KR101779377B1 - A composition for bone grafting having enhanced gel stability and strength and a preparation method thereof - Google Patents
A composition for bone grafting having enhanced gel stability and strength and a preparation method thereof Download PDFInfo
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- KR101779377B1 KR101779377B1 KR1020150109097A KR20150109097A KR101779377B1 KR 101779377 B1 KR101779377 B1 KR 101779377B1 KR 1020150109097 A KR1020150109097 A KR 1020150109097A KR 20150109097 A KR20150109097 A KR 20150109097A KR 101779377 B1 KR101779377 B1 KR 101779377B1
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- A—HUMAN NECESSITIES
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
- A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
- A61L27/46—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with phosphorus-containing inorganic fillers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
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Abstract
The present invention relates to a bone graft material composition having excellent gel stability and strength, and a method for producing the same, and relates to a bone graft material composition comprising a gel-forming powder, a calcium phosphate compound or a demineralized bone matrix and an osteogenic protein, To a kit comprising the composition.
The bone graft composition according to the present invention is capable of transplanting into a bone defect site quickly and conveniently by forming a gel within a few minutes and is not spread by blood even when applied to a human body. Therefore, the bone graft material composition has stability to human body, And thus can be customized for unformed bone defects.
Description
The present invention relates to a bone graft material composition having excellent gel stability and strength, and a method for producing the same, and relates to a bone graft material composition comprising a gel-forming powder, a calcium phosphate compound or a demineralized bone matrix and an osteogenic protein, To a kit comprising the composition.
Bone grafting is performed when the bone is destroyed and damaged by trauma, tumor or deformity, the bone is subsequently reinforced, or bone marrow is promoted by promoting bone regeneration. The most common methods of bone grafting are autografting, grafting and grafting of bone from other parts of the body, allograft implanting and transplanting bone of another person, And xenografts for transplantation.
However, the autotransplantation method has a disadvantage in that a secondary operation is required and a new surgical site is created, thereby increasing the risk of infection and blood loss, and making it difficult to obtain a sufficient amount. Because allografts use bone from human carcasses, there is a high probability of disease transmission, immunological rejection, and high cost. Xenotransplantation methods also present a risk of disease transmission and immune reactions.
In order to overcome these problems, there is a need for a synthetic bone graft material that has no possibility of transmission to diseases, is easy to purchase, is cheap, and has a performance capable of replacing existing graft materials.
Accordingly, although synthetic bone graft materials for bone regeneration have been marketed and developed as various biomaterials according to the characteristics of raw materials and purpose of use, currently available bone graft materials are reabsorbed in an excessively rapid time after being implanted in the human body, It takes a long time to absorb the bone cells to provide a suitable environment for adhesion, growth and differentiation of damaged bone cells. It has been designed and developed with biocompatible materials, and various kinds (Korean Patent Registration No. 10-1041784 and Korean Patent No. 10-1398406).
Under these circumstances, the present inventors prepared a bone graft material composition containing a certain amount of a calcium phosphate compound or a demineralized bone matrix, a gel-forming powder and an osteogenic protein, said composition being manufactured in a short time, And strength, and thus has biocompatibility and physical properties suitable for implantation in a bone defect site, thereby completing the present invention.
It is an object of the present invention to provide a bone graft material composition excellent in gel stability and strength.
Another object of the present invention is to provide a method for producing the bone graft material composition.
It is still another object of the present invention to provide a bone graft kit comprising the bone graft material composition and the injection tool.
In one aspect of the present invention, the present invention provides a bone graft material composition comprising:
(A) a gel-forming powder comprising at least one selected from the group consisting of hydroxyethylcellulose, croscarmellose sodium and sodium carboxy methyl starch;
(B) Hydroxyapatite, Carbonated apatite, Tricalcium Phosphate, Calcium Hydrogen Phosphate, Monocalcium phosphate, Dicalcium phosphate Calcium phosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, A calcium phosphate compound, or a demineralized bone matrix; And
BMP-9, BMP-10, BMP-11, BMP-7, BMP-8, , BMP-13, BMP-14, BMP-15, BMP-16, BMP-17 and BMP-18.
