KR101383757B1 - biomaterials using tooth powder and method thereof - Google Patents

Abstract

The present invention is a biomaterial characterized in that the tooth powder and β-TCP (β-tricalcium phosphate) is mixed, more specifically, the ceramic bone graft material mixed with tooth powder and β-TCP (β-tricalcium phosphate) and its It relates to a manufacturing method.

Description

Biomaterials using tooth powder and manufacturing method thereof

The present invention relates to a biomaterial mixed with tooth powder and β-tricalcium phosphate (β-TCP), and a method of manufacturing the same, and more particularly to a ceramic bone graft material and a method of manufacturing the same.

In general, when a bone tissue is damaged due to trauma, malformation, or physiological phenomena, a new bone is formed by filling the bone with the bone. The most common methods for the recovery of bone defects are autografts that are obtained by taking some of their own bone at other sites, autologous bone grafting techniques (allografts), animal bone grafts And xenografts that are chemically treated and transplanted. Although autogenous bone is generally considered to be the best grafting method since autogenous bone has all the properties of bone formation, bone induction, and bone conduction, the autotransplantation method requires secondary surgery and thus increases the risk of infection and blood loss And it is difficult to obtain the necessary amount. Allogeneic transplantation has the potential to cause disease transmission and immune reactions. The xenotransplantation method can also cause an immune response, and the probability of mad cow disease is low. In order to overcome these problems, it is possible to easily obtain a sufficient amount of bone such as calcium phosphate and calcium carbonate salts of bio-glass and various compositions, and there is no possibility of transmission to disease, and biocompatibility There is a demand for a synthetic bone graft material which is excellent and can be appropriately absorbed at the time of implantation and be replaced with a regenerated bone.

Representative synthetic bone substitutes include calcium phosphate ceramics such as hydroxyapatite (HA), tricalcium phosphate (TCP) and biphasic calcium phosphate (BCP), bioglass, calcium carbonate ). Hydroxyapatite and calcium triphosphate (TCP), which are calcium phosphate type ceramics, are typical biomaterials and have attracted attention as synthetic bone substitutes.

Hydroxyapatite is crystallographically and chemically similar to the inorganic constituent of the actual bone and has a property of directly binding to the bone. However, due to low solubility in vivo, the bone bound at the interface can not grow into the inside, There are disadvantages remaining until the end. The property of calcium triphosphate binding directly to bones is similar to that of hydroxyapatite, but it is gradually dissolving in vivo and eventually disappearing. Therefore, sufficient strength must be maintained in order to use the porous calcium phosphate granules as artificial grains.

Calcium triphosphate is a representative biocompatible goal frame that can remove the risk from blood or tissue-borne infection with sterile artificial material. Specifically, calcium triphosphate is a biologically active material that is linked to microparticles and exhibits higher solubility than hydroxyapatite. It exhibits faster absorption than other calcium phosphate ceramics, promotes bone formation, and can be replaced with autogenous bone through elution and absorption processes .

On the other hand, calcium triphosphate has a heterogeneous β-phase and α-phase, unlike hydroxyapatite, which has no homogeneous heterogeneous form. The β-phase has hexagonal crystals in a low-temperature phase. In general, the low- Phase transformation into a high-temperature phase having a monoclinic phase. This high-temperature α-phase reacts violently with water and is therefore unsuitable for use as a living implant. In addition, the phase transition to the? -Phase having a low density on the high density? Causes a fine crack in the sintered body, resulting in a reduction in the strength of the entire material. As the artificial bone material, β -phosphate calcium triphosphate is preferred, and it is essential to obtain a high-density sintered body in order to apply the β-phase of calcium triphosphate having excellent bioabsorbability as a biomaterial.

Biphasic calcium phosphate (BCP) is a bone graft substitute commonly used in dentistry and orthopedic surgery. It is a mixture of two phases, hydroxyapatite and calcium triphosphate. The abnormal calcium phosphate is gradually degraded in the body, and calcium and phosphate ions released into the body fluid cause supersaturation of the body fluid at the localized site, thereby forming biological apatite on the ceramic surface. This apatite layer promotes the attachment of osteoblasts and contributes to the bone conduction of the abnormal calcium phosphate.

