Cryogenic Applications in the Field of Medicine

Submitted by Nikhil V V 11TH10F

Contents
Cryobiology Cryonics Cryopreservation Cryopreservation by vitrification MNT and cryopreservation revival Cryotherapy and surgery Selected References

Cryobiology
-The study of the effects of subfreezing temperatures on biological systems. Technically, cryobiology is the study of living systems at any temperature below the standard physiological range. Some cryobiological terms associated with temperature below 0 C are: a) Cold hardiness and sensitivity in nature: eg:- certain tree twigs, freezable frogs, insects, polar fish etc. b) Freeze-drying cooled so rapidly that ice doesnot form or forms such small crystals that will not damage cells c) Super-Cooling- Whole body cooling with out freezing, cooling in tiny droplets of water to temperatures several degress below the freezing point. Enzyme denaturation and protein folding occurs which is reversible.

d)
e)

f) g)

Cryosurgery- the surgery involving the selective destruction of tissues by freezing them, as with liquid nitrogen. Frostbite- slow freezing process but no cryoprotecting agent available. Cryopreservation Cryopreservation by vitrification

Cryonics

Cryonics is a legitimate science-based endeavor that seeks to preserve human beings, especially the human brain, by the best technology available. Future technologies for resuscitation can be envisioned that involve molecular repair by nanomedicine, highly advanced computation, detailed control of cell growth, and tissue regeneration. With a view toward these developments, there is a credible possibility that cryonics performed under the best conditions achievable today can preserve sufficient neurological information to permit eventual restoration of a person to full health.

 Cryonics community research in two critical areas: better methods of cryopreservation and reviving of cryopreserved patients. This quest for better cryopreservations continues today, and will continue until some future day when fully reversible cryopreservation becomes possible. The development and application of new technologies to allow the persons body to be restored to complete health. One of these new technologies is MNT-molecular nanotechnology.

Cryopreservation
Today's scientists are faced with the task of genetically stabilizing living cells. Serial subculturing is time consuming and can lead to contamination of genetic drift as smaller and smaller portions of population are selected. However, a population of cells can be stabilized subjecting them to cryogenic temperatures. Stabilizing cells at cryogenic temperatures is called cryo preservation. Advances in cryopreservation technology have led to methods that allow low-temperature maintenance of a variety of cell types. Techniques are available for the preservation of microorganisms, isolated tissue cells, small multicellular organisms, and even more complex embryos.

Steps in Cryopreservation
Cryopreservation steps just starts after the legal death of a patient. Cooling is the first process. The patient is lowered into an ice bath, and as soon as possible, circulation and breathing are artificially restored by a Heart Lung Resuscitator (CPR machine). Intravenous lines are established, and special medications are administered to protect the brain from any lack of oxygen. The patient's blood is circulated through a heart-lung machine. Cryoprotectant gradually replaces the patient's blood. The solution containing glycerol (an antifreeze agent) is to protect against freezing injury. A heat exchanger greatly accelerates the cooling to the freezing point of water. Once perfusion (circulation) of this cryoprotectant solution is complete, the patient is immersed in alcohol for cooling down to -79 degrees Centigrade (dry ice temperature) over the next 48 hours.

 Following initial transport and perfusion, patients are transported at dry ice temperature to the long term care provider of their choice. They are then slowly cooled over a two week period to the temperature of liquid nitrogen (-196 degrees centigrade), and immersed in liquid nitrogen for long term care. At this temperature all biological processes are stopped, and tissue remains unchanged indefinitely. With future advanced medicine, the outer limits of resuscitation could be greatly lengthened. It is for this reason that cryonics is still applied to patients who are far beyond the reach of today's medicine.Many patients whom we would call dead today are likely to be reclassified as seriously ill, but treatable in the future.

Wash out and perfusion process.

Cryopreservation by vitrification
Cryopreservation by vitrification partially replaces water inside cells and tissue with chemicals called cryoprotectants that prevent ice formation. At high enough concentrations, cryoprotectants can prevent freezing. Instead of freezing, the mixture of water and cryoprotectants becomes more and more viscous like syrup during cooling, and eventually solidifies. Solidification without freezing forms the basis of cryopreservation by vitrification. The perfect storage environment for cryopreserved patients is provided using liquid nitrogen.Unfortunately the process of cooling to this very cold temperature tends to cause cryopreserved tissues to fracture. Even with use of cryoprotectants it is not possible to achieve fractureless cryopreservation.

