Cardiovascular homograft (versions), method for preparing homograft, homograft tissue exposure medium (versions)

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine, namely to cardiac surgery, and concerns creating cardiovascular homografts (CVHG) applied as vascular biological prostheses in cardiovascular surgeries. There are presented versions of a homograft representing a great vessel section or right and left ventricular outflow tracks taken from a dead body. The homograft is also characterised by min. 30-minute preparation in a medium containing gentamycin, diflucan and pH stabiliser. At a stage of preparation the homograft is processed with a medium of pH 7.0-7.4 representing a DMEM/F12 solution and containing gentamycin, diflucan, cephalosporin and metrogyl. Further, the homograft is kept in a medium containing the DMEM/F12 solution, gentamycin, diflucan and a phosphate buffer. There are also presented methods for producing the CVHG versions, as well as the media for the homograft treatment at the various stages of producing, keeping and handling.

EFFECT: creating the CVHG of more rigid structures resistant to damaging factors of the treatment, handling and storage media that improves their body functions.

16 cl, 10 ex, 9 tbl, 10 dwg

 

The invention relates to medicine, namely to the surgery of congenital and acquired heart defects, or rather to homografts cardiovascular system (GSSS), used as a vascular biological prostheses during operations on the cardiovascular system (CVS), and technologies of their production.

Now for the prosthetic blood vessels and valves SSS during heart operations use different Bioprocess obtained from animal tissues or cadaverous tissues and prostheses made of synthetic materials, such as artificial heart valves type Bjork-Shiley; Lillehei-Kaster; Star-Edwards, etc. Among bioprocesos the greatest application received GSSS (usually called "vascular homograft") for implantation in blood vessels or in the input (AV) and output (ventricular) the holes in the heart (homograft). Under vascular homografts ("homograft" from lat. homograft, homo -, or, in other interpretations, homogeneus - homogeneous graft - graft prosthesis) understand implantable prosthesis that is fully or partially consists of the inanimate, specially processed human tissue, including, if necessary, heart valves (ru.wikipe-dia.org/wiki).

Advantages GSSS are optimal hemodynamic parameters; the natural functioning of the connective tissue structures surrounding homograft, muscle tissue; no need of receiving indirect anticoagulants; use of children, including newborns; the possibility of "settling" homograft fibroblasts recipient and regeneration of connective tissue components of the matrix homograft in the body of the recipient, providing an increase in size, especially in young children. The term normal functioning of homograft in the aortic position is, on average, 5,0-7,5 years, and during implantation into the output division of the right ventricle (in the correction of complex congenital heart defects) is 2-3 times more.

The disadvantages of the currently used of homografts are complex technological conditions of production and limited shelf life; the possibility of degenerative changes after implantation, limiting the period of use; the originality and uniqueness of each product.

The basis of homograft is a multilayer tube, which is Home Archive Autor Contact RSS feed http://medicinsk.ru/anatomia-chelove-ka/24-118 .php; critical.ru>CardioSchool/content/doctor/1/f) of several shells. The lining (intima), which is about 20% of the wall thickness, covered by a layer of endothelial cells located at the basal membrane facing the lumen of the vessel. Outside intima limited internal elastic membrane. The inner casing m which may form a pocket-type hollows. provinces folds valves. Behind the inner shell is a layer formed by a large number prescribing (fenestrated elastic membrane located concentrically between which are smooth muscle cells, able to produce elastin and collagen, as well as amorphous intercellular substance. Outside the boundary layer is the outer elastic membrane formed mainly circularly arranged smooth muscle fibers as well as collagen and elastic elements and proteoglycans.

To improve the mechanical characteristics of such a tube, if necessary, put in additional tubular frame made of synthetic resin material, receiving the combined homograft (GSSC).

As a material for the manufacture of vascular homografts usually use pork or cadaver vascular tissue subjected to previously or denaturation and subsequent destruction of antigenic determinants, or treated with a solution of glutaraldehyde (NN. Malinowski and other Biological prosthetic heart valves. -M.: Medicine, 1988, pp.24-53; Biocompatibility./Edited Viewactive. -M.: Information centre of Vniigeosistem, 1999; EN 2195879, 2003; 17/00; EN 2197819, 2003). The composition of the biological part of the prosthesis may also include various biological fragment of the CAS - heart valves, fragments, etc.

The disadvantages of homografts received by the first of the above mentioned technologies is a high probability of calcification and perforation of the valves, especially in children and adolescents. Disadvantages of the second technology is a relatively low mechanical characteristics of homografts, the high risk of development of degenerative changes, as well as the impossibility of their long-term storage.

There is a method of preparing vascular homografts (Richard A.Hoprins. Cardiac Reconstructions with Allograft Valves. With Contri-lutions fy V.J.Ferrans, S.L.Hilbert, M.Jones P.L.Lange, L.Wolfinborger, Jr.Springer. Verlag. - 1990, p.44-46), which includes fence vascular complex, its preparation, sterilization, and the introduction of the obtained product in the storage medium. When this preparation is carried out in 0.9% sodium chloride (saline)solution, and then injected into the environment for the treatment of vascular complex, composed of a mixture of antibiotics in physiological solution at a concentration of: cefoxitin 240 g/l; lincomycin hydrochloride 120 g/l; polymyxin b sulfate 100 g/l; vancomycin hydrochloride 50 g/l; amphotericin b 25 g/l Processing tissue should be performed within 48 h at 4°C, after which the resulting product is placed in a medium for storage - medium Needle, in which the drug vascular homografts stored until use in ka is este vascular biological prosthesis operations on the heart, but not more than 28 days.

The disadvantage of the method of preparation and use environments is the lack of protection homograft in terms of mechanical, physical, chemical, and temporal factors. Thus, the sodium chloride in the aqueous solution used in the stages of preparation and processing of antibiotics, and the environment the Needle used for storage, do not form complexes with a protective effect against vascular tissue that does not allow you to prevent the negative environmental impacts generated during the storage of radicals and products of disintegration of cells. When processing complex of antibiotics, in particular a mixture of vancomycin hydrochloride and lincomycin hydrochloride, causing more internal damage to the cells of the vascular tissue due to the impact of these antibiotics on the structure of collagen and elastin proteins.

The closest to the technical nature of the claimed group of inventions is developed earlier by the author (RU 2083105, 1997) is a method of obtaining homografts, including fence vascular complex (SC), its preparation, during which the IC act reagent containing aminopeptide complex (drug "Aminochrome"), used for treatment of cells IC solution containing a mixture of antifungal drug amphotericin b with antibiotics - ceph what Taksim and gentamicin sulfate at a ratio of Cefotaxime, gentamicin sulfate and amphotericin b, respectively, 40-43:5-8:1-2, and the introduction in the storage medium containing the drug "Aminochrome" concentrations of 1.0±0.1 g/l, Cefotaxime and gentamicin sulfate in the following ratio of components, g/l: Cefotaxime 25,0±2,5; gentamicin sulfate 4,0±0,4. Introduction geminiwave complex using "Aminacrine allows you to increase the protection of homograft from oxidation, but long-term storage of its presence may lead to a change in pH and the formation of hard fixed Deposit. In addition, there is a lack of reliable environment for the storage in response to emerging in the biomaterial changes of the connective fibers of the vessels of the trunk type, and heart valve containing glycosaminoglycan, which affects the durability of the implant, and the implant is expressed inflammatory reaction accompanied expressed exudation.

The disadvantage of used nutrient media is their relatively low efficiency during transportation and long term storage, as well as insufficient sterilizing effect when exposed to a spatially complex patterns characteristic of adventitia that makes, as a rule, completely remove adventitia during preparation.

The problem to be solved by the authors, is the creation of GSCs with her is Tami, improve its functioning in the body, as well as the technologies that ensure a long and safe storage.

The technical result was to obtain GSSS with a modified surface, providing more efficient sorption of fibroblasts, the more rigid structures GSS, as well as creating GSS with a modified surface that is more resistant to the effects of damaging factors of environments for handling, transport and storage homograft.

The technical result is achieved in the establishment of the claimed group of inventions, which is based on the creation of GSSS made in the form of a multilayer tube of vascular tissue, comprising an inner layer of the intima layer prescribing membranes and an outer elastic membrane provided with additional outer layer of adventitia thickness of 0.5-3.0 mm, subjected to preliminary processing of the first aqueous solution containing an antibiotic, an antifungal agent, and a pH stabilizer, and then a solution of DMEM/F12 containing antibiotics, antifungal agent, and a pH stabilizer.

In this case, the solution of DMEM/F12 means of industrially produced the environment that are dissolved in water a mixture of inorganic salts, amino acids, vitamins, glucose and phenol red, presterilization the Yu through filters with a pore size of 0.1 microns (http://www.pa-neco.ru/in-dex.php?page=shop.product_details@flypic).

Layer from adventitia subjected to pre-specified two-step process, has a thickness of 0.5 to 3.0 mm and consists of connective tissue, treated first with an aqueous solution containing a mixture of gentamicin and diflucan and components of a phosphate buffer solution, and then a solution of DMEM/F12 containing a mixture of gentamicin, diflucan, cephalosporin, metrogel components and phosphate buffer solution.

The main difference of the proposed GSS from known is the introduction of an additional outer layer of adventitia subjected to pre-treatment with an aqueous solution and a solution of DMEM/F12 containing antibiotics and pH stabilizers. Adventitia wall of the blood vessel is a frame from elastin-collagenous elements, filled with loose connective tissue. This extra layer protects the inner layers of the multilayer tube from the effects of active oxygen during storage. In this pre-treatment of his first aqueous solution containing a mixture of gentamicin and diflucan at pH of 7.2 to 7.4, at concentrations of 0.05-0.5 wt%. gentamicin, 8,0-12,0% of the mass. diflucan and 0.1-0.5% wt. components of a phosphate buffer solution that supports the desired pH and then a solution of DMEM/F12 containing a mixture of gentamicin, diflucan, cephalosporin and metrogel at pH 7.0 to 7.4, with optimal end is Tralach - of 0.05-0.1% of the mass. gentamicin, 3,0-7,0% of the mass. diflucan, 1.5 to 3.0 wt%. of cyclosporine, 0.1 to 0.3% of the mass. metrogel, 0.3-0.5% of the mass. components of a phosphate buffer solution that supports the desired pH, allows full preservation of native connective tissue matrix and allows you to fully solve the problem of reliable sterilization structure with a highly developed surface, to create a "depot" of biocides, which gives additional guarantees sterility of homograft during further handling, transportation and storage. In this case, unlike the above analogy, the effects of antibiotics on cell does not damage the internal tissues of homograft, but on the contrary, influencing patterns of adventitia, creates a more extensive external layer structure that is optimal for the sorption of fibroblasts when placing homograft in the body.

Optimal results are achieved when using an aqueous solution containing 0.3% of the mass. gentamicin, 10.0% of the mass. diflucan, and 0.3% of the mass. components of a phosphate buffer solution, and a solution of DMEM/F12 containing 0.08% mass. gentamicin, 5,0% of the mass. diflucan, and 2.0% wt. of cyclosporine, 0.2% wt. metrogel and 0.4% of the mass. components of a phosphate buffer solution.

Received GSS can be placed in the outer frame of the permitted use in surgery of material non-biological origin, e.g. the measures steel in particular, brand HIM, or pyrocarbon (http://www.splav.kharkov.com/mat_start?name_id=765).