60 to 70% of the bone tissue is composed of inorganic crystals. The inorganic crystals of the bone not only increase the mechanical strength of the bone but physiologically act as a reservoir of inorganic ions such as Ca ^{2 +} and PO ₄ ^{3 -} in the tissue fluid. Inorganic crystals of bone tissue generally have the crystal structure of hydroxyapatite (HA). However, the structure of the ion existing on the surface of the bone crystal is problematic because it is highly reactive, unlike pure hydroxyapatite.

Thus, calcium-sulfate (gymsup) was used more than 100 years ago in an effort to develop artificial bone materials with a structure similar to that of hydroxyapatite present in vivo, but it does not induce sufficient bone growth due to premature rapid absorption I did not. Hydroxyapatite, bio-glass, bio-porcelain, and polymer. However, synthetic compounds similar in physical and chemical properties to those of inorganic crystals have not yet been introduced. (HA) or calcium triphosphate (TCP), which have similar structures to bone, have been introduced, but they have not shown satisfactory results in terms of absorption of the graft material after transplantation.

In the process of replacing ceramics with bone, they react with the surrounding environment or bone by mechanisms such as lysis, precipitation, ion change, deposition, chemotaxis, cell adhesion and proliferation, cell differentiation and extracellular matrix formation. In addition, the biodegradability of calcium triphosphate is caused by physical and chemical dissolution or degradation processes and osteoclast and phagocytic degradation processes. In the early stage of transplantation, mainly physicochemical degradation processes are dominant, The bone reshaping phenomenon occurs. The absorption patterns of α and β-TCP are similar in absorption at 4 weeks, 70% at 40 weeks, 40% at 16 weeks, and less than 10% at 68-86 weeks. It should be placed after 5 ~ 6 months. It is also known from the prior art that hydrolytic-cellular degradation occurs primarily in the osteogenesis process of TCP implanted in experimental animals such as Goettinggen miniature pigs, and bifunctional remodeling occurs in the later 20th week. On the other hand, calcium triphosphate (TCP) is completely absorbed within 12 to 18 months, and is anatomically and functionally replaced by bone.

Initially, the absorption of the goal posture varies greatly depending on the physical and chemical properties of the implant. The decomposition of Ca-P biomass is physically influenced by particle size, pore size, surface area, size and structure of crystals, chemically the pH, Ca / P ratio, pressure and temperature during sintering, It depends on the ion structure. The size of the bone crystal is related to the solubility of the crystal. In particular, the solubility of ultrafine crystals depends solely on the size of the crystals, and even small changes in size will have a sensitive effect on solubility. After the bone crystals are formed, they undergo some chemical and structural changes due to the maturation process. The amount of amorphous ions such as PO ₄ ^{3 -} , CO ₃ ^{2 -} generally believed to be present on the surface of the crystal decreases. Thus, mature bone crystals are less reactive than immature bone crystals.

The reactivity of bioactive ceramics depends on the Ca / P ratio or the ratio of carbonate and PO ₄ ^{3 −} . According to these documents, trisodium phosphate (TCP) is absorbed 12.3 times faster in acid than in hydroxyapatite (HA) and 22.3 times faster in base, and the solubility which affects this absorption is Ca / P ratio, The Ca / P ratio was 1.67 and the Ca / P ratio of hydroxyapatite (HA) was 1.7.

Various attempts have been made to pursue more diversity in the current allograft-based implantable materials market. In particular, the use of autogenous implants and Platelet-Rich Plasma (PRP) blends have helped to increase the diversity of implants . However, as mentioned above, artificial compounds similar in physical and chemical properties similar to those of inorganic crystals of bone have not yet been introduced, and hydroxyapatite (HA) or calcium triphosphate (TCP) It is not satisfactory in terms of absorption.

In order to solve the above problems, the present inventors confirmed that tooth powder is a biomaterial that can substitute for hydroxyapatite (HA) by mixing β-TCP with tooth powder which is a natural biomaterial instead of hydroxyapatite (HA). We also confirmed that it could be used as a new synthetic bone graft.

In the case of tooth powders, the components are similar to those of hydroxyapatite, and various studies have proved their stability and effectiveness, especially biocompatible, bone conductive, absorbed in tissues, and cost different materials. Compared to the advantage of being inexpensive bar, hydroxyapatite (HA) to replace the tooth powder has been used.

An object of the present invention is to provide a biomaterial mixed with tooth powder and β-tricalcium phosphate (β-TCP), more specifically, ceramic bone graft material and a method of manufacturing the same.

