A solution and a way to revitalize and restore the ischemia damaged tissue

 

(57) Abstract:

The invention relates to medicine, namely to the conservation, preservation and restoration of tissues and organs. How is the washing of the body at a temperature of 28 - 37oWith buffer containing saline solution to remove blood and acidotic products and perfusion of the organ at a temperature of 28 - 37oWith buffer containing saline solution, which further comprises means for expansion of blood vessels, trophic factors, means to restore oxidative metabolism and other substances. The proposed method allows to preserve tissues and organs for transplantation for a longer time. 8 C. and 22 C.p. f-crystals, 5 Il., 6 table.

The scope of the invention

The present invention generally relates to the preservation, conservation and restoration of tissues and organs. More specifically, the present invention provides a method by which, using the composition of the present invention, restores the integrity, function and viability of the ischemia damaged tissue or organ.

The background to the invention

The shortage of organs for transplantation OST the plantation remains constant shortage of organs. For example, kidney transplantation is highly dependent on the availability of organs from deceased donors while still working heart. In addition, a large and not yet closed source of organs for transplantation remain the corpses with a broken heart. The corpses with a broken heart are victims of accidents who have died directly from the damage to, or lived a short time after injury. In such cases, the reasons that these bodies are not used, is due to the cessation of cardiac activity, the lack of blood circulation (warm ischemia), which triggers a cascade of damage.

The authority, seriously damaged by warm ischemia, but functionally still saved, cannot move further damage during hypothermia. In hypothermia, which is used for preservation of organs intended for transplantation, lipid bilayer membranes change phase and become similar to gel, with a significant decrease in yield. Hard frozen lipid in cell membranes is not possible to utilize oxygen, even in the presence of high pressure O2. Metabolic consequence of this is glycolysis, which is similar to the condition of hypoxia. Describe the utilization of oxygen makes up approximately 5% from the norm.

Hypothermic storage can also cause vasospasm and subsequent swelling of the body. Have stored in hypothermia bodies may receive swelling of glomerular endothelial cells and the integrity of blood vessels with tubular necrosis; there is a phenomenon inherent in hypothermic conditions. Hypothermia may also inhibit Na/K-dependent ATPase and lead to the loss of the ability of cells to regulate the volume. It is the loss of regulation volume causes swelling and damage to the cells. Sufficient supply of oxygen can actively reduce the degree of swelling. Without adequate delivery of oxygen, hypoxia leads to disintegration of small vessels after a few hours of perfusion. The lack of oxygen and the subsequent depletion of ATP means that anaerobic glycolysis is the main energy source in the traditional terms of conservation. The lack of molecular oxygen for oxidative phosphorylation, which occurs during ischemia, leads to the accumulation of NADP and the depletion of ATP in the mitochondria. Subsequent loss of nucleosides is, apparently, a very important factor in the inability of the tissues subjected to warm ischemia and prolonged predicate supply of oxygen is to rely on routine hypothermia for the conservation of organs.

Thus, ischemia (warm or cold) is a cascade of damage and it can be characterized as prenatally phase and death phase. Proletarka phase damaging effect in three ways: by hypoxia, malnutrition and the inability to remove toxic metabolic wastes. With the cessation of blood circulation stops the flow of molecular oxygen. The onset of hypoxia causes energy depletion, such as the depletion of ATP in the mitochondria. Depletion of ATP leads to cellular changes, including swelling, loss of the normal integrity of the cells and loss of polarity of the membrane. Cellular changes cause lethal phase of ischemia, leading to the accumulation of metabolic waste, activation of proteases and cell death.

Modern perfusion solutions, representing the present state of Affairs in question hypothermic preservation of organs, and delivered for optimal conservation bodies in hypothermia, contain components that prevent tissue swelling, induced hypothermia; metabolites that facilitate the functioning of the organ after transplantation; antioxidants; stabilizers of membranes, colloids, ions and salts (Southard et al., 1 hypothermic induced suppression of metabolism. While it minimizes edema and vasospasm, usually observed during hypothermic storage, it is not intended for use with significantly expanded donor pool.

This is due to the fact that organ or tissue, severely damaged by warm ischemia, but functionally still saved, cannot move further damage during hypothermia. Even if ischemia lasted only 30 minutes, organ function after transplantation may be compromised. For example, when using organs taken from dead bodies with working heart, the frequency of non performance of the bodies immediately after transplantation is approximately 25%, and after 30 minutes of ischemia increases to approximately 60%. Therefore, 60% of kidneys taken from the dead with a broken heart, don't start to work due preleasing ischemic damage. In addition, I believe that the authorities deprived of blood flow for a few hours or less, there is irreversible ischemic damage (Klutz et al., U.S. patent 5395314). Until you have developed new sources of organs for transplantation, the number of transplants will remain constant. In addition, the donor pool cannot be sufficiently expanded, posoltega, damaged by warm ischemia.

Recent attempts have focused on the prevention of ischemic damage by reviving organs of the reperfusion solution immediately after the cessation of blood supply. For example, a protective solution, described in U.S. patent 4415556 used for surgical operations or organs intended for transplantation, to prevent ischemic organ damage. This protective solution is used as perfusion to improve aerobic metabolism during perfusion of the organ. U.S. patent 5395314 describes how to revitalize the brain by circulation after cessation of blood flow through the brain hypothermic preservative solution (about 8-10oS), are designed to lower the metabolism in the body to deliver oxygen and to inhibit free radical damage.

Despite the fact that these methods and preservative solutions are suitable for the prevention of ischemic damage to organs, these benefits are shaded practical and functional deficiencies. First, in order for these methods and solutions were effective to prevent ischemic damage, they must be applied not what you learn, when a donor organ is the victim of an accident, can severely limit the application of such methods and solutions that are practical only in hospitals. Secondly, believed to be irreversible ischemic damage observed in the organs deprived of their blood supply, within minutes (e.g., brain) or within a few hours (heart, kidney). Organ or tissue damaged by warm ischemia, but functionally still saved, cannot move further damage during hypothermic storage prior to transplantation or resumption of blood flow after transplantation. One reason is that the restoration of blood flow after ischemia-reperfusion, paradoxically, may lead to further tissue damage (McCord et al. , 1985, N Engl. J. Med. 312-159-163). The restoration of blood circulation causes reoxygenation damaged tissue. Reoxygenate ischemia damaged tissue can lead to further tissue damage due to the formation of free oxygen radicals, depletion of scavengers of free radicals and the release of chemotactic agents.

Thus, a need exists for a method and solution that could priodol restorative processes in organs or tissues at very early stages of lethal ischemia. Method of induction of ischemia recovery of damaged organs or tissues to such an extent that the dysfunction may be reversible, thus preventing further tissue damage during the resumption of blood flow can lead to a substantial expansion of the donor pool.

