JPH11239609A - Adhesive for blood platelet-containing living body tissue - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an adhesive for a living body tissue having higher wound cure promoting action and a higher tissue conjugating effect. SOLUTION: This adhesive for a blood platelet-containing living body tissue contains a blood platelet in an adhesive for a living body tissue. In this adhesive, the blood platelet is desirably one's own blood platelet. In this adhesive, the adhesive for a living body tissue may also be constituted of a biodegradable material, and the adhesive for a living body tissue may be a fibrin adhesive or a gelatin-based adhesive.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an adhesive for living tissue.

[0002]

2. Description of the Related Art In surgery and the like, closure and joining of wounds such as skin, organs and blood vessels is one of the most basic procedures, and sewing with a thread is now common. However, it is conceivable that the wound is closed and joined quickly using a biocompatible adhesive that can withstand appropriate tension without using sutures. Used clinically. Conventionally, clinically used or developed adhesives for living tissues include cyanoacrylate-based adhesives, polyurethane-based adhesives, gelatin-based adhesives, and fibrin adhesives.

[0003] The cyanoacrylate adhesive is widely used industrially as an instant adhesive, but the adhesive is applied by a direct method of directly applying and bonding between tissues to be joined. There is an indirect method in which a piece of cloth is put on a tissue piece after the tissues are brought into close contact with each other, and the film is adhered. However, cyanoacrylate-based adhesives are excellent in that the bonding speed is high and the bonding strength is large, but the cured product is too hard, so that the hemostasis of the wound surface is likely to be incomplete, and hydrolysis of the cured polymer is generated. Formaldehyde is toxic and inhibits tissue repair (Hajime Kimura: Journal of the Adhesion Society of Japan, 22 (2), p. 77 (1)
986)). For this reason, it cannot be used for a part that directly touches the central nervous system or blood vessels, and there is a limit to the applicable place. In addition, there is also a problem that since the decomposition rate is low, it becomes a foreign substance and hinders wound healing.

As the polyurethane-based adhesive, there is a reactive urethane prepolymer adhesive developed as an elastic adhesive for maintaining the flexibility of a joint after curing.
The reactive urethane prepolymer adhesive has a problem that aromatic diisocyanate, which is a raw material for its synthesis, has biotoxicity. For this reason, attempts have been made to use non-toxic fluorinated aliphatic diisocyanates despite a slight delay in the curing rate (Takehisa Matsuda: Biomaterials, 7).
(1), p. 32 (1989)).

[0005] Gelatin used in the above-mentioned gelatin-based adhesive is a protein obtained by hydrolyzing collagen and has an adhesive action even when used alone. However, when formaldehyde or glutaraldehyde is added as a polymerizing agent, it rapidly gels and contracts. I do. Further, when resorcinol is added to gelatin, the adhesive layer becomes hydrophobic and the adhesive strength is increased. This gelatin-based adhesive uses formaldehyde, which is harmful to tissues, as a polymerizing agent. Therefore, when the adhesive is applied directly to the surgical field, there is a problem in that the formaldehyde comes into contact with the tissues and exhibits biotoxicity.

The above-mentioned fibrin adhesive is based on human fibrinogen at a high concentration, and is composed of thrombin which is an enzyme for converting fibrinogen into fibrin, blood coagulation factor XIII which is a factor for stabilizing fibrin, and which are necessary for these reactions. Aprotinin, which is a fibrinolytic activity inhibitor for protecting calcium ions and the generated fibrin clots from degradation by fibrinolytic enzymes such as plasmin and trypsin to maintain adhesion, is incorporated. At the time of use, the fibrin glue mixes these substances to form a fibrin clot at the point of use.
This fibrin adhesive has many advantages, such as being less likely to be a foreign substance, having no toxicity, not impairing the wound healing process on the wound surface, and working rather quickly, however, because of its flexibility, it has a weak fixing force and adhesive force. There's a problem. In addition, there is a risk of infection because fibrin glue can serve as a culture medium for bacteria as a side effect. In addition, there is also a problem that the operation time is long because the preparation time is long.

As described above, adhesives for living tissues such as cyanoacrylate adhesives, polyurethane adhesives, and gelatin adhesives have a problem of being biotoxic to living tissues and hindering wound healing of the tissues. There is. In addition, fibrin glue does not inhibit the wound healing process on the wound surface, but rather works in an accelerated manner, but has a problem in that the fixation and adhesion are weak. At present, there is no adhesive for living tissue that has an action to promote wound healing more positively, and an adhesive for living tissue that promotes wound healing and has higher adhesive strength is desired.

In recent years, it has become clear that various cytokines and cell growth factors play important roles also in the wound healing process. Recently, administration of fibroblast growth factor or the like has been clinically applied to promote wound healing.
However, since cytokines and cell growth factors form a complex network, administration of a single cytokine or cell growth factor to a living body in a high volume results in increased side effects. In addition, leukocytes, platelets, bone marrow cells, etc. are major cytokine-producing cells, but even if these cells are locally transplanted, they do not stay at the target affected area and spread, resulting in reduced effects and increased side effects. Can cause Administration of collagen or the like as an extracellular matrix does not exert a sufficient healing promoting action when the wound healing power of a living body such as an elderly person is deteriorated. Even with artificial skin, it is effective after leukocyte infiltration and granulation, which are early stages of wound healing, but does not exert a strong effect on promoting healing from the early stage of wounding.

