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EP1635636A1 - Procede et appareil de conservation de l'endothelium dans des organes creux et des vaisseaux biologiques isoles - Google Patents

Procede et appareil de conservation de l'endothelium dans des organes creux et des vaisseaux biologiques isoles

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Publication number
EP1635636A1
EP1635636A1 EP04739802A EP04739802A EP1635636A1 EP 1635636 A1 EP1635636 A1 EP 1635636A1 EP 04739802 A EP04739802 A EP 04739802A EP 04739802 A EP04739802 A EP 04739802A EP 1635636 A1 EP1635636 A1 EP 1635636A1
Authority
EP
European Patent Office
Prior art keywords
perfusion solution
endothelium
solution
perfusion
ions
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP04739802A
Other languages
German (de)
English (en)
Inventor
Stephan Nees
Friderike Nees
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Biotest AG
Original Assignee
Biotest AG
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Biotest AG filed Critical Biotest AG
Publication of EP1635636A1 publication Critical patent/EP1635636A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/10—Preservation of living parts
    • A01N1/14—Mechanical aspects of preservation; Apparatus or containers therefor
    • A01N1/142—Apparatus
    • A01N1/143—Apparatus for organ perfusion
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/10—Preservation of living parts
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/10—Preservation of living parts
    • A01N1/12—Chemical aspects of preservation
    • A01N1/122—Preservation or perfusion media
    • A01N1/126—Physiologically active agents, e.g. antioxidants or nutrients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B17/00—Surgical instruments, devices or methods
    • A61B2017/00969—Surgical instruments, devices or methods used for transplantation

Definitions

  • the invention relates to a method and an apparatus for the treatment and preservation of the endothelium in isolated hollow organs, in particular isolated biological vessels, such as blood vessels and lymphatic vessels, by means of an albumin-containing endothelium-protective perfusion solution or incubation solution, and the use of an endothelium-protective perfusion solution for preparation of hollow organs or biological vessels as a graft for the treatment of organ or vascular diseases, their use for repairing endothelial lesions in isolated hollow organs and / or biological vessels and their use for organ and / or vessel preservation.
  • Closure of blood vessels caused by arteriosclerosis is the cause of a large number of organ diseases in many western countries.
  • a vascular graft i.e. The replacement of diseased, narrowed or closed biological vessels by healthy replacement vessels is therefore of great clinical importance for the treatment of such diseases.
  • a vascular graft i.e. The replacement of diseased, narrowed or closed biological vessels by healthy replacement vessels is therefore of great clinical importance for the treatment of such diseases.
  • a vascular graft i.e. The replacement of diseased, narrowed or closed biological vessels by healthy replacement vessels is therefore of great clinical importance for the treatment of such diseases.
  • 200,000 people die of a heart attack each year as a result of arteriosclerotic occlusion of one or more coronary arteries.
  • peripheral arterial occlusive disease POD
  • patients or their limbs can be treated and saved by bridging the pathologically altered biological vessels with a suitable vascular replacement.
  • the first two vascular replacement materials can be referred to as "biological vascular replacement".
  • the bioprostheses can in turn be distinguished between homologous (obtained from the same species) and heterologous (derived from a foreign species) vascular prostheses.
  • the use of the body's own vessels provides the best results with regard to the so-called "openness rates". This means the number of vessels that are still open after a certain time after a vascular graft.
  • the inventors of the present invention have determined that the decisive reason for the acute occlusion (thrombosis) of biological vessels and a restenosis, which is caused by the intimal wall thickening and the associated failures, is due to the handling which is still common in surgical practice today biological vessels in vascular grafts.
  • the intima consists of a single-layered endothelium and the stratum sub endotheliale (fine, loose connective tissue with the subendothelial pericytes discovered by the inventors) as well as the fenestrated internal elastica membrane and fulfills important biological functions that are essential for maintaining the blood vessel function.
  • the endothelium is the body's actual blood container.
  • the inventors of the present invention have found that a such a conventional treatment procedure leads to a considerable impairment of the endothelial function, which can lead to the complete destruction of the endothelial tissue.
  • the impairment or even the destruction of the endothelial tissue is a reason why biological vascular or organ transplants may close again soon after their implantation.
  • the biological vessels are involved in their treatment, i.e. exposed to strong mechanical stresses during their isolation and implantation, since the vessels are usually pulled over a thick button cannula, similar to a folded tube, and placed under an uncontrollable high pressure with the saline solution and an attached syringe. Then, piece by piece, the vessel is pushed down from the button cannula to find the side branches of the vessel from which liquid is escaping and the side branches are immediately ligated with suitable surgical clips. Finally, the entire length of the vessel segment prepared for transplantation is put under high pressure again to test the tightness, which is connected with a real "inflation" of the insulated vessel. The high pressure to which the vessel is exposed makes it large Part, and often the last remnant, of the luminal endothelial layer torn off and washed away.
  • the inventors of the present invention have found that the luminal endothelial tissue is of considerable importance with regard to the acute maintenance of the vascular function (keeping thromboembolic reactions free for the purpose of an unimpeded fluid conduction) and the long-term keeping of the vascular lumen (prevention of arteriosclerotic stenoses and finally thromboembolism).
  • tissue factor tissue factor
  • a healthy, closed endothelium shields the one expressed by all of the pericytes Tissue factor physiologically completely separates from the blood flowing within the vessels, thus shielding the tissue factor from the blood stream by means of the intermediary intact endothelial tissue, which leads to an acute thrombosis and thus an acute occlusion of the biological vessels prevented.
  • One reason for the destruction of the endothelium when using saline in the treatment of grafts is likely to be that little or no energy and maintenance metabolism can take place in the endothelial tissue. Above all, this makes it clear that the short-term (acute) rate of openness of a biological vessel depends essentially on the condition of the endothelial tissue inside the vessel.
  • restenosis means the recurrent narrowing of vessels or vascular grafts, which is associated with a loss of blood circulation in the affected tissue area.
  • the present invention is therefore based on the object of providing gentle methods and perfusion solutions for the treatment of isolated hollow organs or biological vessels which require preservation, i.e. maintenance and, if necessary, regeneration of the endothelial layer of the vessels, in order to provide reliable and long-term usable organ and vascular grafts.
  • the invention thus relates to endothelium-protective perfusion solutions or incubation solutions, the use of endothelium-protective perfusion solutions described here and methods for endothelium-preserving treatment of hollow organs and / or biological vessels, comprising bringing the hollow organ or vessel into contact with an endothelium. protective perfusion solution.
  • the invention relates to a method for the endothelium-preserving treatment and preservation of isolated hollow organs or biological vessels and vascular systems, in which the hollow organ or the biological vessel is treated with an endothelium-protective perfusion solution (incubation solution) according to the invention.
  • the method for the endothelium-preserving treatment of hollow organs comprises bringing an isolated hollow organ into contact with an endothelium-protective perfusion solution, the endothelium-protective perfusion solution comprising at least the following components:
  • the treatment leads to the preservation and / or repair of the endothelial tissue in the lumen of the hollow organ.
  • the native albumin in the endothelium-protective perfusion solution is replaced by 1-10% by volume homologous hemolysin-free serum or autologous serum.
  • the native albumin in the endothelium-protective perfusion solution is replaced by 2.5% by volume homologous hemolysin-free serum or autologous serum. In a further preferred embodiment, the native albumin in the endothelium-protective perfusion solution is replaced by 5% by volume homologous hemolysin-free serum or autologous serum.
  • the native albumin in the endothelium-protective perfusion solution is replaced by 10% by volume homologous hemolysin-free serum or autologous serum.
  • the native albumin in the endothelium-protective perfusion solution is replaced by a homologous anti-coagulated blood plasma preparation which comprises human plasma proteins, anti-coagulant factors and immunoglobulins and in which the pro-coagulant factors, isoagglutinins and unstable components of the blood plasma have been removed.
  • the anti-coagulated blood plasma preparation contains sodium ions, potassium ions, calcium ions, magnesium ions, chloride ions, human serum proteins, albumin and immunoglobulins.
  • the anti-coagulated blood plasma preparation has the following composition: about 100-170 mM sodium ions, about 1-15 mM potassium ions, about 1-6 mM calcium ions, about 0.1-4 mM magnesium Ions, about 50-200 mM chloride ions, human serum proteins, thereof about 25-45 g / l albumin, 3-15 g / l IgG, 1-10 g / l IgA and 0.2-3 g / l IgM immunoglobulins , at a pH of about 7.3 to about 7.8 and an osmolarity of about 200-350 mosmol / kg.
  • the nutrient substrate in the endothelial protective perfusion solution is L-glutamine.
  • the concentration of L-glutamine in the endothelium-protective perfusion solution is between 0.5 to 10 mM, preferably 2.5 mM.
  • the concentration of L-glutamine in the endothelium-protective perfusion solution is 5 mM. In a further preferred embodiment, the concentration of L-glutamine is
  • the physiological electrolyte solution comprises the following components: 100-150 mM NaCl; 1-15 mM KCl; 0.1-4 mM MgSO 4 ; 0.5-2 mM KH 2 PO 4; 24-48 mM histidine-Cl and 1 -3 mM CaCl 2 .
