KR102316879B1 - Scaffold for regenerating periodontal tissue containing horse bone-derived nanoceramics and PCL and its manufacturing method - Google Patents

Abstract

The present invention relates to a support for periodontal tissue regeneration comprising horse bone nanoceramics and PCL, a manufacturing method thereof, and a periodontal tissue regeneration inducing material for transplantation comprising the support.
The PCL 3D scaffold for periodontal tissue regeneration containing the horse bone nanoceramic of the present invention promotes adhesion with bone tissue, prevents the soft tissue from penetrating into the bone loss site through the support, and promotes bone tissue regeneration. It can be usefully used in industry.

Description

Scaffold for regenerating periodontal tissue containing horse bone-derived nanoceramics and PCL and its manufacturing method

The present invention relates to a support for periodontal tissue regeneration comprising horse bone nano-ceramics and PCL, a manufacturing method thereof, and a periodontal tissue regeneration inducing material for transplantation comprising the support.

Guided Bone Regeneration (GBR) is a type of Guided Tissue Regeneration (GTR), and is a technique known as a method of promoting regeneration of damaged tissues by preventing contact of damaged tissues with other tissues. In particular, in the case of periodontal tissue, since the tissue between the tooth root and the alveolar bone is complicated, it is known as a representative tissue to which GBR treatment is applied. Absorbable periodontal tissue regeneration inducing material is a thin film-type material that is applied to the wounded area to prevent different tissues from adhering to each other.

Currently commercialized membranous scaffolds use ePTFE (expanded polytetrafluoroethylene) or titanium-based metal materials, so they are not biodegradable. In addition, the existing membrane is in the form of a sponge having no pores or the size of the pores is not controlled, so it is difficult to exchange materials, and additionally, it is difficult to control the thickness, so there is a disadvantage that the space maintenance function is poor (Laura T et al., 2010) . Accordingly, recent attempts to overcome the limitations of the prior art through 3D printing technology have been reported (Dawood N et al., 2015).

Under this background, the present inventors studied to develop a membrane for GBR treatment that maintains the size of pores and has excellent space maintenance ability using a biodegradable polymer, and in order to increase bone tissue regeneration ability and adhesion with bone tissue, xenogeneic bone The present invention was completed by manufacturing a membrane maintaining mechanical strength by mixing horse bone-derived nanoceramics, a type of graft material.

It is an object of the present invention to provide a support for periodontal tissue regeneration comprising horse bone nanoceramics and polycaprolactone (PCL).

Another object of the present invention is to provide a method for preparing a scaffold comprising the horse bone nanoceramic and polycaprolactone (PCL).

Another object of the present invention is to provide a filament for periodontal tissue regeneration comprising the horse bone nanoceramic and polycaprolactone (PCL).

Another object of the present invention is to provide a periodontal tissue regeneration inducing material for transplantation comprising the support.

This will be described in detail as follows. On the other hand, each description and embodiment disclosed in the present invention may be applied to each other description and embodiment. That is, all combinations of the various elements disclosed herein fall within the scope of the present invention. In addition, it cannot be considered that the scope of the present invention is limited by the specific descriptions described below.

In addition, 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. Also, such equivalents are intended to be encompassed by the present invention.

One aspect of the present invention for achieving the above object provides a support for periodontal tissue regeneration comprising a horse bone nanoceramic and polycaprolactone (PCL).

The support for periodontal tissue regeneration of the present invention refers to regenerating periodontal tissue and assisting in tooth formation and alveolar bone cell differentiation, and angiogenesis. The support includes horse bone nanoceramic and polycaprolactone (PCL), and may mean that polycaprolactone (PCL) containing the horse bone nanoceramic is printed by a 3D printer, but is not limited thereto.

The "horse bone nanoceramic" of the present invention has a structure similar to that of human bone mineral, and has the most similar chemical composition and crystal structure to hydroxyapatite among calcium phosphate-based materials. This indicates that it is an excellent graft material for biocompatibility and bone regeneration.

The horse bone-derived ceramic can be used when the volume of alveolar bone is insufficient as a heterogeneous bone graft material, and has an effect of rapid bone growth and bone replacement.

The horse bone nanoceramics of the present invention are those obtained by completely removing organic matter from horse bones and obtaining only bone minerals through a high-temperature heat treatment method. The heat treatment may be performed at a temperature of 600 to 1500 °C, specifically 800 to 1200 °C, but is not limited thereto. Heat treatment at this high temperature can prevent further infection.

"Polycaprolactone (PCL)" of the present invention refers to a polyester-based biodegradable polymer polymerized from caprolactone (CL). It has a low melting point of about 60°C and a glass transition temperature of about -60°C. The PCL can be used in various medical fields, such as a support for cell attachment for regeneration of living tissue and an insert for regeneration of bone tissue.