GSSS obtained by removal of tissue components CAS (TC) at the corpse on clean protectorship during the forensic examination of the corpse. Thus sterilize the skin of the breast of the corpse for at least 5 min, abundantly wetting the surface of the chest, neck and abdomen with antiseptic, designed to handle the surgical field. The exposure time after termination of treatment should be no less than 5 minutes the Skin of the chest of the corpse is covered with a sterile protective film. An incision in the midline from the jugular notch to epigastria. Sternum cut along the line of the rib cartilage and subclavian-sternal joints. After opening the chest to expose the pericardium. Tightening vessels for the transverse sinus, indicate the anatomical landmarks of the great vessels. Allocate the ascending aorta, aortic arch and brachiocephalic vessels 1-2 cm distal to their mouths. Distinguish the left and right pulmonary artery. Heart for the top "dislocate" from the wound. Allocate vessels, preventing damage to the esophagus, trachea or lung parenchyma. Cross the inferior Vena cava, pulmonary vein and superior Vena cava, and then additionally secrete pulmonary artery prior to their division into equity artery. The distal cut left l the gotschna artery, the left subclavian artery, the left carotid artery, right carotid artery, right subclavian artery, right pulmonary artery, thoracic aorta. Extract from the chest of the corpse's heart with major vessels. Seized TC (lots of great vessels and plots the output paths of the right and left ventricles) are placed in a sterile plastic container and pour the environment for processing GSCs.

The method of preparation of the drug GSS consists of the following main stages:

- above the fence TC;

- pre-treatment TC aqueous solution containing 0.05-0.5% wt. gentamicin sulfate, 8,0-12,0% of the mass. diflucan and 0.1-0.5% wt. components of a phosphate buffer solution at pH of 7.2 to 7.4 for at least 30 min;

- preparation of TC in the environment with a pH of 7.0 to 7.4, therefore, to save the layer adventitia thickness of 0.5-3.0 mm, and processing the obtained product mixture containing as a source of salts and nutrients DMEM/F12, and as bactericidal drugs - a mixture of diflucan, antibiotic of the group of cephalosporins, gentamicin sulfate and metrogel;

- the location of the product in the storage medium containing DMEM/F12, gentamicin sulfate and diflucan at pH 7.0 to 7.4. Differences between the proposed method from analogues are as follows.

1. Creating at the stage of preparation at the expense of two-stage processing bio is idname preparati protective layer from adventitia, having an extensive surface, guaranteed sterile and simultaneously increase the speed of sorption of fibroblasts on the surface GSSS, resulting in a reduction of the integration period GSSS when the surgical treatment.

2. Use a new source of salts and nutrients, and preventing sedimentation in the conditions of production and storage GSSS.

3. Use at the stage of preparation of a new set of bactericidal drugs, including metrogylum with a bactericidal action regarding a wide range of anaerobic microorganisms, as well as protozoa and some gram-positive bacteria, and provides, along with guaranteed sterility GSSS, modified its outer surface. However, unlike their foreign counterparts, are not used enzyme preparations, causing significant damage to the connective tissue matrix.

4. The implementation stages of production and storage GSSS in the presence of a phosphate buffer solution at neutral pH values. This eliminates the possibility of formation of acidic or alkaline radicals that can damage the connective tissue fibers and glycosaminoglycans, which are components of the vascular trunk type and heart valves, which significantly affects long is enosti implants. In particular, there is a possibility of retention of the implant within 120 days at 4°C or 30 years at a temperature of(130)°C.

5. Using a specially designed environment for handling TC, containing, unlike commonly used for these purposes, distilled water, a solution of gentamicin sulfate and diflucan, which increases guarantees the sterility of the implant during transport and prevents leaching "biocidal depot" from the tissues.

6. Use in the storage medium as a source of salts and nutrients DMEM/F12, and a mixture of antibiotics, along with gentamicin sulfate, diflucan with antifungal activity.

One of the essential differences of the proposed method is the use in the implementation of the original set of environments, including environments that are applied at the stage of preparation at the stage of preparation and storage medium GSSS.

Environment for pre-treatment GSSS and transportation (if necessary) is an aqueous solution containing 0.05-0.5% wt. gentamicin sulfate, 8,0-12,0% of the mass. diflucan, 0.1 to 0.5% of the mass. components of a phosphate buffer at pH of 7.2 to 7.4. Environment use of this structure allows you to start the process of moderate osmotic decellularization (passive osmosis from hypotonic solution widebody to penetrate into the cell, increasing it osmotic pressure); to prevent the multiplication of microorganisms; to ensure the best preservation of the connective tissue matrix.

Environment for processing GSCs at the stage of preparation contains DMEM/F12, which is dissolved a mixture of diflucan with antibiotics group of cephalosporins, gentamicin sulfate and metrogram and components of a phosphate buffer solution in the following ratio of ingredients (% wt.):

diflucan3,0-7,0
gentamicin sulfatethe 0.05-0.1
the cephalosporin1,5-3,0
metrogylumof 0.1-0.3
components of a phosphate buffer solution0,3-0,5
DMEM/F12 in the form of a solutionrest

The best results are achieved when using a medium containing a mixture of 5.0% of the mass. diflucan; 0,08% . gentamicin sulfate, and 2.0% wt. of cyclosporine, 0.2% wt. metrogel and components of a phosphate buffer solution (FB) (mixture KN2PO4and KN2PO4in the ratio 1:4), ensure the maintenance of pH 7,3±0,3.

Environment impact n the fabric GSSS during storage has a pH of 7.0-7.4 and represents dissolved in DMEM/F12 gentamicin sulfate, diflucan and components of a phosphate buffer solution in the following ratio of ingredients (% wt.):

gentamicin sulfate0,05-0,15
diflucan8,0-12,0
components of a phosphate buffer solution0,1-0,5
DMEM/F12 in the form of a solutionrest

The best results are achieved when using a medium containing 0.1% of the mass. gentamicin sulfate; 0.015% wt. Diflucan in solution DMEM-F12 at pH of 7.2.

If necessary cryopreservation GSSS (prior to storage) is placed in a sterile polymeric package, designed for storage of biological tissues, for example in a sterile plastic bag with a perforated edge firm "Nasco WHIRL-CANCER" (USA), and bring in the specified package cryoprotective mixture of albumin - 20 wt. -%, dimethyl sulfoxide - 10% vol., DMEM/F12 in the form of a solution - else), cooled to a temperature of (2±2)°C (100 ml of a mixture of one GSSS). The package is hermetically sealed and placed in a refrigerator with a temperature of(150±2)°C or cryocontainers containing vapors of liquid nitrogen, where cryopreservation GSSS. Stand package with GSS in holodilny the e or in the container for a period of not less than 120 minutes

The above environment is obtained by dissolving predetermined amounts of ingredients in a solution of DMEM/F12 or distilled water.

Figure 1 shows the overall structure of the tube of the biological part of the claimed GSSS in section, figure 2 - General structure of the tube of the claimed combined homograft (GSSC) (in section). It uses the following symbols: 1 - the lumen of the vessel; 2, basal membrane; 3 - intima; 4 - internal elastic membrane; 5 - layer prescribing (fenestrated) elastic membranes; 6 - outer elastic membrane; 7 - adventitia; 8 - outer tube made of steel or pyrocarbon. Figure 3 shows the image of the model combined homograft in contour with the coordinate axes for measurement. Figure 4-10 shows pictures of some homografts: 4. Mitral combined GSS.(CC Mi. C). 5 is a combined Pulmonary GSSS (GSK LEC). 6 is Oralsyaranasy (GSK Ao K). 7. - Aortic-mitral GSSS (GSK, Aomi 3.1.). Fig - with Pulmonary bifurcation GSSS (GSK Le 2.2.). Fig.9. - Aortic GSSS (GSK Ao 1.1.). Figure 10. Pulmonary GSSS (GSK Le 2.1.).

Upon receipt GSSSC multilayer tube of the vascular tissue is placed inside the outer tube (W)generally coaxial. The greatest protection of vascular tissue from external influences is achieved when its length southwestairline outer tube. Inner diameter W, as a rule, exceeds the outer diameter of the tube from the biological material.

Tube of vascular tissue may be a separate tube from the vessel fragment (fragment artery, a fragment of Vienna)and its combination with other anatomical structural elements (e.g., the aortic root, including the front flap of the mitral valve, ventriculoarterial connection, the aortic valve consists of three aortic valves attached at the lower edges of the sinuses of Valsalva to the wall of the aortic root at the annulus of the aorta and connected with the formation of the three commissures, the fibrous ring of the aorta and the outer walls of the root of the aorta; pulmonary artery from the right and left branches of the pulmonary arteries, including roller cone pontegadea attack with the endocardium, valve pulmonary artery, which consists of three semilunar valves, connected with the formation of the three commissures, the pulmonary artery to the bifurcation and branch pulmonary artery; a fragment of the aorta with a fragment of the mitral valve; fragment artery bifurcation and the like). You can use this as and other similar fragments of the human body, subjected to an appropriate treatment.

The thickness of adventitia received GSS was (1,5±0,5) mm Processing when receiving were generally p and the optimal medium composition.

As shown by tests, GSSS the claimed composition obtained by the present technology provide the best accepted of the prosthesis in the body, due to the almost complete elimination of immunogenicity of epithelial cells and good sorption cells of the host body to the collagen skeleton, and also have the ability to survive without losing their characteristics over 120 days at a temperature (0-4°C and about 30 years old, cryoconservation in liquid nitrogen.

Made on the claimed technology GSSS can be successfully used in surgical correction of congenital and acquired heart diseases and blood vessels, in particular when the plastic pin of the Department of the right and/or left ventricles of the heart; with prosthetic mitral valve, tricuspid valve, aortic valve and the pulmonary artery, combined prosthetic aortic and mitral valves; with prosthetic arteries and veins and other similar operations.

The nature and advantages of the claimed group of inventions are illustrated by the following examples.

Example 1. Getting GSSS

Biological component GSSS was obtained by removal of TC during the forensic examination with subsequent processing environments the optimal composition. To determine the optimal medium composition after completion of the forensic meditsinskoj the study received standard aorta and pulmonary artery size (10×10) mm (vascular areas). Vascular areas were placed in distilled water containing 0.05-0.5% wt. gentamicin sulfate, 8,0-12,0% of the mass. diflucan (Medium a) and 0.1 to 0.5% phosphate buffer solution, and then transported to the laboratory at 4°C.

In the laboratory, in aqueous solution, where the vascular areas, and control the pH and, if necessary, introduced sodium bicarbonate to achieve a pH of 7.3, and then carried out the preparation in such a way as to maintain a layer of adventitia thickness of 0.5 to 3.0 mm is Then obtained after preparation of the vascular samples were placed in a container with a solution of DMEM/F12 containing a mixture of diflucan; gentamicin sulfate, a cephalosporin, metrogel and components of a phosphate buffer solution (FB) (mixture KH2PO4and KH2PO4in the ratio of 1:4, ensure the maintenance of pH 7,3±0,3) (Environment B) for different ratios of ingredients (for 3 samples of aorta and 3 sample pulmonary artery on each of the ratios of ingredients in the mixture). Each of the vascular samples were kept in a specified environment (with a specific ratio of ingredients)containing antibiotics (sterile environment), within 48 hours, then removed from the container with a sterilizing medium and placed in vials.

Variations of the ratios of ingredients in the sterilizing environment is summarized in table 1.

12,0/3,0
Table 1
The composition of the environments a and B, used in the preparation (ingredients)
Number of received medicationThe contents of the ingredients in environments a and B, % mass.
diflucan A/Bgentamicin sulfate (A/Bthe cephalosporinmetrogylumFB
The drug 110,0/5,00,2/0,082,00,20,4/0,3
The product 28,0/3,000,3/0,13,00,10,5/0,2
Drug 312,0/7,00,05/0,052,00,30,5/0,1
The product 410,0/3,00,1/0,052,50,30,4/0,5
The drug 50,5/0,11,50,20,3/0,3
The drug 610,0/5,00,2/0,082,00,20,4/0,3

The study of the influence of medium composition for the preparation of samples of aorta and pulmonary artery were carried out by placing the samples in vials with standard suspension cell lines (FLACH-104, lung fibroblasts of human embryo production OOO "Bilat", Russia) with a concentration of 2.0×106cells/vial. Vials with vascular samples and cell suspension were placed in thermoshake ES-20 and was aged for 14 days under conditions of constant shaking.

