KR20180038331A - Porous brushite-based injectable bone filler for application in vertebroplasty and preperation method of the bone filler - Google Patents

Abstract

Disclosed is brushite-based injectable bone cement. To this end, the brushite-based injectable bone cement contains organic protein foam and has injectability sufficiently high enough that cement slurry can be injected through a needle having a diameter of 10 to 15 gauge, so the brushite-based injectable bone cement can be applied to percutaneous vertebroplasty. The bone filling material of the present invention is used as bone cement, can be applied as a bone substitute, and has a high level of reabsorption.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition kit for an injection-type porous brassiot bone filler and a method for producing the same.

The present invention relates to porous brucite based bone cement or bone filler which is applied to the treatment of spinal compression fractures caused by osteoporosis and the like. SUMMARY OF THE INVENTION The present invention provides a porous bursa filler-type bone filler for percutaneous vertebroplasty.

Osteoporosis is a disease characterized by a structural dysfunction of the bone tissue. This disease of osteoporosis results in a high bone vulnerability with a decrease in the weight of the bone, and exhibits a fracture characteristic easily. The disease progresses without any symptoms or discomfort for years, and sudden fractures can cause the symptoms.

Fractures due to osteoporosis occur mainly in the vertebrae, resulting in severe disruption of the vertebral body and loss of physiological posture. Such vertebral fracture due to osteoporosis results in a problematic limitation of pain and mobility. Minimally invasive techniques such as percutaneous vertebroplasty and kyphoplasty have been applied to treat vertebral compression fractures caused by these osteoporosis.

Percutaneous vertebroplasty (PV) is a technique applied to support and fix a damaged vertebral body, which reduces patient pain and restores mobility and quality of life. This method is mainly based on injecting bone cement through a cannula and is applied to the central part of the damaged vertebral body to restore the original strength of the vertebrae.

The calcium phosphate-based cement was prepared according to H. Monma, T. Kanazawa, "Wet-Process Formation of Non-stoichiometric Hydroxyapatite from Tricalcium Phosphate, Yogyo Kyokaishi , 86, 73-76, 1978, b. &Quot;, which is prepared by mixing a calcium phosphate powder of a specific composition with a solution for dough such as distilled water. For example, The kneaded cement that can be applied or filled in the area is kneaded in a mortar and applied to the damaged area using a syringe, spatula or hand, and then hardened.

Minimally invasive surgery is performed through the body's innervated organs or through the smallest incision. Since all the procedure of the operation must be carried out through the small opening, application of bone cement which can be injected by means such as syringe may be required. For example, in order for bone cement to be delivered to fracture sites such as spinal compression fractures, bone cements with injectable and rapid hardness characteristics may be required.

Currently commercially available injectable cements such as Synthes Norian SRS®, Synthes Norian CRS® and Wright Medical MIG X3® are made readily injectable. However, because of the long curing time, the treating person has to wait for cement hardening, which results in prolonging the operating time.

Another commercially available bone cement product, such as Synthes Fast Set Putty® and Lorenz Mimix®, is fast cured but is not readily injectable through syringes or needles and is therefore considered an unacceptable product for minimally invasive treatment .

Therefore, it is not only necessary to research and develop fast-growing bone cement with stability, but also it is possible to apply it to minimally invasive treatment because it can be easily injected, and it is also possible to provide an ideal working time and hardening time for a treating person, It is urgently required to develop a material capable of reducing the amount of water.

On the other hand, it is known that calcium phosphate cement has remarkable biocompatibility and bone conduction in various studies and promotes bone regeneration. In addition, along with the bone conduction of calcium phosphate cement, the ideal cement should be able to be resorbed at a similar rate so that the bone tissue can grow, and be replaced with a gradually forming new bone tissue. Although the majority of the developed cements have better reabsorption properties compared to the high temperature calcined apatites, they still exhibit slow resorption kinetics and in many cases the cement is intact in the surrounding bone tissue for many years Remains. Conventional calcium phosphate cements do not have macroporosity to the extent of promoting bone growth, although they have micro or nano porosity after curing. This limits the reabsorption of cement and conversion to bone tissue for an appropriate period of time.

