JPS60256460A - Composition for filling bone defficient part and gap part containing fibrin and calcium phosphate - 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 [Industrial Field of Application] The present invention relates to a filling composition for filling bone defects and voids and generating new bone at the filling sites.

[Prior Art] In the field of surgery or orthopedics, bone defects or voids occur due to bone fractures or removal of bone tumors, and in the field of dentistry, loss of jaw bone due to alveolar pyorrhea occurs. Often encountered when local prosthesis is required. Conventionally, in many cases, a method has been used in which the patient's own iliac bone or the like is removed and filled into the bone defect site to fill the bone tissue defect or void and to hasten the recovery and healing of the tissue. however.

This method requires removal of normal bone tissue other than the damaged area, which causes great pain to the patient and requires a great deal of labor during the surgery.

Furthermore, if the bone defect is large, it is not always possible to collect enough autologous bone to fill it, and some kind of substitute must be used to make up for the shortage. As a substitute for this, for example, allogeneic bone and xenogeneic bone are available, but there are still problems such as rejection reactions with the implanted living tissue, and the postoperative course is not always good. It has not yet reached the practical stage.

Various metal alloys and organic substances have been used as hard tissue substitutes for living organisms, but they are said to deteriorate due to dissolution in the living environment, to be toxic to living organisms, and to be accompanied by foreign body reactions. Ceramic materials, which have excellent compatibility with living organisms and do not have the above-mentioned drawbacks, are now being used. Among these ceramic materials, artificial bones and artificial teeth made of sintered bodies or single crystals of alumina, carbon, tricalcium phosphate, or hydroxyapatite, which have excellent biocompatibility, are being developed and are attracting attention.

Attempts have been made to fill bone defects and voids with these sintered bodies or single crystals, but the shapes of bone defects that require actual treatment are not constant and have complex shapes. It is difficult to process these sintered bodies or single crystals to fit the shape, and even if these sintered bodies or single crystals are filled, they are significantly harder than the surrounding bone tissue, making it difficult to fill them. The stimulation causes bone resorption around the material, causing problems such as loosening, and it has not yet reached the stage of practical use.

On the other hand, the sintered body is made into a porous body by mechanical methods or by adding flammable fiber to powder during molding and sintering the molded lumps, and this is used as a filling material for bone defects and voids. However, the porosity of the porous body created by these methods is 1. For example, when attempting to mechanically process a ceramic sintered body to make it porous, the processability is Moreover, it is impossible to introduce a large amount of pores because the material is brittle and easily damaged.

Furthermore, even when a porous body is obtained by adding flammable fiber to ceramic raw material powder and sintering it after molding, it is difficult to add a large amount of flammable fiber to the powder and mold it. I can't get it.

For this reason, there are drawbacks such as insufficient infiltration of biological osteogenic components necessary for the generation of new bone into the filler, and a long period of time required for the filler and bone tissue to become integrated.

[Objectives of the Invention] Therefore, one object of the present invention is to form bone tissue in a particularly short period of time with excellent biocompatibility without any foreign body reaction, to allow the filling material itself to be absorbed and replaced by the living body, and to provide a material that is particularly suitable for the filling composition. An object of the present invention is to provide a composition for filling bone defects and voids that can generate new bone up to the center of an object.

Another object of the present invention is to provide a composition for filling bone defects and voids that promotes the function of ashes in the filling area and particularly quickly repairs and restores the structure and function of bone tissue defects. .

Still another object of the present invention is to provide a composition for filling bone defects and voids in which new bone is generated particularly quickly.

Yet another object of the present invention is to provide a composition for filling bone defects and voids that is easy to mold into a shape that matches the shape of the site to be filled.

The above and other objects of the present invention will become more apparent from the following description.

[Configuration of the Invention] According to the present invention, there is provided a composition for filling bone defects, voids, and gaps, which is characterized by containing fibrin and a calcium phosphate compound.

[Description of the invention] The present invention will be explained in more detail below.

The present inventors first focused on utilizing the osteogenic ability of calcium phosphate compounds, since when a calcium phosphate compound is filled into bone defects and voids, new bone is generated in the areas. Calcium phosphate compounds that can be used in the present invention include tricalcium phosphate, hydroxyapatite, tetracalcium phosphate, oxyapatite, birocalcium phosphate, fluoroapatite, hydroxyapatite in which a portion of the hydroxyl groups are replaced with fluorine ions, and compounds thereof. Among these, one selected from those with a high rate of new bone formation, ie, tricalcium phosphate, hydroxyapatite, fluoroapatite, or tetracalcium phosphate.
It is preferable to use a species or a mixture of two or more species.