A two litre volume of solidified vitrification solution shown (a) just below the glass transition temperature and (b) after further cooling. The solidified solution fractured during further cooling below the glass transition temperature. ______________________________________

Differential thermal contraction of vitrification solution cooled in a test tube causes a dimple to form. The warmer inside of the solution continues contracting after the colder outside has solidified and begun cooling at a slower rate.

Vitrified brain of Alcor patient A-2077 under liquid nitrogen. This brain is almost certainly fractured, yet it remains an integrated whole. Movements between fracture planes appear to remain microscopic provided that tissue stays cold and solid.

MNT and Cryopreservation Revival

Molecular nanotechnology is an emerging technology for manufacturing and manipulating matter at the atomic level. The concept was first suggested by Richard Feynman in 1959, and brought to fruition by Dr. K. Eric Drexler, Dr. Ralph Merkle, and others in the 1980s and 1990. Molecular nanotechnology is the most compelling approach ever put forward for comprehensive repair of cryopreservation injury with maximum retention of original biological information. Molecular nanotechnology is the anticipated future ability to manufacture products by inexpensively arranging atoms in most of the ways permitted by physical law. Medical nanodevices are used to repair the cryopreserved biological structure.

Nanobots replacing old damaged chromosomes with new ones in each cell.

Cryotherapy
Cryotherapy is the ablation of tissue by local induction of extremely cold temperatures.The advent of probes cooled by liquid nitrogen in closed circulation marks the beginning of modern cryotherapy. One of the first applications of this new technology was the transurethral cryoablation of benign prostatic hyperplastic tissue. A significant recent development was the introduction of cryotherapy probes that use argon gas rather than liquid nitrogen. Argon rapidly cools the probe tip to -187C (-304.6F) and can be rapidly exchanged with helium at 67C (152.6F) for an active thawing phase. The use of cryotherapy for prostate cancer(cryosurgery), have included the use of real-time transrectal ultrasonography (TRUS) monitoring of probe placement and freezing.

Cryosurgery
Cryosurgery is the freezing of targeted cancer tissues. During cryosurgery, extremely cold temperatures are applied to targeted cancerous tumors in order to destroy them. This flash freezing process is known to cause all of the following events on the cellular level: osmotic stress, denaturation of macromolecules, the release of lysosomal proteases, membrane disruption, and intracellular ice formation. For the treatment of prostate cancer, cryosurgery involves the use of a unique temperature monitoring system with a six- to eightcryoprobe device, in combination with ultrasound in order to precisely freeze and destroy cancerous tissue. A thin catheter that circulates warm fluid is placed in the urethra to protect it from cold temperatures.

 Slender cryoprobes are inserted through a small incision. The probe is placed on the outside of the tumor. Liquefied argon gas is contained within the cryoprobe tips. Argon is used instead of liquid nitrogen because it allows one to freeze the cancerous cell instantaneously and achieve faster and colder freezing. The cryoprobe reduces the temperature to -40 degrees Celsius, freezing cancer tissue. After about ten minutes, this process is complete. The physician immediately administers another freeze-thaw cycle in order to ensure that all cancer cells are killed.

Cervical Cryotherapy
Cervical cryotherapy is a procedure which involves freezing an area of abnormal tissue on the cervix. This tissue gradually disappears and the cervix heals. One cervical cryotherapy is usually sufficient to destroy the abnormal tissue. Cervical cryotherapy is a standard method used to treat cervical dysplasia, meaning the removal of abnormal cell tissue on the cervix. Cervical cryotherapy is done by placing a small freeze-probe (cryoprobe) against the cervix that cools the cervix to sub-zero temperatures. The cells destroyed by freezing are shed afterwards in a heavy watery discharge. The main advantage of cryotherapy is that it is a simple procedure that requires inexpensive equipment.

Conclusion
Though significant advances have been made in the fundamental cryobiology it is still in a developing stage. Successful cryopreservation techniques and cryosurgical methods are yet to be found out. With the advent of new technologies like MNT revival of cryopreserved patients will also be possible.

Churchills commentary on man: Man will occasionally stumble over the truth, but most of the time he will pick himself up and continue on

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