely
synthetic or processed from natural proteins that serve as infusion
solutions which expand intravascular volume.
RBC Substituents :- these are oxygen carriers but, differ the way they
carry.
•Modified Haemoglobins
•Perflurocarbons
PLATELET SUBSTITUENTS
Platelet substituents have following properties:-
•Effective hemostasis with a significant duration of action
•No associated thrombogenicity
•No immunogenicity
•Sterility
•Long shelf life with simple storage requirements
•Easy preparation and administration
Several different forms of platelet substitute are now under development :
•Infusible Platelet Membranes (IPM)
•Thrombospheres
•lyophilized human platelets
INFUSIBLE PLATELET MEMBRANE
(IPM)
• Produced from outdated human platelets.
• The source platelets are fragmented (1,000 platelets per
megakaryocyte), virally inactivated, and lyophilized; they can then be
stored up to 2 years.
• Although the platelet membranes still express some blood group and
platelet antigens, they appear to be resistant to immune destruction.
Cypress Bioscience Incorporated, manufactures an IPM product that
is currently
• in phase II trials.
• for use in patients who have become refractory to platelet transfusions because of
the formation of antibodies to HLA or platelet antigens.
• Overall, the product appears to be safe.
THROMBOSPHERES

• Thrombospheres are not platelets.

• They are composed of cross-linked human ALBUMIN with human
FIBRINOGEN bound to the surface.
• Experimentally, the thrombospheres appear to enhance platelet
aggregation but do not themselves activate platelets.
• Mechanism of action not clarified.
• No evidence of thrombogenicity.
• A similar product, Synthocytes (Andaris Group Ltd, Nottingham,
UK), is in clinical trials.
LYOPHILISED HUMAN PLATELETS

• This product has been under development since the late 1950s.
PROCESS:-
• Briefly fixing human platelets in paraformaldehyde (Kills microbes)
prior to freeze-drying in an albumin solution ( increase shelf life)
• The adhesive properties of the platelets appear to be maintained.
• This product is currently in animal trials.
RED BLOOD CELL SUBSTITUENTS

• Main function is to carry oxygen, as does natural hemoglobin.

• The use of oxygen-carrying blood substitutes is often called Oxygen
therapeutics to differentiate from true blood substitutes.
• The initial goal of oxygen carrying blood substitutes is simply to
mimic blood's oxygen transport capacity.
There are two basic approaches to constructing an oxygen
therapeutics:
• The first is perfluorocarbons (PFC), chemical compounds which can
carry and release oxygen.
• The second approach is haemoglobin derived from humans, animals,
or artificially via recombinant technology, or via stem cell production
of red blood cells in vitro
PFC

• PFC are biologically inert materials that can dissolve about 50 times
more oxygen than blood plasma.
• They are relatively inexpensive to produce and can be made free form
of any biological materials.
• They have the ability to carry much less oxygen than haemoglobin
based products.
STRUCTURE:-

• Not soluble in water, which means to get them to work they must be
combined with emulsions.
• These PFC emulsions consist in droplets of fluorochemicals in the
range of 0.1 to 0.2 μm diameter coated by a thin film of egg yolk
phospholipids.
• It can perfuse smallest capillaries, where no RBC flow.
• Perfluorocarbon core Surrounded by a phospholipid surfactant that
reduces the surface tension of the liquid in which it is dissolved.
• PFCs have oxygen solubilities of the order of 50 ml O2/100 ml at
37°C and 760 mmHg oxygen pressure.
• These materials carry about 30% more oxygen than blood when
diluted to 60% and with 100% inspired oxygen