Patent Application No. 10-2012-0004242, Brushite bone graft substitute containing mussel adhesive protein Patent Application No. 10-2002-7007307, Brushite Hydraulic Cement Stabilized with Magnesium Salt

An object of the present invention is to provide a composition kit for an injection-type porous brassiere bone filler having excellent flowability in a paste (slurry) state and reducing the risk of complications due to leakage of the cement and enabling more precise injection, Method.

In order to achieve the above object, the present invention provides an injection-type porous brassiere bone filler composition kit comprising: 1) an inorganic solid component comprising calcium phosphate; And 2) a bubbling protein comprising any one selected from the group consisting of gelatin, collagen, albumin, soy protein, ombudsin, and combinations thereof.

The bubbling protein may be contained together with a protein binder and a bioactive protein.

The protein binder may be any one selected from the group consisting of gelatin, collagen, albumin, and a combination thereof. The bioactive protein may be any one selected from the group consisting of soybean protein, ombudsin, and combinations thereof .

The composition kit may further comprise 3) a reaction retarder, and 4) a biocompatible liquid.

In the biocompatible liquid, the solid component, which is the sum of the inorganic solid component and the bubbling protein, may be included in an amount of 3 to 9 parts by weight based on 10 parts by weight of the biocompatible liquid.

The reaction retarder may include any one selected from the group consisting of sodium pyrophosphate, glycollic acid, and combinations thereof.

The biocompatible liquid may comprise an acid component comprising orthophosphoric acid.

As the solid component, any one selected from the group consisting of the soybean protein, the ovomucin and the combination thereof may be contained in an amount of 1 to 5 parts by weight based on 10 parts by weight of the inorganic solid component.

The gelatin may be included in the biocompatible liquid in an amount of 1 to 15% by weight.

The pH of the bone filler paste prepared at the time of mixing may be maintained at a pH of 4 or less within 6 minutes of mixing.

The composition kit can be injected with a force of 12 N or less through a needle having an inner diameter of 0.05 to 0.12 mm when a bone filler paste to be prepared at the time of mixing is injected.

According to another embodiment of the present invention, there is provided a method for preparing an injection-type porous brassite bone filler comprising: 1) an inorganic solid component comprising calcium phosphate; And 2) a solid component comprising any one bubbling protein selected from the group consisting of gelatin, soy protein, obrumuchin, and combinations thereof, 3) a reaction retarder, and 4) a biocompatible liquid, And a mixing step of preparing the formed bone filler paste.

Wherein the mixing step comprises: preparing a solution for dissolving the gelatin in the biocompatible liquid containing the reaction retarder so that the gelatin is contained in an amount of 1 to 15% by weight; 10 parts by weight of the inorganic solid component and 1 to 5 parts by weight of the soy protein, obrumuchin and any combination thereof in an amount of 1 to 5 parts by weight based on 10 parts by weight of the inorganic solid component Solid component preparation; And a bubble generating process of mixing the mixed solution and the solid component to form a bubble-filled bone filler paste.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a porous brucite-based bone cement or bone filler which can be applied to the treatment of vertebral compression fractures caused by osteoporosis and the like. The present invention described herein improves upon the disadvantages described above with currently commercially available products.

Porous bone cement according to the present invention can be injected in the form of a paste (slurry) through a relatively small injection needle-type injection device commonly applied to percutaneous vertebroplasty, Suitability, bone conduction, and the like. In addition, the citrate-free delaying agent is applied so as to inhibit the generation of a citrate intermediate product that inhibits cell adhesion, thereby further improving the biocompatibility, thereby completing the present invention.

In order to achieve the above object, the present invention provides an injection-type porous brassiere bone filler composition kit comprising: 1) an inorganic solid component comprising calcium phosphate; And 2) a bubbling protein comprising any one selected from the group consisting of gelatin, soy protein, ombudsin, and combinations thereof.