A brief description of the nature of the invention

The present invention is directed to the fact that up to the time of occurrence of the present invention was considered to be irreversible ischemic damage to organs or tissues deprived of blood supply. Method and compositions are used after ischemic damage to the induction of ischemia recovery of damaged organs or tissues and prevent further tissue damage during the resumption of blood flow. This distinguishes the method and compositions of the present invention from used in the present methods and compositions designed for use up to ischemic damage, with the aim of preventing or inhibiting such damage. The method of the present invention is a method by which you can restore the integrity and function of the ischemia damaged organ or tissue during krainer and the solution of the present invention are designed to prevent or inhibit further tissue damage, which can occur when the resumption of blood flow in an organ or tissue deprived of blood supply.

The method of the present invention includes washing the body through the arterial system recovery solution of the present invention under conditions of high temperature, approximately 28oWith up to 37oWith, to remove blood and acidotic products that have accumulated in the organ or tissue during the period of absence of blood flow, and perfusion washed organ or tissue recovery solution with the aim (1) to expand the blood vessels, especially spastic microvessels in the organ or tissue; (2) to restore the function of an organ or tissue through the supply of trophic factors; (3) to restore the integrity and function of cells in the ischemia damaged tissue or organ; and (4) to restore oxidative metabolism by rehabilitation ischemia damaged organ or tissue, surviving through anaerobic respiration, to oxygeneration recovery solution; making organ or tissue suitable for transplantation and/or to restore blood circulation.

Brief description of drawings

Fig. 1 is a block diagram showing processes in the body, on AET graph of the function parameter of the body (serum creatinine) from the number of days, since transplantation, allotransplantation of the dog, when using the method and recovery solution according to the present invention.

Fig.3 is a graph of the dependence parameter in the function body (creatinine urine) from the number of days that have elapsed after transplantation, allotransplantation of the dog, when using the method and recovery solution according to the present invention.

Fig.4 is a graph of the dependence parameter in the function body (serum creatinine) from the number of days that have elapsed after transplantation, autologous transplantation in the dog, using the method and recovery solution according to the present invention.

Fig.5 is a graph of the dependence parameter in the function body (creatinine urine) from the number of days that have elapsed after transplantation, autologous transplantation in the dog, using the method and recovery solution according to the present invention.

A detailed description of the preferred embodiments

Definition

"Without blood" is the term used for the purposes of the description and claims, which refers to the cessation of blood circulation through the body or TC the minute circumstances include termination heartbeat for surgical procedures or due to natural causes, such as heart attack.

"Organ or tissue" is the term used for the purposes of the description and claims, which belongs to the kidney, heart, liver, lung, small intestine, pancreas, brain, eye, and skin, but is not limited to them.

"Recovery solution" here is a term applied to the purposes of the description and claims, refers to a physiological buffer solution, providing the means to restore the integrity and function of the ischemia damaged organs are deprived of blood flow, and for preventing or inhibiting further tissue damage that can occur when restoring blood circulation through the body, deprived of blood flow.

The method of the present invention is a method through which can be restored integrity and function of the ischemia damaged body, at least during the phase priletali ischemia, using restorative solution of the present invention. Restore the integrity and function of organs using the method and compositions of the present invention was unexpected, since the time of creation of this izobreteny is whether less irreversibly. In addition, the method and the solution of the present invention are designed to prevent or inhibit further tissue damage that can occur during recovery of blood circulation in the body, deprived of blood flow.

The method and the solution of the present invention provide a means to remove blood and acidotic products that have accumulated during the period of cessation of blood flow in the body; means for recovery of cellular integrity and function, restoring, thus, the function of the body; and means for re-adaptation of the body to an oxygenated environment. The ability to restore organ function after ischemic injury deemed possible, based on the following assumptions: (1) the blood does not coagulate, being in contact with viable vascular endothelial cells and, consequently, ischemia damaged organs can be reperfusion, ensuring the viability and intactness of the endothelium; (2) the restoration of vascular dynamics depends on ensuring adequate vasodilation is dependent on endothelial cells, with the aim of adequate perfusion and oxygenation of tissues and ensure the normal mechanisms of self-regulation; (3 what change, so that you can restore cell integrity; and (4) trophic factors that were lost during ischemia, should be replaced, and the polarity of the cells needed for normal operation should be restored.

The method and the solution of the present invention work together to revitalize the ischemia damaged body in order to restore the functions of the body and re-adaptation of the body to oxygenated environment. Fig.1 is a block diagram showing processes in the body, which operates the recovery process and the solution of the present invention.

The following examples illustrate the preferred options for practical implementation of the present invention. The following options exercise used to illustrate, it is important to take into account the following considerations. The model on the calf and the model on the dog were used to evaluate compositions and methods relating to the transplantation of organs intended for a person, because it has been shown that these models reflect a physiological basis. Thus, while the composition and method of the present invention were evaluated in these experimental models, the composition and method should use is Adela per person is well known to specialists, and includes taking into account differences such as the volumes of the organs and the size of blood flow through the organs (see, for example, Harrison et al., 1977, J. Pharm. Sci. 66:1679-1683). It should be understood that these examples are intended for illustration and not for limitation.

Example 1 - Way

The method of the present invention includes assistance in the absence of blood circulation in the body before you come significant cell death. The specialist will be clear that the period during which you can apply this method differs depending on the type of the processed body. For example, the period of time during which the processed heart using the method of the present invention, may be less than approximately one hour; while the kidney can be treated using the method of the present invention for approximately 4 hours to lack of blood flow. To stop the cascade of ischemic damage, leading to cell death, as shown in Fig.1, the method of the present invention involves the following stages:

(i) washing ischemia damaged body through the arterial system recovery solution of the present invention under conditions of high temperature, approximately 28oWith up to 37o
(ii) restore the cellular environment to physiological pH values;

(iii) adequate expansion microvessels;

(iv) the continuous anaerobic metabolism as the rescue process by providing a high-energy compounds and maintenance of glycolysis with additional substrates, which may include glucose, pyruvate and uridine-5'-triphosphate (UTP), but are not limited to;

(v) the initiation of the transition from anaerobic metabolism to oxidative metabolism by providing metabolic substrates to fill the pool Denisovich connections, maintaining the citric acid cycle and system recovery transport electron pairs, while molecular oxygen is injected slowly to avoid reperfusion injury mediated by oxygen toxicity;

(vi) a mechanism for adequate vasodilation microvascular bed of endothelial cells within highly spastic, edema, ischemia damaged organ, without significant changes in the permeability of the body; and this vasodilation makes possible adequate perfusion of tissue that creates a stable pressure perfusion, stabilin shall (i) provide trophic factors to restore the function of the ischemia damaged organ, providing, thus, the metabolites to restore cell integrity and function; and

2) perfusion ischemia damaged body through the arterial system recovery solution of the present invention under conditions of high temperature, approximately 28oWith up to 37oWith, for

(i) the normalization of oxygenlive, temperature and pH;

(ii) continue to provide a mechanism for adequate expansion of the vascular bed of the body, without significant changes in the permeability of the body, providing, thus, a stable perfusion pressure and stable flow speed of the fluid; and

(iii) continue to provide trophic factors to restore the function of the ischemia damaged body, providing, thus, the metabolites to restore cell integrity and functions, such as the strengthening of intercellular connections and restore the polarity of the membrane.

