This Presentation information content includes:- Description of Blood and its system; Introduction to Artificial Blood; Need; Brief History; Design of Artfiicial Blood; Blood substituents - Platelet Sustituents [ Infusible Platelet Membrane (IPM), Thrombospheres, Lyophilised Human Platelets], Advantages & Disadvantages; RBC Substituents [ Perfluorocarbons, Haemoglobin based Oxygen Carriers], Advantages & Disadvantages; Applications; References.

1. ARTIFICIAL BLOOD & BLOOD
SUBSTITUENTS
ARUNDHATI MEHTA
M.Sc. Biotech III Semester
Department of Biotechnology
Guru Ghasidas Vishwavidhyalaya
© Arundhati Mehta 2017

2. BLOOD
Blood is a fluid connective tissue circulated around the
body through blood vessels by the pumping action of
the Heart.
Composed of Blood Cells suspended in Blood Plasma.
Function:-
 Respiration - Transport of O2 & CO2 (RBCs)
 Trophic – Transport of nutrient materials
 Maintaining Homeostasis & body temperature.
 Protection – Immunity and blood Coagulation.
 Regulation – Hormonal regulation.
 Excretive – metabolites delivered to excretory organs.
Blood
Components
Plasma
Water (92% by
volume)
Others
(Proteins,
Mineral ions,
glucose,
hormones, O2,
CO2)
Blood cells
RBCs
WBCs
Platelets

3. BLOOD
Red Blood Cells contains Haemoglobin which impart bright red
colour to cells & are responsible for “typing” phenomena.
Haemoglobin :-
 Oxygen-transport Metalloprotein
 4 globular proteins tightly associated with non-
protein prosthetic heme group.
 Heme group consists of an iron (Fe) ion (charged atom) held
in a heterocyclic ring, known as a porphyrin.
 Oxyhemoglobin is formed during physiological
respiration when oxygen binds to the heme component of
the protein hemoglobin in red blood cells.
 Carbon dioxide binds to hemoglobin and
forms carbaminohemoglobin.
 A variant hemoglobin, called fetal hemoglobin (HbF, α2γ2),
is found in the developing fetus, and binds oxygen with
greater affinity than adult hemoglobin.

4. ARTIFICIAL BLOOD
Artificial Blood (also called Blood Substitute or Blood Surrogate) is a
substance used to mimic and fulfil some functions of biological blood. It aims to
provide an alternative to blood transfusion, which is transferring blood or blood-
based products from one person into another.
It aims to provide an alternative to blood transfusion, which is transferring blood
or blood-based products from one person into another.

5. NEED
CHALLENGE
WHY WE
NEED ??
Shortage
Short shelf
life
Rare blood
group
demand
Immunologi-
cal
incompatibilit
y
Disease
Transmission

6. HISTORY
YEAR EVENTS
1616 Wiliam Harvey described how blood is circulated throughout the body.
1667 First successful Human Blood Transfusions.
Medical practitioners tried substances like Beer, Urine, Milk, Plant resins,
Sheep blood etc. as substituents
1854
Milk was one of the first used material injected in patients to treat Asiatic
Cholera. Physicians believed that the milk helped regenerate white blood
cells. However, many practitioners remained sceptical so milk injections
never found widespread appeal, so soon discarded.
Salt or saline solutions was another substitute . In experiments done on
frogs, it was found that it could keep frogs alive for some time, if whole
blood is replaced with a saline solution but it was little misleading, so,
Saline was developed as a plasma volume expander.

7. HISTORY
YEAR EVENTS
1868 WWII and Vietnam war ignited the search for blood substituents– Hemoglobin solutions
and Synthetic Oxygen carriers.
1871 Examined the use of animal plasma as substitute.
1883 Creation of Ringer’s solution by Sydney Ringer - a solution composed of sodium,
potassium, and calcium salts. scientists found that the heart could be kept beating by
applying the solution. This solution resembles Blood Serum.
• Led to findings that the reduction in blood pressure caused by a loss of blood
volume could be restored by using Ringer's solution.
• This product evolved into a human product when lactate was added.
Dr. Sydney Ringer

8. DESIGN
Ideal characteristics for Artificial Blood is:-
Safe to use
Compatible in the body- elimination of cross matching
Oxygen carrying capacity, equally or surpassing that of
biological blood
Pathogen and toxin free
Minimal side effects
Long shelf life
Survivability over a wider range of storage temperatures
Viscosity similar to Body blood
No immunosuppression
Cost efficient

9. 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.
They are:-
 Modified Haemoglobins
 Perflurocarbons

10. 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

11. INFUSIBLE PLATELET
MEMBRANE (IPM)
 Produced from outdated human platelets.
 The source platelets are fragmented, 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 antigen or platelet antigens.
 Overall, the product appears to be safe.