The term "bone graft composition" of the present invention means a composition used as a material to be implanted and filled into a bone defect portion of a necessary individual, that is, a bone defect portion supplement. Specifically, in the present invention, the bone graft material composition refers to a synthetic bone graft materials composition comprising a gel-forming powder, a calcium phosphate compound or a demineralized bone matrix, and an osteogenic protein.
The term "gel-forming powder" of the present invention refers to a powder-like substance that can be mixed to form a formulation such as a dough-like form. A gel is formed within a few minutes to facilitate transplantation. As shown in Fig.
For the purpose of the present invention, the gel-forming powder may contain one or more selected from the group consisting of hydroxyethylcellulose, croscarmellose sodium, and sodium carboxy methyl starch. And may further include, but is not limited to, hydroxypropyl methylcellulose (HPMC), Gellan Gum, or a combination thereof.
Also, the gel-forming powder may contain 80 to 100 parts by weight of hydroxyethylcellulose; 0.01 to 15 parts by weight of croscarmellose sodium; And 0.01 to 15 parts by weight of sodium carboxymethylstarch, and more specifically, 85 to 95 parts by weight of hydroxyethylcellulose; 1 to 10 parts by weight of croscarmellose sodium; And 1 to 10 parts by weight of sodium carboxymethyl starch. The gel-forming powders containing hydroxyethyl cellulose, croscarmellose sodium and sodium carboxymethyl starch in parts by weight in the above-mentioned range have a long gel-forming time and a low maximum load or gel strength.
Since the bone graft material is intended to be applied to the bone loss site, there is an advantage that the procedure time is shortened if the gel formation time is saved, and the gel strength is high enough to withstand the physical action of the human body. In one experimental example of the present invention, for example, the gel-forming powder contained 85 to 90 parts by weight of hydroxyethyl cellulose; 5 to 10 parts by weight of croscarmellose sodium; And 5 parts by weight of sodium carboxymethyl starch, it was confirmed that the gel was formed in a short time and that the maximum load and gel strength were suitable as a bone graft applied to the human body (Fig. 1).
The term "calcium phosphate compound" of the present invention means a compound containing phosphoric acid and calcium, and is a component similar to natural bone, which induces bone to be transformed and grown.
For the purpose of the present invention, the calcium phosphate compound may be selected from the group consisting of hydroxyapatite, Carbonated apatite, Tricalcium Phosphate, Calcium Hydrogen Phosphate, Monocalcium phosphate, Calcium phosphate, calcium phosphate, dicalcium phosphate, calcium dihydrogen phosphate, tricalcium phosphate, octacalcium phosphate, and calcium pyrophosphate. And may be at least one selected from the group consisting of hydroxyapatite, but is not limited thereto.
The term "demineralized bone matrix" of the present invention refers to a material which is immersed in an acid to remove an inorganic component in the bone, and exhibits an excellent bone regeneration effect by releasing a bone growth factor. Bone growth factors released from the demineralized bone matrix include, but are not limited to, TGF-beta, PDGF, osteopontin, osteoectin, bone morphogenetic protein (BMP)
In one production example of the present invention, the bone donated from the human body was demineralized by immersing in a 0.6 N HCl aqueous solution, pulverized and neutralized with phosphate buffered saline (PBS) to prepare a demineralized bone matrix.
The calcium phosphate compound or demineralized bone matrix may be included in the bone graft material composition of the present invention in an amount of 20 to 80 parts by weight and the gel-forming powder of the present invention may be contained in 20 to 80 parts by weight, The demineralized bone matrix may be included in an amount of 25 to 75 parts by weight, while the gel-forming powder of the present invention may be contained in an amount of 25 to 75 parts by weight, but is not limited thereto. In a bone graft material composition containing a calcium phosphate compound or a demineralized bone matrix in a weight portion other than the range described above, the gel formation time is long, the maximum load or the gel strength is low, and the gel and calcium phosphate compound or demineralized bone material are separated.
