FR2790641A1 - Liquid for preserving human tissue, especially veins, for use during varicose veins operations contains antibiotic of polymyxin, aminoglycosides or lincosanides family and anti-fungal agent, in physiological solution - Google Patents

Abstract

The liquid composition contains antibiotic of polymyxin, aminoglycosides or lincosanides family and antifungal agent, in physiological solution. The liquid contains, in solution of sodium chloride: (a) at least one antibiotic selected from polymyxin, aminoglycosides or lincosanides family, and (b) at least one anti-fungal agent selected from polyenes group. The liquid may also contain polyethylene glycol (preferably purified and having an average molecular weight 35000). An Independent claim is also included for the use of preserving liquid as claimed at temperature 1-7 (preferably 4) deg C.

Description

1 HUMAN OR ANIMAL TISSUE CONSERVATION LIQUID,

ESPECIALLY OF VEINS.

  The invention relates to a liquid for preserving human or animal tissue.

  It relates in particular, but in a limiting manner, to a liquid for preserving veins, arteries and vessels. In the following description, the invention will be

  more particularly illustrated in relation to the conservation of the saphenous veins located in the lower limb.

  In known manner, these veins are two in number, namely: * the long saphenous vein, which goes up along the external face of the leg to end in the popliteal vein; + the long saphenous vein, which goes up along the internal face of the

  leg, then thigh, to end in the femoral vein.

  These veins, by reacting to an increase in intramuscular pressure, can dilate, forming varicose veins. The treatment of these varicose veins can consist of a surgical operation called saphenectomy, according to which all or part of the saphenous vein is resected.20

  Therefore, the saphenous veins taken during these operations can be used after selection as allografts.

  We are particularly interested in the rectilinear veins, taken from

  so-called tubular varices, which have a relatively solid wall.

  In practice, it is necessary after removal of said vein and its surrounding tissue to locate and isolate the segments of veins capable of constituting the grafts, then to remove the peripheral fatty tissues and finally, to ligate the lateral branches. Once these operations are completed, after leakage control, then measurement of the porosity of the vein retained under slight pressure, the allografts are then presented on a glass mandrel and arranged in a

preservation solution.

  The storage conditions must be such that the allograft, while being devoid of any cellular viability, retains its characteristic properties, namely: elasticity of the wall, hemostatic properties, recolonization power

recipient cells, etc.

  To achieve this objective, physico-chemical treatments of the graft have been proposed, in particular by means of glutaraldehyde. However, glutaraldehyde is toxic and poses a risk of salting out and tans tissues, thereby causing

  loss of their mechanical properties.

  It has also been proposed to keep the venous grafts in frozen form at −196 ° C. in liquid nitrogen. However, it was found that the cryopreserved grafts were subject to stenosis, in particular in the proximal part of the allograft, downstream of the upper anastomosis. It is also pointed out that this type of graft induces an immunological reaction and progresses towards the formation of postanastomotic proximal stenoses.15 Finally, it was proposed in document WO 97/22244 to preserve organs or tissues in a calcium-based preservation solution and of

  nitroglycerin at a temperature between 0.5 and 8 C. However, in the case of the conservation of veins, the endothelium is also preserved, which is not always desired.

  In other words, the problem which the invention proposes to solve is to provide a liquid for preserving human or animal tissue, the composition of which

  and the conditions of use allow the fabric to retain their integrity and their mechanical properties.

  To do this, the invention provides a liquid for preserving human or animal tissue, in particular of veins, characterized in that it comprises, in a solution of sodium chloride: 30 has at least one antibiotic chosen from the group comprising the family polymyxins, the family of aminoglycosides and the family of lincosanides; * and at least one anti-fungal chosen from the group comprising the family

polyenes.

  In other words, the invention resides in the selection of a certain number of families of antibiotics and anti-fungals which, when mixed, exhibit a synergistic effect making it possible to ensure the preservation of the tissues after their collection. .5 In practice, this preservation liquid is used at a temperature between 1 and 7 ° C., advantageously 4 ° C. According to another characteristic of the invention, the antibiotic chosen from

  family of polymyxins is colistin.

  In practice, the concentration of colistin is between 500,000 and 1

  500,000 units per liter of 0.9% sodium chloride solution, advantageously 1,000,000 units per liter.

  For a concentration of less than 500,000 units per liter, the bactericidal effect is not satisfactory. For a concentration greater than 1,500,000

  units per liter, bactericidal activity stabilizes.

  Furthermore and advantageously, the antibiotic chosen from the family of aminoglycosides is gentamycin.

  In practice, the concentration of gentamycin is between 200 and 300 mg per liter of 0.9% sodium chloride solution, advantageously 240 mg

per liter.

  As before, for concentrations below 200 or above 300 mg per liter, the bactericidal effect is not obtained or maximum.

  Likewise, the antibiotic chosen from the lincosanides family is

advantageously lincomycin.

