WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Phadke et al.
World Journal of Pharmacy and Pharmaceutical Sciences
SJIF Impact Factor 2.786
Volume 3, Issue 8, 2146-2155.
Review Article
ISSN 2278 – 4357
ARTIFICIAL BLOOD: A LIFE SAVING TOOL
*N M Phadke1, D G Phadtare2, R B Saudagar2
1
Departmentof Quality Assurance Techniques, R.G.SapkalCollege of Pharmacy, Anjaneri,
Nashik, Maharashtra, India (422213)
2
Department of Pharmaceutical Chemistry R.G.SapkalCollege of Pharmacy, Anjaneri,
Nashik, Maharashtra, India (422213)
Article Received on
10 June 2014,
ABSTRACT
Revised on 05 July 2014,
Accepted on 30 July 2014
the body cells such as nutrients and oxygen and transports waste
Blood is a specialized body fluid that delivers necessary substances to
products away from those cells .Artificial blood is a product made to
act as a substitute for red blood cells. While true blood serves many
*Correspondence for Author
N M Phadke
Departmentof Quality
Assurance Techniques,
different functions, artificial blood is designed for the sole purpose of
transporting oxygen and carbon dioxide throughoutthe body.
Depending on the type of artificial blood, it can be produced in
R.G.SapkalCollege of
different ways using syntheticproduction, chemical isolation, or
Pharmacy, Anjaneri, Nashik,
recombinant biochemical technology.Artificial blood is supposed to
Maharashtra, India (422213)
fulfill some functions of biological blood, especially in humans. The
initial goal of oxygen carrying blood substitutes mimics blood’s
oxygen transport capacity. There is additional longer range research on true artificial RBCs
and WBCs which could theoretically compose a blood substitute with higher fidelity to
human blood. The basic approach to make an oxygen therapeutic is using Perfluorocarbons (a
chemical compound which can carry and release oxygen).The artificial blood serves to be a
good tool for the survival of patients at the time of surgery owing to high blood losses.
Therefore the use of PFC solution as a blood substitute can be used to maintain the
circulating blood volume as well as the need of the patients.
KEYWORDS: Human Blood, Perflurocarbons, blood substitutes, oxygen therapeutics.
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INTRODUCTION
This is a new innovation in the field of blood. In which is an artificial substance that can take
the place of bloodand substance could replace during some surgical procedures. Something
that is so central to human life there can be a large reduction in the diseases and other issues
with current blood transfusions. To understand the process, it helps to know a little about how
real blood works.Blood has two main components plasma and formed elements. Blood
carries, like nutrients, hormones and waste, is dissolved in the plasma, which is mostly water.
Formed elements, which are cells and parts of cells, also float in the plasma. Formed
elements include white blood cells (WBC’s), which are part of the immune system, and
platelets, which help forming clots. The red cells in blood create the bright red color. As little
as two drops of blood contain about one billion red blood cells responsible for the
transportation of oxygen and carbon dioxide throughout the body. They make up more than
90 percent of the formed elements in the blood.RBC is a discshaped that's concave on both
side.Artificial blood is a blood substitute that can be used to provide fluid volume and carry
oxygen in the vessels. Two characteristics that a blood substitute should have are that it
should be thinner than real blood and it should have a low affinity for oxygen so that it can be
delivered easily. There has been a need for blood replacements for aslong as patients have
been bleeding to death because of a serious injury.