The specialist will be understood that one or more positive effects achieved at the stage of washing, will continue during perfusion, because the replacement solution of the present invention can be used throughout the process, i.e., including the stage of primavista recovery solution is slowly injected by injection through the cannula in the main for this particular body artery until until the resulting liquid will no longer contain blood. This way ischemic blood and acidotic products that have accumulated in the blood vessels over a period of time during which the body has been deprived of blood supply, are removed from the vessels. Further, the pH is recovered and delivered fresh substrate to maintain anaerobic metabolism and other pathways of metabolism in the cell, required for cell integrity and function. The specialist will be clear that the number of recovery solution, sufficient for washing, may depend on the type and size of the body, which is washed, and the duration of the period of time during which there was no blood. To illustrate but not to limit, from 200 to 600 ml recovery solution may be sufficient for washing a person's kidneys, which was absent blood flow within 1-3 hours.

For purposes of illustration, but not limitation, at the stage of perfusion sufficient recovery solution slowly perfusion when the systolic pressure, which is suitable for ischemia damaged organ, which enliven, until then, until they reach the speed of a liquid, which is priblizitelino was no blood flow within 1-3 hours you can slowly perfusionist recovery solution at a systolic pressure of <80 mm RT.art., as long as the flow speed of the fluid reaches a >50 ml/min. pH is normalized to the physiological limits by slow introduction of molecular oxygen through the oxygenator or through a substance, transporting oxygen, which is an integral part of the recovery solution. Oxygenation of the body during perfusion, as well as normalization of temperature and pH, there is approximately within the first 15-30 minutes of perfusion. Because the vessels of the body slowly expanding, the perfusion pressure and the flow rate of the fluid begin to stabilize, and the body quickly switch to oxidative metabolism. The specialist will be clear that the length of time required for perfusion depends on the particular type and size perfusing body, and the duration of the period of absence of circulation. However, treatment of ischemia damaged organ with the help of the method of the present invention (washing and perfusion) for approximately 2 hours may be sufficient for the revival of most organs (for example, deprived of blood flow during the period of time from 0.5 to 4 hours) the AET urine, this method can lead to the generation of products of the normal activities of the body.

The method of the present invention was developed for the preservation and revitalization of the ischemia damaged organs ex vivo without the use of traditional hypothermia (4-10oC). This method provides the necessary delivery of oxygen, nutrients for metabolism, oncotic pressure, pH, perfusion pressure and the flow rate of the fluid to maintain the metabolism of the organ ex vivo often within their respective normal values of these quantities in vivo or so. Near-normal metabolic rate here is defined as approximately 70-90% of the normal rate of metabolism. Next, the method of the present invention supports this level of metabolism ex vivo, which provides oxidative metabolism, sufficient to generate normal function of the product of this body. The development of this method, which supports organs ex vivo without the traditional hypothermia, provides the ability to maintain a near normal rate of metabolism and install functionality that can be correlated with postoperative or posttransplantation through the device, including laminar or pulsating pumping system for delivering recovery solution, including tools for security and control of perfusion and perfusion pressure; means for temperature control and means to ensure and control the introduction and removal of respiratory gases. A similar device is described by the author of the present invention in the patent application U.S. 08/246801, which is incorporated herein by reference. Such a device may also include means for testing and/or collection of the perfusion solution, which has already passed through the body to the monitor, and measure one or more functional characteristics, such as pH, pressure, flow speed of the fluid, vascular resistance, various chemical constituents, oxygen therapy, the concentration of carbon dioxide and oxygen. In addition, you can use another device or a second device in connection with the first, to identify and/or collect the product produced by the body, such as urine produced by the kidney; and the subsequent determination of the parameters of this product, produced by the body, may be correlated with the integrity and function of the body during or after application of STRs and perfusion of the organ known as the solutions includes basic solution, which consists of a buffered saline solution, such as saline solution or basic environment, such environment for cell cultures, to which is added a variety of additional substances. In a preferred embodiment, the regenerative solution of the present invention also includes such basic solution containing amino acids, ions, physiological salts, non-penetrating substances, serum proteins and/or factors and sugar. In addition to the main components of the solution, restorative solution of the present invention provides a new combination of additional substances that can be grouped in at least 3 categories of components. The specialist will be clear that the components of each category can be replaced functionally equivalent compound to achieve the same result. Thus, the following types of components each component categories serve the purpose of illustration, but not limitation.

The first category of components, vasodilators, includes a combination of components in a physiologically effective amounts, which provide the means for adequate expansion of krupnorogatogo. In order to ensure the normal permeability of the vascular bed, vasodilatation control conditions based on endothelial cells. This combination of components can include (1) the substrates for vasodilation, mediated by endothelial cells, such as acetylcholine, dopamine, bradykinin and arginine; (2) substrates for the expansion of the microvessels, such as prostacyclin (and analogues, for example, carbacyclin) and mg+; and (3) adenosine (and analogues, for example, cyclohexylbenzene), and verapamil for their combined effects on vasodilatation, mediated by blocking calcium channels (other calcium channel blockers include flunarizin, nifedipine. SNX-11, chlorpromazine and diltiazem). The result of applying this combination of vasodilators is that the bloodstream is well expanded, while maintaining its integrity and normal barrier function. Vasodilators approximately from 1 to 50% (weight/volume) of the volume of the new combination of additional substances, which is added to the basic solution in the preparation of the regenerative solution of the present invention.

The second category of components, chemical energy substrates, which I restore oxidative metabolism, lost during the period of absence of blood flow. During the period of absence of blood flow resulting loss of integrity of the membrane leads to loss of intracellular components, such as ions, the components of the pool Denisovich compounds, citric acid cycle and the transport chain electron pairs. Such chemical energy substrates added in the reducing solution may include pyruvate, glucose, ATP, AMP, coenzyme A, -dinucleotide (NAD+-adenine dinucleotide phosphate (NADP+), flavine adenine dinucleotide (FAD), tiaminpirofosfat chloride (kokarboksilazu), uridine-5'-triphosphate (UTP), chloride, adenosine, magnesium, and combinations thereof. If the supply of energy in the cells of tissues restored to cell death, cellular changes that occurred in the absence of blood flow, may be reversible, and cell volume of the tissue returns to normal. Chemical energy substrates comprise from about 0.01 to 90% by volume of a new combination of additional substances, which is added to the basic solution in the preparation of the regenerative solution of the present invention.