Since the bone graft material is intended to be applied to the bone loss site, there is an advantage that the procedure time is shortened if the gel formation time is saved, and the gel strength is high enough to withstand the physical action of the human body. In addition, when calcium phosphate compound or demineralized bone matrix that helps bone regeneration is well fused with gel-forming powder, bone regeneration can be promoted when applied to the bone loss site. In one experimental example of the present invention, for example, when the bone graft material composition of the present invention contains 50 parts by weight of the calcium phosphate compound or demineralized bone matrix and 50 parts by weight of gel-forming powder, a gel is formed within 10 seconds, And calcium phosphate compound or demineralized bone matrix were formed without being separated (Fig. 2).
The term "osteogenic protein" of the present invention means a protein involved in bone formation, which can be separated from the bone of an individual or prepared through recombinant gene technology.
For the purpose of the present invention, the bone-forming protein is preferably selected from the group consisting of BMP-2, BMP-3, BMP-3b, BMP-4, BMP-5, BMP-6, BMP-7, BMP- , BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17 and BMP-18 or a combination thereof.
The osteogenic protein may be dissolved in purified water or saline, and the purified water or saline may be used in the same meaning as the water for injection.
In addition, the osteogenic protein may be included in the water for injection in an amount of 0.01% by weight to 99.99% by weight, but is not limited thereto.
In one preparation example of the present invention, the osteogenic protein is mixed and dissolved in water for injection, and the osteogenic protein aqueous solution is added to the resultant mixture of the gel-forming powder and the calcium phosphate compound or the demineralized bone matrix, Bone graft material.
According to another aspect of the present invention, there is provided a method of manufacturing a bone graft material composition, comprising:
85 to 95 parts by weight of hydroxyethylcellulose; 1 to 10 parts by weight of croscarmellose sodium; And 1 to 10 parts by weight of sodium carboxymethyl starch to prepare a gel-forming powder (step 1);
25 to 75 parts by weight of calcium phosphate compound or demineralized bone matrix, and 25 to 75 parts by weight of gel-forming powder prepared in step 1 (step 2); And
Mixing the osteogenic protein aqueous solution with the resultant mixture obtained in step 2 (step 3).
Wherein said
The
The
For purposes of the present invention, the gel-forming powder of
In still another aspect of the present invention, the present invention provides a bone grafting kit comprising the bone graft material composition of the present invention and an injection tool.
The injection tool may be a syringe, or a tube, but is not limited thereto.
For the purpose of the present invention, the dough-shaped bone graft material composition of the present invention can be injected into a desired position in an injection tool such as a syringe or a tube. After the injection, the viscoelastic properties of the product can be used to make it into a surgical tool so that it can be densely packed in the defect.
The bone graft material composition of the present invention is capable of transplanting into a bone defect site quickly and conveniently by forming a gel within a few minutes and is not spread by blood even when applied to a human body. Therefore, the bone graft material composition has stability to human body, And thus can be customized for unformed bone defects.
1 is an image showing the appearance of a gel formed according to the composition and content of a gel-forming powder. After the gel was formed, the appearance was observed when 10 minutes passed.
FIG. 2 is an image showing the appearance of the gel formed according to the composition and content of gel-forming powder, calcium phosphate compound particles or demineralized bone matrix. After the gel was formed, the appearance was observed when 10 minutes passed.
Fig. 3 shows the stability of the bone graft material composition when applied to a human body. Fig. 3 is an image showing the state of the gel when the beaker is loaded on or supported with physiological saline and the beaker is turned over.
Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.
Manufacturing example 1. Manufacture of calcium phosphate compound fine particles
The hydroxyapatite powder was dried and then fired at 1200 DEG C to prepare porous fine particles.
