MXPA00002673A - Fibrin sponge - Google Patents

Abstract

The invention relates to a fibrin sponge comprising a residual moisture content of at least 3%, preferably of 3 to 35%, in particular 10 to 20%, and preferably containing a blood clotting activator or proactivator, a method of preparing this fibrin sponge as well as a kit for wound gluing which comprises the fibrin sponge and a component containing a blood clotting factor. The sponge according to the present invention is suitable for hemostasis, tissue adhesion and aiding wound healing.

Description

FIBRINE SPONGE The present invention relates to a fibrin sponge having a defined residual moisture, to the methods of its preparation, to its use for hemostasis, adhesion of tissues and support in the healing of wounds as well as to a set of implements for its application.

In the prior art, the most varied range of hemostatic materials for covering wounds, also based on fibrin, have been described and partially used, for example, fibrin sponges, fibrin membranes or fibrin gels.

So far, the opinion has prevailed that for hemostasis, a combination of fibrinogen and trorabine or the corresponding prothrombin activators is necessary. Thus, for example, collagen-based fleece-like flat materials comprising fibrinogen and thrombin (TachoComb®), flat materials comprising fibrinogen and activated factor X (AT 0 359 653, IMMUNO AG) or flat materials REF; 33087 comprising fibrinogen and thrombin as in a dry preparation (EP 0 748 633, IMMUNO AG) have been known from the prior art. While fleece-like flat materials based on fibrinogen and activated factor X are already soft and flexible, other flat materials require absolutely dry storage in order to prevent a premature reaction of fibrinogen contained therein, with thrombin or with other activators of fibrinogen. blood clotting Bering E. A. Jr. (J. Clin. Invest. Vol. 23, 1944, p. 586) discusses the general developments in the field of fibrin foams and their use as hemostatic agents.

A study of various fibrin-based materials, eg, fibrin powders, fibrin films and fibrin foams, is given, for example, in Gerendas M. "Fibrin Products as aids in hemostasis and wound healing", pp. 277, in K. Laki (ed.) Fibrinogen, M. Dekker, New York, 1968.

In addition, the fibrin lamellae or fibrin membranes, respectively, have been known from the prior art (see e.g. EP 0 485 210-A2). Such materials are extremely thin, non-absorbent and therefore usable in hemostasis, only to a limited extent. In contrast to fibrin sponges, these materials are purposely compressed during their preparation, for example, by the application of weight. Therefore, these materials are characterized primarily by their high density, compared to the foams.

The present invention has, as an objective, to provide a means for hemostasis, tissue adhesion and support in tissue healing based on fibrin which, compared to conventional means, has a good, and particularly improved efficacy, is simple to use with a simultaneous simple composition that allows a cheap preparation, and which also has a wide field of application. Beyond this, it must be possible to easily supply the fibrin material with other auxiliary and active substances.

According to the invention, this objective is achieved by providing a fibrin sponge having a residual moisture of at least 3%, preferably in the range of 3 to 35%, in particular from 10 to 20%, which is stable during the storage. The water content or residual moisture, respectively, is preferably agitated in such a way that its handling and efficiency are substantially improved.

It is known from the prior art that dry preparations containing fibrin, which have a hard and brittle consistency, will become smooth and uniform (flexible) by increasing their residual moisture. However, there is a general prejudice regarding the stability of such protein preparations having increased residual moisture, particularly if they contain a sensitive enzyme, such as thrombin, or another activator or pro-activator of blood coagulation.

It is not without reason that similar sensitive protein preparations are commonly stabilized by lyophilization, residual moisture of less than 3%, and in most cases even less than 1%, has been achieved.

Therefore, it is surprising that a fibrin sponge preferably has a thrombin content and an increased residual moisture or an increased water content, and that it is, however, stable during storage.

