KR101193386B1 - Method for preparing hydroxyapatite from tuna bone - Google Patents

Abstract

In the present invention, the tuna bone is washed with water, and the washed tuna bone is mixed with sodium hydroxide solution and acetone to remove protein, lipids, oils and other organic impurities, and then dried and ground, and the ground tuna bone is 600 to 1200 ° C. The method relates to a method for preparing tuna bone-derived hydroxyapatite which is carried out by heating with a phosphate, wherein the hydroxyapatite prepared according to the method of the present invention is calcined at 600 to 1200 ° C. to completely remove organic substances, and β-tree to bone matrix. Calcium phosphate is absent and the crystal size of HAp increases with increasing temperature.

Tuna Bone, Hydroxide, HAp, Bioceramic

Description

Method for preparing hydroxyapatite derived from tuna bone {Method for preparing hydroxyapatite from tuna bone}

The present invention relates to a method of preparing tuna bone-based hydroxyapatite, and more particularly, to wash tuna bone with water and mix the washed tuna bone with sodium hydroxide solution and acetone to remove proteins, lipids, oils and other organic impurities. After that, dried and pulverized, and pulverized tuna bone is carried out by heating to 600 to 1200 ℃ relates to a method for producing a tuna bone-derived hydroxyash.

Bones have a hierarchical structure of organic and inorganic mixtures. 10-20% of the bone matrices are water, 67% of the dry weight consists of inorganic mineral salts (primarily hydroxyapatite), 30-40% of collagen and the rest (about 5%) of non-collagenic proteins (NCP). ) And carbohydrates. Replacing bone lost by bone defects due to accident / injury and / or inflammation or chronic disease remains the goal of orthopedic surgery, and autografts and allografts are widely used in the bone transplantation process.

However, such transplants have their own drawbacks, and autografts often result in an inadequate supply of wounds to ameliorate the complications of wound healing, additional surgery, donor pain and gaps. Allografts have problems such as the risk of developing immune responses and diseases that can be transmitted by tissues and fluids (AIDS and hepatitis). These limitations and concerns have become a real concern for developing artificial materials as bone graft substitutes or exenterate. HAp has been the most widely used artificial force for bone grafts over the past 30 years. HAp can be obtained from natural or synthetic sources. However, synthetic sources have some disadvantages. Chemical synthesis is a time consuming process (eg sol-gel synthesis) that requires steps such as additional time, gelatin, ripening, drying time, sintering time, and also precisely controlled reaction conditions and large groups of individual units. It is required to construct and β-calcium phosphate is formed. Such prior art is to add a gelatin solution in the β-tricalcium phosphate (β-TCP) spherical granules manufacturing method for use as a bone graft material is disclosed in Korea Patent No. 10-2007-10919.

Tuna (Thunnus Obesus) is a commercially important fish species in tropical and subtropical waters of the Pacific, Atlantic and Indian Oceans. Recently, tuna waste has become a serious problem in Korea. The simplest method for reducing pollution and selectively separating HAp from tuna bone is not only to increase its potential use but also to reduce environmental pollution due to the disposal of unused by-products. In conclusion, microstructured HAp has recently been obtained by heat treating fish bones.

Although many studies have explored techniques for separating HAp from natural sources, this has been primarily temperature dependent and little is known about the exact separation temperature. We investigated the thermal properties of tuna bones. The optimal conditions for HAp separation have not been reported so far. Therefore, in the present invention, the optimum conditions for separating pure HAp were established, and the present invention was confirmed by confirming that the separated compound is useful for bone tissue engineering.

It is therefore an object of the present invention to provide a method for preparing hydroxyapatite from tuna bone.

Another object of the present invention to provide a hydroxyapatite bioceramic prepared by the above method.

An object of the present invention as described above is to prepare a hydroxyapatite (HAp) bioceramic by heat-treating the tuna ( Thunnus Obesus ) bone at 600 to 1200 ℃, its properties at various temperatures of 200 to 1200 ℃, TGA Results and X-ray diffraction results were achieved by confirming that the crystallinity increased with increasing calcining temperature in the bone matrix, that the HAp crystals were stable up to 1200 ° C., and that the nanostructured HAp was formed at various temperatures.

