Patent Application
20230364305
The present invention relates to a bone graft composition and a preparation method thereof, and more particularly, a bone graft composition provided in the form of a putty formulation by mixing calcium phosphate compound particles with hydrogel, having excellent physical properties, which is easy to inject, and which maintains its structure even in an in vivo environment after implantation, thereby enabling sustained release of a drug loaded therein.
Bone graft biomaterials developed in the initial stages depended on a characteristic that they are inert in vivo, but the use thereof was significantly limited due to infection and inflammatory reaction which occur in the surrounding tissue after implantation. Since then, with the rapid development of biomaterial technologies based on metals, ceramics, and polymer materials, materials were designed and developed that are biocompatible rather than bioinert, leading to the development of bioactive scaffolds for bone tissue regeneration, which vary depending on the site and purpose of use. It is required that such bioactive scaffolds for bone tissue regeneration have different physical properties depending on the location of graft placement, that they not be toxic to the surrounding tissue, and that they have relatively high mechanical properties compared to other artificial organs. Such bioactive scaffolds for bone tissue regeneration have been marketed and developed as various biomaterials depending on the properties of the raw materials and the intended use thereof.
All materials that are to be grafted into the human body, particularly polymer materials for bone tissue regeneration, should have good processability and moldability or have good in situ polymerization properties so as to be suited to wounds. These materials are required to provide a suitable environment for the adhesion, growth, and differentiation of cells, and degradation products thereof are also required to be biocompatible. In particular, if compressive strength and yield value of a bone graft material are too low, it will be difficult to maintain the abilities of the bone graft material to fix its location and keep its external shape in the closure or implant placement stage after injection or dense filling of the bone graft material. In addition, if adhesiveness of a bone graft material is too high, it will easily stick to a surgical tool during surgery, and thus it will be difficult to easily fill in bone defects, resulting in a decrease in workability.
Accordingly, there is a need for the development of a bone graft composition that has biocompatibility and physical properties suitable for grafting in bone defects and has the property of maintaining the formulation during a specific period after implantation.
Meanwhile, hydroxyapatite, which is a calcium phosphate-based ceramic, is a component found in teeth and bones. Since hydroxyapatite has excellent biocompatibility, it has attracted attention as a graft material for implantation into the body, such as fillers or artificial implants to replace damaged bones. However, since hydroxyapatite itself has a ceramic particle-type formulation, molding thereof is impossible, and thus it is difficult to apply to a narrow site. There is also a disadvantage in that it is difficult to fill hydroxyapatite in a specialized material intervertebral body fusion cage used in lumbar interbody fusion surgery.
The present inventors have also tried to provide a bone graft composition including a calcium phosphate compound, which is injectable into the body, as disclosed in a prior patent (Korean Patent No. 10-1443814). However, the structure of the composition is easily disintegrated in an in vivo environment where blood flow exists, resulting in release of all drugs or physiologically active substances loaded therein within a short time. Thus, it is difficult to anticipate their continuous effects.
Accordingly, the present inventors have made intensive efforts to develop a formulation in which the above-described disadvantages of the existing bone graft composition including a calcium phosphate compound are improved, and as a result, they found that when the content of the calcium phosphate compound to be used is increased while controlling a size distribution of the particles to be used, its structure is maintained even in an environment in contact with a fluid, like that in a living body, and thus the sustained release of a physiologically active substance or a drug loaded therein is possible, thereby completing the present invention.
An object of the present invention is to provide an injectable bone graft composition including more than 55 wt % (% by weight) and 80 wt % or less of calcium phosphate compound particles; and 20 wt % or more and less than 45 wt % of biodegradable hydrogel.
Another object of the present invention is to provide a kit for bone implantation, the kit including the bone graft composition and an injection tool.
A further object of the present invention is to provide use of composition including more than 55 wt % (% by weight) and 80 wt % or less of calcium phosphate compound particles; and 20 wt % or more and less than 45 wt % of biodegradable hydrogel for bone graft.
Each description and embodiment disclosed in this disclosure may also be applied to other descriptions and embodiments. That is, all combinations of various elements disclosed in this disclosure fall within the scope of the present disclosure. Further, the scope of the present disclosure is not limited by the specific description below.
Further, those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Further, these equivalents should be interpreted to fall within the scope of the present invention.
In addition, throughout this specification, when a part is referred to as “including” an element, it will be understood that other elements may be further included rather than other elements being excluded unless content to the contrary is specially described.