The calcium phosphate contained in the solid phase component may be a salt of phosphoric acid and calcium and may be applied to a composition such as bone cement and bone graft material. Specifically, hydroxyapatite (Ca ₁₀ (PO ₄ ) ₆ OH) ₂ ), α-Tricalcium phosphate (α-TCP), β-Tricalcium phosphate (β-TCP), Biphasic calcium phosphate (BCP) TCP mixed with various HA / β-TCP ratios), potassium dihydrogen phosphate, monocalcium phosphate monohydrate (MCPM), dicalcium phosphate (DCP), calcium pyrophosphate (CPC) anhydrate, dicalcium dehydrate (DCPD), amorphous caclum phosphate, tetracalcium phosphate (TTCP), and combinations thereof. _{Specifically, TTCP (Ca 4 (PO 4} ) 2 O), alpha-TCP (Ca 3 (PO 4) 2), MCPM (Ca (H 2 PO 4) 2 · H 2 O), beta-TCP, Ca 2 One selected from the group consisting of P ₂ O ₇ , DCPD (CaHPO ₄ .2H ₂ O), DCPA (CaHPO ₄ ), HA (Ca ₁₀ (PO ₄ ) ₆ (OH) ₂ ) , Which are more suitable to be applied to have in vitro dissolution or neutral pH values. More specifically, in the inorganic solid component, the inorganic solid component is selected from the group consisting of tetracalcium phosphate, dicalcium phosphate anhydrous, dicalcium phosphate dehydrate, and combinations thereof Any one can be included, and β-TCP can be included, and in this case, calcium brucite phosphate can be efficiently formed.

The bubbling protein may generate bubbles (or bubbles) when mixed with the inorganic solid component in a solvent, and a biocompatible organic protein component may be applied to enhance human fitness. Specifically, the bubbling protein may include any one selected from the group consisting of gelatin, collagen, albumin, soybean protein, ovomucin, and combinations thereof. have.

The bubbling protein is preferably a protein binder and a bioactive protein. In this case, bubbles (or bubbles) can be abundantly formed by mixing while utilizing natural proteins.

The protein binder may be any one selected from the group consisting of gelatin, collagen, albumin, and a combination thereof. The bioactive protein may be any one selected from the group consisting of soybean protein, ombudsin, and combinations thereof .

The protein binder and the bioactive protein can be applied in a weight ratio of 1: 0.2 to 2.

The inorganic solid phase component and the bubble generating protein may be contained in a weight ratio of 10: 0.5 to 8, specifically 10: 1 to 5, more specifically 10: 1.8 to 3. When the inorganic solid phase component and the bubble generating protein are mixed together at a ratio of these contents, a bone filler having a strength and porosity suitable for application to a bone filler, particularly a spinal column, can be produced.

Here, the bone filler is a brassite calcium phosphate cement and has excellent resorbability compared to hydroapatite cement. The bone filler can make cells and nutrients permeable to porosity by adding proteins that generate bubbles during the mixing process, as compared to typical Brussite-based cements.

The composition kit may further comprise 3) a reaction retarder, and 4) a biocompatible liquid.

The reaction retarder may be used in combination with other components to control the curing reaction of the bone filler so as to have an initial curing time that is suitable for application to minimally invasive surgery and a total curing time that minimizes the operation time Specifically, it may include any one selected from the group consisting of sodium pyrophosphate, glycolic acid, and combinations thereof.

When sodium pyrophosphate or glycolic acid is used as the reaction retarder, it is possible to apply a citrate-free reaction retarder to inhibit the production of a citrate-based intermediate product that inhibits cell attachment, It is possible to impart cell adhesion ability excellent enough to allow cell attachment within 24 hours.

The reaction retarder may be contained in the biocompatible liquid at 100 to 400 Mm, more specifically 200 to 250 mM. When this content is applied together with other components, the curing time of the bone filler paste can be appropriately controlled.

The biocompatible liquid may include any liquid selected from the group consisting of water, salt water, buffer solution, aqueous solution of carboxylic acid, aqueous solution of propionic acid, and combinations thereof.

