The solution for the conservation of living organs

 

(57) Abstract:

The invention relates to medicine, in particular to the conservation of organs. The solution for the conservation of living organs contains the following components, mol/l: sodium chloride 100,0-117,0; potassium gluconate 12,0-17,0; calcium gluconate 1,2-2,0; magnesium sulfate 8,0-32,0; trihydroxystilbene 2,0-3,0; local anesthetic 0,33-1,66; derived fenotiazina 0,01-0,04; purine derivative 1,0-2,0; poliglyukin else. The solution allows to increase the duration of the conservation of organs and eliminates irreversible damage during reperfusion.

The invention relates to medicine, in particular for solution for conservation of living organs.

Known methods of conservation of living organs and tissues used in medicine and which consists in placing them in a special preservative solutions with subsequent storage in hypothermia. Particularly difficult is the need for conservation in cardiac surgery and transplantation of such intense working of the body, like the heart in relation to the intensity of what is happening in its metabolic processes and energy consumption at its reductions. If even a few years ago, surgeons were able p is Wai and chemical cardioplegia, we offer the Royal Institute and hospital of St. Thomas (UK), it is possible to preserve the heart during the 4 o'clock On the basis of cardioplegic solution proposed by the hospital of St. Thomas, created the drug plegia (commercial product) (Abbott Laboratories, Chicago, USA), which is widely used in clinical practice. As a result, improved cellular protection during open-heart surgery, which requires a long time, create additional conditions for the improvement of surgical techniques, increase the preservation time required for the heart transplant.

However, the conventional preservation time of the heart (4 h) not enough for shipping over long distances, in particular, at any point of the globe, where there is a necessity for his transplant recipient. The increase in the duration of preservation experiencing body must be combined with improved quality and reliability of the protection of cells from ischemic and reperfusion damage.

The basis of the invention is by changing the qualitative and quantitative composition to develop a solution conservation of living organs with increased duration cosnova damage.

The problem is solved by the fact that the solution for the conservation of organs containing sodium chloride, potassium salt, magnesium salt, calcium salt, a pH stabilizer and a solvent, according to the invention additionally contains a local anesthetic, a derivative fenotiazina, purine derivative as salts of potassium and calcium gluconate contains potassium and calcium as magnesium salt is magnesium sulfate as a stabilizer pH trihydroxypyrimidine, and as a solvent - poliglyukin, in the following ratio of components, mmol/l:

Sodium chloride 100,0-117,0

Gluconate potassium 12,0-17,0

Calcium gluconate 1,2-2,0

Magnesium sulfate 8,0-32,0

Trihydroxysilyl - aminomethan 2,0-3,0

Local anesthetic 0,33-1,66

Derived pheno - thiazine 0,01-0,04

Purine derivative 1,0-2,0 Poliglyukin Else

The inventive solution has the following advantages compared to plagiola (prototype). Use poliglyukina dramatically increase the osmolarity of the solution (320 mo m/l to 595 mo m/l and above) and to prevent the abrupt swelling of mitochondria and intracellular edema. The use of salts of potassium and calcium in the form of gluconate, and magnesium salts in the form of magnesium sulfate instead of chlorides can reduce the content is mpany excitable cells, for example myocardial. A local anesthetic, which can be used, for example, lidocaine, procaine, and others added to the solution to stabilize membranes. Derived fenotiazina, which can be used, for example ethacyzin, is used to map the intracellular proteins of the type of calmoduline, in addition, this drug has a pronounced antiarrhythmic and antiischemic effect, a purine derivative, which can be applied, for example, inosine, is used to replenish energy reserves of the cells due to its ability to penetrate to the myofibrils, to stimulate the synthesis of nucleotides and to increase the activity of some enzymes of the Krebs cycle.