Evaluation of the sorption capacity of the vascular samples was performed using flow cytometry with the use of device Cytomics FC 500 (manufactured by "Beckman Coulter, USA) by counting the number of fibroblasts, remaining in solution. The results (average number of fibroblasts in three samples) are given in table 2.

Table 2
The influence of the composition of the sterilizing medium and the thickness of adventitia to Sorb the ion capacity of the vascular sample
MedicationThe thickness of the layer of adventitia vascular pattern mmThe number of fibroblasts in the solution above the aorta samples (×106cells/vial)The number of fibroblasts in the solution over the samples of the pulmonary artery (×106cells/vial)
Drug - similar (control)01,8±0,1131,7±0,89
The drug (control without adventitia)01,76±0,731,8±0,21
The drug 10,5-1,00,57±0,0128*0,6±0,013*
The drug 11,0-2,00,34±0,051*0,47±0,033*
The drug 12,0-3,00,54±0,072*0,44±0,072*
The drug 15,0±1,00,73±0,0328*0,65±0,074*
The product 21,0-2,01,48±0,01281,52±0,052
Drug 31,0-2,01,38±0,0821,68±0,067
The product 41,0-2,01,42±0,0921,6±0,082
The drug 51,0-2,01,73±0,0681,7±0,044
The drug 61,0-2,01,56±0,0721,6±of 0.081
TC without treatment (control)1,0-2,01,72±0,0821,56±0,049
* - significantly relative to the control (p>0,05)

The best results are achieved when using the Drug 1 at save the Sri layer adventitia vascular pattern from 0.5 to 3.0 mm, the technology which was used in the preparation of the sample and its treatment with antibiotics.

To assess the effect on the adhesive properties of the vascular samples of acid-base condition of a solution have been carried out experiments on the sorption of fibroblasts on these samples at different pH. The results are shown in table 3.

Table 3
The influence of pH on the sorption ability of vascular sample
Acid-base state of the environment, pHThe number of fibroblasts in the solution above the aorta samples (×106cells/vial)The number of fibroblasts in the solution over the samples of the pulmonary artery (×106cells/vial)
7,0-7,40,7±0,232*0,56±0,09*
6,8-7,01,29±0,731,56±0,331
7,4-8,01,59±0,01281,16±0,023
* - significantly relative to the control (p>0,05)

The best results are achieved when using interval sour the IDT is the ground state of the medium (pH) in the range of 7.0 to 7.4.

To assess the role of individual ingredients in solution were carried out histological studies on the basis of Preparation 1. For this standardized plots obtained from one of the main vessel size (10×10) mm, were placed for 48 hours in the following solutions, and then they were fixed in formaldehyde. Based on them were made by histological and studied the effect of decellularization, evaluating the influence of solution composition on the number vacuolation (edematous) cells. The painting was done by van gieson. The increase in 1000. The calculation was made automatic method.

As solutions comparison was taken:

- Solution 1 (prototype-control) aqueous solution containing: the drug "Aminochrome - 1.0 g/l, Cefotaxime - 25,0 g/l, gentamycin sulfate - 4.0 g/l;

A solution of 2 - aqueous solution containing 0.3% of the mass. gentamicin sulfate, 10.0% of the mass. diflucan and 0.3% of the mass. components of a phosphate buffer solution;

A solution of 3 - aqueous solution containing 10.0% of the mass. diflucan and 0.3% of the mass. components of a phosphate buffer solution;

A solution of 4 - aqueous solution containing 0.3% of the mass. gentamicin sulfate and 10.0% of the mass. diflucan;

A solution of 5 - water solution containing 10.0% of the mass. diflucan;

A solution of 6 - water solution containing excessive amounts of biocidal additives - 0.8% of the mass. gentamicin sulfate, 14.0% of the mass. is diflucan and 0.3% of the mass. components of a phosphate buffer solution.

It was performed three series of experiments: for each environment was used by five of the vascular samples. The results are shown in table 4.

Table 4
The influence of medium composition for transportation on the number and morphology of cells of the tissue components of blood vessels
main type
The studied environmentThe number of damaged cells, % fragmented cellsThe number vacuolation cells, % of unmodified cells
Solution 122,421,3
Solution 226,3*78,1*
Solution 348,263,8
A solution of 457,979,2
Solution 562,886,1
Solution 618,735,7

The best results was the obtained by using the solution of 2, in the application of which was achieved by the high value of oncotic pressure in the cells (high percentage of vacuolization), while maintaining the integrity of cells, which can prevent the process of autolysis of tissue.

Example 2. Study of the influence of processing technology TC on biocidal characteristics of vascular tissue

To determine the optimal concentration of biocidal ingredients used in environments on the basis of the microbiological laboratory of children's hospital No. 1 were conducted microbiological studies on impact assessment of the composition of the solutions used in the manufacture of sterile fabric material on their biocidal activity. They counted effectiveness against the following strains: Staphylococcus aureus, Acinetobacter baumannii, Klebsiella pneumoniae, Pseudomonas aeruginosa, Candida albicans, Stenotrophomonas maltophilia, Serratia marcensens, Escherichia coli, Staphylo-coccus epidermidis, Candida nonalbicans, E. cloacae.

When this was investigated following solutions:

- Solution 1 (prototype-control) aqueous solution containing the drug "Aminochrome" at a concentration of 1.0 g/l, Cefotaxime 25,0 g/l: gentamicin sulfate 4.0 g/L.

A solution of 2 - aqueous solution containing 0.3% of the mass. gentamicin sulfate, 10.0% of the mass. diflucan and 0.3% of the mass. phosphate buffer solution.

A solution of 4 - aqueous solution containing 0.3% of the mass. gentamicin sulfate and 10.0% of the mass. of diflucan.

A solution of 5 - water solution, provided the excess amount of biocidal additives 0.8% of the mass. gentamicin sulfate, 14.0% of the mass. diflucan and 0.3% of the mass. phosphate buffer solution.

A solution of 6 - aqueous solution containing a smaller number of biocidal additives is 0.01% wt. gentamicin sulfate, 7.0 percent of the masses. diflucan and 0.3% of the mass. phosphate buffer solution. All study of culture at a concentration of 1 million units were sowed in the respective test tube with 5 ml of the respective solution containing biocidal additives. Sowing was performed by standard technique using a loop. The exposure time was 48 hours at 4°C. the evaluation of the results was performed after reseeding solutions on blood agar and environment Saburo. Response was assessed after 16 days of storage media in thermostat according to the criterion of the presence of gains (+) or absence (-). The results are shown in table 5.

Positive seeding Klebsiella pneumoniae indicates a high resistively hospital strains and against receipt of material after treatment, patients in the hospital. It should be noted the presence of strong synergy when using biocides in conjunction with phosphate buffer solution.

Further studies were performed on samples of vascular tissue, obtained according to the technology described in example 1 in accordance with the methodology of "General hygiene. Manual for laboratory practice: the educational allowance. - Kitsch DE 2009. - 288 S.: ill. "Microbiological monitoring of the working environment" (MU 4.2.734-99)". Tissue samples were placed in a sterile chamber and treated with a stream of outside air with exposure time of 5 min and 39 minutes then did the swabs and samples and placed them in Petri dishes containing agar-agar. The surface was dried for 30 minutes at room conditions (temperature (20±2)°C, relative humidity 50-60%) and were placed in a thermostat at 36±1°C for 24 hours, then under a microscope and counting the number of formed colonies of microorganisms.

The results are presented in table 6, where we have introduced the following notation: ++++ solid overgrown; +++ there are separate colonies of bacteria 10-20 cells;++, there are associations for 3-5 cells; + there is a single cell; 0 - bacterial contamination was not detected.

The results showed that the drug 1 and drug 3 have greater biocidal properties, but the use of the receipt of the drug 3 high amounts of antibiotics makes the drug 1 more promising for further research.

Example 3. Evaluation of the safety of connective tissue matrix.

Assessment of morphological preservation of the connective tissue matrix at different stages of preparation and storage of the drug in the use statement, the aqueous method and the method the closest analogue is carried out by carrying out morphological studies according to the following scheme.

Biological component GSSS was obtained by removal of TC during the forensic examination. After completion of the forensic study received standard aorta size (10×10) mm and investigated the changes in the morphological preservation of the connective tissue matrix under the influence of conditions of transportation and storage, in particular under the influence of ingredients used environments, and then spent the choice of the optimal composition of your environment.

The optimality criterion of the medium composition was the degree of disorganization of the collagen and elastic fibers, which was determined by the method described in the work (Book: Microscopic techniques, 1996, Series: prof/Medicina, http://www.e-reading-lib.org/djvureader. php/131685/391/_ne-hudlit_-_Mikroskopicheskaya_teh-nika.html...). In accordance with the specified methods samples of aorta was treated with standard drugs bacterial collagenase and pancreatitis E staining on van Giesen and resorcin-fuchsin, followed by a qualitative evaluation using light microscopy the intensity of staining and the presence of a destructuring of collagen and elastin fibers. This empirically for each of these proteolytic enzymes in the concentration were set, do not cause damage to the respective fibers in the control (untreated) samples of the aorta.

The evaluation was made by the authors of the invention on the basis of experimental studies point system, where you used the following quantitative and qualitative evaluation criteria: 0 - structure of collagen fibers and elastin saved and the intensity of staining corresponds to the test sample; 1 - fiber structure is retained, the intensity of staining decreased in some areas of histological preparation; 2 - fiber structure is retained, the intensity of staining decreased in all histological preparation, 3 - fiber decomposed into separate sections for histological preparation, the intensity of staining decreased in all histological preparation; 4 - fiber structure is broken into separate sections for histological preparation, the intensity of this color is violated in all histological preparation. The nature of the changes occurring in the vascular tissue in the process of preparation of the drug with the use of the claimed method and the method-analogue was evaluated according to light microscopy (magnification ×1000).

Samples of TC after forensic examination and preparation 1 obtained in example 1, were placed in transport medium different composition for 48 hours and opredelitelei option transport environment. When assessing transport environment we used the following songs:

control-patent-similar - distilled water;

TC - 1: water +0.2% mass. gentamicin sulfate and 0.08% of the mass. diflucan;

TC - 2: water +0.05% mass. gentamicin sulfate and 0.1% of the mass. diflucan;

TC - 3: water +0.5% mass. gentamicin sulfate and 0.02% of the mass. the diflucan. The results are shown in table 7.

Table 7
The influence of the composition of the transport environment on the integrity of the vascular tissue
Stage of researchProcessing technologyThe medium usedThe values of the scoring patterns of connective tissue matrix
The original TCThe drug 14,2
similarthe 3.8
Transport environmentThe drug 1TC 14,1*
The drug 1TC 23.2
The drug 1TC 32,5
The drug 1Water2,7
similarWater2,6
* - significantly compared to control (p>0,05)

The results showed that the best results are achieved when using TS-1, an aqueous solution containing 0.2% of the mass. gentamicin sulfate and 0.08% of the mass. diflucan.

The samples after soaking for 1 hour in the TC-1 were placed in a sterilizing environment of different composition for 48 hours, and then 12 days in a storage medium for different staff to determine the optimal sterilizing environment. The results recorded before being placed in a transport medium (TC-1), after 2 and 12 days of storage in the transport medium after sterilization treatment with antibiotics and after storage in nutrient media (claimed and the environment-the equivalent) for 12 days. Each of the treatment options were tested on three samples of aorta, each of which had 5 slices. The results are shown in table 8.

