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Artificial Blood

A closer look at HBOCs
by

Jose Martinez

on 5 April 2014

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Transcript of Artificial Blood

Are We There Yet?
By Jose Martinez
Artificial Blood
What is Blood?
Blood is a specialized type of connective tissue that is composed of Plasma, Formed Elements and Red Blood Cells.
45% = Erythrocytes
55% = Plasma
<1% = Buffy Coat
Plasma contains dissolved solutes, including nutrients, gases, hormones, wastes and products of cell activity, ions and proteins.

Replace labile and non-labile coagulation factors in massive bleeding from trauma OR clotting factor deficiencies when clotting factors are not available

Used to treat patients with stable clotting factor deficiencies or patients undergoing therapeutic plasmapheresis
Consists of WBCs and Platelets

White Blooc Cells form part of the immune system and have the ability to detect pathogens, form antibodies against them, and destroy them.

Platelets work to keep the body in homeostasis. When a blood vessel is injured, platelets come together along with other coagulation factors to form a clot and stop the bleeding.
Platelets prevent spontaneous bleeding or stop established bleeding in thrombocytopenic patients.

Single Donor Platelets expire 5 Days from Collection.
Storage and Expiration
1 year stored at -18 C
7 years stored at -65 C
Erhythrocytes are Red Blood Cells (RBC) about 7.5 um in diameter. They are shaped like flattened disks with depressed centers. Mature RBCs are bound by a plasma membrane, but lack a nucleus and have essentially no organelles.

They are little more than "Bags" of Hemoglobin (Hgb).

Normal cells show an area of central pallor that is about one third the area of the red blood cell. Central pallor is directly related to the amount of hemoglobin inside the cell.
Packed RBCs are stored at 1-6 Degrees Celsius
Their shelf life is only 42 Days.

Donor cells must be compatible with the recipient before products can be transfused.

Administration of any blood product must be prescribed by a doctor and the recipient must sign a release form.
Why Artificial Blood?
Life Threatening Emergencies
Blood Must Be Donated
Potentially Contaminated
Donated Blood Has Short Shelf Life
There are Rare Blood Types
Clinically Significant Antibodies
Every 2 Seconds someone needs a blood transfusion.
<38% of the population are eligible to donate blood.
Only 7% of the people in the USA are O Negative.
There is a shortage of blood donors.
Blood is safer than ever before.
Donor blood is screened for viruses and infectious diseases but there is always the potential for contamination.
Blood is tested for: HBsAg, anti-HBc, anti-HCV, anti-HIV, and more.
Emergencies occur away from the hospital. Many lives could be saved if patients could be transfused while en route to definitive care.
Packed RBCs have a shelf life of only 42 Days.
Irradiated RBCs have a shelf life of 28 days.
Washed RBC have a shelf life of 24 hours.
Must be stored at 1-6 Degrees Celcius.
By coming into contact with another person's blood you could form antibodies to foreign antigens present on their cells.
Hemolytic Transfusion Reactions are the result.
Some blood types are more rare than others.
Not everyone can receive O Negative blood.
Desired Characteristics
Efficiently Transport Oxygen
Disease Free
Universally Compatible
Manufactured blood will be free of viruses and infectious agents.
Artificially made blood will be antigen free eliminating the need for cross matching and worrying about different blood types. One substance will be truly universally compatible.
In order to compete with allogenic blood, Artificial Blood will have to exchange Oxygen and Carbon Dioxide efficiently.
Stored at Room Temperature
Long Shelf Life
Without risk of Heart Attack or Stroke
Artificial blood must be able to be mass produced and widely available. Also extending its shelf life past the 42 day limit for Donor RBCs will be greatly beneficial.
Early attempts at making artificial blood were stopped because of these serious side effects. A transfusion should not increase the risk of heart attack or stroke.
Artificial blood that can be stored at room temperature can be carried in Ambulances and be easily administered in the battlefield.
Jehova's Witness Patients
Religious beliefs do not permit transfusion of blood components.

Bible = "abstain from ... blood" (Genesis 9:4-5)

In 2010 a Patient was diagnosed with Acute Lymphocytic Leukemia ALL.

She was treated with Hemopure (HBOC-201) a polymerized bovine Hb-derivaed artificial oxygen carrier without use of blood transfusions.
Artificial Oxygen Carriers
AOCs = Synthetic solutions with the ability to bind, transport, and unload oxygen in the body.

Function as alternatives to blood transfusion in an attempt to avoid, reduce, or delay transfusion of allogeneic blood.

Improvement of tissue oxygenation of organs with poor blood supply.
AOCs
HBOCs
PFCs
Human
Animal
Recombinant
Perfluorocarbons are liquid fluorinated hydrocarbon compounds capable of carrying dissolved oxygen and delivering that oxygen under physiologic conditions.
History
Research began in 1960s

Clark and Gollan found that mice submerged
in the substance could exchange Oxygen and
Cabon Dioxide in the fluid.
PFC Solutions
PFCs are modified hydrocarbons which
do not mix well with blood.

They need to be emulsified with lipids or oils
to bring them into solution.
Henry's Law
The amount of oxygen dissolved in PFCs is directly proportional to the partial pressure of oxygen delivered and the solubility of oxygen in the PFCs.

Best results are obtained if the patient is breathing 100% oxygen at the time of infusion
Long Term Effects
PFCs have a prolonged presence in the body.

The president of the fluorocarbon-producing company injected himself with 20 mL of the compound.

He died from chemical pneumonitis 1.5 years later.
hHb is produced by lysis of RBCs obtained from outdated banked blood.

Requires human donors which may be in short supply.
Animal hemoglobin is obtained from cows.

Possible transmission of animal pathogens, specifically bovine spongiform encephalopathy.

Forty units of hemoglobin solution can be obtained per slaughtered cow.
= 40 Units
Recombinant hemoglobin is obtained by inserting the gene for human hemoglobin into bacteria and then isolating the hemoglobin from the culture.

This process allows for the manipulation of the gene itself to create variant forms of hemoglobin. One unit of hemoglobin solution can be produced from 750 L of Escherichia coli culture.
Oxygen Dissociation
Oxygen Dissociation
Native Hgb P50 of 26 to 28 mm Hg

hHb has a P50 of 10 to 16 mm Hg
Polymerized bovine Hb-derived artificial oxygen carrier
On Day 6 of chemotherapy the patient complained of mild dizziness.

She was found to be hypotensive 80/50.


Day 7
Hb = 4.5 g/dL
PLT < 30 x 10 / L
Antifibrinolytic therapy for 13 days
Persistent hypotension and tachycardia

Vaginal bleeding


Hb as low as 3.1 g/dL
Fifteen units of Hemopure were administered over a 12-day period after informed consent was obtained.

Hb level was maintained between 3.6 and 5.3 g/dL

Hemopure was providing adequate tissue oxygen supply until marrow red blood cell recovery.

The patient completed the chemotherapy regimen and was dis- charged in stable condition and has been in remission for 28 months.
9
No fetal heart tones = spontaneous miscarriage.
Thank You
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