Hereinafter, the present invention will be described in detail.
The present invention provides an injectable bone graft composition including more than 55 wt % and 80 wt % or less of calcium phosphate compound particles; and 20 wt % or more and less than 45 wt % of biodegradable hydrogel.
Since the composition of the present invention has a predetermined fluidity, it is advantageous when injected into irregular defects, while having excellent physical properties, and thus its microstructure may be maintained in an in vivo environment after implantation, and the composition may be usefully applied as a bone graft material.
In particular, when the composition is prepared by mixing calcium phosphate compound particles with biodegradable hydrogel, a mixture of microparticles and macroparticles is used as the calcium phosphate compound included in the composition. Therefore, its content is increased at a high ratio of 70% or more, thereby providing a graft material having remarkably improved strength.
Further, since the composition includes hydrogel containing a biodegradable polymer, it may be biodegraded over time after implantation. In addition, since hydrogel containing a predetermined amount of a poloxamer and HPMC in combination is used, a compressive strength and a yield value are high, thereby providing a bone graft composition with excellent volume retention in the body temperature range after bone implantation. In addition, since the composition includes the hydrogel and the calcium phosphate compound particles at a suitable mixing ratio, it may have a formulation such as a putty type resulting from the agglomeration of the hydrogel and the calcium phosphate compound particles. At the same time, the composition has low adhesiveness, and thus it does not stick to a surgical tool during surgery, and it does not stick to a surgical tool when it is filled in bone defects, suggesting that it has an advantage of excellent workability.
In particular, in the case of the existing composition containing only β-TCP fine particles (or microspheres) as the calcium phosphate compound in addition to the hydrogel, it is difficult to prepare a formulation itself when the content of β-TCP fine particles (or microspheres) increases over a predetermined range. When a composition is prepared using the maximum amount capable of preparing the formulation, the desired strength may not be achieved, and thus there is a problem in that its structure may not be maintained in the in vivo environment when transplanted into a bone defect.
As used herein, the term “bone graft composition” refers to a composition for use as bone defect replacement that is grafted in bone defects to fill the bone defects. Specifically, the bone graft composition in the present invention means a synthetic bone graft material (alloplastic) composition based on a calcium phosphate compound.
The bone graft composition of the present invention is mainly composed of two components of calcium phosphate compound particles and hydrogel.
First, the calcium phosphate compound particles are similar to natural bone and functions to induce osteoconduction and bone growth.
As used herein, the term “calcium phosphate compound” may refer to a compound including phosphoric acid and calcium. Specifically, the calcium phosphate compound may be any one or a combination of two or more selected from the group consisting of hydroxyapatite, tricalcium phosphate (TCP, Ca3(PO4)2), tetracalcium phosphate (Ca4(PO4)2O), brushite (CaHPO4.2H2O), dicalcium diphosphate (Ca2P2O7), calcium tripolyphosphate (Ca5(P3O10)2), Mg-containing apatite, Mg-containing TCP, Sr-containing apatite, and fluorapatite. More specifically, tricalcium phosphate, for example, β-TCP and hydroxyapatite, may be used in a mixture, but the calcium phosphate compound is not limited thereto.
In this regard, as the calcium phosphate compound particles, a mixture of porous particles of 45 μm to 100 μm and 200 μm to 6000 μm in mean diameter may be used. Generally, considering that the particle size is not defined by a single size, but the diameters of particles having a certain distribution are statistically averaged, the calcium phosphate compound particles may be a mixture of particles having a size of 45 μm to 6000 μm in diameter, but are not limited thereto.
For example, in a specific embodiment of the present invention, a mixture of microspherical β-TCP and hydroxyapatite macroparticles was used.
In this regard, the β-TCP may be in the form of microspheres, but is not limited thereto. For example, the β-TCP may be obtained by spray-drying β-TCP powder, sintering the dried powder at a temperature of 1050° C. to 1250° C., and classifying the sintered powder in the range of 45 μm to 75 μm. More specifically, the β-TCP may form a spherical shape during spray-drying of the β-TCP powder, and porosity of the powder may be increased by sintering the spherical β-TCP powder at a temperature of 1050° C. to 1250° C. In order to obtain a more uniform bone graft composition, the sintered β-TCP powder may be classified in the range of 45 μm to 75 μm. At this time, the sintering may be performed for 1 hour to 3 hours, and most preferably, for 2 hours.