HISTORY
According to medical folklore, the ancient Incas were responsible for the first recorded blood
transfusions. It is when William Harvey, for the first time described and proved scientifically
that how blood is circulated throughout the body. The first successful human blood
transfusions were done in 1667. Other materials that were tried during the 1800s include
hemoglobin and animal plasma. In 1868, researchers found that solutions containing
hemoglobin isolated from red blood cells could be used as blood replacements. In
1909Landsteiner classified human blood into four different groups: A, B, AB, and
O.Recognized the agglutinins in the blood the development of artificial blood came in 1883
with the creation of Ringer's solution-a solution composed of sodium, potassium, and calcium
salts. In research using part of a frog's heart, scientists found that the heart could be kept
beating by applying the solution. Karl Landsteiner who has been called the father of
immunology, was the only child of LeopoldLandsteiner, a prominent Austrian journalist and
editor, they were primarily interested in the lack of safety and effectiveness of blood
transfusions. Prior to his work, blood transfusions were dangerous and underutilized because
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the donor’s blood frequently clotted in the patient. They also intrigued by the fact that when
blood from different subjects was mixed, the blood did not always clot. He believed there
were intrinsic biochemical similarities and dissimilarities inblood. Using blood samples from
his colleagues, he separated the blood’s cells from its serum, and suspended the red blood
cells in a saline solution. Then he mixed each individual’s serum with a sample from every
cell suspension. Clotting occurred in some cases in others there was no clotting. Landsteiner
determined that human beings could be separated into blood groups according to the capacity
of their red cells to clot in the presence of different serums. He named his blood classification
groups A, B, and O. A fourth group AB was discovered the following year. The result of this
work was that patient and donor could be blood-typed beforehand, making blood transfusion
a safe and routine medical practice. This discovery ultimately earned Landsteiner the 1930
Nobel Prize in physiology or medicine.In 1966, experiments with mice suggested a new type
of blood substitute, perfluorochemicals (PFC). These are long chain polymers similar to
Teflon. It was found that mice could survive even after being immersed in PFC; this gave
scientists the idea to use PFC as a blood thinner. In 1968, the idea was tested on rats. The rat's
blood was completely removed and replaced with a PFC emulsion. In the years to follow,
medical practitioners tried numerous substances such as beer, urine, milk, plant resins, and
sheep blood as a substitute for blood
Artificial Blood Cells
Artificial blood doesn't do all the work of real blood. Sometimes, it can't even replace lost
blood volume. Instead, it carries oxygen in situations where a person's red blood cells, can’t
do it on their own. For this reason, artificial blood is synthesized “oxygen therapeutic” that is
often called as oxygentherapy. Artificial blood or blood surrogates is a substance used to help
mimic and fulfill some functions of biological blood, usually in the oxygen-carrying sense.
The main aim is to provide an alternative to blood transfusion, which is transferring blood or
blood-based products from one person into another.
Figure 1: Composition Of Blood
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Artificial blood does not contain the plasma, red and white cells, or platelets of human blood,
they dissolve less oxygen than pure liquids. It could only functions to transport and deliver
oxygen to the body's tissues.Artificial blood can be produced in different ways using
synthetic production, chemical isolation, or recombinant biochemical technology. Current
blood substitutes are either hemoglobin-based oxygen carriers (HBOCs) or perfluorocarbons
(Perfluorocarbons). While HBOCs utilize hemoglobin, an actual component of red blood
cells, perfluorocarbons rely solely on synthetic chemical processes. Unlike real blood,
artificial blood can be sterilized to kill bacteria and viruses. Doctors can also give it to
patients regardless of blood type. Many current types have a shelf life of more than a year and
don't need to be refrigerated, making them ideal for use in emergency and battlefield
situations. So even though it doesn’t actually replace human blood, artificial blood is
beneficial.
Composition Of Artificial Blood
Perfluoro-octyl bromide - 28%
FO-9982 - 12%
Yolk lecithin - 2.4%
DSPE-50 H - 0.12%
Distilled water - 57.48%
Advantages And Disadvantages
One advantage is that being able to manufacture large quantities of blood will end the
shortage of blood in emergency cases. Another advantage is that there is less of a chance of
spreading diseases because the blood is prepared in a secure place and scientists will know
full history about it. Artificial blood is able to transport and release oxygen where needed,
storable and durable for longer time periods, safe to use, compatible in the human body.
Some disadvantages are that it is currently expensive, but the price to make it is expected to
drop less than in comparison to the cost of transfusion. It may also increase chances of a heart
attack.Body immune systems may sometimes react negatively to the foreign blood that is
inserted into the body.