Manufacturing example 2. Demineralisation Production of Demineralized Bone Matrix (DBM)
After removing the soft tissues of the donated bone from the human body, impurities were removed using a tissue cleaner containing a surfactant, partially dipped in 0.6 N HCl aqueous solution, and then rinsed with distilled water. Thereafter, slicing was performed in the form of a sheet having a thickness of 0.5 mm, and then submerged in 0.6 N HCl aqueous solution again. After demineralization, the demineralized bone matrix was precipitated and then pulverized and neutralized with phosphate buffered saline (PBS), washed with distilled water, and lyophilized to prepare demineralized bone matrix.
Manufacturing example 3. Osteogenic protein
Bone-forming protein BMP-2 was supplied by Daewoong Pharmaceutical Co. (BMP-2 amount: 0.5 mg, purified water: 0.5 g).
Manufacturing example 4. Gel-forming powder and calcium phosphate compound or Demineralisation Mixture of substrates
Examples of the gel-forming powders include hydroxyethyl cellulose (HEC), hydroxypropyl methylcellulose (HPMC), gellan gum, croscarmellose sodium, carboxymethylstarch sodium Sodium Carboxy methyl Starch) was used. HEC and HPMC were purchased from Merck, and gellan gum and croscarmellose sodium were purchased from purified gold.
The gel-forming powders and the calcium phosphate compound or demineralized bone were placed in a powder mixer and uniformly mixed at a speed of 1000 rpm.
Manufacturing example 5. Goals Graft Produce
The osteogenic protein was completely dissolved by mixing purified water or saline. Thereafter, a gel-like bone graft material was prepared by adding the bone-forming protein aqueous solution to the resultant mixture of the gel-forming powder and the calcium phosphate compound or the gel-forming powder and the demineralized bone matrix.
Experimental Example 1. Goals Graft Determination of ingredients and content
Experimental Example 1-1. Selection of composition and content of gel-forming powders
To determine the optimal composition and content of gel - forming powders for bone grafting, the changes in physical properties of gel - forming powders were investigated.
Specifically, the gel-forming powders described in Production Example 4 were mixed according to the compositions and contents shown in Table 1 below.
Sodium starch
Thereafter, a gel was prepared according to Preparation Example 5 using 0.5 g of purified water, 0.5 mg of BMP-2 and 5 g of the gel-forming powder thus obtained, and the gel formation time, strength and appearance were observed.
As a result, in the case of the compositions shown in Table 1, gels were formed within 10 seconds if they were mixed with purified water in which BMP-2 dissolved, but in
Based on the above results, it was confirmed that the gel-forming powder of the present invention can shorten the procedure time by forming a gel within a short time.
Experimental Example 1-2. Calcium phosphate compound particles or Demineralized bone Content determination
In order to select the optimal composition and content of gel - forming powder, calcium phosphate compound particles or demineralized bone matrix used in bone graft materials, the changes of physical properties according to the composition of gel - forming powder were investigated. In this Experimental Example, hydroxyapatite fine particles or beta-tricalcium phospate fine particles were used as the calcium phosphate compound particles.
Specifically, the content of the calcium phosphate compound particles or the demineralized bone matrix was adjusted to the composition of the gel-forming powder No. 4, which was selected at the optimal content ratio, according to the composition shown in the following Table 3, 2 aqueous solution was added to prepare a bone graft material. Then, the strength and appearance of the formed gel were observed.
Hydroxyapatite particles
Beta-tricalcium phosphate microparticles
Demineralized bone matrix
As a result, it was confirmed that the gel was formed within 10 seconds when the composition (gel-forming powder and calcium phosphate compound or demineralized bone matrix) prepared in the composition ratio of Table 3 was mixed with purified water containing BMP-2 dissolved therein. Or demineralized bone matrix was formed without being separated. But. It was confirmed that as the composition ratio of the gel-forming powder decreases, the gel-forming ability deteriorates and the gel-like property is weakened, and the gel strength is also reduced.