Evaluation of the influence of the composition of the storage environment on the safety of GSS was performed using the following mediums for storing homografts:

is similar: cefotaxim 25,0 g/l; gentamicin sulfate 4.0 g/l; aminochrome - rest;

- CX-1, gentamicin sulfate 0.1% wt.; diflucan 0.015% wt., the solution DMEM-F12 - rest; pH 7,2;

- CX-2, gentamicin sulfate and 0.15 wt. -%; diflucan 0.01% wt., the solution DMEM-F12 - rest; pH 7.0;

- CX-3, gentamicin sulfate 0.05% wt.; diflucan 0.02% wt., the solution DMEM-F12 - rest; pH 7.4;

Table 8
The influence of the composition of the storage environment on the integrity of the vascular tissue
Storage conditionsProcessing technologyThe medium usedThe number of cells destroyed and modified bodies
To storageThe drug 1, TS-1-13,4±4,2*
Storage for 12 days at 4°CThe drug 1, TS-1CX-119,6±4,7*
The drug 1, TS-1CX-221,4±3,5*
The drug 1, TS-1SH-320,6±4,2*
similar38,4±6,7
Storage for 90 days at +4°CThe drug 1, TS-1CX-133,6±3,2*
The drug 1, TS-1CX-242,6±3,4*
The drug 1, TS-1SH-335,6±5,1*
The drug 1, TS-1similar52,4±6,1
Hranenie days when
(-150±2)°C
The drug 1, TS-1CX-126,3±4,2*
The drug 1, TS-1CX-232,8±3,6
The drug 1, TS-1SH-335,6±5,1
The drug 1, TS-1similar44,4±6,1
Storage 1 year, cryo-preservation (-150±2)°CThe drug 1, TS-1CX-154,9±19,2*
The drug 1, TS-1 CX-261,7±10,4
The drug 1, TS-1SH-368,9±9,2
The drug 1, TS-1similar87,4±12,3 (see sediment)
• - significantly compared to control (p>0,05)

The results showed that the use of the claimed technology and media contributes to a better preservation of vascular tissue GSCs in comparison with analogues.

Example 4. Comparison of GSS obtained by the present technology and technology is the nearest equivalent.

The study was performed according to the techniques described in example 3. The results recorded at the end of transportation, after treatment with antibiotics and after storage in a storage medium for 12 days, when stored in nutrient media (claimed and the environment-the equivalent) for 12 days. Each of the treatment options were tested on three samples of aorta, each of which had 5 slices.

The use of the claimed technology.

At the stage of transporting the product had the following characteristics (the exposure time 6 hours).

Inner sheath (IN): single-layer thin endothelium with a single intact endothelial the cells, in potentiellen layer weak focal edema with weak mononuclear infiltration, the morphometric study of the intimal thickness: 15 μm to 50 μm.

The average shell (): clear elastic fiber model structure, between which are smooth muscle cells with clear nuclei, single core hypochromic, degenerative changes, the morphometric study thickness: from 450-500 microns.

The outer casing (BUT): the connective tissue with collagen fibers, isolated smooth muscle cells, the presence of a few small vessels. A valve. The endocardium of the model structure is lined by a single layer of flat endothelial with good clear endothelial cells, potentiellen layer stratifying bundles of collagen fibers, connective tissue with focal weak mucoidal swelling. Smooth myocytes and fibroblasts with clear nuclei in single cells marked dystrophic changes: swelling of nuclei, hypochromia. Using technology similar. Inner sheath (IN): single-layer thin endothelium with a single intact endothelially cells in potentiellen layer moderate focal edema with weak mononuclear infiltration. The morphometric study of the intimal thickness of 75 μm. The average shell (): clear elasticise the fiber model structure, between them are smooth muscle cells with clear nuclei, single core hypochromic, degenerative changes, weak focal edema (+). Damage to collagen (+). Damage to engines(+): 3-4 dystrophic, 1 autoresonance the core. The morphometric study a thickness of FROM 500 μm. The outer casing(BUT) presents connective tissue with collagen fibers, isolated smooth muscle cells, the presence of a few small vessels, pronounced swelling, razvlechenie. The morphometric study of the thickness BUT 500 μm. A valve. The endocardium of the model structure is lined by a single layer of flat endothelial with good clear endothelial cells, potentiellen layer stratifying bundles of collagen fibers, connective tissue with focal weak mucoidal swelling. Damage to collagen (++/+++). Damage to the nuclei of 20-30%, dystrophic changes of nuclei (swelling of the nuclei, hypochromia), the rest of the kernel intact. Smooth myocytes and fibroblasts with clear nuclei. The use of the claimed technology. Aging in TC-1, exposure time 24 hours. Inner sheath (IN): single-layer thin endothelium with a single intact endothelial cells in potentiellen layer of focal edema with mild to moderate mononuclear infiltration. The morphometric study of the intimal thickness: from 50 microns is about 110 μm. The average shell (): clear elastic fiber model structure, between which are smooth muscle cells with clear nuclei. The morphometric study thickness from 450-550 microns. The outer casing (BUT): presented by loose connective tissue with a few small vessels. The morphometric study of the thickness HO 150-500 μm. A valve. Single layer of flat endothelial model structure, potentiellen, subendocardialnah layer stratifying bundles of collagen fibers, connective tissue with focal edema, Muhiddin swelling. Smooth muscle fibers and cardiomyocytes skeletal heart with signs of dystrophic changes in: rough fuzzy contours of individual nuclei, swelling of the nuclei, a disorderly collapse of the chromatin. Using technology similar. Inner sheath (IN).: single-layer thin endothelium is maintained at a slight over with a single intact endothelial cells in potentiellen layer of focal moderate edema, swelling of the stroma, mild mononuclear infiltration. The morphometric study of the intimal thickness of 75 μm. The average shell (): clear elastic fiber model structure, between which are smooth muscle cells with clear nuclei, weak razvlechenie collagen framework, paaul is the single gaps due to the swelling. Damage to the collagen framework(+)/++. Damage to engines (+): 4-6 cores and degenerative changes (at HC. ×40). The morphometric study a thickness of FROM 650 μm. The outer casing (BUT) is represented by loose connective tissue with a few small vessels. The morphometric study of the thickness BUT 500-750 microns. A valve. Single layer of flat endothelial typical structure stored on a minor extent in potentiellen, subendocardialnah layer stratifying bundles of collagen fibers, connective tissue with focal edema, Muhiddin swelling. Damage to collagen (+++). Damage to the cores 60% dystrophic, (+++). Smooth muscle fibers with signs of dystrophic changes:

50% non-nuclear cardiomyocytes skeleton of the heart, in the remaining 50% - dystrophic changes in: rough fuzzy contours of individual nuclei, swelling of the nuclei, a disorderly collapse of the chromatin. The use of the claimed technology. When selecting in distilled water. Inner sheath (IN): single-layer thin endothelium with a focal desquamation, potentiellen layer of weak diffuse edema, swelling of nuclei isolated smooth muscle cells, weak diffuse mononuclear infiltration. The morphometric study of the intimal thickness: 40 μm to 100 μm. The average shell (CO): a clear elastic hair is on the model structure with focal weak interstitial edema, diffuse weak mononuclear infiltration, isolated smooth muscle cells are hypochromic, with indistinct contours of the nuclei, with signs of degenerative changes and lysis of the nuclei. The morphometric study thickness from 500-550 microns. The outer casing (BUT): presents loose stratifying connective tissue with severe edema, single vessels with good endothelial cells and perivascular weak lymphocytic infiltration. The morphometric study a thickness of 100 μm. A valve. Focal weak and moderately expressed Mokoena swelling and swelling of the endocardium, myocytes and fibroblasts with clearly konturirovany nuclei in isolated nuclei signs of degenerative changes, mild mononuclear infiltration. Using technology similar. Inner sheath (IN): single-layer thin endothelium with a focal desquamation, potentiellen layer of weak diffuse edema, swelling of nuclei isolated smooth muscle cells, weak diffuse mononuclear infiltration. The morphometric study of the intimal thickness: 40 μm to 100 μm. The average shell (): clear elastic fiber model structure with focal moderate interstitial edema, maloideae swelling, weak diffuse mononuclear infiltration, isolated smooth muscle cells are hypochromic, with indistinct outline the mi cores, with signs of degenerative changes and lysis of the nuclei. Damage to the collagen framework (++). Damage to the nuclei of cells: (++) 30 dystrophic changes of cells/10-15 with autolysis (the remnants of the cores of the granules of chromatin). The morphometric study thickness from 1000 μm. The outer casing (BUT): presents loose stratifying connective tissue with severe edema, single vessels with good endothelial cells and perivascular weak lymphocytic infiltration. The morphometric study a thickness of 100 μm. A valve. Focal weak and moderately expressed Mokoena swelling and swelling of the endocardium, myocytes and fibroblasts with clearly konturirovany nuclei in isolated nuclei signs of degenerative changes, mild mononuclear infiltration. The best preservation of the cores 30% nuclear-free cardiomyocytes, 30-40% degenerative changes Damage nuclei of stroma 30-50%(++)Povrezhdeniya collagen (+/++).

The use of the claimed technology. The biocidal treatment drugs within 48 hours. Inner sheath (IN): moderate intimal edema, swelling of nuclei isolated smooth muscle cells. The morphometric study of the intimal thickness: from 15 to 75 microns. The average shell (): clear elastic fibers typical structure with a moderate focal interstitial dissecting elastic frame edema is m, single smooth muscle cells autoryzowany cores, signs of degenerative changes in 1/3 of the nuclei of smooth muscle cells. The morphometric study thickness FROM 700 to 750 μm. The outer casing (BUT): friable dramatically swollen remnants of thin fibers reticular fibers with disruption and lysis of the collagen fibers. The morphometric study of the thickness BUT 200 μm. A valve. Pronounced swelling, maloideae and fibrinoid swelling of collagen fibers and endocardium, smooth muscle fibers, lysis and necrosis of cardiomyocytes (cell-shade).

Using technology similar. Inner sheath (IN): moderate intimal edema swelling of nuclei isolated smooth muscle cells. The morphometric study of the intimal thickness from 15 to 75 microns. The average shell (): clear elastic fibers typical structure with a moderate focal interstitial dissecting elastic frame edema, isolated smooth muscle cells autoryzowany cores, signs of degenerative changes in 1/3 of the nuclei of smooth muscle cells. Damage to the collagen skeleton ++/+. Damaged kernels: 20-30 dystrophic changes of nuclei (++). The morphometric study a thickness of FROM 500 μm. The outer casing (BUT): friable dramatically swollen remnants of thin fibers reticular fibers with disorganizatsiya lysis of collagen fibers. The morphometric study of the thickness BUT 200 μm. A valve. Pronounced swelling, maloideae and fibrinoid swelling of collagen fibers and endocardium, smooth muscle fibers, lysis and necrosis of cardiomyocytes (cell-shade). 50% nuclear-free stroma. Damaged kernels - 100% degenerative changes. Damage to collagen (+++++).

The use of the claimed technology. Storage 13 days in the environment of the CX-1 (gentamicin sulfate 0.1% wt.; diflucan 0.015% wt., the solution DMEM-F12-rest; pH of 7.2). Inner sheath (IN): pronounced intimal edema swelling and lysis of nuclei isolated smooth muscle cells. The morphometric study of the intimal thickness: from 50 microns to 75 microns. The average shell (CO): marked interstitial edema, maloideae swelling of the disorganization of the elastic frame, degenerative changes and autolysis more1/2smooth muscle cells. The morphometric study thickness WITH 500-550 microns. The outer casing (BUT): marked swelling of the connective tissue, which are represented by thin threads of reticular fibers. The morphometric study of the thickness BUT 500 μm. A valve. Pronounced swelling, fibrinoid necrosis of collagen fibers and endocardium, smooth muscle fibers, lysis and necrosis of cardiomyocytes (cell-shade).