According to the present invention, for stable during storage, it is understood that a fibrin sponge that even after a storage of 6 months, and generally, however, even after a storage of 2 or more years at room temperature, this is still extremely suitable for its application, and that the present optional thrombin activity or blood coagulation activator or proactivator activity remain substantially preserved. Preferably, even more than 70% of the activity remains for more than a period of at least 6 months, preferably 2 or more years, at room temperature. Even at a temperature of 37 ° C the fibrin sponge according to the invention, which has a residual moisture of 15%, has been shown to be stable for at least 3 months, preferably for more than 6 months and the activity of thrombin to remain completely preserved The preparation according to the invention particularly combines the various requirements for a medium of hemostasis, adhesion of tissues and support in wound healing, with the preparation according to the invention, properties such as: - a good and lasting efficacy, ie a fast adhesive effect or haemostasis, no secondary bleeding or delayed detachment of the adhered parts of the tissues, a good tolerance, even in multiple applications (that there is no sensitization), a complete resorption during the wound healing process, a smooth and intact wound healing, as much safety as possible with respect to the transmission of viral pathogens and others (such as, for example, prions, the pathogens of bovine spongiform encephalitis, BSE), a simple use, as well as the that stability has already been mentioned during storage of the ready-to-use product which can, for example, be ensured.

Preferably, the fibrin sponge according to the invention is a fleece-like fibrin flat material or a fibrin foam having a low density and a high absorbent capacity.

According to the invention, it is understood that a fibrin-like fibrin flat material refers to a fibrin sponge having a porous structure, in particular to a fine porous structure, without intermingling largely with air bubbles, as is obtained in according to the invention, by lyophilization of non-foamed fibrin clots. On the other hand, it is understood that a fibrin foam is a material obtained by foaming a solution of fibrinogen, and mixing it with a solution of thrombin and lyophilizing the foamed fibrin clot obtained. Foaming may also occur immediately after the addition of thrombin.

The fibrin sponge according to the invention can be used as such. It may consist essentially of fibrin and of the thrombin required to produce fibrin from fibrinogen, surprisingly, even this embodiment exhibits an excellent hemostatic and adhesive effect.

However, it can be impregnated with more additives, preferably with an activator or pro-activator of blood coagulation. Said additives are preferably homogeneously distributed in the preparation according to the invention.

Preferably, the activator or proactivator of blood coagulation is selected from the group consisting of thrombin, prothrombin, activated factor X, prothrombin complex, activated prothrombin complex, FEIBA, calcium ions and mixtures thereof.

In particular, additional thrombin can be incorporated in the preparation according to the present invention, preferably an amount of between 1 and 300 U / cm 3, and more preferably between 5 and 100 U / cm 3, and more preferably between 10 and 50 U / cm3.

Surprisingly, it has been found that the preparations according to the invention based on solid fibrin and also with an additional content of thrombin are still stable for long periods of storage at room temperature or refrigerant and even at 37 ° C (in an accelerated analysis of the stability ). In particular, the activity of the thrombin content is maintained substantially for long periods of time. The content of thrombin activity can, for example, after extraction with a 1M NaCl solution, be determined according to methods known per se, for example, with a chromogenic substrate (Th-1, Pentapharm).

In addition to substances, such as activators or pro-activators of blood coagulation, the fibrin sponge according to the invention may also contain stabilizers, preservatives, antibiotics, therapeutic agents, antifibrinolytic agents, growth factors, more plasma proteins, enzymes, inhibitors or mixtures of such agents.

As stabilizers, thrombin stabilizers are preferably used, in particular polyols, polysaccharides, polyalkylene glycols, amino acids or mixtures thereof or substances that stabilize the water content, in particular polyscarides or polyols,. The use of sorbitol, glycerol, polyethylene glycol, polypropylene glycol, mono or disaccharides such as glucose or sucrose, or any sugar or amino acid that stabilizes thrombin activity or residual moisture, respectively, is preferred.

Likewise, the desired smoothness of the preparation can be obtained or improved by incorporating substances, for example, hydrostabilizing active substances, such as glycerol. After lyophilization in the presence of such substances, increased levels of residual moisture can be obtained.

A preferred antifibrinolytic agent is selected from the group consisting of aprotinin, aprotinin derivative, (a-2-macroglobulin, a-2-plasmin inhibitor, a-1-plasmin inhibitor, plasminogen activator inhibitor, inhibitor or inactivator of activated protein C, a plasmin-binding substance, or mixtures of these substances.Synthetic substances can also be used, in particular synthetic inhibitors of fibrinolysis, for example, e-amino-caproic acid, tranexamic acid Non-plasmatic antifibrinolytics, of vegetable origin, can also be used according to the present invention.