The third category of components, trophic factors, includes Comodoro or more recovery processes in the cells, in order to restore cellular function, lost in the absence of blood flow. This combination of trophic factors provides a means to accelerate protein synthesis that leads to the restoration of tight intercellular connections and to regenerate the polarity of the membrane, restoring thus the function of cells. Such trophic factors added in the reducing solution may contain high concentrations of amino acids and magnesium (e.g., 2-6 times greater than normal concentrations in plasma), derivatives of nucleic acids and ribonucleoside; and growth factors with potentiators membranes, such as acidic fibroblast growth factor (FGF), basic FGF, heparin and chondroitin sulfate, and combinations thereof. These trophic factors comprise from about 1 to 90% by volume of a new combination of additional substances, which are added to and dissolved in the basic solution in the preparation of the regenerative solution of the present invention.

The specialist will be clear that the components of one or more of the above categories may have additional functions that are desirable for the method of the present invention. For example, magnesium ions (incorporated as part of the connection, steriade trophic factor, and as chemical energy substrate. In addition, in the preferred embodiment, the amino acids contained in the reducing solution, include cystine and cysteine in amounts that, in addition to functioning as trophic factors that also act as antioxidants - selective scavengers of free radicals, which absorb toxic free radicals during the stages of washing and perfusion of the present method. Other antioxidants, such as glutathione, cyclodextrin, superoxide dismutase (SOD), catalase, chlorpromazine and prostacyclin may also include, or be used as functionally equivalent compounds in a reducing solution of the present invention. These antioxidants are approximately 0,000% to 10% by volume of a new combination of additional substances, which are added to and dissolved in the basic solution in the preparation of the regenerative solution of the present invention.

In another embodiment of the present invention, in which the cloth that you want to animate using restorative solution of the present invention is nervous tissue (e.g. brain), recovery the ion channel of the NMDA receptor, for example, Aptiganel and Cerestat; blockers glycine sites of NMDA receptors, for example, ZD 9379 and GV 150-562A), blockers accumulation of nitric oxide (NO) (for example, lubeluzole) and blockers of sodium channels to inhibition of the influx of sodium into the cells, which can trigger the release of glutamate (for example, BW619-C89, fosphenytoin).

In another embodiment of the present invention, in addition to the introduction of molecular oxygen through the oxygenator, the recovery solution contains one or more oxygen-carrying compounds ("oxygen-carrying agents") that supply molecular oxygen for oxidative metabolism in ischemia damaged body. Such carrying oxygen agents known in the art and include hemoglobin, stable derivatives of hemoglobin (made from hemolyzed red blood cells of a person, such as paradoxrecovery hemoglobin), polyoxyethylene conjugates (PHP), recombinant haemoglobin products, performance (PFH) of the emulsion and/or performance micro-bubbles (here the common name "performance agents"), but are not limited to. These oxygen-carrying agents are approximately 0,000% to 50% of the volume novostroitelnaya solution of the present invention; or approximately 0,000% to 20% of the total recovery solution (about./vol.).

Emulsion PFH, suitable as oxygen-carrying agents are described, for example, in U.S. patents 5403575, 4868318, 4866096, 4865836, 4686024, 4534978, 4443480, 4423077, 4252827, 4187252, 4186253, 4110474 and 3962439. Such liquid emulsion PFH include performancebased, perforative, bromberger, perforator, Fluosol DATMF-44E, 1,2-bisphthalocyanine, F-4-metalictastelipitor, performin from 9 to 12 carbon atoms, perpendicular, pertarungan, parttorteneti bicyclo [3, 3, 1]Onan, performancemanagement, performimplemented, but are not limited to. Micro-bubbles PFH, suitable as oxygen-carrying agents are described, for example, in U.S. patents 5409688 and 5393524. Agents PFH, which are described as suitable for making such a microbubble include, but are not limited to, dodecaborane (DDFP), sulphur hexafluoride, pentane, HEXAFLUOROPROPYLENE, OCTAFLUOROPROPANE, freon, Octaver-2-Butin, hexaferrite-1,3-diene, isoprene, OCTAFLUOROCYCLOBUTANE, deceptibot, CIS-2-penten, dimethyldisulfide, atrasin, bremgartner, TRANS-2-penten, 2-chloropropane, geksaftorbenzole, ethyl mercaptan, diethyl ether, ethylenically ether, the IDF, Isopropylamine, methylformate, 2-acetylfuran, telephoned, 1-penten, isopropylacetate, performante, isopentane, vinyl ether, 2-Butin, 1,4-pentadien, tetramethylsilane was, dimethylphosphine, dibromodifluoromethane, 2-chloropropene, diftormetan, acetaldehyde, trimethylboron, 3-methyl-2-butene, 1,1-dimethylcyclopropane, aminoate, vinylboronic, decelerometer, Trichlorofluoromethane, Braverman, trifenilamin, performance and other fluorinated hydrocarbons (U.S. patent 5409688).

In the manufacturing process of the regenerative solution of the present invention to the basic solution is added and dissolved therein a new combination of additional substances, which can be divided into 3 categories, including vasodilators, chemical energy substrates and trophic factors. Despite the fact that the composition of the recovery solution for use in the method of the present invention may vary according to the content of the components and their quantity, as described above, the preferred composition is given in table 1 for purposes of illustration, but not limitation (note that for clarity, the component that can function in more than one of the at least three categories, placed in one category, below).

As mentioned, in another embodiment, the recovery solution can also include more antioxidants and one or more oxygen-carrying agents, as follows (per liter of the basic solution):

Antioxidants - Number

Glutathione - 0.1 mg

Cyclodextrin 500 mg

Oxygen-carrying agent performance agent - 20 about.

Example 3 - Effect of cessation of blood flow

Experiments were carried out, showing the influence of warm ischemia, caused by the cessation of blood flow in the body for approximately 30 minutes. This warm ischemia leads to rapid violation of the integrity of cells. The cascade of ischemic damage begins with the loss of pool Denisovich compounds, which leads to swelling. Loss of integrity of the cells and the appearance of edema leads to vascular collapse and disruption of normal vascular permeability. You can see that in the body, such as kidney, ischemic damage caused by the cessation of blood flow in the kidney only 30 minutes, causing the tsya insufficient for adequate perfusion of the kidneys. High vascular resistance in spastic vessels leads to further deterioration of the secondary hypoxia and thus to the loss of functionality (i.e., to stop production of urine). Table 2 illustrates the comparison of perfusion characteristics (pressure, flow speed of the fluid and vascular resistance) and organ function (urine output) in experimental animal models, including kidney calf, in which the blood flow is not stopped ("normal") and including kidney calf, in which the blood flow was stopped 30 min ("ischemic"). Vascular resistance is the average pressure/medium flow speed of the fluid.