Using technology similar. Inner sheath (IN): marked swelling Inti the s swelling and lysis of nuclei isolated smooth muscle cells, focal desquamation. The morphometric study of the intimal thickness: from 0 μm to 75 μm. The average shell (CO): marked interstitial edema, pronounced Mokoena swelling of the disorganization of the elastic frame, degenerative changes and autolysis more1/2smooth muscle cells. Damage to the collagen framework (+++++). Damage to the elastic frame (+++). Damage to the cores 100% of all degenerative changed and lost core. The morphometric study a thickness of FROM 650 μm. The outer casing (BUT): marked swelling of the connective tissue, which are represented by thin threads of reticular fibers In morphometric study of the thickness BUT 200 μm. A valve. Pronounced swelling, fibrinoid necrosis of collagen fibers and endocardium, smooth muscle fibers, lysis and necrosis of cardiomyocytes (cell-shade). Damage to collagen (+++++) - filamentous net collagen. Damage to the cores 100% degeneration and lysis, 70% nuclear-free stroma.

The use of the claimed technology. When stored 13 days in an environment at acidic pH values. Wednesday gentamicin sulfate 0.1% wt.; diflucan 0.015% wt., the solution DMEM-F12-rest; pH 6.4. Inner sheath (IN): detachment and lysis intima at greater length. The morphometric study of the thickness of the intima: 15-50-100 μm. The average shell (CO): pronounced interstitial is flowed, Mokoena swelling of the disorganization of the elastic frame, smoothness undulating contours of elastic fibers, degenerative changes and autolysis 1/3 of smooth muscle cells WITH closer to BUT. The morphometric study of the thickness WITH more than 750 microns. The outer casing (BUT): marked swelling of the connective tissue, which are represented by thin threads of reticular fibers. The morphometric study of the thickness BUT 500 μm. A valve. Pronounced swelling, fibrinoid necrosis of collagen fibers and endocardium, smooth muscle fibers, lysis and necrosis of cardiomyocytes (cell-shade). Using technology similar. Inner sheath (IN): detachment and lysis intima at greater length. The morphometric study of the thickness of the intima 15-100 μm. The average shell (CO): the relatively good preservation of cell nuclei, mild interstitial edema, maloideae swelling of the disorganization of the elastic frame. Damage to the collagen skeleton (+/++). Damaged kernels (++) - 20 dystrophic changes of nuclei. Preferential damage to the nuclei closer to BUT. The morphometric study a thickness of from 500 μm. The outer casing (BUT): marked swelling of the connective tissue, which are represented by thin threads of reticular fibers. The morphometric study of the thickness BUT 500 μm. A valve. Pronounced swelling, fibre oigny necrosis of collagen fibers and endocardium, smooth muscle fibers, lysis and necrosis of cardiomyocytes (cell-shade). Damage to collagen (+++++) - filamentous net collagen. Damage to the cores 100% degeneration and lysis, 60% nuclear-free stroma. The use of the claimed technology. After cryopreservation (freezing at -(150)°C and thawing). Inner sheath (IN): detachment and lysis intima at greater length. The morphometric study of the intimal thickness of 50 μm. The average shell (CO): marked interstitial edema, maloideae swelling of the disorganization of the elastic frame, smoothness undulating contours of elastic fibers, degenerative changes and autolysis 1/3 of smooth muscle cells WITH closer to BUT. The morphometric study of the thickness WITH more than 750 microns. The outer casing (BUT): detachment the most part, HOWEVER, marked swelling of the connective tissue, which are represented by thin individual filaments reticular fibers. The morphometric study of the thickness BUT 500 μm. A valve. Pronounced swelling, fibrinoid necrosis of collagen fibers and endocardium, smooth muscle fibers, lysis and necrosis of cardiomyocytes (cell-shade).

Using technology similar. Inner sheath (IN): detachment and lysis intima at greater length. The morphometric study of the intimal thickness of 50 μm. The average shell (CO): pronounced interstitial is flowed, Mokoena swelling of the disorganization of the elastic frame, smoothness undulating contours of elastic fibers, degenerative changes and autolysis 1/3 of smooth muscle cells WITH closer to BUT. Profound damage to the collagen framework (++++), the fiber breaks. Kernel-clear, relatively intact. Damaged kernels (++) - 25 dystrophic changes of cells/7 lysed cells. The morphometric study a thickness of from 750 microns. The outer casing (BUT): detachment the most part, HOWEVER, marked swelling of the connective tissue, which are represented by thin individual filaments reticular fibers. The morphometric study of the thickness BUT 500 μm. A valve. Pronounced swelling, fibrinoid necrosis of collagen fibers and endocardium, smooth muscle fibers, lysis and necrosis of cardiomyocytes (cell-shade), 70-80% of a nuclear-free stroma. Damage to collagen (++++/+++++).

Comparison of morphological characteristics of the LC and homografts at various stages of processing TC showed that the proposed technology promotes better retention of TC cells and elements of the CCC included in homograft.

Example 5. Evaluation of mechanical properties GSSSC conducted by filling homograft saline and measuring the change in diameter of homograft at the level of the insertion of the commissures of the valve. The results are shown in tab is .9.

Table 9
The effect of the nature of the frame on the degree of preservation of the mechanical properties of the wall combined homograft
Description GSSSThe diameter of homograft
After filling with salineTo fill saline
GSSS aortic (polymer frame)116±278±6
Saar aortic (frame made of pyrocarbon)98±278±6
Saar pulmonary (frame of steel grade KM).96±371±7
SG arterial (a skeleton of steel grade HN.).97±275±5

The findings suggest that the use of a skeleton of steel or pyrocarbon significantly increases the rigidity of homograft.

Example 6. On the technology described in examples 1-3 were obtained following homograft (list not exclusive).

The list of models obtained homog is avtov:

- homograft vascular laparostomy aortic (GSK Ao. 1.1);

- homograft vascular laparostomy with aortic ascending aorta and arch of aorta (GSK Ao.1.2);

- homograft vascular laparostomy aortic modified (GSK AOM);

- homograft vascular laparostomy with aortic ascending aorta modified (GSK AOM);

- homograft vascular laparostomy aortic combined (GSK AOC);

- homograft vascular laparostomy pulmonary (GSK Le);

- homograft vascular laparostomy with pulmonary bifurcation (GSK Le);

- homograft vascular laparostomy pulmonary modified (GSK LEM);

- homograft vascular laparostomy pulmonary combo (GSK LEC);

- homograft vascular laparostomy aortic-mitral (GSK Aomi);

- homograft vascular laparostomy aortic-mitral combined (GSK Omic);

- homograft laparostomy mitral modified (ha MIM);

- homograft laparostomy mitral combined (ha MIC);

- homograft vascular arterial (GSA);

- homograft vascular arterial with bifurcation (GSA);

- homograft vascular venous (GSW);

- homograft vascular venous with bifurcation (GSV).

p> Characteristics of a number of homografts below.

Homograft vascular laparostomy aortic (GSK Ao. 1.1)

General configuration: the root of the aorta, the proximal base wall of the myocardium at the level of 5-10 mm proximal to the virtual ring formed at the connection points of the basal sagging flaps, and mitrale-aortic fibrous contact; distal base - signatureline connection; the mouth of the coronary arteries legirovanyh. The type, size and anatomical relationship of the structural elements: the front flap of the mitral valve; ventriculoarterial connection: cushion endocardial infarction with, attached to the fibrous ring of the aorta, the thickness of (4±1) mm, a length of 5-10 mm in the direction from the virtual ring formed at the connection points of the basal sagging flaps; the aortic valve consists of three aortic valves attached at the lower edges of the sinuses of Valsalva to the wall of the aortic root at the annulus of the aorta and United at the top of each other with the formation of the three commissures; the fibrous ring of the aorta; the outer walls of the aortic root.

The sizes. The thickness of the layer of adventitia 1,0±0,5 mm inner Diameter: aorta at the level of the annulus 4-40 mm Increment of the diameter of the aorta 1 mm Length of the ascending aorta 10-100 mm length Increment of the ascending aorta 1 mm Homograft vascular clap ostergade pulmonary (GSK Le).

The General configuration of a fragment of the pulmonary artery, the proximal base wall pontegadea cone of the myocardium at the level of 5-10 mm proximal to the virtual ring formed at the connection points of the basal sagging flaps; distal base - virtual ring formed by the tops of the commissures semilunar valves.

The type, size and anatomical relationship of the structural elements: the roller pontegadea cone attack with endocardial thickness (4±1) mm, 10 mm proximal to the virtual ring formed at the connection points of the basal sagging valves; valve pulmonary artery, which consists of three semilunar valves, United at the top of each other with the formation of the three commissures; section of pulmonary artery, a limited sinus Valsalva.

Sizes

The thickness of the layer of adventitia of 1.5±0.5 mm internal Diameter, pulmonary artery at the level of the virtual ring formed at the connection points of the basal sagging shutters, 4-40 mm Increment of diameter 1 mm, Length of pulmonary artery 10-100 mm Increment of length 1 mm Homograft vascular laparostomy with pulmonary bifurcation (GSK Le).

General configuration of the pulmonary artery on the right and left branches of the pulmonary artery: proximal base wall pontegadea cone of the myocardium at the level of 5-10 mm proximal to virtualnogo the ring, formed at the connection points of the basal sagging flaps; distal base - level division of the branches of the pulmonary arteries in equity pulmonary artery.

The type, size and anatomical relationship of the structural elements: the roller pontegadea cone attack with endocardial thickness (4±1) mm, 10 mm proximal to the virtual ring formed at the connection points of the basal sagging valves; valve pulmonary artery, which consists of three semilunar valves, United at the top of each other with the formation of the three commissures; pulmonary artery to the bifurcation; the branches of the pulmonary artery.

Sizes

The thickness of the layer of adventitia of 2.5±0.5 mm internal Diameter, pulmonary artery at the level of the virtual ring formed at the connection points of the basal sagging shutters, 4-40 mm, pulmonary artery at the level of the mouth of its branches - the left 3-30 mm, right - 3-30 mm Increment of diameter 1 mm, Length of pulmonary artery 10-100 mm, branches of the pulmonary artery right - 1-40 mm, left - 1-40 mm length Increment, 1 mm.

Homograft vascular laparostomy aortic-mitral (GSK AAMI)

The General configuration of a fragment of the aorta with a fragment of the mitral valve: the proximal base wall of the myocardium at the level of 5-10 mm proximal to the virtual ring formed at the connection points of the basal sagging zaslona is, and the fibrous ring of mitral valve; distal base - arch of the aorta at the level of the left subclavian artery; the mouth of the coronary arteries legirovanyh; continuous seam joining the pericardial space with papillary muscles.

The type, size and anatomical relationship of the structural elements: the mitral valve, consisting of two wings - front and rear, tendinous chords, papillary muscles and the fibrous ring of mitral valve; unification papillary muscles with stitched area of the pericardium; ventriculoarterial connection: cushion endocardial infarction with, attached to the fibrous ring of the aorta, the thickness of (4±1) mm, a length of 5-10 mm in the direction from the virtual ring formed at the connection points of the basal sagging flaps; the aortic valve consists of three aortic valves attached at the lower edges of the sinuses of Valsalva to the wall of the aortic root from the fibrous rings of aorta and United at the top of each other with the formation of the three commissures; the fibrous ring of the aorta; the outer walls of the aortic root; ascending aorta; aortic arch with lots of brachiocephalic vessels; the plot of the pericardium.