The proposed solution contains all three components, which in large quantities can be independent cardioplegic agents (ions K+the ions of Mg2+and a local anesthetic), but the quantity of each in the proposed solution is considerably lower than that needed for cardioplegia, if each of them would be used independently (for example, 30-40 mmol/l for potassium chloride and magnesium; 10 mmol/l or more for a local anesthetic, depending on which of the COI is to provide a reversible stabilization of excitable membranes of cardiomyocytes and other cells and do not cause irreversible damage from prolonged global ischemia and reperfusion.Lower concentrations do not allow to achieve the expected effect in conservation and cardioplegia, large concentrations may cause irreversible damage to the membranes.

Application of the proposed solution allows to increase the duration of the successful conservation of living organs, particularly the heart, up to 48 hours In the study of the prior art was not found solutions for the conservation of living organs with the proposed composition of the components.

Thus, we can conclude that the claimed solution to the conservation of living organs has novelty.

In addition, because the proposed solution is not necessary for the expert in the obvious way from the prior art, it can be concluded that the claimed invention meets the criterion of "inventive step".

The proposed solution is get a simple mixing of the components.

The proposed solution was tested in experiments with preservation of a dog's heart donor with an onward connection to his dog to the recipient in the experiments with preservation of the kidney with subsequent transplantation outbred white rats. Just been delivered 45 experiments.

P R I m e R 1. The proposed solution contains components in the following ratio, mmol/l: sodium Chloride 110,0 Gluconate potassium 16,0 magnesium Sulfate 16,0 Gluconate kaldewey solution prepared immediately before surgery by a simple mixing of its components. Osmolarity was 595 OS m/s, the pH is maintained at 7.8. The solution was ozonirovaniya for 30 min to Rho2400 mm Hg.

Donor - purebred dog, a female weighing 15 kg After intravenous pentobarbital sodium at a dose of 25 mg/kg dog was transferred to artificial respiration 12 per minute volume of 200 ml Median incision in 3-4 intercostal space revealed chest on the right and left. Allocated vessels. Superior Vena cava and the aorta kanglida. All the arteries and veins is taped and intersect. Cardio-pulmonary drug is excreted from the body. Trachea reintubated and cardio-pulmonary drug is transferred into a cuvette. Laundering is proposed cardioplegic solution, flowing from the heart, the liquid collects in the tank. Heart stops declare cardioplegic solution for 1 minute, After which the heart is allocated and transferred to a capacity of 300 ml, filled cardioplegic solution, and placed in the refrigerator at t = 7aboutWith 48 hours

After 48 h, the heart is removed from the refrigerator and filed the dog to the recipient in a heterotopic position. Recipient-the dog is purebred, male, weighing 11 kg Highways of veins and arteries are connected. Smenivshiesya soon ventricular fibrillation. Ventricular fibrillation charged defibrillation discharge. Recorded blood pressure and electrocardiogram taken biopsy material for electron microscopic examination.

At postmortem examination of canned within 48 h and the transplanted heart of the dog identified the following changes. In General good state of preservation, there is a slight change of electron-microscopic picture of myocardial tissue; consisting in marble oedema or more global with moderate divergence Krist mitochondria or a certain degree of enlightenment of the mitochondrial matrix, a moderate degree of hyperchromatism or normal concentration of nuclear chromatin with retraction in some cases the nuclear membrane. Changes are insignificant edema myocardial fibres with small foci of divergence of the myofibrils. After reperfusion for 30 min heart donor blood recipient described above minor changes found in canned heart, completely disappeared. Thus, myocardial tissue was preserved solution from damage total ischemia within 48 h and posledowatel/l: sodium Chloride 117,0 potassium Gluconate 17,0 calcium Gluconate 1.6 magnesium Sulfate 32,0

Trihydroxysilyl - aminomethan 2.5 Lidocaine 1,66 Ethacyzin 0.02 Inosine 2.0 Polyglucin Else

The proposed solution is prepared and injected as described in example 1.

Donor - purebred dog, female weight 5,4 kg General Anesthesia: intravenous pentobarbital sodium at a dose of 25 mg/kg of the operation is similar to that described in example 1.