As described above, in wound healing, there is a demand for an adhesive for living tissue which can further promote wound healing. Japanese Patent Publication No. 7-505076 proposes a bioadhesive rich in platelet factor as having such a wound healing action. However, in this method, platelet-rich plasma is used as a starting material plasma, and a cold precipitate that is a component of the bioadhesive is recovered to prepare a bioadhesive. There is a problem that these may be lost or deactivated.

[0010]

SUMMARY OF THE INVENTION The present invention has been made to solve the above problems, and an object of the present invention is to provide a living tissue adhesive having a higher wound healing promoting action and a higher tissue bonding effect. Is to do.

[0011]

The platelets include platelet-derived growth factor (PDGF) and vascular endothelial cell growth factor (VE).
It is known that various cell growth factors such as GF) and epidermal growth factor (EGF) are contained, and it is known that platelets play an important role in these wound healing processes in the healing process. ing. The present inventor has found that by containing platelets in a conventional adhesive for living tissue, it is possible to obtain a living tissue adhesive having a higher wound healing promoting action and a higher tissue bonding effect, and Completed the invention.

That is, the invention according to claim 1 is a platelet-containing biological tissue adhesive characterized in that the adhesive for biological tissue contains platelets.

According to a second aspect of the present invention, there is provided the platelet-containing biological tissue adhesive according to the first aspect, wherein the platelets are autologous platelets.

[0014] The invention according to claim 3 is the adhesive for platelet-containing biological tissue according to claim 1 or 2, wherein the adhesive for biological tissue is composed of a biodegradable material.

According to a fourth aspect of the present invention, there is provided the platelet-containing biological tissue adhesive according to the third aspect, wherein the biological tissue adhesive is a fibrin adhesive or a gelatin-based adhesive.

Hereinafter, the present invention will be described in detail. The platelet-containing adhesive for living tissue of the present invention contains platelets in the adhesive for living tissue. The above-mentioned content indicates, for example, that platelets are embedded or adhered to the biological tissue adhesive.

As the adhesive for living tissue used in the present invention, any of those conventionally used as an adhesive for living tissue can be used, and there is no particular limitation. The described fibrin glue,
Gelatin-based adhesives and cyanoacrylate-based adhesives are exemplified. In particular, when the adhesive base material is a biodegradable material, it is more useful and preferable for tissue bonding because there is no need to remove the adhesive by peeling off even after tissue bonding has progressed. Examples of such an adhesive substrate include fibrin, gelatin (collagen), chitin, chitosan and the like. When the adhesive base material is fibrin, collagen, chitin, chitosan or the like, the effect of promoting healing is exhibited by itself, so that the promotion of healing is synergistically exhibited.

The platelets used in the present invention are not particularly limited, such as those derived from humans and animals. Platelets can be collected and separated by known methods. For example, a method of separating from blood collected with sodium citrate using a centrifugal separation method, a centrifugal separation method in a density gradient solution, a gel filtration method, or the like may be used. Moreover, what was cultured may be sufficient. As the platelets, in particular, autologous platelets, that is, platelets of the subject to whom the adhesive for platelet-containing living tissue of the present invention is applied are most preferable. If they are your own platelets,
This is because there is no host rejection reaction to platelets after transplantation, and wound healing is promoted safely and tissue conjugation is favorably promoted.

The amount of platelets to be contained in the platelet-containing biological tissue adhesive of the present invention is not particularly limited and may be appropriately selected in a wide range.
Preferably, it is in the range of 10 ³ to 10 ¹² .

The adhesive for a platelet-containing living tissue of the present invention may contain cytokines, cell growth factors, stimulants and the like, if necessary. When a stimulant is included, platelets are easily activated.

In the platelet-containing biological tissue adhesive of the present invention, platelets may be used, if necessary, in combination with cells such as fibroblasts and epidermal cells used in known artificial skin. Good.

[0022]

In the adhesive for platelet-containing living tissue of the present invention, various healing promoting factors such as cell growth factors are released and produced from platelets at the junction, thereby promoting healing. In addition, since platelets are contained in the adhesive for living tissue, the local retention property is excellent.

[0023]

Example 1 Heart blood was collected from 7-week-old Wistar rats using sodium citrate as an anticoagulant, and platelets were collected from the blood by centrifugation. The platelets were suspended in a calcium / magnesium-free phosphate buffer at a concentration of 5 × 10 ⁷ per ml to prepare a platelet suspension. The above-mentioned platelet suspension was applied to fibrin glue (Beriplast P, manufactured by Hoechst) consisting of solution A in which fibrinogen powder and blood coagulation factor XIII were dissolved in an aprotinin solution, and solution B in which thrombin powder was dissolved in a calcium chloride solution. The platelet-containing adhesive for living tissue was obtained by adding the liquid by the following method. That is, the platelet suspension was added to and mixed with the liquid B of the fibrin glue so that the ratio of the platelet suspension was 10 μl per 1 ml of the liquid B. Then, the obtained platelet-added liquid B and the liquid A were mixed and mixed. The adhesive for platelet-containing living tissue of the present invention was used. In the performance evaluation described later, 0.1 g of the platelet-containing adhesive for living tissue was applied to the cut wound immediately after preparation, and the adhesive was applied to the cut wound.