The objects and advantages of the invention will become apparent from the following detailed description, claims and drawings.

In order to achieve the above object, an embodiment of the present invention provides a biomaterial and a method of manufacturing the same, characterized in that the tooth powder and β-TCP (β-tricalcium phosphate) are mixed.

At this time, the mixing ratio of the tooth powder and β-TCP (β-tricalcium phosphate) used in one embodiment of the present invention may be 3: 1 weight, but is not necessarily limited thereto. In addition, the biomaterial according to an embodiment of the present invention may be used as a ceramic bone graft material, in particular a synthetic bone graft material.

On the other hand, when the biomaterial according to an embodiment of the present invention is used as a bone graft material, using a method such as compression, compression, pressing contact, packing, pressing, hardening, putty, pistist, moldable strip, block, It can be molded and used in the form of chips, and can be used in the form of gels, powders, pastes, tablets, pellets, etc. using chemical additives, and can also be used as it is in powder form.

Biomaterial manufacturing method according to an embodiment of the present invention comprises the first step of selecting a tooth without tooth caries or filling; A second step of washing the tooth selected in the first step with 5 (w / v)% hydrogen peroxide aqueous solution and then ultrasonically washing three or more times with distilled water; A third step of heat-treating the teeth having passed through the second step at 1230 ° C. or more for 3 hours; A fourth step of grinding the heat treated tooth; And a fifth step of mixing β-TCP (β-tricalcium phosphate) with the tooth powder obtained in the fourth step.

In addition, the biomaterial manufacturing method according to an embodiment of the present invention by applying the following process can produce a biomaterial, in particular, ceramic bone graft material. Biomaterial manufacturing method according to an embodiment of the present invention comprises the steps of selecting a tooth without tooth caries or filling; Washing the tooth selected in the selection step with 5 (w / v)% hydrogen peroxide aqueous solution, and then ultrasonically washing three or more times with distilled water; Heat-treating the treated teeth at 1230 ° C. or more for 3 hours or more; Grinding the heat treated tooth; And mixing β-TCP (β-tricalcium phosphate) with the tooth powder obtained in the grinding step.

More advantageously, it is preferable to grind the hot-treated teeth to 500 μm or less.

Incorporating one or more therapeutic molecules, such as molecules for preventing or treating a pathological condition selected from, for example, cancer and osteoporosis, into a biomaterial according to one embodiment of the invention.

In addition, it may be considered to introduce adipose tissue or any other tissues or cellular preparations taken from a patient intended for the biomaterial into the biomaterial according to an embodiment of the present invention, wherein the adipose tissue or a preparation thereof may be prepared in advance. Or suspended in plasma or physiological saline.

In addition, natural or synthetic growth factors may be introduced into the biomaterial according to one embodiment of the present invention. It is also possible to consider the presence of biomarkers or contrast agents that facilitate visualization by resorption of biomaterials and medical imaging of their fate in the organism.

According to the size and arrangement of bone bonds, filling with biomaterials according to one embodiment of the present invention is necessary for affected tissues during bone restoration at the site of implantation of the biomaterials according to one embodiment of the present invention. It can be combined with temporary osteosynthesis (osteosynthesis) application that confers mechanical strength. As confirmed by the present inventors, the implantation of the biomaterial according to one embodiment of the present invention could promote bone tissue formation in a short period of time (weeks).

The biomaterial according to an embodiment of the present invention, more specifically, the implant material according to the embodiment of the present invention can compensate for the shortcomings of the prior art and obtain bone of excellent quality in terms of angiogenesis and hardness to be obtained. In addition, the method for producing a biomaterial according to an embodiment of the present invention is simple and easy to implement, does not require a multi-step process for the individual to be treated, and is relatively inexpensive compared to the prior art methods.