The recipient is a purebred dog, a female weighing 12 kg the course of the transplant donor hearts dog-recipient similar to that described in example 1 except that the heart is transplanted into normolipidemia position. Heart regained its activity after 10 minutes of slow perfusion of the blood of the recipient. When reviving the hearts of the observed group atrial and ventricular extrasystoles, episodes of ventricular tachycardia. Recorded indicators of electrocardiogram and blood pressure showed no life-threatening disruptions of heart. Taken biopsy material for electron-microscopic examination of myocardial tissue. At postmortem examination of myocardial tissue canned within 24 h and transplanted to the recipient of the heart revealed changes similar to that observed in the example is cipient. During recovery from anesthesia the next day, the dog feels normal; organs and systems no changes, except a separate atrial and ventricular premature beats in the electrocardiogram.

P R I m e R 3. The proposed solution contains components in the following ratio, mmol/l: sodium Chloride 100,0 Gluconate potassium 12,0 calcium Gluconate 2.0 magnesium Sulfate 8,0

Trihydroxysilyl - aminomethan 3.0 Lidocaine 0,33 Ethacyzin 0.04 Inosine 1.5 Polyglucin Else

The proposed solution is prepared as described in example 1.

Donor - purebred white rat, male, weight 260 g General Anaesthesia essential. Produced median laparotomy on the white line of the abdomen. The intestine is shifted to the right, freeing the operating field. Carefully otrabativayutsya renal artery, renal vein throughout. The ureter is also uprepared, then superimposed microsurgical clamp on the renal artery and vein, below which they intersect. The donor kidney is allocated from the operating wounds, renal artery onuliruetsya. Laundering kidneys proposed solution is the introduction of it into the renal artery at a rate of 1 ml per minute until receipt of transparent liquids is alwaysa in the refrigerator where the kidney is stored for 26 hours at a temperature of 6aboutC.

Recipient - purebred white rat, male, weighing 260, Is described above, the operation of removing a kidney. Then the intestine is removed, freeing the surgical field for the donor kidney transplantation. Carefully allocated renal vessels and ureter. On the renal artery, renal vein and ureter superimposed mikrozajmy below which they intersect. The second kidney is allocated and removed from the surgical field. From the refrigerator canned kidney is transferred to the operating field, usually located renal artery, renal vein and ureter. Then anastomosis between the renal artery of the donor and recipient single interrupted sutures, ligature 10,0 8. Is the anastomosis of the renal vein continuous suture. Removed the clamps and restore circulation in the transplanted organ; the kidney is filled evenly, there is a slight stenosis of the wall of the renal veins. The anastomosis is expanded by puncture of the inferior Vena cava on the needle. Superimposed seam 3 on the ureter. Performing anastomosis has slowed due to the active excretion of urine. Urine is excreted dropwise. The wound is sewn in layers. The next day, at McAskie departure OK, on other organs and systems with no violations. From the tail vein taken blood for analysis. The dynamics of kidney weight; the content of adenosine triphosphate decreased by 15% less than in the control experiment with kidney, canned using cardioplegic solution of the hospital of St. Thomas.

The SOLUTION FOR the CONSERVATION of LIVING ORGANS, containing sodium chloride, potassium, magnesium, calcium, pH stabilizer and a solvent, characterized in that it further comprises a local anesthetic, a derivative fenotiazina, purine derivative as salts of potassium and calcium gluconate contains potassium and calcium as magnesium salt is magnesium sulfate as a stabilizer pH - trihydroxypyrimidine, and as a solvent - poliglyukin in the following ratio of components, mm/l:

Sodium chloride - 100,0 - 117,0

Gluconate potassium - 12,0 - 17,0

Calcium gluconate - 1,2 - 2,0

Magnesium sulfate - 8,0 - 32,0

Trihydroxystilbene - 2,0 - 3,0

A local anesthetic is 0.33 - 1,66

Derived fenotiazina - 0.01 to 0.04)

Purine derivative - 1,0 - 2,0

Poliglyukin - the Rest is up to 1 L.

 

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