12. THROMBOSPHERES
 Thrombospheres (Hemosphere, Irvine, Calif) 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 elucidated.
 No evidence of thrombogenicity.
 A similar product, Synthocytes (Andaris Group Ltd,
Nottingham, UK), has just entered into clinical trials in
Europe.

13. 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.

14. 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 merely 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 specific PFC usually used is
either perfluorodecalin or dodecafluoropentane emulsion (DDFPe).
 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

15. PERFLUOROCARBONS
 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
devoid of any biological materials.
 Emulsion particles are 0.2 micron in diameter → Can
perfuse smallest capillaries, where no RBC flow.
 Not soluble in water, which means to get them to work they
must be combined with emulsions.
 They have the ability to carry much less oxygen than
haemoglobin based products.
 STRUCTURE:- Perfluorocarbon core Surrounded by a
phospholipid surfactant that reduces the surface tension of
the liquid in which it is dissolved.
 PRODUCTION PROCESS :-
 Water, salts, and phospholipids surfactant,
antibiotics,vitamins,nutrients are added and emulsified
through high-pressure homogenization.
 Purified through high temperatures of steam.

16.  EXAMPLES :-
PERFLUOROCARBONS

17.  ADVANTAGES :-
 Do not react with oxygen
 Inexpensive
 Allow easy transportation of the oxygen to the body
 They allow increased solubility of oxygen in plasma
 minimize the effects of factors like pH and temperature in blood circulation.
 DISADVANTAGES :-
 Often causes flu-like symptoms
 Unable to remain mixed as aqueous solutions
 A decrease in blood platelet count.
 PFC products cannot be used by the human body, and must be discarded.
 PFCs absorb oxygen passively, patients must breathe at a linear rate to ensure
oxygenation of tissues.
 The problem with Fluosal-DA was that they dissolve less oxygen than pure liquids
PERFLUOROCARBONS

18. HAEMOGLOBIN BASED
OXYGEN CARRIERS
 Hemoglobin-based Oxygen Carriers were created as a mechanism to mimic the
oxygen-carrying role of hemoglobin in the body, while still reducing the need for
real human hemoglobin.
 Native HB dissociates into
monomer and diamer.

19. HAEMOGLOBIN BASED
OXYGEN CARRIERS
 To avoid such spontaneous dissociation native Hb is modified by intramolecular
cross-linking between alpha and beta Chains, polymerization, pyridoxylation, or
conjugation to larger molecules, such as Albumin or Polyethyleneglycol
("pegylation"),encapsulation of hemoglobin into a liposome or polymer structure.

20. HAEMOGLOBIN BASED
OXYGEN CARRIERS

 PRODUCTION PROCESS :-
 HB synthesis
 Synthetically produced Hb: E.coli(P678-54)
 Isolated Hb: human or animal blood(bovine blood)
 Seed Tank
 Fermentation
 Final processing

21. HAEMOGLOBIN BASED
OXYGEN CARRIERS
 EXAMPLES :-

22. HAEMOGLOBIN BASED
OXYGEN CARRIERS
 ADVANTAGES :-
 Available in much larger quantities.
 Can be stored for long durations.
 Can be administered rapidly without typing or cross matching blood types.
 Can be sterilized via pasteurization.
 DISADVANTAGES :-
 Short half-life
 Disrupts certain physiological structures, especially the gastrointestinal tract and
normal red blood cell haemoglobin.
 They release free radicals into the body
 Availability and cost

23. APPLICATIONS
 POTENTIAL CLINICAL APPLICATIONS:-
 Blood substitutes :
 hemorrhagic shock; hemorrhage (war, surgery); anaemia.
 Whole-body rinse out : acute drug intoxication; acute hepatic failure.
 Local Ischemia : acute Myo-cardial infection ; evolving MI; cardiac failure;
brain infarction; acute arterial thrombosis and embolism.
 General Ischemia : CO intoxication.
 Aid for organ recovery : acute renal failure; acute hepatic failure;acute
pancreatitis.
 Adjuvant therapy : radiotherapy; chemotherapy.
 Perfusional protection of organs during surgery – cardiopulmonary bypass.
 Preservation of donor organ.
 Drug carrier - drug-conjugated haemoglobin and perfluorochemicals.

24.  NON CLINICAL APPLICATIONS :-
 Culture medium
 Chemical examination - oxygen sensor; standard solution for oxygen calibrator
 Bioreactor
 PARADOXICAL UTILISAIONS (of high-oxygen affinity)
 Oxygen absorbent
 Oxygen pulse therapy for malignant tumour in combination with radiotherapy or
chemotherapy.
APPLICATIONS

25. REFERENCES
L. Kresie, Artificial blood: an update on current red cell and platelet substitutes,
Proc (Bayl Univ Med Cent). 2001 Apr; 14(2): 158–161; PMCID: PMC1291332
S. Sarkar, Artificial blood, Indian J Crit Care Med. 2008 Jul- Sep; 12(3):140-144;
PMCID: PMC2738310.
American Chemical Society News
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