Example 4 Effects of the method of recovery and recovery solution.

Experiments were conducted to demonstrate the ability of the method of recovery and the recovery of the solution of the present invention to overcome, and not only to inhibit the effects of warm ischemia in organs, and to support the recovery process to such an extent that it is possible restoration of disturbed functions of the body. Kidneys were removed from dead calves cattle. After 30 or 60 minutes of a lack of blood flow to the kidney was removed median times the control kidney was deprived of blood flow for 30 minutes, being thus, in this period of ischemic damage. After removal of the control kidneys were washed in 100 cm3the primary environment for cell cultures at a temperature of 32oWith, so that the kidneys were washed from the blood remaining in the bloodstream. Each experimental kidney was deprived of blood flow for 60 minutes, being, thus, in this period of ischemic damage. After removal of the experimental kidney was washed 100 cm3restorative solution of the present invention at a temperature of 32oC. After washing through these buds were pumped using a modified system for the conservation of organs during transport MOX-100TM. The control kidney was pumped when the 32oUsing previously known techniques for the conservation of organs with the help of technology, warm preservation, using as a perfusion solution of a basic environment for cell cultures. The experimental kidney was pumped, using the method and recovery solution of the present invention, when the 32oC. Comparison of perfusion characteristics (pressure, flow speed of the fluid and vascular resistance) and organ function (urine output) in the control group (30 minutes of ischemia) and in asasa ischemic damage in one hour, which were then animated using the method and recovery solution of the present invention, showed perfusion characteristics (pressure, flow speed of the fluid and vascular resistance) and organ function (urine output) functional limits of normal kidneys, are presented in table 2. Thus, the ability of the method and recovery solution of the present invention to overcome, and not only to inhibit the effects of warm ischemia in organs, and to support the recovery process to such an extent that it is possible to recover the function of an organ.

Example 5 - the Effectiveness of different duration period damage

Experiments were carried out to assess the effects of the method and recovery solution of the present invention in the bodies, which were deprived of blood flow for periods of time more than 1 hour. Kidneys were removed from dead calves cattle through different periods of time of absence of blood flow, including 60 minutes, 90 minutes, 2 hours or 4 hours. Prior to the removal of the kidneys did not take any effect, including the introduction of anticoagulants. Each kidney is then washed 100 cm3recovery solution neither in the bloodstream. Blood appeared in the remaining liquid, since she was in contact with viable endothelium of blood vessels. After washing these kidneys was pumped for several hours, using the method and recovery solution of the present invention, when the 30oC. Comparison of mean perfusion characteristics (pressure, flow speed of the fluid and vascular resistance) and organ function (creatinine concentration in the urine - creatinine clearance; histology) in the kidneys, deprived of blood flow for 60 minutes (60'), in the kidneys, deprived of blood flow at 90 minutes (90'), in the kidneys, deprived of blood flow for 2 hours (120'and kidney deprived of blood flow for 4 hours (240') are shown in table 4.

The results indicate that the method and recovery solution of the present invention can revive the ischemia damaged body, deprived of blood flow for the period at least up to 4 hours. For example, when the kidney undergoing warm ischemic damage within 60 minutes, pumped for 2 hours, using the method and recovery solution of the present invention, perfusion characteristics equivalent to those observed in normal kidney, as shown in the table and, the morphology and integrity of the well preserved.

When the deprivation of blood flow at 90 and 120 minutes of the kidneys reflect more extensive cellular damage (i.e., increased diastolic pressure and reduced velocity of a liquid) compared with the kidneys, deprived of blood flow for 60 minutes. However, despite such damage cells, these buds were developed urine, and histologically they seemed well-preserved. In addition, these kidneys was not observed necrosis.

In the kidney, deprived of blood flow for 4 hours, at a significantly reduced rate of power fluid and a concomitant increase in diastolic pressure, which includes a spasm of the vascular bed. However, it is important to note that these kidneys are still functioning. Urine was developed with creatinine 23,5 mg/DL. Histologically in these kidneys was observed the first signs of pitting early necrosis. Adjacent to the foci point tubular necrosis was observed signs of mitotic activity, indicating the start of active rehabilitation process. Thus, even after 4 hours of deprivation of blood flow was shown by the ability of the method and recovery solution this is the SS recovery to such an extent, when the dysfunction may be reversible.

It is important to note that control kidney (without processing method and recovery solution of the present invention) were evaluated histologically after 2 or 4 hours of the absence of blood flow in order to determine the relative advantages of this method and recovery solution. Histological assessment of control kidneys subjected to 2-hour warm ischemia, showed early diffuse tubular necrosis. After 4-hour warm ischemia in the control kidney was observed diffuse destruction of the cells of the tubules. In contrast, in the kidney subjected to 2-hour warm ischemia, and then processed using the method and recovery solution of the present invention, there has been a restoration of the integrity of the cells. Further, in the kidney subjected to 4-hour warm ischemia, and then processed using the method and recovery solution of the present invention, was observed only point tubular necrosis, compared with widespread damage to the tubules in the control kidney. Histological evaluation subsequently confirmed the significant effectiveness of the method and recovery solution of the present invention in tonirovannogo body in vivo

A. Allotransplantation

Body reanimated by using method and recovery solution of the present invention, was evaluated for function in vivo after resuscitation. Was performed allotransplantation dog; kidney took the dog to a donor after 60 minutes after the post mortem of circulatory arrest; washed the body restorative solution of the present invention; and perfesional it within 2 hours, using the method and recovery solution of the present invention, at a temperature of 30-32oC. After recovery kidney then put the dog to the recipient with simultaneous bilateral nephrectomy kidney recipient. Thus, the survival of dogs recipient depended on reanimated kidneys. During the post-transplant period the dog-recipient shown in Fig.2 and 3.

Kidney well repertoires and worked out the urine within two hours after transplantation. Bud continued to produce urine during the entire period of post-transplantation monitoring. As shown in Fig.2, the recipient was observed a small increase in serum creatinine above 2 mg/DL within 24 hours after transplantation. The serum creatinine level returned to normal C the day after transplantation, when there was an acute rejection of the transplanted organ (dog-recipient has received insufficient immunosuppressive therapy). As shown in Fig.3, the level of creatinine in the urine increased quickly and reached normal limits of about 70 mg/DL within 48 hours after transplantation. This is a very minor episode of acute tubular necrosis (REL) observed initially, quickly underwent reverse development through 48 hours after transplantation. Reversible acute tubular necrosis, along with the ability of the transplanted kidney to maintain long-term survival of the recipient, showed the viability and function in vivo transplanted organ, which has undergone a period of warm ischemia lasting more than 60 minutes. Thus, the body reanimated by using method and recovery solution of the present invention, after the animation may not work in vivo.