Sizes

The thickness of the layer of adventitia 1,5±0,5 mm inner Diameter: aorta at the level of the virtual ring formed at the connection points of the basal sagging flaps of 15-30 mm, the annulus mi the Central valve 15-40 mm. The increment of the diameter of the aorta 1 mm Length of the ascending aorta 10-100 mm length Increment of the ascending aorta 1 mm.

Homograft vascular laparostomy aortic combined (GSK OK)

General configuration: combined structure in the form of a frame with stiffeners, inside of which is fixed to the aortic valve; the proximal base annulus of the aorta at the level of 5 mm proximal to the virtual ring formed at the connection points of the basal sagging flaps, and mitrale-aortic fibrous contact; distal base - virtual ring formed by the tops of the commissures semilunar valves.

The material from which made GSSSC: textile components allogeneic post-mortem human valve material: the aortic valve consists of three aortic valves attached at the lower edges of the sinuses of Valsalva to the wall of the aortic root at the annulus of the aorta and United at the top of each other with the formation of the three commissures; the fibrous ring of the aorta. The fixing frame separate and continuous sutures; the thickness of the layer of adventitia 1,0±0,5 mm; frame - steel KM.

Homograft vascular laparostomy pulmonary combo (GSK LEC)

General configuration: combined structure in the form of a frame with stiffeners, inside of which is fixed pulmonary valve; proximal founded the e - wall pontegadea cone of the myocardium at the level of 5 mm proximal to the virtual ring formed at the connection points of the basal sagging flaps; distal base - virtual ring formed by the tops of the commissures semilunar valves.

The material from which made GSSSC: roller pontegadea cone attack with endocardial thickness (2±1) mm, 5 mm proximal to the virtual ring formed at the connection points of the basal sagging valves; valve pulmonary artery, which consists of three semilunar valves, United at the top of each other with the formation of the three commissures; the thickness of the layer of adventitia of 1.5±0.5 mm; frame material - pyrocarbon.

Homograft laparostomy mitral combined(ha MIC)

General configuration: combined structure in the form of a support ring, on the outer surface of which is fixed to the fibrous ring of mitral valve fragment of the mitral valve: the proximal base of the fibrous ring of mitral valve from the left atrium, which is described relative to the circumference of the corresponding base support ring; the distal base of the fibrous ring of mitral valve from the left ventricle;

The material from which made GSSSC: textile components allogeneic post-mortem human what about the valve material; mitral valve consisting of two wings - front and rear, tendinous chords, papillary muscles and the fibrous ring of mitral valve; unifikasiyaya papillary muscles with stitched area of the pericardium; fixing a separate and continuous welds along the line of contact between the bearing rings with a circumference of the annulus; a continuous seam joining the pericardial space with the papillary muscles, the thickness of the layer of adventitia 1,0±0,5 mm; frame - steel.

Example 7. Sick Hours, 2 years old, was admitted for a routine examination and treatment at the Department of cardiac surgery, St. Petersburg state budgetary institution of health "Children's city hospital №1" (children's hospital №1). Complaints cyanosis of the lips.

Performed echocardiography (EchoCG). Diagnosis: Congenital heart disease (CHD). Tetralogy Of Fallot. Atresia of the pulmonary artery. Ventricular septal defect. Collateral blood flow in the pulmonary circulation.

The survey recommended palliative surgery: univocality pulmonary blood flow, implantation biodisposition of homograft between the right ventricle and pulmonary artery.

The operation produced using the claimed GSS model: homograft vascular laparostomy aortic modified, type "freshly prepared" (G Is To ASV). The diameter of the tube at the level of the annulus 12 mm, tube length 45 mm (GSS-1). During the operation, performed a median sternotomy. Native pulmonary artery with a diameter of 5 mm giperplazirovanne. Selected two collaterals left lung root with diameters of 4 mm and the collateral right 3 mm in diameter with a solitary lesion. The selected vessel, extending from the thoracic aorta, at the level of the Atria divisible by collaterals, going to the right and left lung. Connected to the cardiopulmonary bypass (CPB). Patient: hypothermia 28°C. the Mouth of collateral vessels tied up. Collaterals to the lower lobes of the lungs anastomoziruet in the rear wall of the pulmonary artery at the level of bifurcation. Two left collaterals anastomoziruet between a side-to-side and sewn into the left pulmonary artery. The collateral from the upper lobe of the right sewn into the right pulmonary artery. Anastomoses performed continuous suture (8-0 Prolene). In the front wall of the pulmonary artery is sewn into the distal end biodisposition pulmonary homograft (GSS-1). Made cardioplegia. Stopped heart. In the outflow tract of the right ventricle sewn proximal end of the aortic homograft with a diameter of 12 mm. Restored cardiac function. Produced modified ultrafiltration. Hemostasis. Completed layer stitching wounds.

Postoperative care the patient is satisfactory. The results of Echocardiography: on the 10th, 30th and 90th day noted that transylavania gradient on implantirovannomu GSSS corresponds to the age norm.

The patient was discharged on the 14th day after surgery for outpatient treatment under the supervision of a cardiologist.

Three months after surgery due to identification during the sensing of the heart cavities availability ortolani collaterals recognized the need for a second surgery for ligation ortolani collaterals to the left.

According to the comprehensive clinical examination (Echocardiography, ECG, radiography, spiral computed tomography, angiography, performed within six months once in two months: chambers of the heart correspond to the sizes, the sizes of the vessels of a small circle close to the age norm.

In connection with the formation (in the previously executed operations) vessels of the pulmonary circulation in the form of a single channel, the opportunity to carry out a radical correction of the UPU: the closure of the ventricular septal defect. The specified operation is performed with the use of declared GSS model: homograft vascular laparostomy aortic type "freshly prepared" (GSK ASV). The internal diameter of the tube at the level of the annulus 16 mm, tube length 40 mm (HSS-2). During the operation, re Regina sternotomy. The heart is separated from the binding of cautery.

Visual inspection GSS-1 (implanted during palliative surgery): signs of calcification is not identified; noted the sprouting of blood vessels that feed the wall GSS-1. The right pulmonary artery with a diameter of 6 mm is Connected AIK. Patient: hypothermia 28°C. Parijata aorta, made cardioplegia in aortic root. Opened right atrium. The left atrium is drained through the mouth of the pulmonary veins. Revealed: the aorta 2/3 shifted to the right; ventricular septal defect increased. Ventricular septal defect (subaortalnom, diameter 2×3 cm) closed patch from xenopericardial continuous suture (Prolene 6-0). Made prevention of air embolism. Removed the clamp from the aorta, the marked recovery of cardiac activity. Removed previously implanted GSS-1. Isolated and cut off the mouth of the left pulmonary artery. Between the mouths of the pulmonary arteries and right ventricle sewn GSS-2 (Prolene 7-0). The proximal end HSS-2 is connected with the right ventricle (Prolene 7-0). Produced by closure of the right ventricle. Disabled AIK. Made manometry: PJ = 65/0, radial artery = 66/34 mm Hg HSS-2 cut and wedge-shaped removed portion of its wall to prevent tipping GSS-2. The results of the second manometry: PJ = 65/0, radial artery = 85/44 mm Hg Filed the electrode into the right ventricle. Hemosta is. Drainage behind the sternum. Produced suturing wounds of the chest with stable hemodynamics, with minimal inotropic support.

According to the results of microscopic examination remote GSS-1 revealed the presence of blood vessels that feed the wall GSS-1, and fibroblast recipient, "occupying" wall HSS-1.

The patient was discharged in satisfactory condition on the 10th day after the operation. During the observation time within one year after radical correction UPU violations of GSS-2 was not observed. Signs of hemodynamic and circulatory failure is not marked.

Example 8. Patient S., age 12, he entered the Department of cardiac surgery children's hospital No.1 for urgent indications in connection with the deterioration in the background of progressive cardiovascular disease caused by stenosis of the mechanical prosthesis in the tricuspid position. The Diagnosis Of Wilms Tumor. Condition after bacterial endocarditis. Condition after surgical removal of vegetation from the cavity of the right ventricle. After implantation of the mechanical prosthesis in the tricuspid position (aged three years). Stenosis mechanical prosthesis. Set indications for urgent surgical treatment.

Performed the surgery to replace mechanical prosthesis stated GSS model: homograft laparostomy mitral, type "SV is uprightly" (ha MISW). The diameter of the ring of mitral valve 26 mm (HSS-3). During the operation, removed stenotic mechanical prosthesis. Pad chords GSSS-3 fixed two separate " " U" joints to the wall of the right ventricle. The valve ring is fixed to the atrioventricular opening continuous suture. Performed hydraulic test showed the absence of regurgitation on the leaves. During intraoperative trancedevotee Echocardiography pathology implanted GSSS-3 have been identified.

The patient was discharged on the 7th day after surgery to outpatient observation.

The patient was surveyed three months after surgery. In the future, the survey was conducted annually for three years. The results: signs of circulatory failure and hemodynamic changes were not found.

Example 9. Patient W., age 17, he enrolled at the Department of cardiac surgery children's hospital No.1 routinely for surgical treatment with a diagnosis of Abnormality of Ebstein. Condition after prosthetic tricuspid valve mechanical prosthesis (aged five years). Treatment is indicated in connection with stenosis mechanical prosthesis and the inability to continue receiving indirect anticoagulants (warfarin) due to increased levels of transaminases in two times in comparison with normal values. Postavlennyheyu for urgent surgical treatment.

Performed the surgery to replace mechanical prosthesis stated GSS model: homograft vascular laparostomy pulmonary combo of "freshly prepared" (GSK LECs). The diameter of the tube at the level of the annulus 28 mm, tube length 20 mm (GCSS-4). During the operation, removed stenotic mechanical prosthesis. Valve ring HSS-4 fixed to the atrioventricular opening continuous suture. Performed hydraulic test showed the absence of regurgitation on the leaves. During intraoperative trancedevotee Echocardiography pathology implanted GSS-4 have been identified.

Indications for use of indirect anticoagulants not.

The patient was discharged on the 11th day after surgery to outpatient observation.

The patient was surveyed three months after surgery. According to the survey results: liver function is normalized, the transaminase levels began to comply with the age regulations. According to the annual surveys within 3.5 years after surgery: signs of circulatory failure and hemodynamic changes were not found.

Example 10. Declare GSSS in the amount of 124 units were used during the surgical treatment of 117 patients with congenital and acquired heart defects (CHD and PPP) aged from 10 days to 56 years. Treatment osushestvljali is based on the following institutions: children's hospital №1; State budget-funded health care institution "Republican cardiology clinic" (, Ufa, Russia), Samara regional clinical cardiology dispensary (, Samara); State budgetary institution of the Republic of Komi "Cardiologic dispensary (, Syktyvkar);

Federal State budget-funded health care institution "Federal centre for cardiovascular surgery of the Ministry of health of the Russian Federation", Penza, and Astrakhan.

In the course of operations have been applied, in particular, vascular cleanstream homograft aortic, aortic modified and combined aortic; pulmonary, pulmonary modified and combined pulmonary; cleanstream homograft mitral etc. All patients produced a comprehensive examination in dynamics, including ultrasound of the heart (at discharge: on the 12th - the 40th day, and after one and three years after surgery) and the sensing cavities of the heart (according to the testimony or three years after surgery). The effectiveness of the treatment was performed using criteria such as: growth translating gradient (three years after surgery compared with baseline at discharge) and the degree of regurgitation on the valves homografts (three years after surgery). Accordingly, the analysis of the results of clinical observations produced the respect implanted GSSS with a period of observation of at least three years after the operation.