Among them, hydroxyapatite can be said to be the most preferable because it has the fastest rate of new bone formation. Among hydroxyapatites, temperatures of 500°C or higher, particularly preferably 70°C
Hydroxyapatite obtained by heat treatment at 0° C. or higher is particularly preferable because nova bone formation is quick.

The upper limit temperature of the heat treatment is not particularly limited, but since bitroxyapatite starts to decompose, it should be lower than the decomposition temperature. In addition, the calcium phosphate compound that can be used in the present invention may be artificially synthesized by a known manufacturing method such as a wet method, a dry method, or a hydrothermal method, or it may be a natural product obtained from bones etc. It's okay.

On the other hand, fibrin used in the present invention is prepared using fibrinogen obtained from human or animal plasma as a raw material. From the viewpoint of biocompatibility, when the target is a human, it is preferable to use fibrinogen obtained from human plasma as the raw material, and when the target is an animal, it is preferable to use fibrinogen obtained from the plasma of that animal as the raw material. As such fibrinogen, medical dry fibrinogen manufactured in accordance with the Biological Product Standards (1979, pp. 201-203) supervised by the Pharmaceutical Affairs Bureau of the Ministry of Health and Welfare can be used, and this commercially available product is available under the trade name "Fibrinogen". Nogen-Midori J [[Midori Jujiko powder is available. This is made by adding coagulating proteins to dry fibrinogen, sodium citrate as a stabilizer, and monosaccharides such as glucose, fructose, and mannitol. Before use, it is dissolved in distilled water for injection or a low salt concentration buffer with a pH of 6 to 7.The low salt concentration buffer includes (1,01 to 0.0
A 3 molar citrate buffer is preferred. Melting temperature is 32
It is preferable to maintain the inside of the bottle containing fibrinogen under reduced pressure during dissolution. Under these dissolution conditions, dry fibrinogen powder is approximately 2
Soluble within the range of ~IOW/V%. Further, fibrinogen may be used in powder form. As detailed below, fibrinogen is solidified into fibrin by adding thrombin and calcium chloride as anabolic agents when preparing the composition for filling bone defects and voids of the present invention.

In preparing the composition for filling bone defects and voids of the present invention, the above-mentioned calcium phosphate compound powder or granules (hereinafter collectively referred to as powder or granules) or granules made from the powder may be used. When using a porous body of a calcium phosphate compound, for example, in one method of wet method, since it is obtained as a precipitate, it is separated from the solution by a method such as filtration or centrifugation. By drying and then pulverizing, a powder of calcium phosphate compound is obtained. In addition, in the case of a dry method and a hydrothermal method, the material is pulverized as necessary. When using a natural calcium phosphate compound such as bone, a similar pulverization process is performed. In any case, in the case of granular materials, the maximum size depends on the size of the defect, but
It is preferred to use up to II11 size in forming the filling composition of the present invention.

The calcium phosphate compound powder thus obtained can be used as it is as the filling composition of the present invention, but it is also possible to improve the crystallinity of the particles and improve biocompatibility.
In addition, in order to prevent infection by bacteria and rejection by organic matter, heat sterilization should be carried out sufficiently. In the case of calcium phosphate compounds obtained by wet synthesis, the temperature should be 500°C or higher, preferably 700°C or higher. After firing at a high temperature, it is preferable to carry out appropriate pulverization treatment if necessary to form a powder.

The powder and granules obtained by each of the above synthesis methods and the powder and granules obtained by further calcination are further added with a liquid, such as water or physiological saline, using a transfer granulator or the like. It can also be made into granules.

The particle size of the granules is not particularly limited, but is preferably about 0.1 to 5 m in order to manufacture the filling composition of the present invention and to prevent the granules from flowing out to other parts of the body. .

In either case, a mixture of a fibrinogen solution or powder and the powder or granules of the calcium phosphate compound described above is prepared. Separately, a mixed solution is prepared in which thrombin and calcium chloride are dissolved in physiological saline or citrate buffer as an anabolic agent for solidifying fibrinogen into fibrin. Usually thrombin 1-50
ONIH units/l1lllQ and calcium chloride 10~
It is preferable to include 100 mmol/mQ; if it is less than the lower limit, the assimilation of fibrinogen may be insufficient; on the other hand, if it is added above the upper limit, the assimilation effect will not change much in many cases. A protease inhibitor may be added to the mixed solution to prevent fibrin from being dissolved depending on the site to be filled with the composition for filling bone defects and voids of the present invention. As the protease inhibitor, a plasmin limiting agent can be preferably used, and examples thereof include aprotinin, epsilon aminocaproic acid, planexamic acid, soybean trypsin limiting agent, and the like. The amount of protease inhibitor added is usually 100 to 5000 KIE units/mQ.
That's about it.