Century Autotransplantation

In another embodiment, the body reanimated by using method and recovery solution of the present invention, was evaluated for function in vivo after resuscitation. Using two dogs, performed autologous transplantation by removal of the left kidney, which was then confirmed by warm ischemia in the bath with instruments and washed their restorative solution of the present invention, and then perfesional them for 2 hours, using the method and recovery solution of the present invention, at a temperature of 30-32oC. After recovery kidney was autotransplantation with simultaneous nephrectomy untreated, contralateral kidney. Thus, the survival of each dog-recipient depended entirely on the reanimated kidneys. During the post-transplant period in dogs-recipients shown in Fig.4 and 5.

Kidney well repertoiremaps and worked out through urine hours after transplantation. The kidneys continue to produce urine during the entire period of post-transplantation monitoring. As shown in Fig.4, both dogs were observed a small increase in serum creatinine level, consistent with REL. In each case, the peak serum creatinine was observed on the third day after transplantation and was 3.5 mg/DL and 2.8 mg/DL, respectively. However, the levels of serum creatinine returned to normal in 10 days after transplantation. Biochemical parameters serum remained normal during the remaining part of the post-transplant period. As shown in Fig. 5, the levels of creatinine in the urine increased quickly and reached normal the histological study showed nearly normal kidney. These studies are revealing in terms of regeneration of the epithelium of the tubules. Reversible acute tubular necrosis, along with the ability of the transplanted kidney to maintain long-term survival of the recipient, showed the viability and function in vivo transplanted organ, which has undergone a period of warm ischemia lasting more than 2 hours. Thus, the body reanimated by using method and recovery solution of the present invention, after the animation may not work in vivo.

Example 7 is Compared with a known preservative solutions

The bodies were recovered, using the method and recovery solution of the present invention or a known preservative solution (basic culture medium RSM-210TMor VIA-SPANTM), and then compared the function and histology of organs. Each group of kidneys who underwent a 60-minute cessation of blood flow, washed with the 32oWith an appropriate solution, and then was perfesional within 2 hours of the respective solution: VIASPANTMat 4oWith or RSM-210TMor restorative solution of the present invention when 30-32oWith using the same method of recovery. Comparison of mean perfusion ceatinine in the urine; histology) in the kidneys, deprived of blood flow for 60 minutes and treated with a solution shown in table 5.

The results presented in table 5, show that the replacement solution of the present invention has the ability to restore vascular characteristics, and functions of the ischemia damaged organs, the superior abilities VIASPANTMand RSM-210TM. For example, in the kidney, washed and perfuziruemah restorative solution of the present invention in the process of recovery, was observed reduced vasoconstriction and higher speed of a liquid compared to kidneys, washed and perfuziruemami RSM-210TMor VIASPANTM. Also, kidney, washed and perfezione restorative solution of the present invention, were the only who have recovered their function, which resulted in the development of urine with concomitant secretion of creatinine. Histologically, only the kidneys, washed and perfezione restorative solution of the present invention or RSM-210TMwere restored and quite safe, as evidenced by the well-preserved architecture of renal tissue. On the contrary, kidney, washed and perfezione fabularny necrosis. Demonstrated excellent ability in comparison with known solutions specialists, fashion and recovery solution of the present invention to overcome the effect of warm ischemia on organs and to support the recovery process to such an extent that the violation of the functions of the body can become reversible.

Example 8 delivery of oxygen during the process of recovery

As discussed in detail above, in one embodiment of the present invention in the composition of the recovery solution include as a component of one or more oxygen-carrying agents. Assessed the effects of various oxygen-carrying agents as components of the recovery solution, and their relative role in the delivery of molecular oxygen; and the ability of the method and recovery solution of the present invention to support ongoing oxidative metabolism. Recovery solution that does not contain oxygen-carrying agent (table 6 labeled "BP"); recovery solution containing the washed erythrocytes (table 6 labeled "BP-ER"; 15 vol.%); restorative solution containing the purified hemoglobin (table 6 oboznacheniyu emulsion (table 6 labeled "BP-PFH"; 20% vol.) used for the recovery of the kidney who underwent 60 minutes of warm ischemia. Each group of kidneys who underwent a 60-minute cessation of blood flow, washed at a temperature of 30-32oWith using the appropriate solution, and then was perfesional at 30-32oWith over 2 hours of the respective solution, using the same method of recovery. Comparison of mean perfusion characteristics (pressure, flow speed of the fluid and vascular resistance) and organ function (creatinine concentration in the urine; histology) in the kidneys, deprived of blood flow for 60 minutes and treated with a solution shown in table 6.

The results presented in table 6, show that with increasing concentration of molecular oxygen in the recovery solution through more effective oxygen-carrying agent, as a component of recovery solution, the function body is improved as a result of the recovery process. For example, an effective oxygen-carrying agents, performance or purified hemoglobin, provide a higher concentration of molecular oxygen delivered to the process of recovery. Renal function treated with restorative Rast is subjected to reductive solution, not containing added oxygen-carrying agent or containing less effective oxygen-carrying agent. For example, when using recovery solution with purified hemoglobin or performics.com agent according to the present invention, the average concentration of creatinine in urine was 18 mg/DL and 41.8 mg/DL, respectively. On the contrary, when the recovery process is not used oxygenator and recovery solution does not contain carrying oxygen agent, or contains a low concentration of washed erythrocytes, the average concentration of creatinine in the urine is 8.4 mg/DL and 8.3 mg/DL, respectively. Demonstrated another option for remediation of the solution in which adding one or more effective oxygen-carrying agents as a component of the solution has improved the function of the ischemia damaged organ processed using the method of the present invention.

Example 9

Method and recovery solution of the present invention can be applied to overcome the action of warm ischemia on liver, devoid of blood flow, and maintain the recovery process to such an extent that it is possible vosstanovlenie the lack of blood flow, treatment of ischemia damaged liver by using method (washing and perfusion) of the present invention for about 2 hours may be sufficient for recovery of most of these bodies (for example, deprived of blood flow during the period of time from about 0.5 to 4 hours) to restore organ function. The overall function of the liver, as well as certain aspects of the physiology of the liver can be assessed by determining the concentration of components in the circulating perfusion solution, as well as in the secret of the liver (bile). The functional characteristics of the liver can be assessed by determination of some parameters, including, but not limited to, the concentration in the bile of bile salts, cholesterol, alkaline phosphatase; pH of bile and the flow rate of fluid through the vessels of the liver, oxygen consumption and glucose utilization (as measured in the perfusion solution). Thus, according to the method and solutions, as shown in examples 1 and 2, ischemia damaged liver can handle, and then evaluate its prospects in relation to metabolic functions.