The comparison group consisted of 140 patients with EPS and PPS in age from 5 days to 62 years, in whom the treatment was carried out with the use of previously developed by the author GSSS adopted for the nearest equivalent (147 GSSS). Operations were performed on the basis of children's hospital No. 1, the establishment of health care Leningrad regional clinical hospital and the establishment of health care of the Arkhangelsk region "First city clinical hospital. Was" (Arkhangelsk). The treatment efficiency was evaluated similarly claimed GSSS.

Comparative results of the effectiveness of surgical treatment using the inventive GSSS and GSSS taken the closest analogue, showed that the growth translating gradient through 3 years after surgery was to declare GSSS - (12,7±7,9) mm Hg, for GCSS-analog - (22,6±7,5) mm Hg and the degree of regurgitation, valve GSSS 1 degree for the claimed GSSS was achieved in 46 patients (39.3 per cent), in the case of the use GSS-analog - 22 patients (15.7 percent), and 1 and 2 degrees, respectively, to 59.8% patients against 46,4%, which clearly indicates that degenerative changes in the leaves of the valve claimed GSSS slower than GSSS adopted for the nearest equivalent.

As shown by tests, the claimed GSSS provide the best accepted of the prosthesis in the body, due to the almost complete exclusion of the immunogenicity of cells EP is Telia and good sorption cells of the host body on the treated adventitia, and can be stored more than 90 days at a temperature (0-4°C and above, with the cryoconservation in liquid nitrogen.

Declare GSSS can effectively be used in the correction of congenital and acquired heart diseases and blood vessels, such as atresia of the pulmonary arteries, transposition of the great arteries with stenosis of the pulmonary artery, common arterial trunk, etc., correction of valvular heart disease and other pathologies CCC.

1. Homograft cardiovascular system, which are taken from the corpse of a section of the main vessels or output paths of the right and left ventricles of the heart, characterized in that it is subjected to pre-processing environment, which is an aqueous solution containing 0.05-0.5% wt. gentamicin sulfate, 8,0-12,0% of the mass. diflucan and 0.1-0.5% components of a phosphate buffer solution, for at least 30 minutes; and then at the stage of preparation environment with a pH of 7.0 to 7.4, representing a solution of DMEM/F12 containing 3,0-7,0% of the mass. diflucan, 0.05 to 0.1% of the mass. gentamicin sulfate, 1.5 to 3.0 wt%. of cyclosporine, 0.1 to 0.3% of the mass. metrogel, 0.3-0.5% of the mass. components of a phosphate buffer solution; and then stored in an environment that represents a solution of DMEM/F12 containing 8,0-12,0% of the mass. diflucan, 0.05 to 0.15% of the mass. gentamicin sulfate and 0.1-0.5% wt. components of a phosphate buffer solution.

2. homograft cardiovascular system according to claim 1, characterized in that the section of the main vessels or output paths of the right and left ventricles of the heart contains the outer layer of adventitia thickness of 0.5 to 3.0 mm

3. Homograft cardiovascular system, containing taken from the corpse of a section of the main vessels or output paths of the right and left ventricles of the heart, placed in a tube of material non-biological nature, characterized in that it is subjected to pre-processing environment, which is an aqueous solution containing 0.05-0.5% wt. gentamicin sulfate, 8,0-12,0% of the mass. diflucan and 0.1-0.5% components of a phosphate buffer solution, for at least 30 minutes; and then at the stage of preparation environment with a pH of 7.0 to 7.4, representing a solution of DMEM/F12 containing 3,0-7,0% of the mass. diflucan, 0.05 to 0.1% of the mass. gentamicin sulfate, 1.5 to 3.0 wt%. of cyclosporine, 0.1 to 0.3% of the mass. metrogel, 0.3-0.5% of the mass. components of a phosphate buffer solution; and then stored in an environment that represents a solution of DMEM/F12 containing 8,0-12,0% of the mass. diflucan, 0.05 to 0.15% of the mass. gentamicin sulfate and 0.1-0.5% wt. components of a phosphate buffer solution.

4. Homograft cardiovascular system according to claim 3, characterized in that the section of the main vessels or output paths of the right and left ventricles of the heart contains the outer layer of adventitia thickness of 0.5 to 3.0 mm

5. G is MoGraph cardiovascular system according to claim 3, characterized in that the tube of material non-biological nature is made of pyrocarbon.

6. Homograft cardiovascular system according to claim 3, characterized in that the tube of material non-biological nature is made of steel.

7. Homograft cardiovascular system according to claim 6, characterized in that the tube of material non-biological nature is made of steel grade HIM.

8. The method of producing homograft cardiovascular system, including the sampling of tissue components, their preparation when exposed to the resulting product with a solution of salts and nutrients and the mixture bactericidal drugs, containing an antifungal drug, an antibiotic of the group of cephalosporins and gentamicin sulfate, and introduction to storage medium containing solutions of salts and nutrients, a mixture of antibiotics, including gentamicin sulfate, wherein the pre-tissue components of the cardiovascular system treated with an aqueous solution containing gentamicin sulfate, and diflucan, and preparation carried out in such a way as to maintain a layer of adventitia thickness of 0.5-3.0 mm while on the tissue components of the cardiovascular system is affected by the solution containing as a source of salts and nutrients DMEM/F12 and a mixture of antibacterial preparations containing Ni is sustained fashion metrogylum, and storage is carried out at pH 7.0-7.4 in a medium containing as a source of salts and nutrients in a solution of DMEM/F12, and a mixture of antibiotics diflucan.

9. The method of producing homograft cardiovascular system of claim 8, wherein the tissue components of the cardiovascular system treated with an aqueous solution containing 0.3% of the mass. gentamicin, 10.0% of the mass. diflucan and 0.3% of the mass. components of a phosphate buffer solution, and a solution of DMEM/F12 containing 0.08% mass. gentamicin, 5,0% of the mass. diflucan, and 2.0% wt. of cyclosporine, 0.2% wt. metrogel and 0.4% of the mass. components of a phosphate buffer solution.

10. The method according to claim 8, characterized in that the storage mode of the cryopreservation homograft cardiovascular system previously placed in a sterile polymeric package with cryoprotective mixture containing 20% of the mass. albumin, 10% vol. dimethyl sulfoxide and DMEM/F12 in the form of a solution, cooled to a temperature of (2±2)°C, placed in a refrigerator with a temperature of (150±2)°C and maintain the service for a period of not less than 120 minutes

11. Environment effects on tissues of homograft cardiovascular system after their selection and/or transportation containing a mixture of antifungal drug with antibiotic group of cephalosporins and gentamicin sulfate, characterized in that it further comprises DMEM/F12, metrogylum and to mponent phosphate buffer solution in the following ratio of ingredients (% wt.):

diflucan3,0-7,0
gentamicin sulfatethe 0.05-0.1
the cephalosporin1,5-3,0
metrogylumof 0.1-0.3
components of a phosphate buffer solution0,3-0,5
DMEM/F12rest

12. Environment effects on tissues of homograft cardiovascular system after their selection and/or transportation in claim 11, characterized in that it contains the solution in DMEM/F12 mixture, containing 5.0% of the mass. diflucan, 0.08% mass. gentamicin sulfate, and 2.0% wt. of cyclosporine, 0.2% wt. metrogel and components of a phosphate buffer solution, providing maintenance of pH 7,3±0,3.

13. Environment effects on tissue components of the cardiovascular system after their separation and during transportation, which is an aqueous solution containing 0.05 to 0.15% of the mass. gentamicin sulfate, 8,0-12,0% of the mass. diflucan and 0.1-0.5% wt. components of a phosphate buffer solution at pH 7,2-7,4.

14. Environment effects on tissue components of the cardiovascular system after their separation and their transport is rouke indicated in paragraph 13 characterized in that the aqueous solution contains 0.2% of the mass. gentamicin sulfate and 0.08% of the mass. diflucan.

15. Environment effects on tissues of homograft cardiovascular system when storing, containing solutions of salts and nutrients and a mixture of antibiotics, including gentamicin sulfate, characterized in that it has a pH of 7.0-7.4 and additionally contains components of a phosphate buffer solution, as a source of salts and nutrients in a solution of DMEM/F12, and a mixture of antibiotics diflucan in the following ratio of ingredients (% wt.):

gentamicin sulfate0,05-0,15
diflucan8,0-12,0
components of a phosphate buffer solution0,1-0,5
a solution of DMEM/F12rest

16. Environment effects on tissues of homograft cardiovascular system during storage at 13, characterized in that it contains 0.1% of the mass. gentamicin sulfate, 0,015% of the mass. diflucan in solution DMEM-F12 at a pH of 7.2.



 

Same patents:

FIELD: medicine.

SUBSTANCE: correcting the neurotoxic action of administering the preparation dapsone is ensured by using alpha tocopherol acetate as a correcting agent. Alpha tocopherol acetate is administered per os in a combination with dapsone to non-linear male and female white rats in a dose of 5 mg/kg once a day for 21 day.

EFFECT: using the method enables providing more effective correction of the neurotoxic side actions of dapsone with no side effects.

2 tbl

FIELD: medicine.

SUBSTANCE: allogenic multipotent mesenchymal stromal cells in the number of 6 million cells/kg are introduced intravenously in a dose of 3.0 Gy into rats suffered from radiation exposure. Additionally, haemopoietic stem cells recovered from umbilical blood in the number of 300 thousand cells/kg are introduced. The cells are transplanted in such a manner 60 min after radiation exposure.

EFFECT: higher mitosis count that leads to a substantial cryptal epithelium increase.

3 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to the field of biochemistry, in particular to clostridial neurotoxins with a modified persistence. Claimed is a polypeptide, containing HC-domain, the first and, at least, one additional LC-domain with amino acid sequences, at least, 90% identical to the respective sequences of a neurotoxic component of botulotoxin of a serotype A, B, C1, D, E, F or G. Also claimed are nucleic acid, an expression vector and a host cell, intended for the expression of the said polypeptide. Also claimed are a method of obtaining and application of the said polypeptide, including as a component of a pharmaceutical composition, for treatment of a condition, associated with hyperactive cholinergic innervations of a muscle or a exocrine gland, and for cosmetic procedures, associated with wrinkles.

EFFECT: invention makes it possible to controllably vary a period of activity of clostridial neurotoxins.

12 cl, 1 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: invention can be used to identify a high risk of developing impaired glucose tolerance in patients with stable effort angina with underlying administering beta-adrenergic blocking agents with no additional vasodilating properties. Therapy is preceded by conducting 2 exercise tests on the same day to achieve a threshold load power according to the same protocol, initially and 2 hours after administering a single dose of the beta-adrenergic blocking agents. If observing an interval gain of 120 seconds and more from the beginning of the load to the angina attack and/or reduction of an ischemic ST segment on the electrocardiogram not less than 1 mm at the 2nd load as compared to the 1st load, a risk of impaired glucose tolerance is considered to be high. A glucose tolerance test is carried out in these patients 4-5 weeks after the scheduled administration of the beta-adrenergic blocking agents. If impaired glucose tolerance is detected, administering the beta-adrenergic blocking agents is withdrawn. If the 2nd load as compared to the 1st load shows an interval to the angina attack and/or reduction of the ischemic ST segment on the electrocardiogram at a depth not less than 1 mm increasing less than by 120 seconds, a risk of developing impaired glucose tolerance is considered to be negligible. Treatment of these patients with the beta-adrenergic blocking agents is continued without the glucose tolerance test required.

EFFECT: method provides preventing carbohydrate metabolic disorders by the early identification of the high risk of developing impaired glucose tolerance in the given patients by detecting a compensatory increase of the glucose consumption with insulin resistance and a lower availability of free fatty acids to provide myocardial energy needs.

6 ex

FIELD: medicine.