Therefore, it was judged that No. 2, No. 5 and No. 8 were suitable as the composition capable of gel-like deformation when applied to unformed bone defects without separation of formed gel, calcium phosphate compound or demineralized bone matrix. The results are summarized in Table 4, and the appearance observed after gel formation is shown in FIG. The numbers shown in Fig. 2 refer to the respective gels. Are the same as those described in the item.
Based on the above results, it was confirmed that the bone graft material composition of the present invention can promote bone regeneration when applied to the bone loss site because the calcium phosphate compound or demineralized bone matrix and the gel-forming powder are well fused, It can be confirmed that it can be filled tightly in the defect part.
Experimental Example 2. Goals Graft Confirm stability
From the results of Example 1, it was confirmed that the bone graft material compositions No. 2, No. 5 and No. 8 selected by the present invention retained the shape of the bone graft material when the human body was applied.
The bone grafts prepared in the physiological saline solution at 37 ° C for about 1 hour were carried, and then the morphology was observed.
As a result, it was confirmed that all the compositions of No. 2, No. 5 and No. 8 retained the shape of the bone graft material. In addition, when the beaker was turned over after removing the physiological saline, it was confirmed that the bone graft materials of the compositions No. 2 and No. 5 did not fall on the beaker (FIG. 3). The numbers given in Fig. 3 refer to the respective gels. Are the same as those described in the item.
Based on the above results, it was confirmed that the bone graft material composition of the present invention is not spread by blood even when applied to a human body, and applied to a bone defect part well.
Claims (12)
(A) 85 to 95 parts by weight of hydroxyethylcellulose, 1 to 10 parts by weight of croscarmellose sodium, and 1 to 10 parts by weight of sodium carboxy methyl starch. Gel forming powder;
(B) Hydroxyapatite, Carbonated apatite, Tricalcium Phosphate, Calcium Hydrogen Phosphate, Monocalcium phosphate, Dicalcium phosphate Calcium phosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, calcium pyrophosphate, A calcium phosphate compound, or a demineralized bone matrix; And
BMP-9, BMP-10, BMP-11, BMP-7, BMP-8, , BMP-13, BMP-14, BMP-15, BMP-16, BMP-17 and BMP-18.
Wherein the gel-forming powder further comprises hydroxypropyl methylcellulose (HPMC), Gellan Gum, or a combination thereof.
Wherein the gel-forming powder comprises 85 to 95 parts by weight of hydroxyethylcellulose; 1 to 10 parts by weight of croscarmellose sodium; And 1 to 10 parts by weight of sodium carboxymethyl starch.
Wherein said composition comprises 25-75 parts by weight of a gel-forming powder and 25-75 parts by weight of a hydroxyapatite or demineralized bone matrix.
Wherein the osteogenic protein is dissolved in purified water or saline.
85 to 95 parts by weight of hydroxyethylcellulose; 1 to 10 parts by weight of croscarmellose sodium; And 1 to 10 parts by weight of sodium carboxymethyl starch to prepare a gel-forming powder (step 1);
25 to 75 parts by weight of calcium phosphate compound or demineralized bone matrix, and 25 to 75 parts by weight of gel-forming powder prepared in step 1 (step 2); And
Mixing the osteogenic protein aqueous solution with the resultant mixture obtained in step 2 (step 3).
Wherein the gel-forming powder of step 1 is prepared by further mixing hydroxypropylmethylcellulose, gellan gum, or a combination thereof.
The calcium phosphate compound of step 2 may be at least one selected from the group consisting of hydroxyapatite, Carbonated apatite, Tricalcium Phosphate, Calcium Hydrogen Phosphate, Monocalcium phosphate, Calcium phosphate, calcium phosphate, dicalcium phosphate, calcium dihydrogen phosphate, tricalcium phosphate, octacalcium phosphate, and calcium pyrophosphate. At least one of the first and second layers comprises at least one.
BMP-3, BMP-3b, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP- 11, BMP-12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17 and BMP-18.
Wherein the injection tool is a syringe, or a tube.
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