Preferred growth factors are, for example, cytosines. As other plasma proteins, preferably albumin, factor XIII or fibronectin are provided.

Furthermore, according to the invention, specific proenzymes, enzymes or enzyme inhibitors can also be mixed with the fibrin sponge for example, in order to regulate the absorption of the fibrin sponge in the body, ie to accelerate it or inhibit it. These substances include, for example, plasminogens, collagenase, plasminogen activators, plasminogen activator inhibitors, plasmin inhibitors or elastase inhibitors.

According to the invention, the preparations as such already have a good absorbent capacity. Preferably, a small amount of a tensing agent, such as Tween®80 (sorbitan polyoxyethylene monooleate) is further incorporated, wherein the absorbent capacity of the preparation according to the invention can be further increased.

In this way, the preparation according to the invention is characterized in particular by the following properties: It has a great softness or adaptability so that it can be applied without any problem to an uneven surface of the wound. In addition, it has a high absorbent capacity, so that the emerging blood can be absorbed without any delay.

Preferably, the fibrin sponge according to the invention has a liquid absorption capacity of at least twice, preferably more than 4 times its own weight.

In a preferred embodiment, the material has a homogeneous distribution of its coagulation promoting properties, of such a level, that according to the present invention, the absorbed blood coagulates in the preparation in a few seconds, preferably within less than 30 seconds. .

The present invention also relates to a method for preparing the fibrin fibrin sponge, which is characterized by the following steps: preparing a fibrin clot by mixing fibrinogen and thrombin and incubating the mixture at a temperature, preferably 20 at 37 ° C for a period of time sufficient for the formation of the fibrin clot and in particular to complete the reticulated form of the fibrin, preferably for 10 minutes to 24 hours, (rapid freezing and) freeze-drying the fibrin clot, and adjust a residual moisture of at least 3%, preferably from 3 to 35%, and in particular from 10 to 20%.

In a preferred embodiment, the adjustment of the water content is effected by lyophilization. The residual moisture can, however, also be adjusted to a desired level by being incubated in a humid chamber after lyophilization.

When a preparation according to the invention is produced, preferably a washing and / or incubation step is also provided. Therefore, in particular, the soluble components, such as, for example, the salts, can be removed, while the thrombin content, which is explained in the preparation method, is surprisingly retained to a large extent.

The washing can be carried out with distilled water, although solutions containing active substances or additives can also be used. In this way, it is possible to vary or adjust the composition of the final product.

In particular, it has been found that thrombin containing the fibrin clot can practically not be eluted when washed with an isotonic solution. For example, after washing for 4 hours with 10 times the volume of the isotonic saline solution, approximately only the 4% of the total thrombin is found in the wash solution. On the other hand, other plasma proteins, such as, for example, albumin, elute comparatively rapidly under the same conditions (approximately 25% for 4 hours, approximately 50% for 8 hours).

Finally, the surprisingly high stability of the activator or pro-activator of blood coagulation, respectively, which according to the invention is preferably distributed homogeneously in the preparation, can be further increased by the stabilizers known per se, such as, for example, sugars, alcohols of sugar, polyols, amino acids, etc., or mixtures thereof.

In another preferred embodiment, as already mentioned above, during the course of its preparation, the fibrin sponge is washed and / or impregnated with other substances. Preferably, the fibrin clot is impregnated before lyophilization. However, it is also possible to carry out washing or incubation, respectively, after lyophilization. this then after another lyophilization step.

Fibrin foam can be prepared by foaming the fibrinogen solution before or immediately after mixing with thrombin, but before coagulation occurs. All the common methods of the prior art can be used for this foaming. For example, these can be chemical, physical and / or mechanical methods or means. The gases can be introduced or generated, in particular the air is introduced by a mechanical and intense agitation treatment (Ultraturrax treatment or similar). As an example of a physical method, the application of a vacuum can be mentioned.