Example 2 A platelet suspension prepared in the same manner as in Example 1 was added to a gelatin / resorcinol mixture in gelatin paste (trade name: GRF glue, manufactured by EHS (France)).・
10 μl of platelet suspension per 1 ml of resorcinol mixture
1 and mixed. In the performance evaluation described below, 0.1 g of this solution was applied to a cut wound, and several drops of a formaldehyde-added solution were applied to the cut wound.

Example 3 A platelet suspension prepared in the same manner as in Example 1 was applied to a cyanoacrylate-based adhesive for living tissue (trade name: Ar, manufactured by Sankyo).
On Alpha A), the platelet suspension was mixed at a ratio of 10 μl with respect to 1 ml of the cyanoacrylate-based adhesive for living tissue to obtain an adhesive for platelet-containing living tissue of the present invention. In the performance evaluation described below, 0.1 g of the platelet-containing adhesive for living tissue was applied to the cut wound immediately after preparation, and the adhesive was applied to the cut wound.

Comparative Example 1 Liquid A and liquid B of fibrin glue (trade name: Beriplast P, manufactured by Hoechst) were mixed to prepare an adhesive for living tissue.
In the performance evaluation described below, 0.1 g of the adhesive for living tissue was applied to the cut wound immediately after preparation, and the adhesive was applied to the cut wound.

Comparative Example 2 Gelatin glue (trade name: GRF glue, manufactured by EHS (France)) was used as an adhesive for living tissue. In the performance evaluation described later, 0.1 g of the adhesive liquid for living tissue was applied to a cut, and a few drops of a formaldehyde-added solution were applied to the cut to administer the cut.

Comparative Example 3 As a biological tissue adhesive, a cyanoacrylate-based biological tissue adhesive (manufactured by Sankyo Co., Ltd., trade name: Aron Alpha)
A) was used. In the performance evaluation described below, 0.1 g of the adhesive for living tissue was applied to the cut wound.

Performance Evaluation The performance of the platelet-containing adhesive for living tissue of Examples 1 to 3 and the adhesive for living tissue of Comparative Examples 1 to 3 were evaluated by the following methods.

The back of a 10-week-old Wistar rat was shaved and a 2-cm long cut was made on the back skin with a scalpel in a direction perpendicular to the midline of the back. After suturing the cut portion at three locations at equal intervals, the adhesives of Examples 1 to 3 and Comparative Examples 1 to 3 were administered as described above. Three days after the treatment, the rats were sacrificed.
A strip of skin with a width of cm was cut out. The tensile strength (g / cm) of the wound was measured using a rheometer. For each Example and Comparative Example, five rats were used, and the tensile strength (g / cm) was an average of the values obtained for these rats. The results are shown in Table 1.

[0031]

[Table 1]

From Table 1, it can be seen that the group to which the adhesive for platelet-containing living tissue of Example 1 was administered exhibited a higher tensile strength than the group to which the adhesive for living tissue of Comparative Example 1 was administered. The same can be said for the relationship between Example 2 and Comparative Example 2 and between Example 3 and Comparative Example 3.

[0033]

According to the first aspect of the present invention,
As described above, according to the present invention, a biological tissue adhesive having a higher wound healing promoting action and a higher tissue bonding effect is provided. The configuration of the invention described in claim 2 of the present invention is as described above, and according to the present invention,
The present invention provides an adhesive for living tissue, which has no host rejection to platelets after transplantation, has a higher wound healing promoting action, and has a higher tissue bonding effect.

The structure of the invention according to claim 3 of the present invention is as described above. According to the present invention, it is not necessary to remove the adhesive by peeling off even after the tissue joining has progressed.
The platelet-containing biological tissue adhesive according to claim 1 or 2, which has a higher tissue bonding effect.

According to a fourth aspect of the present invention, the adhesive for a living tissue is the adhesive for a platelet-containing living tissue according to the third aspect, wherein the adhesive for a living tissue is a fibrin adhesive or a gelatin-based adhesive. According to this, an adhesive for platelet-containing living tissue having a higher tissue bonding effect is provided.

Further, in view of the high wound healing promoting effect, it is presumed that the inventions according to claims 1 to 4 alleviate the toxicity seen in the conventional adhesive for living tissue.

Claims (4)

[Claims] 1. A platelet-containing adhesive for living tissue, wherein the adhesive for living tissue contains platelets. 2. The adhesive according to claim 1, wherein the platelets are autologous platelets. 3. The adhesive for a platelet-containing biological tissue according to claim 1, wherein the adhesive for a biological tissue is made of a biodegradable material. 4. The adhesive for a platelet-containing biological tissue according to claim 3, wherein the biological tissue adhesive is a fibrin adhesive or a gelatin-based adhesive.