  • the physiological electrolyte solution contains energy substrates, preferably 2-10 mM glucose and / or 1-10 mM pyruvate.
  • the physiological electrolyte solution contains 0.1-0.6 U / ml heparin and / or in each case 50-100 ⁇ M uric acid and / or ascorbate.
  • the pH in the physiological electrolyte solution in atmospheric air is 7.4 +/- 0.04.
  • the endothelium-protective perfusion solution additionally contains antibiotics.
  • the antibiotics are 50-400 U / ml penicillin and / or 0.1-0.4 mg / ml streptomycin.
  • the endothelium-protective perfusion solution is a blood plasma preparation freed of coagulation factors and isoagglutinins.
  • the blood plasma preparation comprises the following constituents: 100-170 mM sodium ions, 1-15 mM potassium ions, 1-6 mM calcium ions, 0.1-4 mM magnesium ions, 50-200 mM chloride ions.
  • the blood plasma preparation was treated with ⁇ -propiolactone and UV radiation to inactivate the virus.
  • the perfusion solution contains one or more endothelium-promoting growth factors.
  • the growth factor is selected from the group consisting of epidermal growth factor (EGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF) and stem cell factor (SCF).
  • EGF epidermal growth factor
  • FGF fibroblast growth factor
  • VEGF vascular endothelial growth factor
  • SCF stem cell factor
  • the perfusion solution contains flavonoids such as quercetin or rutosides such as trihydroxyethyl rutoside or derivatives thereof.
  • the perfusion solution contains vasodilatory substances such as papaverine, adenosine or cardioplegic KCl concentrations.
  • the hollow organs are the heart, intestine, uterus, kidney, bladder, lung, liver, spleen.
  • the hollow organs are biological vessels or vascular systems.
  • the biological vessels are blood vessels or lymphatic vessels.
  • the endothelium-protective perfusion fluid is passed through the hollow organ with the aid of an apparatus according to the invention.
  • the present invention further relates to an endothelium-protective perfusion solution which comprises at least the following components:
  • the native albumin is replaced by 2.5% by volume homologous hemolysin-free serum or autologous serum.
  • the native albumin is replaced by 5% by volume homologous hemolysin-free serum or autologous serum.
  • the native albumin is replaced by 10% by volume homologous hemolysin-free serum or autologous serum.
  • the concentration of L-glutamine is
  • the concentration of L-glutamine is 5 mM.
  • the concentration of L-glutamine is
  • the physiological electrolyte solution comprises the following components: 100-150 mM NaCl; 1-15 mM KCl; 0.1-4 mM MgSO 4 ; 0.5-2 mM KH 2 PO 4; 24-48 mM histidine-Cl and 1-3 mM CaCl 2 .
  • the physiological electrolyte solution contains energy substrates, preferably 2-10 mM glucose and / or 1-10 mM pyruvate.
  • the physiological electrolyte solution contains 0.1-0.6 U / ml heparin and / or in each case 50-100 ⁇ M uric acid and / or ascorbate.
  • the pH in the physiological electrolyte solution in atmospheric air is 7.4 +/- 0.04.
  • the endothelium-protective perfusion solution additionally contains antibiotics.
  • the antibiotics are 50-400 U / ml penicillin and / or 0.1-0.4 mg / ml streptomycin.
  • the perfusion solution is an anti-coagulated and non-agglutinating blood plasma preparation which comprises human plasma proteins, anti-coagulant factors and immunoglobulins and in which the pro-coagulant factors, isoagglutinins and unstable components of the Blood plasma has been removed.
  • the anti-coagulated blood plasma preparation contains sodium ions, potassium ions, calcium ions, magnesium ions, chloride ions, human serum proteins, albumin and immunoglobulins.
  • the anti-coagulated blood plasma preparation has the following composition: about 100-170 mM sodium ions, about 1-15 mM potassium ions, about 1-6 mM calcium ions, about 0.1-4 mM magnesium Ions, about 50-200 mM chloride ions, human serum proteins, thereof about 25-45 g / l albumin, 3-15 g / l IgG, 1-10 g / l IgA and 0.2-3 g / l IgM immunoglobulins , at a pH of about 7.3 to about 7.8 and an osmolarity of about 200-350 mosmol / kg.
  • the blood plasma preparation was treated with ⁇ -propiolactone and UV radiation to inactivate the virus.
  • the perfusion solution contains endothelial growth factors.
  • the growth factors are selected from the group consisting of epidermal growth factor (EGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF) and stem cell factor (SCF).
  • the perfusion solution contains flavonoids such as quercetin or rutosides such as trihydroxyethyl rutoside or derivatives thereof.
  • the perfusion solution contains vasodilatory substances such as papaverine, adenosine or cardioplegic KCl concentrations.
  • the invention further relates to an apparatus for the endothelium-preserving treatment of isolated biological vessels, comprising an chamber (1), an axially movable plunger (6), a cannula (5), a reservoir (7) which contains endothelium-preserving perfusion fluid and a sealing device (3), the cannula being connected to the axially movable plunger (6) so that the cannula can be moved into the chamber with the plunger, and wherein the sealing device (3) can enclose one end of the vessel and the cannula with can be connected to the other end of the vessel, so that the endothelium-protective perfusion solution from the reservoir (7), preferably under a pressure gradient, can be guided selectively into the biological vessel.
  • the sealing device comprises sealing disks which are arranged in a stack shape in a knurled screw.
  • the sealing washers have a diameter of 1-10 mm and / or a thickness of 0.3-3 mm.
  • the apparatus additionally contains a thermostat for heating the perfusion liquid.
  • the perfusion solution which is used in the apparatus is one of the previously mentioned and defined solutions.
  • the invention also relates to the use of one of the aforementioned endothelium-protective perfusion solutions for the preservation of isolated hollow organs or biological vessels, the perfusion solution creating suitable conditions for maintaining and / or renewing the endothelial tissue in the lumen of the hollow organs or biological vessels.
  • the invention further relates to the use of the endothelium-protective perfusion solution for maintaining and / or repairing the endothelial tissue in isolated hollow organs or biological vessels.
  • the invention also relates to the use of the endothelium-protective perfusion solution for the therapy and / or prevention of vascular occlusions in isolated hollow organs or biological vessels.
  • the present invention further comprises combinations of the compositions of the perfusion solutions used according to the invention described here according to their individual embodiments.
  • the composition of the endothelium-protective perfusion solution can vary depending on the area of application and intended use and can contain additional constituents which are important for maintaining the function of the endothelium tissue.
  • endothelium or “endothelial tissue” is understood to mean a single-layer cellular lining of the biological vessels and the serous cavities.
  • endothelial cell denotes both differentiated endothelial cells organized in the association and endothelial progenitor cells that have not yet been differentiated.
  • endothelium-protective perfusion solution is understood to mean a solution with which the endothelial tissue can be treated in isolated hollow organs or biological vessels, while maintaining the consistency of the endothelial tissue. Detachment or destruction of the endothelium is achieved by using the Endothelium-protective perfusion solution is prevented.
  • the endothelium-protective perfusion solution according to the invention maintains the ability of the endothelial cells to divide and the regeneration ability of the tissue.
  • the perfusion solution has the property of penetrating even the smallest vessels and vessel sections.
  • endothelium-protective is understood to mean the property that the endothelial tissue is maintained (preserved), regenerated and / or strengthened, ie that the endothelium retains or builds up its tissue architecture and structure. This includes all endothelium-protecting and Measures that have a beneficial effect on the endothelium (endothelial tissue), including the maintenance of the cell structure of the endothelial tissue, the maintenance of inter-cellular communication and the maintenance of cell-cell connections Endothelial cells for division and the increase in endothelial cell mass per area.
  • perfusion solution includes a solution for the preservation of isolated hollow organs and vessels.
  • a perfusion solution can be understood in a synonymous way as an “incubation solution”.
  • physiological electrolyte solution is to be understood as an aqueous solution of electrolytes which are found in whole blood in a physiological manner.
  • physiological should not be seen as limiting the concentrations of the individual ions used. "Unphysiological" concentrations, ie concentrations that do not naturally occur in the organism, also fall within the scope of the present invention.
  • physiological therefore includes isotonic (synonymous with “isotonic”) solutions which ensure a uniform osmotic pressure and a constant molecular concentration.
  • Preferred perfusion solutions are isotonic with the blood or plasma at about 290 mosm / kg H2O.
  • bringing into contact is understood to mean the incubation, insertion, rinsing or other way treated organ or vessel with the perfusion solution for a time which is sufficient for the maintenance and / or renewal of the endothelium in the vessels.
  • the term “hollow organ” is used to refer generally to internal organs and organ vascular systems in the heart, intestine, uterus, kidney, urinary bladder, lungs, liver, spleen, etc. be understood.
  • the term “hollow organ” also includes well-known biological vessels or vascular systems, such as blood vessels (arteries and veins) and lymphatic vessels.
  • biological vessel or “vessel” is understood to mean all of the body's own (autologous) vessels lined with endothelium, which transport body fluid.
  • this means blood vessels, such as arteries and veins, and lymphatic vessels.