  In practice, the concentration of lincomycin is between 50 and 150 mg per liter of 0.9% sodium chloride solution, advantageously 102 mg

per liter.

  Finally, as already said, the preserving liquid also comprises an anti-fungal agent chosen from the family of polyenes, advantageously amphotericin B. In practice, amphotericin B is used at a rate of 10 to 90 mg per liter of 0.9% sodium chloride solution, preferably 50 mg per liter. To avoid the formation of tissue and cellular edema, the preservation liquid of the invention also comprises polyethylene glycol (PEG)

  to guarantee oncotic pressure.

  Advantageously, the polyethylene glycol used is a purified polyethylene glycol of average molecular weight equal to 35,000.

  In the following description and claims, by purified polyethylene glycol is meant a polyethylene glycol powder free of any molecule

  of molecular weight less than 15,000.

  It has in fact been found that PEGs of molecular weight less than 8,000

  caused intoxication similar to that of ethylene glycol, while derivatives of high molecular weight greater than 15,000 were non-toxic.

  In practice, the concentration of polyethylene glycol in the preservation liquid is between 0.1 and 0.3 mM per liter of chloride solution.

  0.9% sodium, preferably 0.25 mM.

  The invention and the advantages which result therefrom will emerge more clearly from the following exemplary embodiments.

Example 1

  A preservation liquid is prepared, the composition of which is reproduced in the table below.

  Trade name Laboratory Active ingredient Concentration per liter of CINa 0.9% Colimycin Bellon RPR Colistin 1,000,000 U / L Gentalline Schering-plow (Gentamicin 250 mg / L Lincocin Upjohn Lincomycin 100 mg / L Amphotericin B Sigma Amphotericin B 50 mg / L As already mentioned, saphenous vein samples are exclusively taken by surgeons in operating rooms.

  collection is made available to the sampling service, this case containing 8 sterile bottles filled with storage fluid of the invention, tubes for blood samples as well as pre-numbered labels for identification.

  The sterile bottles are kept at a temperature of 4 ° C and the case containing the allografts is returned to the bank a maximum of 15 days after

his expedition.

  In practice, the saphenous veins removed are used to form vascular approaches for hemodialysis or for revascularization of the

inferior member.

Example 2

  The activity of the preservation liquid of the invention was studied on the following strains: * Staphylococcus epidermidis 459 924.54 IE / ml * Saccharomyces cerevisiae 760.89 IE / ml * Bordetela bronchiseptica 78557.71 IE / ml

  The advantages of the invention appear clearly from the description above. Note in particular the originality of the combination of antibiotics and anti-fungals

  from specific families, thus ensuring the conservation of tissues and in particular of veins, and thus maintaining their integrity and their properties

mechanical.

Claims (8)

  1 / Preservative liquid, in particular of veins, characterized in that it comprises in a sodium chloride solution: * at least one antibiotic, chosen from the group comprising the family of polymyxins, the family of aminoglycosides and the family of lincosanides ; e and at least one anti-fungal, chosen from the group comprising the family polyenes.   2 / Preservative liquid according to claim 1, characterized in that the antibiotic chosen from the family of polymyxins is colistin.   3 / Preservative liquid according to one of the preceding claims,   characterized in that the antiobiotic chosen from the family of aminoglycosides is gentamycin.   4 / Preservative liquid according to one of the preceding claims, characterized in that the antibiotic chosen from the lincosanides family is lincomycin.   / Preservative liquid according to one of the preceding claims, characterized in that the anti-fungal agent chosen from the family of polyenes is   amphotericin B.25 6 / Preservative liquid according to claim 2, characterized in that the colistin concentration is between 500,000 and 1,500,000 units per liter of 0.9% sodium chloride solution, advantageously 1 000,000 units per liter. 30 7 / Preservative liquid according to claim 3, characterized in that the concentration of gentamycin is between 200 and 300 milligrams (mg) per liter of 0.9% sodium chloride solution advantageously 240 mg per liter. 35 8 / Preservative liquid according to claim 4, characterized in that the concentration of lincomycin is between 50 and 150 milligrams (mg) per   liter of 0.9% sodium chloride solution, preferably 102 mg per liter.   9 / Preservative liquid according to claim 5, characterized in that the concentration of amphotericin B is between 10 and 90 milligrams (mg)   per liter of 0.9% sodium chloride solution, preferably 50 mg per liter.   O10 / Preservative liquid according to one of the preceding claims, characterized in that it also comprises polyethylene glycol.   11 / Preservative liquid according to claim 10, characterized in that the polyethylene glycol is purified and has an average molecular weight of 35,000.   12 / Preservative liquid according to the preceding claim, characterized in that the concentration of polyethylene glycol is between 0.1 and 0.3   millimoles (mM) per liter of 0.9% sodium chloride solution, preferably 0.25 mM.   13 / Use of the preservation liquid according to one of claims 1 to   12 at a temperature between 1 and 7 C, advantageously 4 C.