In addition, however, it has been found that in contrast to the methods described hitherto to prepare the fibrin sponges, which after the lyophilization are produced absorbent preparations and particularly fine pores, without a substantial contraction of the fibrin clots that have been prepared without foaming.

Before freezing and lyophilization, preferably the fibrin clot has a low salt content or a low ionic strength, respectively, preferably less than 0.15, it is preferably substantially free of salts.

By avoiding frothing, the method of preparation is simplified and substantially facilitated, particularly if the latter is to be carried out under sterile conditions.

In another preferred embodiment, the fibrin sponge, i.e., in particular the fibrin flat material or the fibrin foam, is then cut from its respective preparation. Preferably, it is cut such that, after being cut, the cut area that can later be used as a wound cover is as large as possible. It has surprisingly been found that immediately after lyophilization, the absorbent capacity of these cut areas is even greater than that of the uncut, or so to speak, "original" surfaces.

Good tolerance even after multiple application, in particular to avoid allergic reactions, is preferably achieved by the exclusive use of human proteins as well as by avoiding methods, for example, sterilization methods, by which proteins can be denaturalize (for example, such as, by irradiation-? In a particularly simple embodiment, the preparation of the wound cover according to the invention is carried out from a sterile raw material, by maintaining the sterile conditions until the final product.

Preferably, the preparation according to the invention is prepared from fractions of human plasma protein which are treated according to suitable methods of viral inactivation for sensitive plasma proteins.

The inactivation treatment is preferably ensured by a heat treatment and / or tensile agents, for example, by a solid state heat treatment, in particular a steam treatment according to EP 0 159 311, EP 0 519 901 or EP 0 674 531.

In a preferred embodiment, the heat treatment is carried out as a steam heat treatment with a residual water content between 6 and 15%, preferably about 12%. When thrombin was present in the sponge, it was surprisingly found that even under these conditions almost all thrombin activity is maintained.

Other steps to inactivate viruses also include treatment with chemical or chemical-physical methods, for example, with chaotropic substances according to the WO 94/13 329, DE 44 34 538 or EP 0 131 740 (solvents) or photoinactivation.

Preferably, two independent methods of viral inactivation are carried out.

Nanoinfiltration also constitutes, within the scope of the present invention, a preferred method for decreasing viruses.

Another possibility to increase viral safety is to initially start the preparation exclusively from - to the greatest extent possible - virus-free raw materials and then exclude all risks of contamination during the course of the preparation process. The greatest possible safety in terms of transmission of viral or other pathogens in the preparation from human plasma is ensured in an additional way, through the selection and careful verification of plasma donors, analysis of the primary plasma for pathogenic viruses (preferably analysis). single donation), and in particular by means of PCR methods.

Even, in a basic way, the preparation according to the invention can also be prepared from recombinant or transgenic material, where the necessary safety measures must also be met.

Preferably, the preparation is carried out under sterile conditions, starting from sterile intermediary products (sterile filtered solutions) to the final product packaged by sterile methods, so that a subsequent sterilization in the final container is no longer necessary, such as for example , by irradiation- ?.

Complete absorption during the wound healing process as well as uniform healing and without wound interruptions are achieved through the use of fibrin and optionally other plasma proteins, such as, for example, fibronectin, as a matrix, the formation of fibrin takes place preferably with an approximate physiological ionic strength, at an approximate physiological pH and in the presence of factor XIII, so that the fibrin clot is obtained in a reticulated form having a physiological fibrin structure which is essential for the germination of the fibroblasts and therefore for a rapid and uniform healing of the wound (see also Redi et al., Med. Welt, 36, pp. 769-776, 1985).

The width and length of the fibrin sponge according to the present invention can be freely chosen depending on the type of use. Its thickness is preferably in a practical range of between 1 and 20mm, depending on the indication, and thicknesses of between 3 and 15 or between 5 and 10mm are preferred, respectively.

The fibrin sponge according to the invention can also be presented in combination with other materials.

In a specific embodiment the sponge is made of several layers so that a multi-layer material is formed. Preferably at least one layer has hemostatic properties. The layers can be constructed with the same material or they can differ in the material and / or its composition.