  • a “blood plasma preparation” is understood to mean the liquid portion of the blood which has not been clotted and which remains after removal of the blood cells (by centrifugation) and which, in contrast to the serum, contains no coagulation factors.
  • agglutination is to be understood as the gluing of erythrocytes by specific antibodies (agglutmine-mediated).
  • hemolysin-free or autologous serum is understood to mean a serum in which there are no antibodies (isoagglutinins) which could bind the red blood cells of a recipient. This prevents complement activation and consequently lysis (dissolution) of the blood cells autologous serum, that is to say in a serum originating from the patient himself, naturally does not have such hazard potentials.
  • nutrient substrate is understood to mean an amino acid or protein that is processed by the cell to generate energy in the energy metabolism.
  • Nutrient substrates are usually sugars, fats or amino acids that are burned by the cell and are thus used to generate energy.
  • vascular diseases is understood to mean diseases or pathological conditions of blood vessels, such as, for example, angiopathy, vasculitis, in particular arterial and venous stenoses, angina pectoris, myocardial infarction, (heart attack), apoplexy (stroke), sudden hearing loss, aneurysm, arteriosclerosis, thrombosis, varicose veins , Thrombophlebitis, claudication, smoking leg and gangrene.
  • angiopathy vasculitis
  • vasculitis in particular arterial and venous stenoses
  • angina pectoris myocardial infarction
  • stroke apoplexy
  • sudden hearing loss aneurysm
  • arteriosclerosis arteriosclerosis
  • thrombosis varicose veins
  • Thrombophlebitis varicose veins
  • claudication smoking leg and gangrene.
  • the present invention is based on the finding that the endothelial layer of biological vessels is a decisive factor for the preservation of the vessels - even after the transplantation.
  • all biological vessels and hollow organs carrying endothelium can be treated in a targeted manner, so that the structure and function of the endothelial tissue and thus the function of the vessel or hollow organ are retained.
  • the treatment of isolated hollow organs and biological vessels with the endothelium-protective perfusion solutions and their modifications described in more detail below enables the preparation of organ and vascular grafts before their implantation in the body (e.g. in animals or humans) for an organ and vascular replacement (prosthesis or Bypass) or is suitable for repairing lesions (ie injuries) of the endothelial tissue in such hollow organs or biological vessels.
  • organ and vascular replacement prosthesis or Bypass
  • lesions ie injuries
  • the endothelium in the vessels can be preserved and even repaired by keeping the endothelial cells in the cell structure, stabilizing them and stimulating them to proliferate. This increases the lifespan of the hollow organs if they e.g. can be used as a graft during operations in patients.
  • the endothelium-protective perfusion solutions described here are also suitable for use in the preservation of organs and vessels of mammals (organ and vessel preservation) by incubating these organs or vessels in the perfusion solution.
  • organ and vessel preservation Such preservation of organs or vessels is particularly desirable, for example, in transplant surgery.
  • Preservation can also be necessary to protect organs or vessels for transport and to keep them functional.
  • the aim of the method according to the invention and the perfusion solution used for this purpose is to ensure the function, performance and durability of the organ or vessel by maintaining or even regenerating the luminal endothelial cell layer.
  • a functional and structurally intact endothelial tissue enables a long lifespan of vascular prostheses, since this prevents the vessels from occluding, for example by thrombosis or skier-rotating restenosis.
  • the invention also relates to an apparatus with which an isolated biological vessel can be flushed with the endothelium-protective perfusion solution.
  • biological vessels can be easily and efficiently checked for leaks.
  • all side branches of the biological vessel can be found and ligated unambiguously when flushing with suitable ligation aids, such as clamps or micro clips.
  • vascular diseases or pathological conditions are, for example, angiopathy, vasculitis, in particular arterial and venous stenoses, angina pectoris, myocardial infarction, (heart attack), apoplexy (stroke), sudden hearing loss, aneurysm, arteriosclerosis, thrombosis, varicose veins, thrombophlebitis, smoking and claudication, claudicatio.
  • the endothelium-protective perfusion solution is furthermore suitable for the repair of endothelial lesions in the organ vascular system of hollow organs and / or in biological vessels or for the preservation of entire organs or vessels or parts thereof for the purpose of preservation during surgery, transplantation, storage or transport.
  • the hollow organs or biological vessels treated with the endothelium-protective perfusion solution according to the invention have an undamaged endothelium and improved perfusability of the vessels compared to untreated or with conventional solutions, for example physiological saline (Brine) solution, Bretschneider solution, University of Wisconsin (UW) solution , Carolina Rinse solution and HTK solution rinsed or incubated hollow organs or vessels.
  • conventional solutions for example physiological saline (Brine) solution, Bretschneider solution, University of Wisconsin (UW) solution , Carolina Rinse solution and HTK solution rinsed or incubated hollow organs or vessels.
  • the inventive use of the perfusion solutions described here prevents early thrombosis (closure) of the vessel in question or at least delays it.
  • the isolated hollow organs or biological vessels treated with the method according to the invention therefore have the advantage over the hollow organs or biological vessels previously used in the field of organ and vascular transplantation that the endothelial tissue of the treated hollow organ or vessel remains intact and even remains in one damaged endothelial tissue regenerates (e.g. by stimulating the proliferation of endothelial cells).
  • endothelium When the endothelium is treated with a perfusion solution according to the invention, renewal of the endothelial tissue is thus also found.
  • the endothelial cells retain their ability to divide and are even able to repair endothelial lesions of the hollow organs or biological vessels, which is achieved by stimulating the proliferation of the endothelial cells. Vessels that are untreated or treated with conventional physiological saline in surgical practice, on the other hand, lead to a lesion or to complete detachment of the endothelium within the first hour after treatment.
  • the basic composition of the endothelium-protective perfusion solution according to the invention comprises a physiological electrolyte solution, at least 0.1% by weight of native albumin and a nutrient substrate, such as e.g. L-glutamine (2.5-10 mM).
  • a physiological electrolyte solution at least 0.1% by weight of native albumin and a nutrient substrate, such as e.g. L-glutamine (2.5-10 mM).
  • the physiological electrolyte solution contains as electrolytes at least magnesium ions (Mg 2+ ), chloride ions (Cl " ) and calcium ions (Ca 2+ ) within a physiological concentration range.
  • the physiological electrolyte solution preferably contains one or more of the following ions: potassium -Ions (K + ), sodium ions (Na + ), sulfate ions (SO 4 2 “ ) and, phosphate ions (PO 4 3 " ).
  • the effectiveness of the endothelium-protective perfusion solution according to the invention can be achieved by the combination of albumin, the salts of the electrolytic solution, especially due to the presence of calcium (Ca 2+ ) and a nutrient substrate, calcium is necessary to maintain the function of the endothelial tissue because the structure of its glycocalyx is maintained in the presence of Ca 2+ This structure creates the necessary micro-environment on the endothelial surface, which is important for the physiological function and activity of many endothelial enzymes, membrane transporters, ion channels and receptors.
  • the added amino acid glutamine (L-glutamine or D-glutamine) is used as a nutrient substrate in the main composition of the perfusion solution according to the invention.
  • glutamine primarily serves the purpose that especially endothelial cells, which constantly produce highly reactive oxygen metabolites such as hydrogen peroxide, oxygen radicals and nitrogen monoxide, can better protect themselves against these oxidants in an as yet unexplained but clearly demonstrable manner if glutamine is available.
  • Albumin is also an essential component of the perfusion solution according to the invention and i.a. responsible for the endothelium-preserving effect of the solution.
  • albumin lowers the "shear forces" on the surface of the endothelium when flushing (blood) vessels and the surface tension, which would otherwise favor bending instead of spreading of the endothelial cells. Both physical quantities (forces) are for a possible detachment of Endothelial cells responsible.
  • Albumin therefore acts in a certain way in the vessels like the "lubricating oil" in machines.
  • crystalloid protein-free
  • cardioplegic because of high potassium content
  • a combination of nutrient substrate (eg glutamine) and albumin for the purpose of maintaining the endothelium is necessary simply because an increased shear stress on the endothelium in albumin-free solution means a higher energy conversion and makes energy consumption of the endothelium necessary, since more energy has to be used to hold the endothelial cells on the vessel wall.
  • nutrient substrate eg glutamine
  • albumin for the purpose of maintaining the endothelium is necessary simply because an increased shear stress on the endothelium in albumin-free solution means a higher energy conversion and makes energy consumption of the endothelium necessary, since more energy has to be used to hold the endothelial cells on the vessel wall.
  • plasma derivatives that do not have coagulation factors and agglutinating factors e.g. Contain agglutinins (e.g. agglutinating antibodies, hemagglutinins, lectins, hemolysin, phythaemagglutinins), complement factors, inflammation mediators or blood group-specific antibodies, which are particularly preferred for the treatment of isolated hollow organs or biological vessels.
  • agglutinins e.g. agglutinating antibodies, hemagglutinins, lectins, hemolysin, phythaemagglutinins
  • complement factors e.g. agglutinating antibodies, hemagglutinins, lectins, hemolysin, phythaemagglutinins
  • complement factors e.g. agglutinating antibodies, hemagglutinins, lectins, hemolysin, phythaemagglutinins
  • Blood plasma preparations can also be used as endothelium-protective perfusion solutions for preserving the endothelium in the endothelium-protective method according to the invention or for the uses according to the invention and are particularly preferred.