The present invention provides a method for preparing hydroxyapatite from tuna bone.

The method according to the invention comprises the steps of washing the tuna bone with water; Mixing the washed tuna bone with sodium hydroxide solution and acetone to remove proteins, lipids, oils and other organic impurities; Drying and grinding the tuna bone; The pulverized tuna bone is heated to 600 to 1200 ° C to prepare a hydroxyapatite.

In another aspect, the present invention provides a tuna bone fish sashimi, which is prepared according to the above method, characterized in that the organic material is completely removed, and β-tricalcium phosphate is not present in the bone matrix.

The present invention also provides a bioceramic composition for implants containing the tuna bone-derived fish sashimi as an active ingredient.

In the present invention, the term "active ingredient" means to include a main ingredient as a substance or a group of substances that are expected to express their efficacy or effect directly or indirectly by intrinsic action.

In the present invention, pure hydroxyapatite was isolated from tuna bone calcined at 600 ° C. or higher. Fish bone calcined at 600 to 1200 ° C. forms pure hydroxyapatite. As a result of the characterization, organic matter was completely removed from fish bones above 600 ℃. Also, the crystallinity of hydroxyapatite was directly proportional to the calcination temperature. There was no β-tricalcium phosphate in the bone matrix when calcined above 1000 ° C. This result indicates that the separated hydroxyphosphate is more stable at higher temperatures in bovine bone. The crystal size of HAp increased with temperature. The crystal size was small at low temperatures, and the crystal size gradually increased with increasing temperature. It was confirmed from the XRD results that the isolated HAp complies with JCPDS 090432. Based on the results of the present invention, it can be estimated that the desired properties can be obtained by appropriately adjusting the temperature. As shown in the present invention, the potential value of fish bone waste for use with ceramic-like HAp has great potential for use as a practical and economical implant material for implant applications.

The phosphorus hydroxide prepared according to the method of the present invention is calcined at 600 to 1200 ° C. to completely remove organic substances, β-tricalcium phosphate is not present in the bone matrix, and the crystal size of HAp increases with increasing temperature. It has the great potential to be used as a practical and economical implant material for implant use as a ceramic-like HAp.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these examples.

Example

Tuna (Thunnus Obesus) bones were obtained from the coast of Busan, South Korea, hydroxide nathrum and acetone materials were purchased from ducksan pure chemical company (Ducksan pure chemical company, Korea).

Example 1: Preparation of hydroxyapatite from tuna bone

Tuna bone was washed with water and mixed with 1.0% sodium hydroxide solution and acetone to remove protein, lipids, oils and other organic impurities. The washed tuna bone was dried at 160 ° C. for 48 hours and ground to 200 size. In the pyrolysis process, 2 g of the tuna bone powder was placed in an open silica crucible and heated for 5 hours at various temperatures in an indirect heating furnace to prepare a hydroxyapatite sample.

Organic removal was observed at various temperatures with bone color changes. The color of the raw fish bone was pale yellow. As the bone was calcined at 300, 400, 500 and 600 ° C., the color of the sample changed to black, yellowish brown and white, respectively. At higher temperatures, fish bones turned white. This shows that all organic matter has been completely removed from the bone. Various colors appearing below 700 ° C were found to be related to organic matrix.

Experimental Example 1 XRD Diffraction Analysis

The phase composition of the sample prepared in the above example was measured with an X-ray diffractometer (Philips-XPert-MPD type) using copper Kα X-ray radiation. The wavelength used for the X-ray was 1.5410 A ° and the diffraction pattern was measured at a temperature in the range of 5 to 80 ° C., an angle of 4 ° ratio and a step size of 0.0.02 °.