The material of the sponge can be characterized by the specific weight of the material or by each layer. In a preferred embodiment, the specific weight is between 0.005 and 0.15 g / cm3, more preferably between 0.01 to 0.09 g / cm3, and more preferably between 0.02 to 0.05 g / cm3.

In another preferred embodiment, the layers are composed of the same material but differ in their physical properties, for example, their specific weight. By varying the physical properties of the layers, a sponge can be obtained which is characterized in that one side of the sponge is smooth, with a high absorbent capacity while the other side of the sponge is more compact and compressed. Another way of varying the properties of the different layers can be achieved by varying the amount or concentration of the substances forming the layer or of those substances in which each layer is impregnated. For example, the concentration of thrombin in the layers may be different.

The use of fibrin foam, for example, is such that it is applied to the bleeding wound and slightly pressed into it for a short period of time (approximately 30 seconds). The sponge absorbs the blood very quickly and the absorbed blood coagulates inside the sponge. Therefore, firm attachment of the fibrin sponge to the wound surface is achieved, which is at least the basis of its hemostatic and adhesive effect.

The preparation can also be used in the form of so-called dry adhesive, preferably as a relatively thin sponge, to adhere two (soft) parts of bleeding tissue, a suitable piece of "dry adhesive" is applied to the wound surface. and subsequently the second surface of the wound is quickly adapted and pressed.

According to the invention, the medical field indicated for the fibrin sponge is quite large. Not only can the sponge be used to stop an oozing hemorrhage, it can also be used to stop bleeding in the case of areas of heavy bleeding with high blood pressure. According to the invention, the following internal and external surgical procedures can be carried out successfully by using the fibrin sponge: general surgery, for example, surgery on parenchymal organs (liver, kidney, spleen, etc.), cardiovascular surgery, thorax surgery, graft surgery, orthopedic surgery, surgical operations in the fields of bone surgery and plastic surgery, ear, nose and throat surgery, operations in the field of neurosurgery, operations in the urological and gynecological tracts , as well as generally for hemostasis as well as to treat conventional wounds.

According to yet another aspect of the present invention, a set of implements for adhering the wounds is also provided, characterized in that it comprises a fibrin sponge according to the invention with a component that contains a blood coagulation factor.

Preferably, this blood coagulation factor is fibrinogen, factor XIII, fibronectin, thrombin, or mixtures of these factors, a fibrinogen component preferably in a storage stable form, as is particularly preferred, a commercially available product. of a tissue adhesive based on fibrinogen.

The. Sponge according to the present invention, may also be useful as a cell culture biological assistant, especially for mammalian cells, for example, keratinocytes, epithelial cells, epidermal cells, fibroblasts, chondrocytes or the like.

Therefore, according to the present invention, a set of implements comprising according to the present invention is also provided, the fibrin sponge in its sterile form for use as a biological helper for cell cultures.

Other additives such as antibiotics, various amino acids and / or various growth factors can be present inside the sponge or can then be added to the medium used.

According to the present invention, by using the fibrin sponge as support for the growing cells, it is possible to form tissue replacements, for example, of skin, bone, cartilage, and / or nerves, respectively.

The invention will be explained in more detail by the following example, however, without restricting it to this: Example 1: The fibrin-like fibrin-like flat material of reticulated? Irrira having a (relatively low) thrombin content. 1. 1 Preparation A commercially available tissue adhesive preparation, freeze-dried and inactivated to viruses by a steam treatment (Tissucol®, IMMUNO AG) served as the raw material for preparing a solution of fibrinogen having a content of factor XIII and fibronectin. According to the manufacturer's instructions, the preparation was dissolved with an aqueous solution of aprotinin (3000 KlU / ml) and further diluted 1: 4 with distilled H20 to produce a fibrinogen concentration of approximately 20 mg / ml. 20 ml of this solution was quickly mixed with 20 ml of a solution of thrombin-CaCl 2 (human thrombin, approximately 10 U / ml 5 mM CaCl 2) and poured into a Petri dish (ID = 8.4cm), without touching at room temperature environment for approximately 30 minutes and then incubated for approximately 16 additional hours in a humidity chamber at 37 ° C. The clot in the form of a round disc having a thickness of approximately 7 mm was then placed in rapid freezing and lyophilized. The lyophilized form (flat material similar to fleece) has a light appearance and fine pores that substantially retains the shape of the clot prior to lyophilization. Although the material was still too brittle for optimal application.