  • Many special proteins in normal blood plasma additionally perform certain functions that can have a beneficial effect on endothelial functions: transferrin and ceruloplasmin, for example, act as transport proteins for iron or copper ions, and hormone transport proteins favor essential enzyme functions on the endothelium or signal transduction processes in endothelial cells. Growth hormones are also an important component, which promote cell division as a prerequisite for repairing and covering endothelium-damaged vessel walls.
  • Blood plasma preparations of this type show an even greater endothelium-preserving effect than the basic solution mentioned above and its special embodiments, which is probably due to additional factors present in the blood plasma, such as growth factors, for example. which has a favorable effect on the preservation of the endothelial tissue in the lumen of the vessels.
  • Blood plasma naturally consists of approximately 90% water, approximately 8% proteins such as albumin (approximately 40 g / l), globulin ( ⁇ 1-globulin, ⁇ 2-globulin, ⁇ -globulin, ⁇ -globulin), transport vehicles such as hormones and bilirubin as well as coagulation factors.
  • those blood plasma preparations are preferred in which the aforementioned factors, such as coagulation factors, agglutinating factors, such as e.g. Agglutinins, complement factors, inflammation mediators or blood group-specific antibodies are no longer available and are free of toxic lipids and germs such as viruses or bacteria.
  • factors such as coagulation factors, agglutinating factors, such as e.g. Agglutinins, complement factors, inflammation mediators or blood group-specific antibodies are no longer available and are free of toxic lipids and germs such as viruses or bacteria.
  • an electrolyte solution corresponding to the physiological inorganic salt components of healthy human blood plasma is preferably used as the electrolyte solution.
  • the bicarbonate portion of conventional electrolyte solutions is replaced by equimolar histidine chloride.
  • the electrolytic solution according to the invention has the following composition: 100-150 mM NaCl; 1-15 mM KCl; 0.1-4 mM MgSO 4 ; 0.5-2 mM KH 2 PO 4 ; 24-48 mM histidine-Cl and 1-3 mM CaCl 2 .
  • a preferred composition of the electrolyte solution comprises: 140 mM NaCl; 4.5 mM KCl; 1.2 mM MgSO 4 ; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2.5 mM CaCl 2 .
  • the histidine-Cl can also be replaced by 24 mM bicarbonate.
  • the calcium concentration in the solution can be reduced isotically to a value of 0.01-0.1 mM.
  • the pH is adjusted before the addition of CaCl 2 as the last added component of the solution.
  • the pH can be adjusted, for example, by means of a buffer which contains one or more acids and / or bases. Examples of such buffer substances are lactate and / or bicarbonate.
  • energy substrates preferably 2-10 mM glucose and / or 1-10 mM
  • these energy substrates alone enable a sufficient supply of metabolic energy for the endothelial tissue, even under almost hypoxic conditions (p ⁇ 2 ⁇ 10 mm Hg).
  • Preferred concentrations of the energy substrates are 8 mM glucose and / or 2 mM pyruvate.
  • the physiological electrolyte solution contains heparin or other anticoagulants, such as heparin, heparinoids, coumarin (vitamin K antagonist), in an anticoagulant concentration.
  • Usual anticoagulant concentrations for high molecular weight heparin are 0.2-0.6 U / ml, preferably 0.4 U / ml in the case of human blood.
  • Another embodiment of the invention uses low molecular weight heparin, such as that from Pharmacia Ltd. provided heparin, to 100 ⁇ l / 100 ml. 50-100 ⁇ M uric acid and / or ascorbate can also be added as antioxidants as exogenous reducing agents against reactive oxygen compounds.
  • Native albumin means that the albumin is in its natural (native) form and has preferably been purified by chromatographic methods and not by application of heat. Preferred is human albumin.
  • the tightness of the endothelial lawn can be measured quantitatively by determining the hydraulic conductivity (Lp [cm / s / cm H 2 O]).
  • the endothelial layers treated in saline or in a "Bretschneider” solution lose their tightness completely, because the individual cells detach spherically from the surface very quickly.
  • Another embodiment of the endothelium-protective perfusion solution of the invention contains the addition of homologous hemolysin-free or autologous serum instead of Albumin. In this case, no additional native albumin needs to be added, since this is already present in the serum.
  • a stabilization of the native tightness of the endothelial cells that does not decrease over several days can be achieved if homologous hemolysin-free serum or autologous serum is added to the electrolyte solution.
  • the serum is preferably free of lipoprotein.
  • Undiluted autologous serum has an albumin concentration of at least 6% by weight which, according to the perfusion solution according to the invention, can take over the functions of the native albumin even with a greater dilution.
  • human serum contains numerous growth factors in effective concentrations (e.g.> 0.1 ng / ml bFGF, TGF, VEGF).
  • a concentration of 1-10% by volume homologous hemolysin-free serum or autologous serum is added to the endothelium-protective perfusion solution.
  • the serum concentration is preferably 5-10% by volume. Even with a concentration of 1% by volume of autologous or homologous hemolysin-free serum, a noticeable improvement in the stabilization of the endothelial tightness can be determined.
  • the L-glutamine contained in the endothelium-protective perfusion solution can have a concentration of 2.5 mM, 5 mM or 7.5 mM, a concentration of 2.5 mM L-glutamine being preferred.
  • the endothelium-protective perfusion solution additionally contains antibiotics in bactericidal concentrations.
  • antibiotics for example, 100-400 U / ml penicillin and / or 0.1-0.4 mg / ml streptomycin are preferred, with 200 U / ml penicillin and 0.2 mg / ml streptomycin being most preferred.
  • a preferred perfusion solution that can be used for endothelial preservation and endothelial renewal has the following composition: Physiological (isotonic) electrolyte solution (127 mM NaCl; 4.6 mM KCl; 1.1 mM MgSO 4; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2 mM CaCl 2 (pH to 7.40 before adding CaCl 2 ); 0.1% albumin and 2.5 mM L-glutamine, 2 mM Na-pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution) and additionally each 50 ⁇ M uric acid and ascorbate.
  • Another very effective solution tested in the examples for use in the processes according to the invention or for the use according to the invention has the following composition:
  • an effective amount of one or more flavonoid compounds can be added to the perfusion solution according to the invention, which prevents microcirculatory complications (e.g. endothelial lesions), which are caused by release products of simultaneously activated granulocytes or thrombocytes, in the vascular lumen.
  • Preferred flavonoids tested in the process according to the invention are quercetin (approx. 50-250 ⁇ M) and rutosive compounds, in particular trihydroxyethyl rutoside (approx. 50-250 ⁇ M) in effective concentrations.
  • a particularly preferred solution for endothelial preservation and for the regeneration of the endothelial layer in the case of microcirculatory complications has the following composition: isotonic electrolyte solution (127 mM NaCl; 4.6 mM KCl; 1.1 mM MgSO 4; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2 mM CaCl 2 (pH 7.40 before adding CaCl 2 ); 0.1% albumin and 2.5 mM L-glutamine, 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution) and additionally 100 ⁇ M freshly added quercetin, 100 ⁇ M trihydroxyethyl rutoside can also be added instead of quercetin.
  • vasodilators such as papaverine or adenosine.
  • 50-200 ⁇ M papaverine are preferred.
  • a concentration of 100 ⁇ M papaverine is particularly preferred.
  • adenosine is preferably added in a concentration of 0.5 to 2 mM, 1 mM being particularly preferred.
  • a particularly preferred solution used in the methods and uses according to the invention has the following composition: 127 mM NaCl; 4.6 mM KCl; 1.1 mM MgSO 4 ; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2 mM CaCl 2 (pH 7.40 before adding CaCl 2 ); 0.1% albumin and 2.5 mM L-glutamine, 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution) and additionally 100 ⁇ M papaverine (or 1 mM adenosine).
  • cardioplegic solutions are asotonic as possible, i.e. they should have the same molecular concentrations in order to avoid damage to the vessel walls or the functionality of the organ.
  • a decrease in the contraction of the heart is already observed at a potassium concentration of more than 6 mM. At a concentration of 8 mM, the heart becomes almost completely paralyzed. This immobilization of the organ enables easy handling by the surgeon during the implantation.
  • the finished solution can be stored for months at a temperature of 4 ° C and in complete darkness without loss of effectiveness. Sterilization to keep germs in the solution free can be carried out, for example, using sterile filters.
  • a blood plasma preparation can be used in the endothelium-protective method for the treatment of isolated hollow organs or biological vessels of the present invention.
  • the inhibitory and modulating properties of such a serum-like solution are obviously conducive to maintaining endothelial cell integrity.
  • the basic composition of the Endothelium-protective perfusion solution which contains a physiological electrolyte solution, a nutrient substrate and albumin and which already shows good endothelial sensitivity (see examples)
  • can have an effect on the preservation and preservation of the endothelium in hollow organs by using a homologous anti-coagulated blood plasma preparation instead of the albumin can be further increased.