SUBSTANCE: method involves intravenous allogeneic transplantation of multipotent mesenchymal stromal cells recovered from the placenta in an amount of 6 mln cells/kg. Additionally, haemopoietic stem cells recovered from umbilical blood in an amount of 300 thousand cells/kg are introduced.

EFFECT: method enables reducing the number of cytogenetically changed cells, promotes activating myeloid tissue regeneration in old laboratory animals.

4 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method involves providing an anaesthetic support with using the preparations: midazolam, fentanyl, propofol by bolus injections and infusions, artificial pulmonary ventilation by gas mixture O2:N2O=1:2 inhalations. Postoperative nausea and vomiting are prevented by intravenous administration of ondansetron 8 mg and dexamethasone 8 mg at the stage of induction, and by intravenous administration of clonidine 0.0016±0.0002 mg/kg/h in infusions or 0.02-0.025 mg in bolus doses up to 0.0016±0.0002 mg/kg after intubation of the trachea within the first hour of operation with having blood pressure and heart rate controlled.

EFFECT: method is safe and high-effective for preventing postoperative nausea and vomiting.

1 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely surgery, and may be used for preventing peritoneal adhesions. That is ensured by administering Licopid 0.01 g once a day orally for 6 days from the first postoperative day without regard to the extent of operation.

EFFECT: method enables reducing the postoperative adhesion formation with no side effects and reducing the length of treatment.

FIELD: medicine.

SUBSTANCE: invention refers to medicine and can be used in treating chronic inflammatory diseases accompanied by immune deficiencies. To do this, having a clinical-ambulatory immune deficiency stated, the therapeutic stage of a primary disease is followed by introducing to a patient of one-group fresh umbilical blood in an amount of three millilitres mixed with one gram of a broad-spectrum antibiotic, and one millilitre of a local anaesthetic. The above is injected three times into an infrascapular region every two days.

EFFECT: using the given invention enables providing the manifested and stable effect of increasing values of the humoral component of the immune system enabling preventing manifestations of secondary infections.

10 tbl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: what is presented is an agent for poly(ADP-riboso)polymerase inhibition. The agent represents 7-methylguanine(2-amino-7-methyl-1H-purin-6(7H)-one)- a purine derivative of formula (I) The agent has shown the efficacy higher than that in 7-methyl-xanthine and is non-toxic for the human body.

EFFECT: agent can be used in treating conditions caused by necrotic cell death: stroke, myocardial ischemia, diabetes and complications thereof, shock, neurotrauma, arthritis, colitis, allergic encephalomyelitis and other inflammations.

1 dwg, 2 ex

FIELD: biotechnology.

SUBSTANCE: composition is proposed which comprises stem cells of human amniotic fluid with the phenotype CD73+/CD90+/CD105+/CK19+, nutrient medium, erythropoietin, epidermal growth factor, and collagen taken in an effective amount.

EFFECT: invention enables to increase the proliferative potential and viability of the cells, while simultaneously providing cytoprotective effect on the cells of the transplant and stimulation of migration and proliferation of patient's own cells, and also to reduce significantly the concentration of injectable cells and to activate vascularisation and regeneration at the defect site and can be used in therapy for elimination of congenital and acquired defects of soft tissue arising as the result of injuries, after removal of tumors, congenital diseases, age-related changes or other damages.

2 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical formulations and is applicable for providing bactericidal efficacy. A pharmaceutical formulation contains two various antibiotics as active ingredients in the form of a synergetic combination of a fixed dose in a parenteral dosage form. The first antibiotic represents carbapenem or its pharmaceutically acceptable salts, while the second antibiotic represents aminoglycoside which represents etimycin or its pharmaceutically acceptable salts. The above first antibiotic and the above second antibiotic are found in weight ratio of 6:1 to 13:1. Besides, the pharmaceutical formulation contains one or more additives specified from a group of synthetic/natural amino acids/vitamins/stabilisers/polymers/antioxidants/micronutrient elements.

EFFECT: pharmaceutical formulation used in the very low concentrations, provides higher clinical effectiveness in the patients suffering from or sensitive to mixed multibacterial lethal infections, with a low tolerance to drugs and disease, and having a risk of potential toxicity, wherein potential toxicity caused by high doses provides a cause for concern.

8 cl, 3 tbl, 6 dwg, 1 ex

FIELD: medicine.

SUBSTANCE: method of treating apical periodontitis involves necrectomy from root canals, mechanical treatment thereof, root canal widening with multiple administration of a solution therein, insertion of a drain sponge wetted with the solution into the root canals left until the next doctor's appointment, root canal orifice closure with a dressing, root canal opening 48 hours later, dressing removal from the root canals, processing with 2% aqueous sodium hypochlorite, and gutta-percha filling. Mechanical treatment involves multiple introduction of a butol solution; mechanical treatment is followed by inserting drain sponges wetted in the butol solution into the root canals, or by introducing mixed zinc sulphate cement with butol with using a root canal filling instrument.

EFFECT: provided haemostatic effect in the root canal, optimal drug diffusion into the root canal tissues, and an anti-inflammatory action.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely medicine, and may be used for treating urethral syndrome in females with using the antibacterial preparation gentamycin. Gentamycin is introduced into a urethral wall in the amount of 80 mg dissolved in anaesthetic solution 2 ml. The above preparation is introduced in parallel with the urethral wall. The anaesthetic is presented by 0.25% Novocaine. That is followed by a vulvar exposure to an infrared laser light at wave length 0.82 mcm, frequency 3000 Hz, and power 10 mWt. A procedure length makes 5-6 minutes; the therapeutic course is 8-10 procedures.

EFFECT: method is well tolerated, accessible and high effective by providing the high gentamycin concentration in the area of inflammation and stimulating repair processes.

1 ex, 2 tbl, 1 dwg

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to neurosurgery, resuscitation and neurology, and may be used for treating the permanent unconscious vegetative state. That is ensured by intratympanic streptomycin treatment with underlying common drug therapy of the state. The completion of the above is followed by the electric transcranial exposure on the brain. For the intratympanic treatment, streptomycin 1 g is dissolved in physiologic saline 1 ml. The preparation is introduced once a day from one side, and on the other day from the opposite side. The therapeutic course is 5-10 days. The electric transcranial exposure is presented by transcranial DC micropolarisation of intensity 200-400 mcA for 30-40 minutes daily for 16-24 days. The exposure is performed through 3-4 electrodes places on the areas of temporo-caudal projections and posterior associated cortical areas from both sides. For the first 6 days, such exposure covers the temporo-caudal projections, while on the other days the posterior associated cortical area projections are exposed. If required, the above therapeutic courses are repeated not earlier than in 3 months until the clear consciousness is reached.

EFFECT: method provides an immediate and convincing clinical effect without brain invasiveness, an ability to recover the clear consciousness in these patients, as well as a significant reduction of the pathologically increased muscle tone and as a result, achieved satisfactory level of the patient's self-service.

2 ex

FIELD: medicine.

SUBSTANCE: combination contains fulvic acid or a salt thereof, and an antibiotic representing oxacillin or gentamycin, or a combination thereof. The invention also refers to a method of treating or inhibiting a bacterial infection in an individual that involves administering an effective amount of a composition containing the above combination.

EFFECT: invention provides the synergetic effect of the combination of fulvic acid and antibiotics.

16 cl, 1 tbl, 7 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine and aims at treating amnestic syndrome. A local anaesthesia is applied by infiltration of the tissues surrounding the external auditory canal with 0.5% novocaine along an anteromedial surface of the mastoid process bypassing the eardrum directly into the ear. The therapeutic effect is implemented by administering 1 g of streptomycin dissolved in 1 ml of physiological saline, intratympanic from one or both sides once a day every 2-3 days, with the manipulations performed 2 to 5 times per a course during 10-20 days. The transcranial exposure is generated by direct electric current at the intensity of 200-600 mA in the mid-temporal area of the head for 20-40 minutes daily for 8-10 days, repeated 2-3 times every 1-2 months. The electric exposure is generated by applying lead electrodes with each electrode having an area of 400-600 mm.

EFFECT: method enables improving associative cortex activity and improving the clinical effectiveness.

3 cl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to veterinary medicine, namely to medicines for treating mastitis in animals; a medicine for treating mastitis in animals contains the antibiotic tylosin, a gelling agent, water, with the gelling agent presented by carbopol and triethanolamine; the medicine further comprises Siberian fir essential oil and glycerol; an additional aseptic ingredient is plumepoppy juice prepared by pressing a vegetative part of a plant brought in a blossom time, in a step-loaded mechanical press with the first pressing force of 25-30 kN/cm2, and the exposure of 5 minutes, the second pressing force of 55-60 kN/cm2, and the exposure of 2.5 minutes; and the third pressing force of 80 kN/cm2, and the exposure of 1 minute; the ingredients are related as, %; tylosin 0.05-1.0; Cok plumepoppy juice 0.05-1.0; glycerol 5.0-10.0; carbopol 0.15-0.20; triethanolamine 0.02-0.03; Siberian fir essential oil 0.1-0.3; water up to 100.

EFFECT: agent for treating mastitis in animals has a combined therapeutic and repellent effect and a prolonged shelf life.

4 tbl

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular to ophthalmology, and is intended for treatment of inflammatory and trophic cornea diseases. For this purpose silicone-hydrogel soft contact lens, covered from inner surface by silico-dried amnion, is placed on cornea. Amnion with diameter smaller than lens diameter is used. Amnion is fixed on lens by means of polyacrylamide film, saturated with medication. As such means, used is antibacterial, anti-inflammatory, regenerating medication.

EFFECT: method ensures elimination of amnion dislocation under lens and reduction of time of corneal defect epithelisation, which increases treatment efficiency.

3 cl, 3 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to traumatology, oncology, pulmonology and surgery, and can be applied for treatment of pleurisy. For this purpose immediately after removal of exudate from pleural cavity into it introduced is wide-spectrum antibiotic and intravenously introduced is amikacin, after 10 minutes patient does 5 forced inhalations and exhalations with 3-minute intervals. After 10 minutes chest is subjected to electrophoresis with potassium iodide solution, which after day is alternated with solution of calcium chloride, with anode being placed into the space between scapular and midaxillary line at the level of 4-th rib to lower edge of rib arc. After finishing each session strips of capsicum plaster are stuck on back around shoulder-blades from both sides for one day, not less than 3 times.

EFFECT: method makes it possible to improve results of treatment and reduce development of complications by fast arrest of disease with adequate anti-inflammatory therapy and sanitisation of pleural cavity.

FIELD: medicine.

SUBSTANCE: invention refers to medicine, specifically dermatovenerology. Baneocin ointment is used as an agent for topical treatment of infiltrative-suppurant trichophytosis capitis for 48 hours with underlying conventional systemic antimycotic therapy. The agent shows manifested antimicrobial, anti-inflammatory, resolving and regenerative effect.

EFFECT: invention extends the range of topical products for treating infiltrative-suppurant trichophytosis capitis.

2 ex

FIELD: medicine.

SUBSTANCE: presented are engineered multilayered vascular tubes, comprising at least one layer of differentiated adult fibroblasts, at least one layer of differentiated adult smooth muscle cells. Further, any layer comprises differentiated adult endothelial cells. The said tubes have the following features: a ratio of endothelial cells to smooth muscle cells makes approximately 1:99 to approximately 45:55; the tube is deformable; an internal diameter of the tube is approximately 6 mm or smaller; the length of the tube is up to approximately 30 cm; the thickness of the tube is substantially uniform along the tube section. It is also provided that the vascular tube is free of any pre-formed frame. What is also described is a method of forming the above tubes.

EFFECT: engineering the therapeutically acceptable alternative vascular tubes withstanding physiological pressure, for transplantation into a patient's body and for use in testing cardiovascular drugs and devices.

29 cl, 4 ex, 3 dwg, 2 tbl

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