JPS60204725A - Manufacture of tissue adhesive - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for preparing tissue adhesives based on human or animal proteins containing fibrinogen and factor XI, optionally containing plasmin inhibitors, antibiotics and cytostatic agents. It is something.

The use of coagulants for hemostasis and wound dressing is known. The first suggestion in this regard was that fibrin tampons and fibrin platelets were used. Methods for preparing fibrinogen and factor X tissue adhesives are disclosed in U.S. Pat.
No. 598 and No. 4,377,572.

In addition, collagen 1 has a porous structure and is used to cover wounds.
It is also known to use tissue adhesives based on this, in which non-woven fabrics made of collagen fibers are applied to the wound.

For fixation of the collagen nonwoven, a fibrinogen-thrombin mixture is applied to the wound area or to the inner surface of the collagen nonwoven and the nonwoven is pressed against the wound. but,
This method has the disadvantage that the fibrinogen is Ml'd very rapidly by thrombin and is therefore unable to penetrate into the collagen. Furthermore, when a fibrinogen/thrombin mixture is used, it is extremely difficult to perform the fixation operation at the optimum time.

In addition, wound treatment materials (West German Publication No. 291482
No. 2), this material has a fibrous structure (eg of collagen or synthetic polymers) in which factor XI is fixed. This material is not suitable as a wound adhesive because it can function only with the cooperation of coagulation activators present in the wound area. This is because these factors exist in trace amounts and are not sufficient.

The present invention seeks to overcome the above-mentioned drawbacks and difficulties by providing a tissue adhesive which can be used without restriction and which can be used for hemostasis, wound dressing and tissue bonding by providing the method as described above. This makes it possible to produce one which does not require any special preparation of the wound area for its application and which ensures tight wound coverage or bonding.

According to this invention 1, the above object is a tissue compatible material,
In particular, an aqueous mixture containing collagen, gelatin or polysaccharides, as well as fibrinogen and factor This is accomplished by lyophilizing the matrix.

According to a preferred embodiment 1, the mixture and its shaping into a planar structure are carried out by impregnating a porous collagen nonwoven fabric with an aqueous solution containing fibril and factor X.

It is appropriate to use a prefabricated porous collagen nonwoven fabric.

As the porous collagen nonwoven fabric, it is preferable to use a nonwoven fabric obtained by freeze-drying a layer of collagen aqueous solution.

Advantageous Embodiment 1 Accordingly, in order to make a multilayer planar material,
The mixture is shaped into a planar structure and freeze-dried at least once.

Hereinafter, this invention will be explained in detail with reference to Examples.

Example 1 Frozen fresh human plasma is heated to 2°C, and a frozen precipitate containing factor X■ and fibrinogen is obtained by centrifugation.

Cold-soluble proteins are removed from the frozen precipitate by extraction with a buffer.

The remaining protein was purified by aprotinin (250 OKIU), heparif (2QIU) and amikacin sulfate (2000 OKIU).
100ml of citric acid/glycine buffer containing
! Dissolve at 37°C and sterile filter. Solution F contains at least 1000 units of Factor XI and at least 75 oomg of fibrinogen.

The reprinted collagen nonwoven fabric is divided into flat pieces of size approximately 70 cr/l and each flat piece is treated with a solution containing factor X11 and fibrinogen under sterile conditions.

Thereafter, the flat pieces are frozen, and the swollen pieces are freeze-dried and packaged aseptically.

Instead of using pre-fabricated collagen slabs or non-woven materials, they can be made in situ by freeze-drying them in petre dishes, again with a factor X and fibrinogen containing solution. The impregnation step is carried out in the same manner as above.

Example 2 Frozen fresh human plasma is heated in a 2°C oven, and a frozen precipitate containing factor x■ and fibrinogen is obtained by centrifugation.

Cold-soluble proteins are removed from the frozen precipitate by extraction with a buffer.

The remaining proteins were treated with aprotinin (2509 KIU), heparin (201 U) and amikacin sulfate (2000 KIU).
100 mf of citric acid/glycine buffer containing fng)!
and sterile filter. The p-fluid contains at least 1000 units of factor X and at least 750 units of fibrinogen.
Contains 0 ■.

Add 10% of a sterile 1% collagen solution to this mixture under aseptic conditions.
Add 0m+'. The solution is then divided into 20 rne portions and placed in Petri dishes to form a layer with a thickness of 2 to 5 mm. The liquid in the Petri dish is then frozen and lyophilized to create a porous planar structure. These are placed in a Petri dish as the final container. In use, it is removed from the Petri dish and applied to the wound area.

Example 3 In the same manner as in Example 1, a cryoprecipitate was obtained from frozen human fresh plasma, and cold-soluble proteins were separated from this to obtain a protein-based material containing factor X and fibrinogen, and a C0-inactivating agent ( 35PU), heparin (20IU
) and glutamine sulfate (2500 μl) and sterile filter.

In addition to the lysis buffer, no more than twice the natural content of Part XII.
Factor I may also be added, which is preferred if antibiotics are included.

To this mixture, under aseptic conditions, add an equal amount of 3.5% reprint gelatin solution. The mixture is then molded into a flat shape in a Petri dish, and the swollen pieces are lyophilized and aseptically packaged as described in Example 1.

Example 4 A protein-based material containing factor x■ and fibrinogen was produced in the same manner as in Examples 1 and 2.
Dissolution with glycine buffer and sterile filtration are performed in the same manner.

The same amount of 6% hydroxyethylene starch is mixed with the p-liquid, the mixture is divided into Petri dishes, formed into flat structures, frozen, lyophilized, and aseptically packaged.

Patent applicant: Immuno Akchengesellschaft Für Hemisumedeitzinitzie Producte

Claims (7)

[Claims] (1) Method for producing tissue adhesives based on human or animal proteins I
In this case, fibritoma (IJ)
A method comprising the step of freeze-drying the impregnated flat material having lux. (2) The method according to claim 1, wherein the tissue-compatible planar material consists of a nonwoven fabric or sheet based on a material selected from collagen, gelatin or polysaccharides. (3) The solution to be impregnated into the tissue-compatible flat material is
2. The method of claim 1, comprising at least one of a plasmin inhibitor, an antibiotic, or a cytostatic agent. (4) In a method for producing a tissue adhesive based on human or animal protein, a step of freeze-drying an aqueous collagen solution to produce a flat collagen material so as to obtain a porous collagen flat material; impregnating with a solution consisting of fibrinogen and factor X III;
and lyophilizing said impregnated collagen planar material having a coherent matrix of said collagen planar material. (5) A process for producing tissue adhesives based on human or animal proteins, from freeze-drying an aqueous solution of fibrinogen, collagen and factor Xl such that a flat material containing fibrinogen, factor X and collagen is formed. How to become. (6) The method according to claim 5, wherein the aqueous solution contains at least one of a plasmin inhibitor, an antibiotic, or a cell growth inhibitor. (7) Furthermore, the lyophilization of fibrinogen, collagen and factor The material consists of lyophilizing an additional amount of the aqueous solution to produce a material. The method according to claim 5.