As a result, the β-TCP particles finally obtained as described above may be microspherical particles having a diameter of 45 μm to 75 μm. Moreover, the final β-TCP particles may have porosity of 60% or higher as a result of performing the spray-drying and sintering processes as described above.
Meanwhile, the hydroxyapatite may be a granule having a broad size distribution of several tens of μm to several mm, but is not limited thereto.
Further, the calcium phosphate compound particles may be porous particles having a three-dimensional pore connectivity of 90% or more and/or porosity of 60% or more. For example, by using the porous calcium phosphate compound particles, a drug and/or a bone morphogenetic protein may be loaded in the pores thereof, as needed, thereby exhibiting two or more effects at the same time or achieving a synergistic therapeutic effect. Furthermore, since the porous structure facilitates penetration of newly formed tissues, it thereby promotes tissue regeneration.
The hydrogel which is the second component of the bone graft composition of the present invention is a gel formed by dispersing a polymer having a sol-gel transition property in water, and is a means that agglomerates the calcium phosphate compound particles to form a formulation suitable for bone grafting.
The hydrogel may include one or more selected from the group consisting of a poloxamer, collagen, hyaluronic acid, gelatin, a PEG/PPG/PEG block copolymer, and cellulose. The hydrogel, which is a material having a non-crosslinked structure, may be a material without a swelling property, and may be a material that decomposes within several months. In this regard, the hydrogel may include the above-described components at a concentration of 15 wt % to 35 wt %, but is not limited thereto. If the concentration of the hydrogel is less than 15 wt %, it may be difficult to have sufficient strength, and if the concentration is more than 35 wt %, its adhesiveness is high, and thus a large amount thereof may remain in a container for manufacturing and/or storage or in a tool for transport and/or injection.
In the present invention, the hydrogel may further include 0.5 parts by weight to 2 parts by weight of hydroxypropyl methylcellulose (HPMC), based on 100 parts by weight of the hydrogel.
In the present invention, a poloxamer and hydroxypropyl methylcellulose (HPMC) may be used as polymers which are biodegradable, which have a sol-gel transition temperature lower than the body temperature, and which may maintain the gel state in the body temperature range, in order to provide a bone graft composition that has excellent biocompatibility and to have an excellent ability to maintain its formulation after implantation.
As used herein, the term “poloxamer” refers to a triblock copolymer (PEO-PPO-PEO) having two polyethylene glycol (PEG) chains bonded to a central chain of polypropylene glycol (PPG). Generally, a ratio of PEG/PPG in a poloxamer may vary in the range from 1:9 to 8:2. A molecular weight of a poloxamer may be in a wide range from 1,100 g/mol to 14,000 g/mol. A poloxamer is a temperature-sensitive polymer. In the present invention, the poloxamer functions to impart injectability and moldability to the bone graft composition and to enable the bone graft material to be degraded rapidly after filling in bone defects so as to allow only the calcium phosphate-based bone graft material component to remain. In order to maintain the ease of injection and moldability in the room temperature range and formulation stability during storage and transport at room temperature, a high-molecular-weight poloxamer having a relatively low sol-gel transition temperature and high viscosity is preferably used. Preferably, a poloxamer that has a sol-gel transition temperature of 4° C. to 35° C. so as to be able to maintain the gel state at the body temperature of about 37° C. may be used in the present invention. Specifically, poloxamer 407, having an excellent ability to maintain the gel state at the body temperature of about 37° C., may be most preferably used in the present invention.
As used herein, the term “hydroxypropyl methylcellulose (HPMC)” refers to a semi-synthetic, inert, viscoelastic polymer, also called hypromellose. In the present invention, HPMC functions to improve the elasticity of the hydrogel. In particular, as the viscoelasticity of the hydrogel increases, the ability to fix the location of the bone graft material when filling in bone defects becomes better so that the leakage of the bone graft material to the outside may be advantageously minimized.
The viscosity of HPMC may be preferably 1,000 cps to 100,000 cps, and most preferably 100,000 cps. HPMC is added in a trace amount in order to induce high viscosity and high elasticity. As the viscosity thereof increases, the ability to fix the location of the bone graft material and the density of filling of the bone graft material may be increased, and the adhesion of the bone graft material to a surgical tool and gloves may be minimized Thus, it is most preferable to use HPMC having a viscosity of 100,000 cps, which shows the highest viscosity when being added in a trace amount.
In the composition of the present invention, the hydrogel including HPMC in addition to a specific amount of a poloxamer is used, thereby providing a bone graft composition that is better in terms of compressive strength, yield value, and adhesiveness as compared with a hydrogel including a poloxamer alone or a hydrogel outside of the above-described ranges.