1 is a flow chart illustrating a method of manufacturing a biomaterial (ceramic bone graft material, etc.) according to an embodiment of the present invention.
Figure 2 is a photograph of the bone defects formed on the head bones of the experimental rabbit.
3 is a photo of the bone graft treated with the bone graft material according to the present invention after the bone defect is formed in the head bone portion of the experimental rabbit.
Figure 4 is a scanning electron microscope (SEM) picture of the tooth powder according to an embodiment of the present invention. The magnifications of A, B, C, and D are X100, X500, X1000, and X3000, respectively.
5 is a scanning electron microscope (SEM) photograph of β-TCP according to an embodiment of the present invention. The magnifications of A, B, C, and D are X100, X500, X1000, and X3000, respectively.
FIG. 6 is a spectrum photograph of ATR-FTIR (Attenuated Total Reflectance-Fourier transform infrared) of tooth powder and β-TCP according to an embodiment of the present invention.
7 is a micro-CT picture of the control group 4 and experiment 1, 2, 3 group. A, B, C, D are photographs of the control group, experiment group 1, experiment group 2, and experiment group 3, respectively.
FIG. 8 is a micro-CT photograph of week 8 control group and experiment 1, 2, 3 group. A, B, C, D are photographs of the control group, experiment group 1, experiment group 2, and experiment group 3, respectively.
Figure 9 is a bone sample photograph containing bone defects of the control group 4 weeks.
Figure 10 is a bone specimen photograph including bone defects of the fourth week of experiment 1 group.
FIG. 11 is a bone specimen photograph including bone defects of Week 2 group 2.
12 is a bone sample photograph including bone defects of the third group of the fourth experiment.
Figure 13 is a bone specimen including bone defects of the 8 week control group.
Figure 14 is a bone sample photograph containing bone defects of the eighth week experiment group.
FIG. 15 is a bone specimen photograph including bone defects of the 8th week experiment group 2.
16 is a bone sample photograph including bone defects of the third group of the eighth experiment.

Hereinafter, some embodiments of the present invention will be described in detail with reference to exemplary drawings. It should be noted that, in adding reference numerals to the constituent elements of the drawings, the same constituent elements are denoted by the same reference symbols as possible even if they are shown in different drawings. In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

In describing the components of the present invention, terms such as first, second, A, B, (a), and (b) may be used. These terms are intended to distinguish the constituent elements from other constituent elements, and the terms do not limit the nature, order or order of the constituent elements. When a component is described as being "connected", "coupled", or "connected" to another component, the component may be directly connected to or connected to the other component, It should be understood that an element may be "connected," "coupled," or "connected."

The biomaterial according to an embodiment of the present invention relates to an implant mixed with tooth powder and β-TCP and shows excellent bone formation rate.

Hereinafter, a biomaterial and a method of manufacturing the same according to an embodiment of the present invention, and more specifically, a ceramic bone graft mixed with tooth powder and β-TCP and a method of manufacturing the same will be described in detail.

Biomaterials according to an embodiment of the present invention, more specifically, ceramic bone grafts were prepared by mixing tooth powder and β-TCP, and then implanted into bone defects to evaluate the degree of bone formation of the grafts.

Example

Laboratory Animals and Materials

Experimental animal

Twelve new zealand rabbits of about 2.5 to 3.0 kg (15 weeks) of body weight were used as experimental animals.

Experimental material

In this experiment, we used human tooth powder (Tooth powder, Chosun university dental hospital, Korea) and β-Tricalcium Phosphate (TCP, Junsei, Japan) in a 1: 1, 1: 3, 3: 1 weight ratio.

Manufacture of implant materials

Healthy teeth without tooth caries or fillings were selected from the extracted teeth (S110), washed with 5 (w / v)% hydrogen peroxide solution, and then distilled water with ultrasonic cleaner (Ultra Sonic 4020, KODO, Korea) at least 3 times. Ultrasonic cleaning (S120). After washing, the tooth was treated with an electric furnace (Furnace MF-21G, JEIO TECH, Korea) at 1230 ° C. or more for 3 hours to remove all of the organic substances with infection and toxicity, and only pure minerals were obtained (S130). The high temperature treated teeth were ground to 500 μm or less using a grinder (Mixer Mill mm400, Retsch, Germany) (S140). Then, the pulverized tooth powder and β-TCP were mixed in a weight ratio of 1: 1, 1: 3, and 3: 1, respectively (S150).

in In vivo experiment (IRB approval number: CDMDIRB-1009-A67)