The biocompatible liquid may be applied to an acidic solution, more specifically, an acidic solution containing a mineral acid (inorganic acid) may be applied. As the mineral acid, orthophosphoric acid may be applied, and the inorganic acid may be further added, so that the curing reaction of the bone filler (bone cement) proceeding by mixing at the time of application can be stably performed.

It is preferable that the acid solution contains an acid component at a concentration of 5 M or less, specifically at a concentration of 3 M or less. In this case, when mixing with solid components, an appropriate chemical reaction is induced and a proper viscosity and biocompatibility A filling material paste can be obtained.

The solid component, which is the sum of the inorganic solid component and the bubbling protein, may be included in an amount of 3 to 9 parts by weight based on 10 parts by weight of the biocompatible liquid, 4.5 to 7.5 parts by weight, and 5 to 7 parts by weight, , 4.75 to 5.5 parts by weight. With such an amount, it is possible to obtain a filler having proper viscosity and injection ability and self-hardening property at an appropriate time.

The composition kit may contain, as a solid component, any one selected from the group consisting of the soy protein, the ovomucin and combinations thereof in an amount of 1 to 5 parts by weight based on 10 parts by weight of the inorganic solid ingredient, In the biocompatible liquid, from 1 to 15% by weight, in particular from 3 to 13% by weight, more particularly from 5 to 10% by weight. In this case, it is possible to facilitate the injection with a proper flow rate, but also to suppress the occurrence of leakage and to impart appropriate porosity to the hardened bone filler.

The composition kit may be such that the pH of the bone filler paste prepared at the time of mixing can be maintained at a pH of 4 or less within 6 minutes of mixing, and the pH is maintained at a range of 2 to 4 to 5 to 6 minutes after mixing. When the initial acid condition is applied, the curing reaction for forming the brassite based bone filler in the bone filler paste can proceed effectively.

The composition kit may be such that the bone filler paste to be prepared at the time of mixing may be injectable with a force of 12 N or less through a needle having an inner diameter of 0.05 to 0.12 mm. This can be achieved by mixing the powder phase with the liquid phase to produce a cement slurry having a viscosity sufficiently low enough to be injected through a needle having a diameter of 10-15 G (gauge number) applied to percutaneous vertebroplasty .

According to another embodiment of the present invention, there is provided a method for preparing an injection-type porous brassite bone filler comprising: 1) an inorganic solid component comprising calcium phosphate; And 2) a solid component comprising any one bubbling protein selected from the group consisting of gelatin, soy protein, obrumuchin, and combinations thereof, 3) a reaction retarder, and 4) a biocompatible liquid, And a mixing step of preparing the formed bone filler paste.

The content of the inorganic solid phase component, bubbling protein, solid component, reaction retarder, biocompatible liquid, etc., including calcium phosphate, constituting the above-mentioned bone filler, and their specific content and application ratio are the same as those described above, It is omitted.

Wherein the mixing step comprises: preparing a solution for dissolving the gelatin in the biocompatible liquid containing the reaction retarder so that the gelatin is contained in an amount of 1 to 15% by weight; 10 parts by weight of the inorganic solid ingredient and 1 to 5 parts by weight of the soybean protein, obrumuchin and any combination thereof in an amount of 1 to 5 parts by weight based on 10 parts by weight of the inorganic solid ingredient Solid component preparation; And a bubble generating process of mixing the mixed solution and the solid component to form a bubble-filled bone filler paste.

The method of mixing and the like can be generally applied to the preparation of a bone filler, so that a detailed description thereof will be omitted.

In the bubbling process, mixing may be applied at 100 rpm or more, 100 to 500 rpm may be applied, and when the mixing conditions are applied, bubbles may be formed and retained at a level sufficient to form a porous structure .

The composition kit for a porous brucite bone filler of the present invention and the method of manufacturing a bone filler have excellent flowability in a paste (slurry) state, reduce the risk of complications due to cement leakage, and enable more precise injection . It is also non-toxic, has better porosity than conventional products, has excellent cell adhesion and a high level of resorbability.