Example 10

Method and recovery solution of the present invention can be applied to overcome the action of warm ischemia on pagesmore restoration of disturbed functions. Although the time required for perfusion may be dependent on the duration of the period of absence of blood flow, treatment of ischemia damaged pancreas by using method (washing and perfusion) of the present invention for about 2 hours may be sufficient for recovery of most of these bodies (for example, deprived of blood flow during the period of time from about 0.5 to 4 hours) to restore organ function. The overall function of the pancreas, as well as certain aspects of the physiology of the pancreas, can be assessed by determining the concentration of components in the circulating perfusion solution, as well as in the pancreas. The functional characteristics of the pancreas include the concentration of pancreatic enzymes such as amylase, lipase; insulin; pH, sodium and potassium pancreatic secretions, and the flow rate of fluid through the vessels of the pancreas, oxygen consumption and glucose utilization (as measured in the perfusion solution). Thus, according to the method and solutions, as shown in examples 1 and 2, ischemia damaged pancreas can handle, and then evaluate its prospects in Ameritania can be applied to overcome the action of warm ischemia on the heart, deprived of blood flow, and maintain the recovery process to such an extent that it is possible to recover its function. Although the time required for perfusion may be dependent on the duration of the period of absence of blood flow, treatment of ischemia damaged heart by using method (washing and perfusion) of the present invention for about 2 hours may be sufficient for recovery of most of these bodies (for example, deprived of blood flow during the period of time from about 0.5 to 4 hours) to restore disturbed functions of the body. The overall function of the heart, as well as certain aspects of the physiology of the heart, can be assessed by determining the concentration of components in the circulating perfusion solution, as well as in heart. The functional characteristics of the heart include, but are not limited to, mechanical and electrical work, the enzymes of the heart, such as transaminases (aspartate aminotransferase, ACT), lactate dehydrogenase (LDH), fructose-1,6-diphosphate aldolase (ALD), malatdegidrogenaza (MDY), glutathion reductase (GR), CPK (CPK), hydroxysteroiddehydrogenase (HBB); the flow rate of fluid through the vessels of the heart, oxygen consumption and util is but in examples 1 and 2, ischemia damaged heart can handle, and then to assess its prospects in relation to metabolic functions.

Example 12

Method and recovery solution of the present invention can be applied to overcome the action of warm ischemia in the small intestine, deprived of blood flow, and maintain the recovery process to such an extent that you can restore their impaired function. Although the time required for perfusion may be dependent on the duration of the period of absence of blood flow, treatment of ischemia damaged small intestine by using method (washing and perfusion) of the present invention for about 2 hours may be sufficient for recovery of most of these bodies (for example, deprived of blood flow during the period of time from about 0.5 to 4 hours) to restore organ function. The overall function of the small intestine, as well as certain aspects of the physiology of the small intestine, can be assessed by determining the concentration of components in the circulating perfusion solution, as well as in the small intestine. The functional characteristics of the small intestine can be assessed by determining some of the parameters that is produced in the stomach, and tests on suction, using labeled molecules; the flow rate of fluid through the vessels of the small intestine, oxygen consumption and glucose utilization (as measured in the perfusion solution). Thus, according to the method and solutions, as shown in examples 1 and 2, ischemia damaged small intestine can be treated, and then to assess its prospects in relation to metabolic functions.

Example 13

Method and recovery solution of the present invention can be applied to overcome the action of warm ischemia on easy, deprived of blood flow, and maintain the recovery process to such an extent that you can restore their impaired function. It may be desirable to first treat pulmonary graft surface-active substance immediately before perfusion (see, for example, Erasmus et al., 1996, Am. J. Respir. Crit. Care Med. 153:665-670). Although the time required for perfusion may be dependent on the duration of the period of absence of blood flow, treatment of ischemia damaged lung by using method (washing and perfusion) of the present invention for about 2 hours may be sufficient for recovery of most of these bodies (the AI body. Overall lung function, as well as certain aspects of the physiology of the lung, can be assessed by determining the concentration of components in the circulating perfusion solution, such as surfactant protein a (SS-A). The functional characteristics of the lung can be estimated by defining some parameters, including, but not limited to, FVC (forced vital capacity), FEV1 (forced expiratory volume in 1 second), DEDICATED (peak volumetric expiratory flow), AO (mean alveolar volume), OEL (total lung capacity) and JI (transfer of carbon monoxide). Thus, according to the method and solutions, as shown in examples 1 and 2, ischemia damaged easy you can handle, and then to assess its prospects in relation to metabolic functions.

It should be understood that the described ways of practical implementation of the present invention and examples serve only the purposes of illustration, and not limitation, and any changes or modifications which will be obvious to an experienced specialist from the above description and the accompanying drawings are included in the scope of the attached claims and their equivalents.

1. The method of restoring the body to transplantations be reversible, this method includes washing the body at a temperature of approximately 28 - 37oWith buffer containing saline solution to remove blood and acidotic products that have accumulated in the body over a period of absence of blood flow, and perfusion of the organ at a temperature of about 28 - 37oWith buffer containing saline solution, which additionally includes means for expanding the blood vessels in the body, and trophic factors in order to restore cell integrity and cell function, reducing thus the function of the body, and means to restore oxidative metabolism in the body when re-adaptation of the body to oxygenated environment.

2. The method according to p. 1, which contains the buffer saline solution for perfusion of the body additionally includes vasodilators and antioxidants as a means to prevent tissue damage during the resumption of blood flow in the body.

3. The method according to p. 1 in which the means to restore oxidative metabolism include the introduction of molecular oxygen by adding oxygenator or oxygen-carrying agent.

4. The method according to p. 1, wherein the perfusion implementing the corresponding buffer saline; said device further includes means for providing and controlling perfusion and perfusion pressure, means for temperature control and means to ensure and control the introduction and removal of respiratory gases, and means for testing or collection for testing buffer containing saline solution, which has already passed through the body to the monitor, and measure one or more functional characteristics, such as pH, pressure, flow speed of the fluid, vascular resistance, various chemical constituents, oxygen therapy, the concentration of carbon dioxide and oxygen.

5. The method according to p. 4, wherein the perfusion is performed using a second device connected to the first device for testing or collection for product testing by the authority, and the subsequent determination of the parameters of the product produced by the body, is related to the integrity and function of the body.