Particularly, the bone graft composition of the present invention may have a yield value ranging from 1500 g/cm2 to 4000 g/cm2. When the composition of the present invention has a yield value within the above-described range, it will exhibit excellent viscoelasticity, and thus may be easily filled in bone defects during grafting, suggesting that it shows physical properties suitable for use as a graft material.
As used herein, the term “compressive strength” is used interchangeably with “strength”, and means the strength at which the external shape of the bone graft composition is changed by an external force. The term “yield value” is a physical property value related to the elasticity of a finished product, and means the maximum strength at which the composition is not deformed by an external force. Thus, as the compressive strength and yield value increase, it is possible to maintain the abilities of the composition to fix its location and maintain its external shape in the closure or the implant placement stage after injection or dense filling of the bone graft material.
As used herein, the term “adhesiveness” means the property of adhering to stainless steel. It is a force acting in a direction opposite to that of compressive strength, and (−) means only direction. The higher absolute value of the adhesiveness means that a greater force is required to detach the bone graft composition formulation that stuck to stainless steel, and it may also mean the degree of adhesion of the bone graft composition formulation not only to a surgical tool made of stainless steel, but also to gloves made of resin.
Compressive strength, yield value, and adhesiveness, which are physical strengths that are measured in the present invention, may be measured using a common rheometer and/or UTM (universal testing machine).
The bone graft composition of the present invention includes a hydrogel filled between calcium phosphate compound particles close to each other. After implantation of the bone graft composition into bone defects, the hydrogel is degraded and released, the calcium phosphate compound particles are maintained in the close state, and bone grows into the space between the calcium phosphate compound particles after release of the hydrogel. Therefore, it is necessary to have the ability to maintain its shape for a predetermined period of time.
The bone graft composition of the present invention may further include a physiologically active substance. The physiologically active substance may be loaded in pores of the porous calcium phosphate compound particles included in the composition. The physiologically active substance may be one or more selected from the group consisting of bone morphogenetic proteins (BMPs), bone morphogenetic peptides, extracellular matrix proteins, and tissue growth factors.
For example, the bone morphogenetic proteins may include BMP-2, BMP-3, BMP-3b, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, or a combination thereof, but are not limited thereto.
For example, the tissue growth factors may include VEGF, FGF-2, IGF-1, TGF-β, or the like, but are not limited thereto.
For example, the bone graft composition of the present invention may be used in bone grafting, maxillary sinus lifting, lumbar interbody fusion, cervical interbody fusion, or upper & lower extremity fracture fusion, but is not limited thereto. For example, the bone graft composition may be used in lateral lumbar interbody fusion (LLIF), oblique lumbar interbody fusion (OLIF), or anterior lumbar interbody fusion (ALIF), but is not limited thereto. In particular, the bone graft composition of the present invention is a putty formulation which may be injected regardless of the shape of the site to be transplanted, and has excellent strength, and thus may be usefully applied to the bone damage site having an unspecified shape.
The composition of the present invention may be provided in a putty formulation.
Another aspect of the present invention provides a kit for bone implantation, the kit including the bone graft composition and an injection tool.
The kit of the present invention may be a bone graft material of a putty formulation. Such a graft material of the putty formulation may be injected into a desired site by putting it in an injection tool such as a syringe, tube, etc. After injection, the bone graft material may be packed by a surgical tool so that it is densely filled into bone defects due to a viscoelastic property of the product.
To this end, the kit of the present invention may further include an injection tool, wherein the injection tool may include a mixing syringe or a vial transport device, but is not limited thereto.
As described above, the kit for bone implantation of the present invention may be used in bone grafting, maxillary sinus lifting, lumbar interbody fusion, cervical interbody fusion, or upper & lower extremity fracture fusion, e.g., lateral lumbar interbody fusion (LLIF), oblique lumbar interbody fusion (OLIF), or anterior lumbar interbody fusion (ALIF), but is not limited thereto.
A further aspect of the present invention provides use of composition including more than 55 wt % (% by weight) and 80 wt % or less of calcium phosphate compound particles; and 20 wt % or more and less than 45 wt % of biodegradable hydrogel for bone graft.