  • a perfusion solution is therefore preferred in which the native albumin in the basic composition of the endothelium-protective perfusion solution is replaced by an anti-coagulated blood plasma preparation which comprises human plasma proteins, anti-coagulant factors and immunoglobulins and in which the pro-coagulant factors, Isoagglutinins and unstable components of the blood plasma have been removed.
  • anti-coagulated blood plasma preparation which comprises human plasma proteins, anti-coagulant factors and immunoglobulins and in which the pro-coagulant factors, Isoagglutinins and unstable components of the blood plasma have been removed.
  • unstable factors include lipoproteins and other toxic lipids.
  • Heparinized (i.e., heparinized) blood plasma is preferred because of the maintenance of the physiological calcium concentration.
  • a blood plasma preparation is understood to mean a blood plasma solution which has been produced from the blood plasma of isolated whole blood and which is an isotonic 5% by volume solution of human serum proteins.
  • the blood of mammals is preferred as whole blood, in particular human whole blood from healthy donors (e.g. a pool of healthy 1000 donors).
  • healthy donors e.g. a pool of healthy 1000 donors.
  • inorganic electrolytes e.g sodium, potassium, calcium, magnesium, chloride ions
  • a blood plasma preparation also contains nutrient substrates such as amino acids, sugars and fatty acids and native albumin in the sense of the present invention usable.
  • the plasma also contains many important transport proteins (e.g. transferrin, coeruloplasmin, etc.).
  • Such a blood plasma preparation is preferably a non-agglutinating and anti-coagulated blood plasma preparation in which blood coagulation is inhibited by the addition of anticoagulant substances or no or only a few coagulation factors are still present.
  • anticoagulant substances are, for example, antithrombin III, together with heparin and heparinoids.
  • Blood plasma preparations freed with oxalic acid, EDTA or citrate and free Ca ions are also included for use in endothelial maintenance in isolated organs or vessels within the meaning of the invention.
  • blood plasma preparations are also preferred in which unstable components, such as toxic lipids or lipoprotein, as well as factors triggering an immune reaction (eg blood group-specific antibodies) and germs, such as viruses and bacteria, have been removed by appropriate measures known in the art are.
  • a typical blood plasma preparation used according to the invention preferably comprises the following ionic constituents:
  • Such a blood plasma preparation further comprises natural albumin, e.g. in concentration ranges of about 25-45 g / l, usually about 30-40 g / l, most preferably about 32 g / l.
  • the blood plasma preparation usually contains immunoglobulins, where the content of immunoglobulin G can be about 3-15 g / l, IgA 1-10 g / l, and IgM about 0.2-3 g / l.
  • the blood plasma preparation contains about 7 g / l IgG, 1.7 g / l IgA and about 0.5 g / l IgM.
  • the pH of the blood plasma preparation corresponds to about 7.3-7.8 and is preferably 7.4.
  • the blood plasma solution preferably has an osmolarity of 200-350 mosmol / kg, preferably about 288 mosmol / kg.
  • a particularly preferred embodiment of the blood plasma preparation used according to the invention comprises the following composition: 150 mM sodium ions, 3.65 mM potassium ions, 1.97 mM calcium ions, 1 mM magnesium ions, 107 mM chloride ions, 32 g / l albumin, immunoglobulins IgG 7 g / l, IgA 1.7 g / l, and IgM 0.5 g / l and 51 g / l total protein and natural nutrients and energy suppliers contained in the blood plasma.
  • a particularly preferred embodiment of the blood plasma preparation is an isotonic 5% solution of human serum proteins, made from the plasma of healthy donors.
  • the preparation contains complete and biologically intact immunoglobulins in stable form, albumin and transport and inhibitor proteins in a natural concentration ratio.
  • the preparation is preferably lipoprotein-free, hemolysin-free and free of coagulation factors.
  • a preferred embodiment of such a preparation comprises the following composition: about 3.65 g / l sodium ions, 0.16 g / l potassium ions, 0.08 g / l calcium ions, 0.02 g / l magnesium Ions, 3.65 g / l chloride ions, 50 g / l human serum proteins, thereof approx.
  • the blood plasma preparation as a perfusion solution is pyrogen-free and was treated by virus and / or bacterial inactivating measures (treatment with ⁇ -propiolactone and UV light, heating to 37 ° C.).
  • the blood plasma solution preferably no longer exhibits any anti-complementary, coagulatory or agglutinating activity.
  • the method comprises the provision of blood plasma (for example from a mammal such as horse or cattle, preferably from humans).
  • the human blood plasma can come from several different donors and is collected as a mixed preparation (plasma pool).
  • the cryoprecipitate is preferably separated from the liquid blood plasma components using centrifugation at a temperature of approximately 5 ° C.
  • the remaining blood plasma is treated with ß-propiolactone, ie ß-propiolactone is added to the blood plasma pool in liquid form.
  • the plasma treated with ⁇ -propiolactone is treated with ultraviolet (UV) light with the aid of a UV irradiation apparatus.
  • UV ultraviolet
  • coagulation-active (ie coagulatory) proteins are separated by anion exchange chromatography, for example with the addition of a DEAE Sephadex column. Deep and sterile filtration can also be carried out, which enables the plasma protein solution to be stored at a temperature of 22 ° C. for a longer period (more than 21 days).
  • Subsequent treatment with Aerosü® causes the adsorption of stable proteins and lipids. This treatment also removes any viruses that may still be present.
  • This is followed by slide and ultrafiltration, which enables the protein solution to be adjusted.
  • the pH is adjusted and sterile filtration takes place, which makes it possible to store the intermediate product at a temperature of 37 ° C. for about 30 days.
  • This blood plasma preparation is pyrogen-free and virus-free and can be used in the method according to the invention for the endothelium-preserving treatment of isolated hollow organs or biological vessels or for the uses mentioned, ie for the treatment or preservation of organs or vessels for preparation as organ or vascular grafts or for the repair of injuries to the endothelial tissue, are used.
  • the perfusion solutions or incubation solutions can be used in modified forms.
  • the perfusion solution can differ in its salt and protein composition.
  • solutions that contain either naturally occurring or artificially added amino acids, lipids, carbohydrates, nucleic acids (RNA, DNA) 5 peptides such as gelatin preparations, hormones and plant substances such as dextrans, liposomes for the transfer of lipophilic active substances, hormones or nutrients to the effect of Enhancing or specifying perfusion solution can be used in the methods according to the invention or for the uses of the perfusion solution as long as they contain the above-mentioned basic constituents which constitute the endothelium-preserving effect.
  • Modified solutions of the perfusion solutions according to the invention can also contain growth factors or other proliferation-promoting substances in effective concentrations which are useful for the maintenance and / or for the proliferation of the endothelial tissue by stimulating the division of the endothelial cells or promoting the tissue cohesion of endothelial cells with one another.
  • the growth factors or growth hormones such as hydrocortisone or similar factors
  • the growth factors or growth hormones also reduce the generation time of cell division, which is conducive to a rapid recovery of the endothelial turf.
  • the recovery of the endothelium or the stimulation of the division of endothelial cells in an injured endothelial tissue (lesions) of a biological vessel or hollow organ is therefore an area of application in which growth factors or hormones can be used in the endothelium-protective solution.
  • growth factors examples include epidermal growth factor (EGF), vascular endothelial growth factors (e.g. VEGFl, VEGF, VEGF3), fibroblast growth factor (FGF) and stem cell factor (SCF) or equivalent factors.
  • EGF epidermal growth factor
  • vascular endothelial growth factors e.g. VEGFl, VEGF, VEGF3
  • FGF fibroblast growth factor
  • SCF stem cell factor
  • growth factors include VEGF 121, VEGF 165, VEGFl 89, bFGF, PlGF, PDGF, GM-CSF and G-CSF or equivalent factors.
  • Such growth factors promote the specific maintenance or proliferation of endothelial tissue in vessels by binding to specific surface receptors of this tissue, thereby initiating signal transduction mechanisms which, as a result, cause the expression of growth-promoting genes or gene groups.
  • the gene products can in turn cause the division of the endothelial cells in a complicated interplay.
  • a particularly preferred solution containing growth factors and used according to the invention has the following composition:
  • Isotonic electrolyte solution (127 mM NaCl; 4.6 mM KCl; 1.1 mM MgSO 4 ; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2 mM CaCl 2 (pH to 7.40 before adding CaCl 2 ); 0.1% albumin and 2.5 mM L-glutamine, 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution), 0.1-1 ng / ml epidermal growth factor (EGF), 0.2-2 ng / ml basic fibroblast growth factor (bFGF) and hydrocortisone 0.2-3 ⁇ g / ml.
  • EGF epidermal growth factor
  • bFGF basic fibroblast growth factor
  • TNF, TGF, IFN herbal active ingredients, such as berberine (isolated from Berberis airistata), to the perfusion solution can also be useful in order to maintain or regenerate the endothelial cell layer in order to prevent early thrombosis or to prevent them.
  • berberine isolated from Berberis airistata
  • endothelial cell-specific factors can be added which stimulate the expression of genes which code for cell surface receptors.