FIG. 1 is a scanning electron microscope (SEM) photograph of a bone filler sample of Comparative Example in which no bubbling protein was observed, as observed in Example 1 of the present invention. FIG.
FIG. 2 is a scanning electron microscope (SEM) photograph of the bone filler sample of the sample in which bubbling proteins are observed, as observed in Example 1 of the present invention. FIG.
Figure 3 is an X-ray diffraction spectrum showing the phase analysis results of the injectable bone filler after being stored in a wet state in a simulated body fluid (SBF) for 7 days at 37 ^° C in Example 2 of the present invention. Results (BC: Comparative Example, ISBC: Examples).
4 is an experimental photograph showing the possibility of injection of the bone filler prepared in Production Example 3 in Example 3 of the present invention.
FIG. 5 shows the results of in vitro evaluation of cell adhesion and proliferative capacity of injectable bone filler in Example 4 of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. In the following, the percentages or percentages set forth without specific recitation refer to weight percentages and weight ratios.

Manufacturing example

Optimal conditions of bone cement are prepared as follows.

1) 200 to 250 mM delay sodium pyrophosphate claim (sodium pyrophosphate of retardant) and a protein binder of gelatin (5 to 10% by weight content), from 40 to 50 ^o the acidic solution preheated to C (3 to 5 M, orthophosphoric acid aqueous solution ) To prepare a mixed solution.

2) Mixing a solid component containing a bioactive protein (soybean or ombudsin, 15-20 parts by weight) and betaine ginseng calcium (β-TCP, 80-85 parts by weight, purchased from Innobon Co., Ltd.) with the above mixed solution , Mixed at 100 to 500 rpm and foamed to prepare a bone filler paste. At this time, L / P ration (weight ratio) indicating the application ratio of the solution component (L) and the solid component (P) was varied in the range of 0.3 to 0.7 in 0.05 unit.

As a comparative example, a sample in which betaine ginseng calcium was applied as a solid component without applying the bubbling protein was prepared in the same manner as above.

3) The bone filler paste thus prepared was collected and placed in a syringe.

In the prepared bone filler paste, the following reaction proceeds to form brucite-type dicalcium phosphate dihydrate [beta-Tricalcium phosphate (beta-TCP) + orthophosphoric acid (OCP) - dicalcium phosphate dihydrate (brushite)].

[Reaction Scheme]

Ca ₃ (PO ₄ ) ₂ + H ₃ PO ₄ + 6H ₂ O ↔ 3CaHPO ₄ .2H ₂ O

Porous brassite-based bone filler was prepared for implantation using a syringe and the main input was below 10 N or below 8 N.

4) The initial curing time at which the entire volume of the bone filler paste having the L / P ratio of 0.5 to 0.65 was injected was within 5 minutes or within 3 minutes.

5) It was confirmed that the application of a sample having an L / P ratio of 0.5 in an open wet field had the best low leakage resistance and improved biocompatibility.

Example  One

The surface of the bone filler prepared according to the above Preparation Example was observed using a scanning electron microscope and the results are shown in FIGS. 1 and 2. FIG.

FIG. 1 is a photograph of a comparative sample prepared without applying the bubbling protein component in the above procedure, and FIG. 2 is a photograph of an example sample prepared by applying the bubbling protein component according to the above procedure . Both FIGS. 1 and 2 observed the filler surface morphology after being stored in soaking (simulated body fluid) for 7 days at 37 ^° C (SBF).

The scanning electron microscope photographs shown in Figures 1 and 2 show Brushite crystals in flower form (see below, high magnification photo). The micro flakes confirmed that beta-tricalcium phosphate (beta-TCP), which is a major component of the bone filler of the present invention, started to change to calcium deficient hydroxide (CDHA). Also, comparing the photographs of FIGS. 1 and 2 with the low magnification photographs (the above photographs), it can be seen that there is a distinct micropore structure in the sample of the example than the sample of the comparative example.