The composition of the present invention is designed to compensate for the disadvantages of the existing formulation, for which it is difficult to improve physical properties, by using a microparticle-type calcium phosphate-based compound and increasing the content thereof. When a bone graft material is prepared by mixing the calcium phosphate-based compound with hydrogel, a mixture of microparticles and macroparticles is used as the calcium phosphate-based compound to increase the content of the calcium phosphate-based compound by 70% or more based on the total composition. Thus, the composition may maintain its shape for a long period of time under in vivo mimetic conditions, and enables sustained release of a drug loaded therein, and therefore, it may be usefully applied as a graft material for regeneration of injured bone tissues by loading physiologically active substances such as bone morphogenetic proteins, etc.
Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to exemplary embodiments. However, these exemplary embodiments are for illustrative purposes only, and the scope of the present invention is not intended to be limited by these exemplary embodiments.
Pure β-TCP powder (Cerectron Co., Korea) was spray-dried to prepare a spherical shape. Then, the spherical β-TCP powder was sintered at 1050° C., and the sintered particles were classified in the range of 45 μm to 75 μm.
Calcium phosphate compound particles having a distribution in the range of 200 μm to 6,000 μm were prepared with reference to the method disclosed in Korean Patent No. 10-0401941.
First, HPMC and poloxamer 407 were mixed using a high-speed vacuum mixer to produce hydrogel, and then the β-TCP powder prepared according to Preparation Example 1 was uniformly mixed therewith to obtain a hydrogel complex.
Subsequently, the prepared hydrogel complex was mixed with the hydroxyapatite ceramic granules having a size of 0.6 mm to 6 mm prepared according to Preparation Example 2 to prepare a bone graft material of a putty formulation. The mixing was performed using a specialized mixing syringe so that the hydroxyapatite granules were pulverized.
Particle-type calcium phosphate-based compounds having a size in the range of less than 100 μm, 600 μm to less than 1,000 μm, 1,000 μm to less than 3,000 μm, and 3,000 μm to 6,000 μm were prepared using the samples of Comparative Examples 1 to 4, respectively.
Calcium phosphate-based compound particles having a size of less than 100 μm and hydrogel were mixed at a weight ratio of 30:70, 50:50, and 70:30 to prepare bone graft compositions of Comparative Examples 5 to 7, respectively.
Calcium phosphate-based compound particles having a size of 1,000 μm to less than 3,000 μm and hydrogel were mixed at a weight ratio of 50:50 to prepare a bone graft composition of Comparative Example 8.
The shapes of the calcium phosphate-based compound particles of Comparative Examples 1 to 4 were observed with the unaided eye and photographed, and are shown in
Further, appearance and features of the bone graft compositions of Comparative Examples 5 to 8 and Example 1 are shown in
As described, the bone graft composition of Example 1, which was prepared by including the high 70% content of the calcium phosphate-based compound, was formulated into various preparations, and the clinical applicability thereof was tested. The test results are shown in
To examine properties of the bone graft materials under in vivo mimetic conditions, each of the compositions of Comparative Examples 4 to 8 and Example 1 was put in a cage and immersed in physiological saline at 37° C. After 5 minutes and 24 hours of immersion, their shape retention was examined. The results are shown in
As shown in
As shown in
The same force (shear stress) was applied using a rheometer to the graft material composed of the composition of Comparative Example 6, which is a commercially available product, and the graft material composed of the composition of Example 1, in which calcium phosphate-based compound macroparticles were additionally included to increase the content thereof. At this time, the shear strain of each formulation was measured and shown in
In order to examine changes in the strength of the composition of Example 1 in which the content of calcium phosphate-based compound was increased by additionally including macroparticles in addition to calcium phosphate-based compound microparticles, the composition of Example 1 in which the content of the calcium phosphate-based compound was increased to 70% by including both microparticles and macroparticles, the composition of Comparative Example 6 in which the content of the calcium phosphate-based compound was 50% by including only microparticles, and the composition of Comparative Example 8 in which the content of the calcium phosphate-based compound was 50% by including only macroparticles were measured for compressive strength, and the results are shown in
Based on the above description, it will be understood by those skilled in the art that the present disclosure may be implemented in a different specific form without changing the technical spirit or essential characteristics thereof. Therefore, it should be understood that the above embodiment is not limitative, but illustrative in all aspects. The scope of the disclosure is defined by the appended claims rather than by the description preceding them, and therefore all changes and modifications that fall within metes and bounds of the claims or equivalents of such metes and bounds are therefore intended to be embraced by the claims.
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2020-0127278 | Sep 2020 | KR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/KR2021/001110 | 1/27/2021 | WO |