  • receptors are CD34, CD133, KDR (VEGFR-2), VE-Cadherin, E-selectin, ⁇ v ß 3 , endothelial progenitor cells specific lectins or other receptors which are specific for endothelial cells and / or endothelial progenitor cells.
  • those compounds or factors can be added to the base solution of the endothelium-protective perfusion solution which promote the spread and proliferation of endothelial cells or endothelial progenitor cells.
  • Such substances are, for example, fibrin, fibronectin, laminin, gelatin or collagen to promote wall anchoring of endothelial cells or purine and pyrimidine compounds such as adenosine, inosine, hypoxanthine or thymidine, uridine and cytosine to promote energy and nucleic acid metabolism and signal transmission of endothelial cells.
  • Certain medications and nutritional supplements can also promote endothelial functions and seal the endothelium.
  • a large number of flavonoid compounds such as quercetin, rutosi compounds or their glycosides (eg quercetin glucuronate), as shown in the examples, promote the tightness of the interendothelial cell joints by the relax the contractile apparatus of the endothelial cells.
  • Antioxidant substances such as uric acid, vitamin E or flavonoids protect the endothelium against the influence of oxidants.
  • Vasodilatory agents such as adenosine or papaverine can relax the smooth vascular muscles and prevent spasms, which can also damage this tissue due to extreme folding of the endothelium. For example, the vascular muscles of the bypass vessels are relaxed and vascular spasms avoided, which could otherwise only be broken through the use of high intravascular expansion pressures, resulting in massive endothelial lesions.
  • Modifications of the endothelium-preserving perfusion solution according to the invention include fresh frozen plasma. This corresponds to human blood plasma that was frozen at or below a temperature of -18 ° C within a few hours.
  • the fresh frozen plasma contains plasma protein (like the necessary albumin), coagulation factors, Von Willebrand's factor and colloids.
  • the fresh frozen plasma does not contain any erythrocytes, leukocytes or thrombocytes.
  • anticoagulation of the fresh frozen plasma should be avoided (e.g. by adding anticoagulants such as heparin, citrate, EDTA etc.).
  • perfusion solution used according to the invention examples include e.g. Plasma expander solutions (containing gelatin or dextran) or modified human albumin preparations, which have been enriched with additional nutrient and energy substrates.
  • Plasma expander solutions containing gelatin or dextran
  • modified human albumin preparations which have been enriched with additional nutrient and energy substrates.
  • the invention also encompasses all combinations of the solution components and additives described here.
  • the invention should in no way be limited to a specific embodiment.
  • the person skilled in the art recognizes that the combination of individual components described here with their individual effects in the sense of endothelium preservation and preservation of vessels and organs is possible.
  • the person skilled in the art will select the further components of the perfusion solution and their concentrations depending on the use and type of the hollow organ, the basic components of physiological or isotonic electrolyte solution, nutrient substrate and above all albumin being contained in all the solutions described here.
  • the person skilled in the art will also recognize that the constituents described here can also be replaced by factors or substances having the same effect in order to achieve the particular effect (for example growth factors having the same effect on EGF or bFGF).
  • the apparatus according to the invention for the endothelium-preserving treatment of isolated biological vessels comprises a chamber (1), an axially movable plunger (6), a cannula (5), a reservoir (7) which contains an endothelium-preserving perfusion fluid and a sealing device (3), the cannula being connected to the axially movable plunger (6) so that it can be moved into the chamber with the plunger, and wherein the sealing device (3) can enclose one end of the biological vessel and the cannula with can be connected to the other end of the vessel, so that the endothelium-protective perfusion solution from the reservoir (7) can be guided selectively, preferably under a pressure gradient, into the biological vessel.
  • the sealing device comprises deformable sealing disks which are arranged in a stack shape under a knurled screw.
  • the sealing disks arranged in a stack in the knurled screw can preferably be pressed on and separated by perforated intermediate disks, for example by steel disks.
  • the steel discs preferably have a thickness of 0.5-2 mm.
  • the stacked sealing washers should be kept at a distance by the perforated steel washers.
  • the diameter of the washer openings should preferably be 1-2 mm smaller than the diameter of the respectively selected sealing washers.
  • the sealing disks preferably have a diameter which is adapted to the size of the vessel to be sealed.
  • the diameter of the vessel is larger than the diameter of the sealing washers, so that the vessels are closely enclosed by the sealing washers in the passage area, but the lumen of the vessels which are filled under pressure and thus widened is not closed.
  • the diameter of the sealing washers can be adapted to the outer diameter of the vessel by pressing the deformable sealing washers together.
  • the diameter of the sealing washers is preferably 1-10 mm and / or has a thickness of 0.3-3 mm.
  • the vessels correspond to the supplying arteries or the draining veins of the organ vascular system in the respective hollow organ.
  • the preferred material of the sealing washers is silicone or any other deformable material that can be used for sealing.
  • the perforated steel discs are preferably made of steel, for example V2A steel.
  • a preferred embodiment of the apparatus of the invention shown in FIG. 1 contains a chamber (1) and silicone sealing disks (3) with a central perforation, which are arranged in a knurled screw.
  • the chamber is preferably cylindrical. Due to the perforation in the sealing washers, one end of a blood vessel (1) (e.g. artery or vein) is initially only pulled through a short distance and closed with a clamp. Since the other end of the vessel is connected to the cannula (5) of the stamp part.
  • the cannula is preferably connected to a hose, which in turn is connected to the storage container (7).
  • the storage container contains the endothelium-protective perfusion solution according to the invention and can be, for example, a Boyle Mariott bottle.
  • the perfusion solution is passed from the storage container into the cannula and thus into the blood vessel at a defined and constant pressure.
  • a pressure gradient ( ⁇ p) existing between the storage container and the chamber.
  • the simplest case for generating a natural pressure gradient is a hydrostatic height difference, for example due to the Position the storage bottle at a height of approximately 1.30 m above the chamber. Then the axially movable plunger (6) is inserted into the interior of the chamber in a sealed manner with the upstream blood vessel.
  • the perfusion fluid may flow out through the side branches of the vessel.
  • the spraying of liquid from the side branches of the vessel in the interior of the chamber decreases rapidly due to the back pressure building up there.
  • By further screwing in the knurled screw in the sealing area it is largely possible to seal the interior of the cylinder in the area where the vessel is carried out, so that the outflow via the side branches located in this area soon stops.
  • perfusion solution is selectively sprayed out of all newly emerging side branches (8), which can then be ligated immediately with ligation aids (9).
  • This surgically essential leak test of biological vessels can be carried out very gently for the endothelium and at a defined and constant pressure. You can also work very cleanly, i.e. Floods as with the use of saline described above, do not occur. Finally, the vessel is pulled out of the vessel and checked again for leaks over its entire length by applying perfusion pressure (e.g. 250 mm Hg). It can then be placed in the protective perfusion solution until transplantation, preferably at a temperature of 37 ° C.
  • perfusion pressure e.g. 250 mm Hg
  • the perfusion liquid according to the invention is additionally passed over a flow spiral of a thermostat system, so that the perfusion liquid can preferably be heated to a temperature of 37 ° C.
  • the apparatus can additionally contain one or more breather screws, which are attached to the chamber and are used to vent or empty the contents of the chamber.
  • the apparatus according to the invention is suitable for all biological vessels, such as blood vessels, that is to say arteries and veins, or lymphatic vessels.
  • An application of the apparatus for the endothelium-preserving treatment of blood vessels, particularly with regard to the Provision of vessels for vascular transplantation or bypass surgery is preferred here.
  • the apparatus according to the invention advantageously enables a constant pressure gradient across the vessel wall, which prevents the vessel from collapsing and accompanying destruction of the endothelial tissue.
  • the contact pressure on the vessel segment can be varied and thus the tightness can be regulated.
  • the stamp preferably moves axially, i.e. through the chamber towards the sealing device.
  • the apparatus according to the invention enables a precise and uniform treatment of the vessel with the perfusion liquid according to the invention.
  • the apparatus enables the endothelial cells to be supplied with perfusion fluid evenly. This enables the exchange of biologically important metabolic molecules that are necessary for the maintenance or proliferation of the endothelial cells.
  • the apparatus is particularly suitable for the leak test of biological vessels.
  • a vessel segment is pulled out over the sealing washers and checked for leaks bit by bit.
  • the vascular outlets identified can then be removed using suitable ligation aids, e.g. Clamps or micro clips.
  • suitable ligation aids e.g. Clamps or micro clips.
  • even the smallest side branches can be easily discovered by spraying perfusion liquid and can be ligated efficiently under pressure control.
  • the entire vessel pulled out of the apparatus can be checked again in its entirety for leaks at an adequate pressure ( ⁇ p at least 180 mm Hg).
  • the perfusion solution according to the invention can therefore be used in a method for the endothelium-preserving treatment of hollow organs or biological vessels.
  • the hollow organs or biological vessels are brought into contact with the perfusion solution according to the invention.
  • the contacting can be the rinsing Insert, or include the complete or partial treatment of the hollow organ or vessel with the perfusion solution.
  • the method for the endothelium-preserving treatment of hollow organs preferably comprises the use of the apparatus according to the invention with which the perfusion liquid is passed through the hollow organ.