Blood Substitutes
Research for blood substitutes began in the1960’s. Iron-rich hemoglobin compounds were
used to carry oxygen to tissues but they caused kidney toxicity.There are a few major
Advantages of Blood Substitutes over conventional blood. Probably the most important is the
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universal compatibility which allows the transfer of any blood type without tests. Also of
great importance knows that the substitute is free of any disease, has a much greater shelf life,
and very predictable outcomes. Blood substitutes can be broken down into 2major categories:
volume expanders and Oxygen therapeutics. Volume expanders simply increase the blood
volume and consist of 2 smaller categories:
1. Crystalloid-based
2. Colloid-based.
Figure 2: The Different Forms Of Blood Substitutes
Perflurocarbons (PFC)
In 1966, scientists synthesized “oxygen therapeutic” that was often called oxygentherapy.Per
fluorocarbons are derived from a group of hydrocarbons in which the hydrogen atoms are
replaced by fluorine atoms. Perfluorocarbons are chemically inert due to the strength of the
carbon-fluorine bonds, used to create artificial blood during surgeries. Many perfluorinated
carbons are chemically and biologically inert but are able to dissolve a large amount of gas.
One of the problems with perfluorocarbons is that they are an oil-like fluid that does not mix
well with water and cannot carry water-soluble salts and metabolic substrates. Now a day’s
most of the Perfluorocarbons oxygen carriers are mixtures of perfluorocarbons with
emulsifying agents. Emulsifying agents are substances that help stabilize two seemingly
unbendable things. Perfluorocarbons oxygen carriers utilize Puronic-68, egg yolk
phospholipids and triglycerides.
Figure 3: Pfc Dimensions
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Advantages
1. Perfluorocarbons do not react with oxygen.
2. Perfluorocarbons allow easy transportation of the oxygen to the body.
3. They allow increased solubility of oxygen in plasma.
4. Perfluorocarbons minimize the effects of factors like pH and temperature in blood
circulation
Disadvantages
1. This is often caused by phagocytosis of the per fluorocarbonemulsion by the recipient
organism’s immune system.
2. Often causes flu-like symptoms
3. Unable to remain mixed as aqueous solutions –thus, they must be prepared as emulsions
for use in patients
4. PFC products cannot be used by the human body, and must be discarded
this takes
approximately 18-24 months
5. Because PFC absorbs oxygen passively, patients must breathe at a linear rate to ensure
oxygenation of tissue.
Current Therapeutics
Perfluorocarbons-based blood substitutes are completely man-made; this provides advantages
over blood substitutes that rely on modified haemoglobin, such as unlimited manufacturing
capabilities, ability to be heat-sterilized and Perfluorocarbons efficient oxygen delivery and
carbon dioxide removal. Perfluorocarbons in solution act as an intravascular oxygen carrier to
temporarily augment oxygen delivery to tissues.
Table 1: Some Perfluorocarbons Based Terms
Sr. No.
1.
Terms
Perftoran
2.
Oxygent
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Description
Approved for Russian clinical application in 1996.Registered in
Mexico as PERFTEC, distributed by KEM Laboratory
(Mexico).Facilitates oxygen delivery together with remaining red
blood cells at blood replacements and will have wider area for
application than just a blood substitute.
Oxygent is currently approved for Phase II Trials in US and Europe. It
is developed to reduce the need for donor blood Pharmaceuticals
during surgery. Perfluorocarbons are surrounded by a lecithin
surfactant in a water-based solution. The lecithin is digested
intracellular.
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Hemoglobin-based Oxygen Carriers (HBOCs)
Hemoglobin-based Oxygen Carriers were created as a mechanism to mimic the oxygencarrying role of hemoglobin in the body, while still reducing the need for real human
hemoglobin. The hemoglobin used was found to have enterocyte membrane stromal lipids as
well as bacterial endotoxins. Hemoglobin is a tetramer with two alpha and two beta
polypeptide chains; each is bound to an iron heme group which successively binds to an
oxygen molecule Hemoglobin’s heme bond allows it to have a higher affinity for oxygen,
thus making it an excellent source of blood substitutes. Currently, HBOCs represent an
interesting class of blood substitutes, which are undergoing advanced clinical trials. The
therapeutic goal of these compounds is to avoid or reduce blood transfusion in different
surgical and medical situations of acute Hb. Synthetic hemoglobin-based product are
produced from hemoglobin harvested from an E. coli bacteria strain. The hemoglobin is
grown in a seed tank and then fermented.