After injecting zoletil (Zoletil, Virbac S. A, France) and Xylazine (Rompun, Bayer, Korea) into the experimental animals to induce general anesthesia, the surgical site of the head was disinfected with povidone iodine. The surgical site was infiltrated with 2% lidocaine and an incision was made along the central part from the anterior to the posterior part of the frontal bone. After raising the periosteum and exposing the skull, four circular bone defects were formed using an outer diameter 8 mm trephine bur while being careful not to damage the meninges (Fig. 2). Gentamicin (Gentamycin, Donghwa Pharm, Korea) was injected to prevent infection until the day and 2 days after surgery. Four bone defects were formed in 6 rabbits out of 12 rabbits, and the control group (empty defect) in the four bone defects, the experimental group 1 (tooth powder: β-TCP = 1: 1 weight ratio), 0.27 g of bone graft material was injected into each of the experimental group 2 (tooth powder: β-TCP = 3: 1 weight ratio) and the experimental group 3 (tooth powder: β-TCP = 1: 3 weight ratio) (Fig. 3). Four weeks later, six rabbits underwent the same procedure, and eight weeks after the first experiment, four and eight weeks of sacrifice were simultaneously performed.

In other words, the present inventors prepared the tooth powder by heat-treating the teeth as described above, and implanted the implant material mixed with the tooth powder and the existing excellent ceramic biomaterial β-TCP to the rabbit skull defect 4 The extent of skull recovery was reviewed over weeks and eight weeks. The structure and composition of the implants were measured by scanning electron microscopy (SEM) and infrared spectroscopy (ATR-FTIR) as follows, and micro-CT and histological examination were performed to measure the degree of bone filling and the distribution of new bone. Skull samples were analyzed.

Assessment Methods

Surface shape observation

The surface shape of the implant was observed with a critical scanning electron microscope (FE-SEM, Hitachi, S4800, Japan) by coating the surface of the implant with platinum.

ATR - FTIR  analysis

Samples prepared to observe the interchain linkage structure of the implant were analyzed by infrared spectroscopy using ATR-FTIR (Attenuated Total Reflectance-Fourier transform infrared, Nicolet 6700/8700, ThermoFisher Scientific).

Histological observation

After 8 weeks of experiment, bone specimens including bone defects were collected, fixed with 10% formalin for 24 hours, and deciphered for 12 hours using Calci-Clear RapidTM (National Diagnostics, Atlanta, GA, USA). The tissues were washed in running water, and then paraffin-treated using Hypercentre XP tissue processor (Shandon, Cheshire, UK) and cut into 4-5 μm thick and stained with hematoxylin-eosin and Goldner's trichrome. Samples were prepared and measured new bone density of the localized area using a MagnaFire digital camera system (Image & Microscope Technology, Daejeon, Korea).

Micro - CT  shooting

Rabbit skull samples were taken with Micro-CT (Skyscan 1176, Belgium). The tube voltage was 60kV, the tube current was 167uA, 0.5mm aluminum filtration was used, and the pixel size was 9um. The captured images were converted into Bmp files using Nerecon Reconstruction (1.6.4 version, Skyscan, Belgium) and reconstructed as 2D images.

Evaluation of Physical and Chemical Properties of Implants

Surface shape

In order to analyze the surface state of the tooth powder and β-TCP, the surface crystal of the implant was observed by scanning electron microscope. As a result of photographing at a high magnification of the tooth powder, the surface showed an angular crystallinity (FIG. 4), and in the case of β-TCP, it showed a fine grain shape (FIG. 5).

Surface chemical properties ( ATR - FTIR )

6 shows the polymer binding form of the implant through infrared spectroscopy. The results, tooth powder and β-TCP is the peak of the similarity ^{Phosphate ions (1050cm -1 ~ 1150cm -1} ) and ^{Carbonate ions (1400cm -1 ~ 1500cm -1} ) appears on the components of the conventional hydroxyapatite (HA) sequence appear. This confirmed that the tooth powder is not significantly different from the hydroxyapatite-based ceramics.

Micro - CT  result

As can be seen in Figure 7, the control group was found to be almost unfilled, and showed a better bone formation rate than the control group as a whole, especially in the experimental group 2, not only around the bone defect center of bone defects It was confirmed that the bone regeneration progressed to the part (Fig. 7). Similarly, as shown in FIG. 8, the control group and the experimental group at 8 weeks showed similar results to the control group and the experimental group at 4 weeks (FIG. 8).

Table 1 below shows the mixing ratio (weight ratio) of the tooth powder and β-TCP of the control group and the experimental group (Group 1, 2, 3) and the degree of new bone formation according to such mixing ratio (weight ratio). * Indicates that the statistically significant p value (p <0.05) appeared.