Next, when the mixture of fibrinogen and calcium phosphate compound and the mixed solution are mixed and stirred, 1
Thrombin and calcium chloride insolubilize fibrinogen into fibrin in about 0 to 30 minutes, and the powder or granules of the calcium phosphate compound are dispersed in the colloid buiplin for filling bone defects and voids of the present invention. A sample of composition is obtained.

It takes about 10 to 30 minutes for the assimilation of fibrinogen, thrombin, and calcium chloride, so a mixed solution of thrombin and calcium chloride is added to the fibrinogen solution or powder, and then the powder or granules of the calcium phosphate compound are added. The composition for filling bone defects and voids of the present invention can also be obtained by adding and stirring and mixing.

As mentioned above, the composition for filling bone defects and voids of the present invention can also be prepared using a porous body of a calcium phosphate compound. In this case, for example, the above-mentioned calcium phosphate compound powder is suspended in a liquid such as distilled water to form a slurry, and the slurry of the calcium phosphate compound is applied to a spongy organic porous material having continuous pores in a three-dimensional network structure. A porous body of a calcium phosphate compound can be prepared by impregnating it, drying it, and burning and extinguishing the organic porous body by heating. Next, the calcium phosphate compound is immersed in the above-mentioned fibrinogen solution or powder, and then the mixed solution containing thrombin, calcium chloride, and optionally a protease inhibitor is added dropwise to solidify the fibrinogen to form a calcium phosphate compound porous material. The composition for filling bone defects and voids of the present invention, in which the continuous pores of the fibrin are filled with fibrin, is obtained.As mentioned above, the reaction of fibrinogen with thrombin and calcium chloride takes some time. Adding a mixed solution containing thrombin and calcium chloride to the fibrinogen solution or powder,
The composition for filling bone defects and voids of the present invention can also be obtained by immediately immersing a calcium phosphate compound porous body or dropping it into the porous body.

The content of the calcium phosphate compound in the composition for filling bone defects and voids of the present invention is preferably 5 to 95 vol% in the final product composition, with the remainder containing fibrin. Calcium phosphate compound is 5v0
If the fibrin content is less than 1%, the formation of new bone may be slowed down, while if it exceeds 95vol%, the fibrin content will be low and new blood vessels will be less likely to form.

[Effects] Calcium phosphate compounds have a cremation effect, and when filled into bone defects or voids, new bone is formed. On the other hand, fibrin is a hard protein obtained by the action of fibrinogen and thrombin, and has the function of adhering the wound surface. Due to this adhesion, fibroblasts are generated on the wound surface, which then become fibrocytes and fix the tissue. At the same time, blood vessels are generated within the tissue. Therefore, in the present invention, the calcium phosphate compound is held in a fixed state by the agglutinating action of fibrin, and the ashes action of the calcium phosphate compound is promoted, and angiogenesis is promoted to form new bone and repair of living tissue is accelerated. be able to.

In particular, when the composition for filling bone defects and voids of the present invention is filled, new blood vessels are formed and extended to the center of the composition, and new bone is rapidly formed.

The composition for filling bone defects and voids of the present invention is prepared under aseptic conditions by using physiological saline or a low-temperature concentration buffer solution with a pH of 6 to 7.
For example, store it in a 0.01-0.03M citrate buffer, and cut it into pieces that match the shape of the bone defect or cut into sections of approximately 1 m to 15 marks as needed. can be used to fill bone defects or voids.

[Example code] Next, the present invention will be explained with reference to its embodiments.

Fibrinogen 60 extracted and purified from rabbit blood.
Thrombin 4 NIH units/mQ, aprotinin 3'0OO
1 m of 40 m M calcium chloride solution containing KIE units/-
A composition containing Q (hereinafter abbreviated as fibrin composition) was prepared. Next, the fibrin composition was added to hydroxyapatite powder (calcined at 900°C), tricalcium phosphate powder (calcined at 1150°C), and tetracalcium phosphate powder (135°C), respectively.
0°C firing) were mixed at a volume ratio of 1:1 and solidified to prepare a composition for filling bone defects and voids of the present invention.
It was cut out into a size of 4 x 3 mm and placed in 0.01 M 9 enoic acid buffer.