3 days
Seed tank
E.coli
Fermentation
Manufacturing Procedure
The production of hemoglobin-based products, thisinvolves isolation or synthesization of
hemoglobin, molecular modification then reconstitution in an artificialblood formula. PFC
products involve a polymerizationreaction.
Steps Forhemoglobin Synthesis
Step 1
To start the fermentation process, a sample of the pure bacteria culture is transferred to a test
tube that contains all the nutrients necessary for growth. This initial inoculation causes the
bacteria to multiply. When the population is great enough, they are transferred to a seed tank.
Step 2
A seed tank is a large stainless steel kettle that provides an ideal environment for growing
bacteria. It is filled with warm water, food, and an ammonia source which are all required for
the production of hemoglobin. Other growth factors such as vitamins, amino acids, and minor
nutrients are also added. The bacterial solution inside the seed tank is constantly bathed with
compressed air and mixed to keep it moving. When enough time has passed, the contents of
the seed tank are pumped to the fermentation tank.
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Step 3
The fermentation tank is a larger version of the seed tank. It is also filled with a growth media
needed for the bacteria to grow and produce hemoglobin. Since pH control is vital for optimal
growth, ammonia water is added to the tank as necessary. When enough hemoglobin has been
produced, the tank is emptied so isolation can begin.
Step 4
Isolation, it begins with a centrifugal separator that isolates much of the hemoglobin. It can
be further segregated and purified using fractional distillation. This standard column
separation met hood is done by the principle of boiling a liquid to separate one or more
components and utilizes vertical structures called fractionating columns. From this column,
the hemoglobin is transferred to a final processing tank.
Step 5
Ensuring the stability of hemoglobin; When hemoglobin is left outside a cell, it has the
tendency to break into its individual parts, instead of remaining as an entire hemoglobin
protein Methods have been discovered to retain the stability of hemoglobin including.
Figure 4: Stability Of Hemoglobin Prepared Artificially
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Advantages
1. Available in much larger quantities.
2. Can be stored for long durations.
3. Can be administered rapidly without typing or cross-matching blood types.
4. Can be sterilized via pasteurization.
Disadvantages
1. Reduced circulation half-life.
2. Disrupts certain physiological structures, especially the gastrointestinal tract and normal.
3. Red blood cell hemoglobin.
4. The release of free radicals into the body.
Table 2: Some Hemoglobin Based Terms
Name
Description
Hemopure Hemopure is currently approved for Phase III trials in the United States and was
more widely approved in South Africa. It is Biopure’s first-in-class product for
human use, and is a HBOC solution. It is made of chemically stabilized, crosslinked bovine (cow) haemoglobin in a salt solution
Hemospan Hemospan is currently in Phase II trials in the United States. It is produced by
the company Sangart, which was found by Dr. Robert M. Winslow in 1998. It is
produced in powder form, which can then be mixed into liquid form and
transfused immediately, regardless of a patient’s blood type. This technology
relies on coupling with polyethylene glycol (PEG) to eliminate the toxicity
associated with free haemoglobin.
Hemotech Hemotech, a human blood substitute developed in 1985 by researchers, Mario
Feola, MD and Jan Simoni, PHD, DVM from the Texas Tech University Health
Sciences Center. It has been able to identify and nullify the source of toxicity
issues associated with previous blood substitute candidates.
CONCLUSION
Artificial blood can be regarded as a blood substitute for providing increase in fluid volume
and oxygencarrying capacity of vessels. The main purpose of artificial blood is to lessen the
demand for human blood supplies and to give immediateresponse without triggering a
rejection in cases of massive blood loss. Synthetic chemical compounds called
perfluorocarbons are currently being studied as a substitute for red blood cells. These blood
substitutes are not stable enough to form a clot. Currently artificial blood technology is
limited to short-term blood replacement applications. In the years to follow researchers are
focusing onstudy of numerous substances such as beer, urine, milk, plant resins, and sheep
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blood as a substitute for blood.In the future, it is expected that new materials to carry oxygen
in the body will be made available as blood substitutes.
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