Example  2

The results of the phase analysis of injectable bone cement were measured at 1 day intervals at 37 ^o C while being soaked in simulated body fluid (SBF). The results of XRD at 7 days (Rigaku's Miniflex II model Is shown in Fig.

Referring to FIG. 3, peaks appeared at about 21 °, 29.5 ° and 30.5 °, indicating brucite peaks, which could be observed from day 1 after soaking. On the other hand, at about 27.9 ° and 31.5 °, the β-TCP peak disappeared after 5 to 7 days (BC: Comparative Example, ISBC: Example). These results show that the bone filler of the present invention can undergo crystallization in a similar biologic solution and improve bone conduction.

Example  3

FIG. 4 shows an experimental photograph showing the possibility of injection of the bone filler prepared in the production example. The powder phase (solid phase component) and the liquid were mixed for 30 to 60 seconds to prepare a paste type bone filler paste. The bone filler paste was placed on a syringe and injected with a normal injection force of 12 N or less. The entire volume in the syringe was fully injected and no movement or leakage occurred.

Example  4

In vitro performance evaluation of bone filler was made. The bone filler paste was prepared as a sample at a height of 5 mm on a syringe having a diameter of 8 mm. The sample was placed in a 24-well plate and maintained at 37 [deg.] C for 7 days, Lambda stem cells) and media (α-MEM + 10% FBS + 1% P / S). The results are shown in Fig. 5, and cell adhesion and growth were confirmed after 7 days (see arrows in Fig. 5), and it was evaluated that the examples of the present invention had excellent initial cell adhesion and proliferation ability.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, Of the right.

Claims (10)

1) inorganic solid components including calcium phosphate; And 2) a bubbling protein comprising any one selected from the group consisting of gelatin, collagen, albumin, soy protein, ombudsin, and combinations thereof. The method according to claim 1,
Wherein the composition kit further comprises: 3) a reaction retarder; and 4) a biocompatible liquid, wherein the biocompatible liquid comprises 10 parts by weight of the biocompatible liquid, wherein the solid component, which is the sum of the inorganic solid component and the bubbling protein, By weight based on the total weight of the porous brucite bone filler. 3. The method of claim 2,
Wherein the reaction retarder comprises any one selected from the group consisting of sodium pyrophosphate, glycolic acid, and combinations thereof. ≪ Desc / Clms Page number 19 > 3. The method of claim 2,
Wherein the biocompatible liquid comprises an acid component comprising orthophosphoric acid. The composition kit according to claim 1,
A composition for an injection-type porous brassiere bone filler, which contains, as a solid component, any one selected from the group consisting of soybean protein, obrumuchin and combinations thereof in an amount of 1 to 5 parts by weight based on 10 parts by weight of the inorganic solid component Kits. 3. The method of claim 2,
Wherein the gelatin is contained in the biocompatible liquid in an amount of 1 to 15% by weight. 5. The method of claim 4,
Wherein the composition kit is maintained at a pH of 4 or less within 6 minutes of mixing the pH of the bone filler paste to be prepared during the mixing. 8. The method of claim 7,
Wherein the composition kit is injectable with a force of 12 N or less through a needle having an inner diameter of 0.05 to 0.12 mm, wherein the bone filler paste to be prepared at the time of mixing is injectable. 1) inorganic solid components including calcium phosphate; And 2) a solid component comprising any one bubbling protein selected from the group consisting of gelatin, soy protein, obrumuchin, and combinations thereof, 3) a reaction retarder, and 4) a biocompatible liquid, And a mixing step of preparing a bone filler paste to be formed. 10. The method of claim 9,
Preparing a solution of the biocompatible liquid containing the reaction retarder by dissolving the gelatin in an amount of 1 to 15% by weight;
10 parts by weight of the inorganic solid component and 1 to 5 parts by weight of the soy protein, obrumuchin and any combination thereof in an amount of 1 to 5 parts by weight based on 10 parts by weight of the inorganic solid component Solid component preparation; And
And a bubble generating step of mixing the mixed solution with the solid component to form a bone filler paste having bubbles formed therein.

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