Next, make a 5X4X3+in+ hole in the rabbit femur,
The progress after 4 weeks of filling with the above composition of the present invention was observed. Separately, only the fibrin composition, only the hydroxyapatite powder, only the tricalcium phosphate powder, and only the tetracalcium phosphate powder were filled into the holes, and the progress after 4 weeks was observed wA. In this case, as a control, only a hole was made and nothing was filled, and the progress was also observed.

As a result, when the composition for filling bone defects and voids of the present invention was filled, new bone was observed to form even in the center. Among them, the combination of fibrin composition and hydroxyapatite produced the most new bone.

On the other hand, when only the fibrin composition was filled or when only a hole was made, the defect was filled with soft tissue, and only a small amount of new bone was observed around the defect.

Hydroxyapatite, dicalcium phosphate.

When only tetracalcium phosphate was used, although new bone formation was observed inside the filling material, almost no new bone was observed in the center of the filling material. Powder (120
0℃ firing) at a volume ratio of 4,0:1,20:1゜1
:1, 1:20, 1:40 in the same manner as in Example 1, mixed and solidified, cut into 5 x 4 x 3 mm, and placed in the same solution as in Example 1 to prepare each sample. A 5 x 4 x 3 hole drilled in the femur of a rabbit was filled with the solution, and the condition was observed 4 weeks later.

As a result, new bone formation was observed up to the center of the hole with the mixtures with volume ratios of 1:20, 1:1, and 20:1, but with the mixtures with volume ratios of 40:1 and 1:40. No new bone was formed in the center of the hole.

Example 3 A fibrin composition was prepared in the same manner as in Example 1 using human-derived fibrinogen, and hydroxyapatite (granules fired at 800°C, particle size 2.0-1, 0 + n
m) fibrin composition: hydroxyapatite = 1
: 10 (volume ratio) and solidify it, then 5X5X
A sample was prepared by cutting into 5++a and placed in the same solution as in Example 1.

A bone tumor occurring in a human femur was excised and filled with the sample, and the progress after filling was observed using X-rays and bone scintigraphy. As a result of X-ray observation, the formation of new bone was observed from one week after the surgery, and the boundary between the filled sample and the living bone became unclear after 3 to 4 weeks. On the other hand, bone scintigrams showed that the tumor area, which was cold before surgery, became hot after surgery, suggesting that active bone regeneration was occurring.

After immersing a porous hydroxyapatite material (calcined at 1200°C, porosity 80%) in a solution containing fibrinogen used in Example 1 and taking it out, a solution containing thrombin and aprotinin used in Example 1 was taken out. A calcium chloride solution was added dropwise to prepare a composition for filling bone defects and voids of the present invention. A sample was prepared by cutting this composition into a size of 5 x 4 x 3 mm, and the sample was placed in 0.01M citrate buffer.

Next, a hole of 5 x 4 x 3 ma was made in the rabbit femur and filled with the above sample. When the progress was observed after 4 weeks, new bone formation was observed up to the center of the filled sample.

Patent Applicant: Midori Juji Co., Ltd. Continued from page 1 @ Inventor: Mikiya Ono @ Inventor: Hiroyasu Takeuchi [Partner] Inventor: Yoshitaka Okubo 1, 38 Nakayama, Hanno City, 39 Ooukuna, Chichichibu City, Saitama Prefecture Patent Agency Commissioner Manabu Shiga 1. Description of the case 1982 Patent Application No. 109693 2 Name of the invention Composition for filling bone defects and voids containing fibrin and calcium phosphate compound 3 Person making the amendment Relationship to the case Patent applicant Midori Juji Co., Ltd. Mitsubishi Mining Cement Co., Ltd. 4, Agent Address: 1-20-5 Toranomon, Minato-ku, Tokyo 105, Japan Subject of amendment - Sai Section 6 of "Detailed Description of the Invention" of the specification, contents of amendment as shown in the attached sheet K′ Thread i δ＼ In the present specification, the following portions are amended as follows.

10 3 For powder and granule 11 for powder and granule 6 Ultraagent Inhibitor 1 9 1417-18 Low temperature concentration buffer Low salt concentration buffer 1:
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Claims (1)

[Claims] A composition for filling bone defects and voids, characterized by containing fibrin and a calcium phosphate compound.