  • FIG. 1 shows an apparatus for use in the method according to the invention for the endothelium-preserving treatment of isolated biological vessels.
  • Figure 2 shows a cultured endothelial layer from the saphenous vein in the course of an incubation with saline.
  • FIG. 3 shows a cultured endothelial layer from the saphenous vein in the course of an incubation with a basic composition of the perfusion solution according to the invention.
  • FIG. 4 shows a cultured endothelial layer from the saphenous vein in the course of an incubation with a further embodiment of the perfusion solution according to the invention.
  • FIG. 5 shows cultured venular endothelial cells under the action of release products from blood platelets and granulocytes with subsequent treatment of a perfusion solution according to the invention which contains quercetin.
  • FIG. 6 shows cultured venular endothelial cells under the action of release products from blood platelets and granulocytes with subsequent treatment of a perfusion solution according to the invention which contains papaverine.
  • FIG. 7 shows the response of cultured cultured venular endothelial cells from the human saphenous vein to cardioplegic potassium concentrations.
  • FIG. 8 shows the application of the method according to the invention and the effect on endothelial coverage in the vena saphena when treated with isotonic saline.
  • FIG. 9 shows the application of the method according to the invention and the effect on the endothelial coverage in the vena saphena when treated with Bretschneider solution.
  • FIG. 10 shows the application of the method according to the invention and the effect on the endothelial coverage in the vena saphena when treated with a 5% albumin solution as a perfusion solution.
  • FIG. 11 shows the use of the method according to the invention and the effect on endothelial coverage in the saphenous vein when treated with a blood plasma preparation as a perfusion solution.
  • Example 1 Preferred method for the endothelium-preserving treatment of isolated hollow organs or biological vessels.
  • a perfusion solution with the following composition was used when using blood vessels:
  • the perfusion solution further contained 10% by volume lipoprotein-free, hemolysin-free, homologous serum preparation from a pool of blood preparations (from healthy donors), 2.5 mM L-glutamine, 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution) and additionally 50 ⁇ M uric acid and ascorbate.
  • lipoprotein-free, hemolysin-free, homologous serum preparation from a pool of blood preparations (from healthy donors), 2.5 mM L-glutamine, 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100
  • an apparatus which was to reliably ligate the side branches of the vessel intended for the bypass prosthesis.
  • the apparatus essentially corresponds to that of FIG. 1.
  • This consists of a chamber (1) and silicone sealing washers with interposed steel washers made of V2A steel (3), the central perforations of which are arranged axially in a knurled screw.
  • This sealing device the one end of a human saphenous vein was initially only pulled through a short distance and closed with a clamp.
  • the other end of the vessel was connected to the cannula (5) of the stamp part.
  • the cannula was connected to a tube, which in turn was connected to a Boyle Mariott bottle (7).
  • the Boyle Mariott bottle contained the above-mentioned endothelium-protective perfusion solution and was approx. 1.30 m above the chamber to create a natural pressure gradient ( ⁇ p).
  • ⁇ p natural pressure gradient
  • the perfusion solution from the Boyle Mariott bottle was passed through the tube into the cannula and thus into the vein.
  • the axially movable plunger (6) was inserted into the interior of the chamber in a sealed manner with the upstream blood vessel.
  • the endothelium-protective perfusion solution was applied into the vessel under moderate pressure, so that the perfusion fluid could flow out through the side branches of the vessel.
  • Example 2 Regeneration of endothelial tissue of the saphenous vein by treatment with an embodiment of the endothelium-preserving perfusion solution
  • isolated endothelial cells from the human saphenous vein were microscopically observed during the incubation with various embodiments of the perfusion solutions according to the invention after treatment of the cells, and the effect was monitored by series or video time-lapse -Microphotography documented.
  • Endothelial cells from remnants of the human vena saphena that are no longer required by cardiosurgery were selectively detached by collagenase incubation and saturated with water vapor in minimal essential medium (eg “Dulbecco minimal essential medium”, DMEM) with the addition of 10% v / v fetal calf serum Atmospheric air with the addition of 5% v / v carbon dioxide at a temperature of 37 ° C ("incubator conditions”) grown to confluence. Subsequently, selected dishes were placed in an incubation system, which was firmly mounted on the stage of a Zeiss Axiovert microscope and guaranteed the constancy of the growth conditions described above.
  • minimal essential medium eg “Dulbecco minimal essential medium”, DMEM
  • the continuous photographic documentation was carried out with the aid of a computer-controlled Zeiss AxioCam camera, using software developed by Zeiss and using yellow light with the lowest illuminance that is technically just sufficient to image the respective endothelial layer. Between the individual, automatically triggered individual shots, the cultures were shielded from the light of the microscope lamp by an automatically retractable aperture.
  • the endothelial cells prepared in this way were incubated in conventional saline which had previously been used in the surgical field (FIG. 2; solution 1).
  • the saline had the following composition: 154 mM (0.9% by weight) NaCl in distilled water.
  • the images document the condition of the endothelial cells at various times during the incubation with saline.
  • the starting culture (0 minutes) showed an intact endothelial cell layer.
  • spherical detachment of the endothelial cells and death of the endothelial cells occurred within a few minutes. This led to a complete destruction of the tissue structure.
  • treatment of the isolated endothelial cells with the perfusion solution according to the invention (FIG. 3; solution 2), which contained native albumin, resulted in the endothelial cells being retained over the entire observed incubation period. Mitotic activity of the endothelial cells was also avoided when using this solution.
  • the solution 2 in this example had the following composition: Physiological electrolyte solution (127 mM NaCl; 4.6 mM KCl; 1.1 mM MgSO 4 ; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2 mM CaCl 2 ( pH to 7.40 before addition of CaCl 2 ), 0.1% albumin and 2.5 mM L-glutamine
  • the perfusion solution also contained: 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution) and additionally 50 ⁇ M uric acid and ascorbate.
  • This serum-containing solution (solution 5) had the following composition: physiological electrolyte solution with 127 mM NaCl; 4.6 mM KCl; 1.1 mM MgSO 4 ; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2 mM CaCl 2 .
  • the perfusion solution contained 10% by volume lipoprotein-free, hemolysin-free, homologous serum preparation from a pool of blood preparations, 2.5 mM L-glutamine, 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution) and additionally 50 ⁇ M uric acid and ascorbate.
  • solution 5 contains homologous serum instead of albumin. As shown below, this not only has a pronounced structure-preserving effect on the tissue of the endothelial cells, but also increases the ability of the cultured endothelial cells to divide.
  • the perfusion solution according to the invention in comparison to the conventionally used solutions, such as saline solution or Bretschneider solution.
  • the perfusion solution according to the invention leads to the preservation of endothelial cell tissue and on the other hand has the ability to stimulate endothelial cells to divide. This is particularly advantageous with regard to the regeneration of damaged endothelial layers in biological vessels and is desirable in the preparation or manufacture of biological vessels or vascular grafts.
  • the perfusion solution according to the invention also has the outstanding property that the biological vessels can be stored in the perfusion solution for up to weeks without the endothelial layer being damaged or impaired.
  • Example 3 Preferred Perfusion Solution or Incubation Solution Including Quercetin
  • quercetin a naturally occurring, anti-inflammatory flavonoid
  • various isolated human donor hearts were flushed with the basic solution of the perfusion solution according to the invention in the presence and absence of quercetin.
  • This solution had the following composition: Isotonic electrolyte solution (127 mM NaCl; 4.6 mM KCl; 1.1 mM MgSO 4; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2 mM CaCl 2 (pH on 7, 40 before addition of CaCl 2 ), 0.1% albumin and 2.5 mM L-glutamine
  • the perfusion solution also contained: 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin , low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution) and additionally 100 ⁇ M freshly added quercetin.
  • the arterioles and venules were proteolytically isolated from the human donor heart and purified by isopycnic centrifugation.
  • the endothelial cells were kept in culture. Endothelial cell cultures were established on polycarbonate filters to measure the hydraulic conductivity.
  • Figure 5 shows an 8-part image sequence of the cultured venular endothelial cells of the human heart and the effect of quercetin on the endothelium:
  • starting culture the cells were treated with the basic solution described above, which contains no quercetin.
  • the starting culture has a dense cell lawn.
  • this cell lawn was strongly contracted and thus leaked by the contractile release products from activated platelets and granulocytes of human blood, which was expressed by open spots in the endothelial lawn.
  • regeneration of the endothelial turf is observed, which has become as dense in the course of the incubation in the perfusion solution containing quercetin as in the starting culture.
  • the endothelial turf treated with this incubation solution shows> 98% tightness of the intercellular joints and a very low hydraulic conductivity ⁇ 1 [cm x 10 "6 x S -1 Cm H 2 O].
  • quercetin in the perfusion solution promotes the regeneration of endothelial lesions, which are caused by the release products of activated and metabolically cooperating granulocytes and thrombocytes, and contributes to the preservation of the endothelium.
  • This solution is therefore for use in the Organ and vascular preservation can be used for the purpose of endothelial preservation in the lumen of the vessels.