To improve the desired smoothness and flexibility, the flat material was incubated for approximately 3 hours in a humid chamber at room temperature, whereby an adaptable and highly absorbent material was obtained. 1. 2 Analysis, In vitro tests: Residual humidity Of approximately 15% Flexibility: > 90 ° to a mean radius of curvature corresponding to half the thickness of the layer Reticulated form of fibrin Chains- ?: 100% Chains-a: approximately 80% Thrombin content: Determination after extraction with 1M NaCl: 1.4 U / cm3 Water absorption capacity: Approximately 6 times its own weight Water absorption rate: A drop of water (50μl) placed on the surface of the flat material is completely absorbed in about 25 seconds (in areas cut off from the flat material under otherwise equal conditions, the water is absorbed practically immediately, it is say, in less than 5 seconds). 1. 3 Efficiency The adhesive and haemostatic effect of the preparation prepared according to 1.1 was analyzed in a rabbit with the so-called renal pole resection model.

For this, a kidney is exposed in the test animal under anesthesia with ketamine and xylazine (with a postdosing of pentobarbital), the renal pole is removed with the scalpel. A circular area is formed that bleeds with a uniform intensity and that has a diameter of approximately 1 to 1.5 cm. The preparation of the analysis is placed in the bleeding area, making light pressure to it by means of the finger during exactly 30 seconds, then it is released. The adhesion of the preparation is observed, and the time of complete hemostasis is measured. The blood possibly filtered is taken with cotton gauze and the blood loss is determined colorimetrically after the extraction with ammonia, the analysis material and the cotton gauze.

The analyzes with the preparation obtained according to 1.1. They were carried out in two rabbits.

Results: In both cases, the preparation adheres well to the surface of heavy bleeding and complete hemostasis was achieved almost immediately (in less than 30 seconds). No further bleeding occurred during a secondary observation period of one hour.

The total blood loss (absorbed in the analysis material, without other blood absorbed) had a total of 0.10 and 0.18 ml, respectively.

In comparison to this, with conventional hemostasis by gauze the blood loss in this model is about 5 ml with a bleeding time of 3 minutes, and about 27 ml with a bleeding time of 9 min without any treatment.

In this way, the substantially improved efficiency of the preparation obtained according to 1.1 has been impressively demonstrated as compared to conventional methods.

Claims (27)

Claims

1. A stable fibrin sponge during storage, characterized in that it has a residual moisture of at least 3%, preferably in the range from 3 to 35%, and in particular from 10 to 20%.

2. A fibrin sponge according to claim 1, characterized in that it comprises an activator or proactivator of blood coagulation.

3. A fibrin sponge according to claim 2, characterized in that the activator or proactivator of blood coagulation is selected from a group consisting of thrombin, prothrombin, activated factor X, activated prothrombin complex, FEIBA, calcium ions and mixtures of these.

4. A fibrin sponge according to claim 2 or 3, characterized in that it comprises from 1 to 300 U / cm 3 of thrombin, preferably from 5 to 100 U / cm 3 and more preferably from 10 to 50 U / cm 3.

5. A fibrin sponge according to any one of claims 1 to 4, characterized in that it comprises stabilizers, preservatives, antibiotics, therapeutic agents, antifibrinolytic agents, growth factors, other plasma proteins, enzymes, inhibitors or mixtures of these agents.

6. A fibrin sponge according to any of claims 1 to 5, characterized in that it has a lipid absorption capacity of at least 2 times, preferably 4 times its own weight.

7. A fibrin sponge according to any of claims 1 to 6, characterized in that it is inactivated of virus.

8. A fibrin sponge according to claim 7, characterized in that the inactivation of virus has been carried out in two independent steps of viral activation.

9. A fibrin sponge according to any one of claims 1 to 8, characterized in that it is free of eluitable plasma proteins, in particular, is free of elutionable thrombin.