6. The method of recovery and inhibition of further ischemic damage of the organ for transplantation, in priletali phase of ischemia, when such recovery is still possible, in which violations funkcie: (a) washing of the body containing the buffer saline, which removes the blood and acidotic products that have accumulated in the body over a period of absence of blood flow, and further includes means to restore the physiological pH of the body, means for expanding the microvessels in the body, means for maintaining ongoing anaerobic metabolism, metabolic substrates to fill the pool Denisovich connections, maintaining the citric acid cycle and system recovery transport electron pairs to initiate the transition from anaerobic metabolism to oxidative metabolism, trophic factors to restore the function of the body, thus providing metabolites to restore cell integrity and cell function; and (b) perfusion of the organ containing the buffer saline, which further includes means for normalizing the supply of the body with oxygen and the pH of the body, means for expanding the blood vessels in the body and trophic factors to restore the function of the body, thus providing metabolites to restore cell integrity and cell function.

7. The method according to p. 6, which contains the buffer saline solution for perfusion of the organ further what I resume blood flow in the body.

8. The method according to p. 6, wherein the means to restore oxidative metabolism include the introduction of molecular oxygen by adding oxygenator or oxygen-carrying agent.

9. The method according to p. 6, wherein the perfusion is performed using the device, including laminar or pulsating pumping system for delivering containing buffer saline; said device further includes means for providing and controlling perfusion and perfusion pressure, means for temperature control and means to ensure and control the introduction and removal of respiratory gases, and means for testing or collection for testing buffer containing saline solution, which has already passed through the body to the monitor, and measure one or more functional characteristics, such as pH, pressure, flow speed of the fluid, vascular resistance, various chemical constituents, oxygen therapy, the concentration of carbon dioxide and oxygen.

10. The method according to p. 9, wherein the perfusion is performed using a second device connected to the first device for testing or collection for tiemogo body correlated with the integrity and function of the body.

11. Recovery solution for recovery and inhibition of further ischemic damage of the organ for transplantation, in priletali phase of ischemia, when such recovery is still possible, in which violations of the functions of the authority be reversible in the body, deprived of blood flow, specified recovery solution includes a buffer containing saline solution and further includes vasodilators in a physiologically effective amount to expand the blood vessels of the body, the chemical energy substrates in a physiologically effective amount to restore oxidative metabolism, discontinued in the absence of blood flow in the body, and trophic factors in a physiologically effective amount to accelerate one or more recovery processes in the cells in order to restore cellular function, lost in the absence of blood flow.

12. Recovery solution on p. 11, in which the vasodilators include a combination of components selected from the group consisting of substrates for vasodilation, mediated by endothelial cells, the subst what about p. 12, in which substrates for vasodilation, mediated endothelial cells include acetylcholine and arginine, the substrate for the expansion of microvessels include prostacyclin and mg+and calcium channel blockers include adenosine and verapamil.

14. Recovery solution on p. 11, in which chemical energy substrates include a combination of components selected from the group consisting of the components of the pool Denisovich compounds, components of the citric acid cycle and components of the transport chain electron pairs.

15. Recovery solution on p. 14, in which chemical energy substrates are selected from the group consisting of pyruvate, glucose, ATP, AMP, coenzyme A, -dinucleotide (NAD+), flavinadeninnukleotid (FAD), tiaminpirofosfat chloride (cocarboxylase), UTP, chloride, magnesium, and combinations thereof.

16. Recovery solution on p. 11, in which trophic factors are selected from the group consisting of amino acids, magnesium, derivatives of nucleic acids, ribonucleotides, acidic fibroblast growth factor (FGF), basic FGF, heparin and chondroitin sulfate and combinations thereof.

17. Recovery solution on p. 11, Toyama of antioxidant, oxygen-carrying agent and combinations thereof.

18. Recovery solution on p. 11, which further includes a pharmacologically effective amount of a neuroprotective drug.

19. Method of recovering organs for transplantation, in priletali phase of ischemia, when such recovery is still possible, in which the dysfunction of the body is reversible, this method is carried out at a temperature of about 28 - 37oWith and includes washing the body and organ perfusion recovery solution on p. 11.

20. Method of recovering organs for transplantation, in priletali phase of ischemia, when such recovery is still possible, in which the dysfunction of the body is reversible, this method is carried out at a temperature of about 28 - 37oWith and includes lavage and perfusion of the organ recovery solution on p. 17.

21. The method of recovery and inhibition of further ischemic damage of the organ for transplantation, in presettling phase of ischemia, when such recovery is still possible, in which the dysfunction of the body is reversible, kazanavicienes solution under item 11.

22. The method of recovery and inhibition of further ischemic damage of the organ for transplantation, in presettling phase of ischemia, when such recovery is still possible, in which the dysfunction of the body is reversible, this method is carried out at a temperature of about 28 - 37oWith and includes lavage and perfusion of the organ recovery solution on p. 17.

23. The method of solution for recovery and inhibition of further ischemic damage of the organ for transplantation, in priletali phase of ischemia, when such recovery is still possible, in which the dysfunction of the body is reversible, which includes adding to the buffer containing saline combination of additives, including vasodilators in a physiologically effective amount to expand the blood vessels of the body; chemical energy substrates in a physiologically effective amount to restore oxidative metabolism, discontinued in the absence of blood flow in the body, and trophic factors in a physiologically effective amount to accelerate one or more vosstanovitelya specified pH of the solution was adjusted to values in the range of about 6.5 to 7,5, and its osmolarity greater than 330 mOsm.

24. The method according to p. 23, wherein the vasodilators include a combination of components selected from the group consisting of substrates for vasodilation, mediated by endothelial cells, substrates for the expansion of the microvessels, calcium channel blockers, and combinations thereof.

25. The method according to p. 24, in which the substrates for vasodilation, mediated endothelial cells include acetylcholine and arginine, the substrate for the expansion of microvessels include prostacyclin and mg+and calcium channel blockers include adenosine and verapamil.

26. The method according to p. 23, in which chemical energy substrates include a combination of components selected from the group consisting of the components of the pool Denisovich compounds, components of the citric acid cycle and components of the transport chain electron pairs.

27. The method according to p. 26, in which chemical energy substrates are selected from the group consisting of pyruvate, glucose, ATP, AMP, coenzyme A, -dinucleotide (NAD+), -nikotinamidadenindinukleotida (NADH), flavinadeninnukleotid (FAD), tiaminpirofosfat chloride (cocarboxylase), UTP, chloride, is the present from amino acids, magnesium derivatives of nucleic acids, ribonucleotides, acidic fibroblast growth factor (FGF), basic FGF, heparin and chondroitin sulfate and combinations thereof.

29. The method according to p. 23, wherein the additive substances further include a physiologically effective amount of a component selected from the group consisting of an antioxidant, oxygen-carrying agent and combinations thereof.

30. The method according to p. 23, wherein the additive substance additionally include a pharmacologically effective amount of a neuroprotective drug.

 

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