Histological observation

As can be seen in FIG. 9A, in the fourth week control group, new bone was observed at the edge of the defect, and the center was filled by fibrosis. Looking at the circled portion of Figure 9A it can be seen that the formation of new bone in the bone defect is limited. In FIG. 9B, red arrows indicate missing edges, and white arrows show new bone formed.

Referring to FIG. 10A, in the first week of Experiment 4, new bone was mainly formed around the bone defect margin (circled portion of FIG. 10A), and an implant (arrow in FIG. 10A) and fibrosis were observed in the center portion. In FIG. 10B, red arrows indicate defect edges, black arrows indicate implant particles, and the circled portions show that new bone is formed.

Referring to FIG. 11A, in the second week of Experiment 2, new bone was formed around the graft due to bone defect margin and some centripetality (circled portion of FIG. 11A). The black arrows in FIG. 11B represent new bone formed, and the red arrows represent implants.

12A, in the third week of Experiment 3, new bone was slightly formed around the bone defect (circled portion of FIG. 12A), and the central part of the bone graft and the surrounding fibrosis were observed without formation of the new bone. In FIG. 12B, the white arrows represent the defect edges, the red arrows represent the formed new bone, and the black arrows represent the implant.

Week 8 also did not differ significantly from Week 4. 13A, in the 8th week control group, new bone was observed around the edge of the bone defect (circle of FIG. 13A). In Fig. 13B, the arrow indicates the defect edge and the circled portion shows the new bone formed.

Referring to FIG. 14A, in the first week of Experiment 8, new bone was active around the bone defect margin (circled portion of FIG. 14A), and a small amount of new bone formation was observed around the implant. In FIG. 14B, black arrows indicate defect edges, red arrows show new bone formed, and white arrows indicate implants.

Referring to FIG. 15A, in the second week of Experiment 8, new bone was formed around the graft (circled portion of FIG. 15A) with bone defect margin and some centripetality. In FIG. 15B, white arrows represent new bone formed and black arrows represent implants.

In FIG. 16A, new bone was partially observed in the bone defect margin (circled portion of FIG. 16A) in the 3rd week of experiment 3, and the center was filled with the graft and the surrounding fibrosis with little new bone. In FIG. 16B, the white arrows represent the defect edges, the red arrows represent the new bone formed, and the black arrows represent the implant.

As a result of the experiment, in the experiments 1, 2, and 3 groups containing the three kinds of mixed implants, a better recovery rate was observed than the control group without any one, and especially in the experimental group containing a lot of tooth powder, the recovery rate was better than the other experimental groups. I could see it.

More specifically, i) Micro-CT confirmed that the experimental group was able to confirm better bone recovery than the control group, especially in the Experiment 2 group showed the best recovery rate, ii) the degree of new bone formation of three types of implants The treated group showed better bone formation rate than the untreated group, and it was confirmed that more new bones were formed around the implant in Experiment 2, and iii) the rate of bone recovery over time was 8 weeks in both the control and experimental groups. It can be seen that the 4 weeks group showed a better degree of bone recovery.

When using the implant according to an embodiment of the present invention, it is possible to obtain a bone of a superior quality than conventionally used implants. In addition, the implant according to an embodiment of the present invention is simple and easy to manufacture, does not require a multi-step process to the individual to be treated, there is an effect that the burden is relatively inexpensive to patients.

Claims (5)

Biomaterial, characterized in that the tooth powder and β-TCP (β-tricalcium phosphate) is mixed in a 3: 1 weight ratio. delete The method of claim 1,
The biomaterial is a biomaterial, characterized in that the ceramic bone graft for use as a graft in the bone filling method. A first step of selecting teeth without tooth caries or fillings;
A second step of washing the tooth selected in the first step with 5 (w / v)% hydrogen peroxide aqueous solution and then ultrasonically washing three or more times with distilled water;
A third step of heat-treating the teeth having passed through the second step at 1230 ° C. or more for 3 hours;
A fourth step of grinding the heat treated tooth; And
A fifth step of mixing the tooth powder obtained in the fourth step and β-TCP (β-tricalcium phosphate) in a 3: 1 weight ratio; Biomaterial manufacturing method comprising a. 5. The method of claim 4,
The biomaterial is a biomaterial manufacturing method, characterized in that the ceramic bone graft.

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