10. A fibrin sponge according to any of claims 1 to 9, characterized in that it has been prepared from raw material safe from viruses.

11. A fibrin sponge according to any of claims 1 to 10, characterized in that it is present in an aseptic package in a ready-to-use form.

12. A fibrin sponge according to any one of claims 1 to 11, characterized in that it has a thickness of at least 1 mm, and preferably between 5 and 10 mm.

13. A fibrin sponge according to any of claims 1 to 12, characterized in that it has multiple layers, preferably at least one layer having haemostatic properties.

14. A fibrin sponge according to any of claims 1 to 13, characterized in that it has a specific gravity of 0.005 and 0.15 g / cm3 preferably 0.01 to 0.09 g / cm3, and more preferably 0.02 to 0.05 g / cm3.

15. A set of implements for covering wounds, characterized in that it comprises a fibrin sponge according to any one of claims 1 to 14 and a component that contains a blood coagulation factor.

16. A set of implements according to claim 15, characterized in that the blood coagulation factor is fibrinogen, factor XIII, fibronectin, thrombin or mixtures of these factors.

17. A method for preparing a fibrin sponge according to any of claims 1 to 14, characterized in that it comprises the following steps: a) preparing a fibrin clot by mixing a solution of fibrinogen, which preferably contains factor XIII, with a solution of thrombin, which preferably contains ions of Ca, and which is incubated at a temperature, preferably 20 to 37 ° C, for a period of time sufficient for the formation of the fibrin clot and optionally for the crosslinked form of the fibrin, and preferably for 10 minutes to 24 hours. hours, b) freeze quickly and lyophilize the fibrin clot, and c) adjust to a residual moisture of at least 3%, preferably 3 to 35%, and in particular 10 to 20% d) optionally pack the fibrin sponge in a suitable container, and in particular in an air-tight and / or water-tight container.

18. A method according to claim 17, characterized in that other plasma proteins and / or other active substances and / or additives are contained in the fibrinogen solution and / or in the thrombin solution.

19. A method according to claim 17 or 18, characterized in that the fibrinogen solution or a mixture of these with the thrombin solution is foamed before the coagulation is established.

20. A method according to any of claims 17 to 19, characterized in that the fibrin clot is washed with water or an aqueous solution and / or incubated with another solution containing thrombin and / or other plasma proteins and / or other active substances.

21. A method according to any of claims 17 to 20, characterized in that the other plasma proteins are selected from the group consisting of fibronectin, inhibitor of fibrinolysis, plasminogen and albumin, the other active substances are selected from the group which consists of synthetic or non-plasma inhibitor of fibrinolysis, respectively, and of antibiotics and growth hormones.

22. A method according to any of claims 17 to 20, characterized in that the additives or auxiliary agents are selected from the group consisting of sugar, sugar alcohol, polyol, amino acids, tensile agent, plasticizer and wetting agent, and in particular of glycerol.

23. A method according to any of claims 17 to 22, characterized in that the washing and / or the incubation with a solution containing substances according to claim 20 is carried out after lyophilizing and subsequently freezing rapidly and freeze-drying again.

24. A method according to any of claims 17 to 23, characterized in that the product obtained in lyophilization is cut.

25. A method according to any of claims 17 to 24, characterized in that the preparation is made from a sterile raw material maintaining the sterile conditions until the final product.

26. The use of a fibrin sponge according to claim 1 for hemostasis, tissue adhesion, cell culture assistant and / or wound healing.

27. A set of implements comprising a fibrin sponge according to claim 1 characterized in that it is in sterile form for use as a biological assistant for cell cultures. FIBRINA SPONGE Summary The invention relates to a fibrin sponge comprising a residual moisture content of at least 3%, preferably from 3 to 35%, in particular from 10 to 20%, and preferably containing an activator or proactivator of the blood coagulation, a method for preparing this fibrin sponge, as well as a set of implements for adhering the wounds which comprises the fibrin sponge and a component comprising a blood coagulation factor. The sponge according to the present invention is suitable for hemostasis, adhesion of tissues and aid in wound healing.