  • Example 4 Preferred Perfusion Solution or Incubation Solution Including Rutosides
  • the endothelial dressing was destroyed by the activity of activated granulocytes and platelets.
  • flavonoids can prevent and even treat the destruction of the endothelium in vessels by the release products of granulocytes and thrombocytes in inflammatory reactions.
  • Example 5 Preferred perfusion solution or incubation solution including vasorelaxing papaverine
  • spasms can very often develop in the veins freshly dissected from the lower leg, which the surgeon then usually breaks through by "inflating" the veins with isotonic saline solution injected under great pressure Vascular musculature is broken, but also that of the luminal vascular endothelium, which dies as a result of this treatment and is washed away by adding an effective vasodilator to the basic solution of the perfusion solution according to the invention, the formation of spasms in the vein preparation until implantation can be prevented.
  • the solution used in this example had the following composition: Isotonic electrolyte solution (127 mM NaCl; 4.6 mM KCl; I 1 I mM MgSO 4; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2 mM CaCl 2 ( pH to 7.40 before addition of CaCl 2 ), 0.1% albumin and 2.5 mM L-glutamine
  • the perfusion solution also contained: 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution) and additionally 100 ⁇ M papaverine.
  • FIG. 6 shows the endothelial turf of isolated body vein segments before and 3 hours after the incubation in papaverine-containing medium.
  • a vasodilator such as papaverine in the incubation solution did not compromise the tightness of the vascular endothelium.
  • FIG. 6 shows a photograph of a 630-fold enlargement of the endothelial lawn.
  • the top illustration in FIG. 6 shows the endothelial turf immediately before the addition of papaverine.
  • the figure below shows the condition of the endothelium 3 hours after adding papaverine.
  • the endothelium treated with the perfusion solution described above had excellent tightness (> 98%) of the intercellular joints and a very low hydraulic conductivity ⁇ 1 [cm x 10 "6 xs ' WH 2 O].
  • a vasodilator can be added in a practical way to vascular and organ transplants and help to prevent spasms in addition to preserving the endothelium.
  • relaxing the muscles of an organ or vessel to be transplanted is desirable for manipulation by the surgeon and for the comfortable insertion of the organ into the body.
  • Example 6 Preferred Perfusion Solution or Incubation Solution Including Adenosine Analogous results to the results described in Example 5 were also obtained with adenosine as a means of preventing spasm.
  • Adenosine is a physiologically occurring compound that (like papaverine in Example 5) can strongly relax vessels such as veins.
  • the advantage of adenosine is that this nucleoside can be rapidly broken down in the body into vascular physiologically inactive secondary products, so that no systemic effects on the coronary circulation are to be expected.
  • the solution used in this example had the same composition as described in Example 5 above, but it contained 1 mM adenosine instead of papaverine. In the experiments carried out and the subsequent investigations on cell cultures and isolated vein segments, an excellent unity of the endothelial turf was found.
  • adenosine is used to a large extent for the full physiological build-up of the ATP stores of the vascular endothelium and is therefore available as an energy supplier (corresponding to pyruvate and glucose in another embodiment of the perfusion solution according to the invention).
  • adenosine in the incubation solution showed no impairment of the tightness of the vascular endothelium by the vasodilator, both in cell cultures and in isolated vein segments. This solution is therefore suitable for the preservation of endothelium in vessels and organs while at the same time preventing spasms.
  • Example 7 Preferred Perfusion Solution or Incubation Solution Including Growth Factors ( ⁇ GF, bFGF)
  • a perfusion solution which contained growth factors (bFGF, EGF) and growth hormones (hydrocortisone). These factors have a favorable influence on the proliferation of endothelial cells of various vascular origins.
  • the perfusion solution used in this example had the following composition: Isotonic electrolyte solution (127 mM NaCl; 4.6 mM KCl; 1.1 mM MgSO 4; 1.2 mM KH 2 PO 4 ; 24 mM histidine-Cl; 2 mM CaCl 2 ( pH to 7.40 before addition of CaCl 2 ), 0.1% albumin and 2.5 mM L-glutamine
  • the perfusion solution also contained: 2 mM Na pyruvate, 8 mM glucose, 200 U / ml penicillin and 0.2 mg / ml streptomycin, low molecular weight heparin (Fraxiparin from Pharmacia Ltd .: 100 ⁇ l / 100 ml finished solution)
  • the results of these studies show that the closure of the endothelial cell lawn in this medium was> 98%.
  • the generation time of the cells without growth factors was 62 h ⁇ 4, with growth factors the generation time was 19 h ⁇ 3 (n 8 each).
  • the endothelial cells show a significantly shorter generation time in this medium compared to the control solution, which contained no growth factors.
  • Example 8 Preferred Perfusion Solution or Incubation Solution Including Cardioplegic KCl
  • the physiological potassium concentration in the blood plasma is about 4-5 mM. If the potassium concentrations in the extracellular space are increased to concentrations> 6 mM (cardioplegic solution), an increasing paralysis of the electrical activity occurs, at about 8 mM. A heart can no longer beat. In cardiac surgery, such perfusion solutions are used to stop the heart during surgery (e.g. during a heart transplant).
  • the dialyzed solution was sterile filtered and used for treatment as previously described.
  • FIG. 7 shows the response of cultured endothelial cells of human origin to the high potassium concentrations in the solution over time. There is a clear gap formation in the area of the intercellular areas, but not the detachment of the cells. There were systematic perfusion attempts on isolated cattle hearts after perfusion
  • Example 9 Regeneration of endothelial tissue of the saphenous vein by treatment with a blood plasma preparation as an endothelium-protective perfusion solution
  • the blood plasma preparation used contained the following composition:
  • confocal microscopy was carried out on isolated vein segments.
  • the saphenous vein was treated with the blood plasma preparation for 60 min.
  • Saline or Bretschneider solution served as control solutions.
  • Example 10 Embodiment of the blood plasma preparation as a perfusion solution including papaverine
  • Example 10 The same results as described in Example 10 were obtained with a perfusion solution (blood plasma preparation according to Example 9) which additionally contained 10 ⁇ M papaverine as a vasodilator. In addition to maintaining endothelial turf, the addition of papaverine also reduced or prevented the formation of spasms.
  • Example 11 Embodiment of the blood plasma preparation as a perfusion solution including KCl (cardioplegic perfusion solution)
  • the electrical activity with a cardioplegic perfusion solution (blood plasma preparation according to Example 9) on isolated hearts could also be prevented.
  • the solution used contained 20 mM KCl instead of the KCl concentrations stated in the basic solution of the blood plasma preparation.
  • This solution is ideally suited to protect the endothelium during surgery and to immobilize the graft (e.g. vessel, organ) due to the high potassium content due to the incubation in the cardioplegic perfusion solution.
  • graft e.g. vessel, organ
  • tissue factor located in the deeper wall layers
  • the tissue factor is normally shielded from the blood (and thus the coagulation factors in the plasma) by the intact vascular endothelium. This is exactly the case if the veins were previously incubated with one of the perfusion solutions according to the invention.
  • the wall proved to be prothrombogenic because the intravascular coagulation factors could diffuse through the patchy endothelium into the deeper wall layers and came into contact with the tissue factor there.
  • the present invention offers an excellent possibility for the treatment, production and preservation of isolated hollow organs and biological vessels during operations, transplantations, transport and incubations.
  • the hollow organs or biological vessels treated with one of the perfusion solutions according to the invention described here are therefore suitable for use as vascular or organ prostheses (e.g. bypass) with a long service life and functionality.
  • the still high risk of restenosis of such prostheses is considerably reduced for the benefit of the patient by using a perfusion solution according to the invention.

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Abstract

Procédé et appareil pour le traitement, aux fins de leur conservation, d'organes creux isolés, en particulier de vaisseaux biologiques isolés, tels que des vaisseaux sanguins et des vaisseaux lymphatiques, à l'aide de solutions de perfusion ou plus précisément de solutions d'incubation protégeant l'endothélium qui contiennent au moins 0,1 % en poids d'albumine native et un substrat de nutriment, en particulier de la L-glutamine. La présente invention concerne également l'utilisation desdites solutions de perfusion protégeant l'endothélium afin de préparer des organes creux, ou plus précisément des vaisseaux biologiques en tant que greffons pour le traitement de pathologies desdits organes ou vaisseaux, l'utilisation de ces solutions pour réparer des lésions endothéliales dans des organes creux et / ou des vaisseaux biologiques isolés et leur utilisation pour la conservation d'organes et / ou de vaisseaux.
EP04739802A 2003-06-13 2004-06-11 Procede et appareil de conservation de l'endothelium dans des organes creux et des vaisseaux biologiques isoles Withdrawn EP1635636A1 (fr)

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DE10326764A DE10326764A1 (de) 2003-06-13 2003-06-13 Endothel-protektive Perfusionslösung, eine Apparatur und Verfahren zur Konservierung des Endothels in isolierten Hohlorganen und biologischen Gefäßen
PCT/EP2004/006309 WO2004110145A1 (fr) 2003-06-13 2004-06-11 Procede et appareil de conservation de l'endothelium dans des organes creux et des vaisseaux biologiques isoles

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