TABLE 1

2 shows XRD patterns of raw tuna bone and treated bone. XRD results show that, as shown in Table 1, the induced HAp is stable up to 1200 ° C and the crystals are similar to standard HAp when calcined at 700 ° C, 800 ° C, 900 ° C, and 1000 ° C. I confirmed it. Raw fish bone strength was dispersed by X-ray irradiation, resulting in lower strength and broader peaks due to extracellular matrix and fibrous protein. As the temperature rose to 600 ° C., the intensity of the apatite peak increased likewise. This confirms that organic matter has been removed from the raw bone. The relative intensities at the obtained d-spacing lines, two angles, and various temperatures were compared with standard HAp (JCPDS-09-0432 / 1996) and as a result were close to the standard HAp. The relative strength of the calcined bone at 800 ° C. was very close to the standard HAp compared to other calcination temperatures. The two angles were found to have some change compared to the standard HAp, probably because trace OH radicals were removed. According to Wang and Chaki, dehydrogenation on HAp can result in some peak shifts in XRD results. In the present invention, the XRD 2 of the bone samples calcined at 700 and 1000 ° C. shifted to a total error of 0.031 and 0.056, indicating that the HAp lattice contracted due to the loss of OH radicals. Although no dissolution on the HAp was found in the sample calcined at 1000 ° C., the larger strength planes of calcined fish bones (0 0 2), (2 1 1), (1 1 2), (3 0 0), ( 2 0 2) It should be noted that it can be observed by a simple comparison with the XRD peak positions corresponding to (3 1 0) and (2 1 3).

Experimental Example 2 TGA Analysis

Thermal analysis was performed on a PerkinElmer (USA) TGA 7 analyzer. Samples were scanned within a temperature range of 50-900 ° C. at a ramp rate of 10 ° C./min under continuous air flow.

The natural fish bone and calcination curves at 900 ° C. HAp are shown in FIG. 1 via Thermal Gravimetric Analysis (TGA). The average amount of water and organic phase removed during the heat treatment was calculated. TGA and DTG analysis of the accepted fish bones had two curves at 100.474 ° C and 365.635 ° C and corresponding weight loss 4.7660% and 30.0218%, contributing to the removal of water molecules along with organic matter, The curves correspond to the oxidation of organic materials.

Experimental Example 3 SEM Using EDX Analysis

The surface shape of HAp was observed using field emission scanning electron microscopy (FESEM) (Hitachi, S-2700 model microscope, Japan) with an acceleration voltage of 15.0 KV connected to EDX.

In general, the ratio of ionic calcium to phosphorus in the chemical formula of standard HAp is approximately 1.67. EDX spectra of the treated bones are shown in FIG. 4. Analysis revealed that the material consists of inorganic phases consisting mainly of calcium, phosphorus and oxygen. As shown in the figure, the Ca / P ratios of HAp obtained at 600 ° C., 900 ° C. and 1200 ° C. were 2.04, 1.94 and 1.98. Deviations from these values may be associated with carbonate groups in apatite, rather than from standard values. The Ca / P ratio of the obtained HAp does not vary much, indicating that Ca / P is independent of temperature.

Finally, it is noted that the various temperature processes used produced HAp with significant properties. The yield of HAp at various concentrations varied, as the HAp was degraded to small scale, and the residues at 600. 900 and 1200 ° C. were 62.12%, 59.33% and 57.64%, respectively.

The shape of the apatite obtained using the FESEM apparatus was observed. 3 shows SEM images of bones obtained at 600, 900 and 1200 ° C. The microstructure of raw fish bones appears to be dense due to the presence of organic materials. The shape of the bone calcined at 600 ° C. showed a porous network. As shown in FIG. 3C, a network as in a typical bone was observed in the surface shape of the sample calcined at 900 ° C., and the crystal size increased with temperature. The formation of hydroxyapatite during the heat treatment can be thought of as the tendency of the particles to crystallize and grow at high temperatures.

1 is a diagram showing the TAG curve of the raw tuna bone and tuna bone calcined at 900 ℃.

2 is a diagram showing the XRD results of tuna bone at various temperatures.

Figure 3 shows the SEM results of the raw tuna bone and tuna bone treated at 600, 900 and 1200 ℃.

4 shows EDX results of tuna bones treated at 600, 900 and 1200 ° C.

Claims (3)

In the method for preparing hydroxyapatite by washing the tuna bone with water and removing organic impurities, Mixing the washed tuna bone with sodium hydroxide solution and acetone to remove proteins, lipids, oils and other organic impurities; Drying and grinding the tuna bone; Calcining the ground tuna bone to 1000 ~ 1200 ℃ to prepare a hydroxyapatite free of β-tricalcium phosphate Method for producing hydroxide phosphorus for implants, characterized in that carried out sequentially. delete delete

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