The polycaprolactone (PCL) has excellent biocompatibility and is non-toxic when decomposed. "Biocompatibility" of the present invention means that it does not induce undesirable long-term effects when administered to a living body.

The scaffold for periodontal tissue regeneration of the present invention can use horse bone nanoceramics excellent for biocompatibility and bone regeneration, and to overcome the non-uniformity of the horse bone nanoceramic particles, it is mixed with PCL, a polymer with excellent biocompatibility, and is a more suitable 3D printing material. may have been used as

The PCL/horse bone nanoceramic scaffold of the present invention may have a mass ratio of horse bone:polycaprolactone of 1:5, 1:7, 1:10, 1:15, or 1:20, but is not limited thereto.

The term "periodontal tissue regeneration" of the present invention refers to the attachment of new periodontal ligaments generated by cells derived from periodontal ligaments to new cementum. The scaffold of the present invention can regenerate periodontal tissue by inducing differentiation of osteocytes from alveolar bone damaged by periodontal disease. The support may promote proliferation of pulp-derived stem cells, and may promote alveolar bone differentiation.

In a specific embodiment of the present invention, it was confirmed that the higher the content of horse bone ceramic, that is, the greater the adhesion of cells was in the support having a horse bone content of 10%, and it was confirmed that the most activated after 7 days (Example 5).

In another specific embodiment of the present invention, it was confirmed that as the content of horse bone ceramic increased, calcium mineral accumulation increased and differentiation into osteoblasts was promoted through an increase in the expression level of bone differentiation protein markers (Example) 6).

Another aspect of the present invention provides a method for preparing a scaffold comprising the horse bone nanoceramic and polycaprolactone (PCL).

The method for producing a PCL/horse bone nanoceramic scaffold of the present invention comprises the steps of: (a) extracting minerals from horse bone through high-temperature heat treatment; (b) pulverizing the extracted horse bone to a size of 100 to 1000 nanometers; (c) adding ethanol to the pulverized horse bone nanoceramics and dispersing them with an ultrasonic grinder; and (d) mixing the horse bone nanoceramic and an organic solvent containing PCL and evaporating the solvent.

The heat treatment in step (a) may be performed at 800 to 1200° C., but is not limited thereto.

The ethanol used in step (c) may be 80% or more, specifically 90%, more specifically 99% ethanol, but is not limited thereto. The horse bone nanoceramic mixed in the ethanol may be 1 to 10 g/ml, specifically 1 to 5 g/ml, more specifically 1 to 2 g/ml, but is not limited thereto.

The horse bone nanoceramics mixed with the ethanol may be dispersed for 1 minute to 10 minutes, specifically for 3 minutes to 5 minutes using an ultrasonic grinder, but is not limited thereto.

The organic solvent used in step (d) may be chloroform, dichloromethane, tetrahydrofuran, dimethylformamide, or dimethylsulfoxide, but is not limited thereto. does not

In addition, after mixing the PCL solution and the horse bone nanoceramic in step (d), the PCL/horse bone nanoceramic solution is poured on the plate and the solvent is completely evaporated for 3 to 20 hours, specifically 5 to 10 hours in a hood. can

The method of manufacturing a scaffold for periodontal tissue regeneration of the present invention comprises the steps of: (d), (e) heat-treating the PCL/horse bone nanoceramic at 80 to 150°C; and (f) 3D printing the heat-treated PCL/horse bone nanoceramic at a pressure of 100 to 500 kPa using a nozzle of 10 to 20 g.

The 3D printing may be printed at a head speed of 0.01 to 20 mm/s, specifically 0.1 to 10 mm/s, but is not limited thereto.

"3D printing" of the present invention is a technology for manufacturing tissues or organs by laminating cells in a desired shape or pattern, and biocompatible polymers, natural polymers, biomolecules, cells, or bioactive materials can be used as printing materials. .

The 3D printing has characteristics of biomimicry, small tissue, and autonomous self-assembly. The "biomimicry" refers to the reproduction of artificial organs or cells by applying the characteristics of living organisms to the entire industry, and the "small tissue" refers to the generation characteristics of tissues that become large units of organs by gathering small tissues in the body in 3D. It refers to what is applied to printing, and the "autonomous self-assembly" means that the material constituting the initial cells of the tissue in the developmental stage can autonomously form an ideal structural tissue.

"Polycaprolactone (PCL)" used as a 3D printing material of the present invention is a biodegradable polymer that is decomposed in vivo, has no toxicity to decomposition products, and has excellent mechanical properties, so it is easy to secure physical strength in a desired structure or shape. It has the advantage of being able to control the decomposition rate in the human body.

Another aspect of the present invention provides a filament for periodontal tissue regeneration comprising the horse bone nanoceramic and polycaprolactone (PCL).

"Horse bone nanoceramic", "polycaprolactone", and "periodontal tissue regeneration" of the present invention are as described above.

The "filament" of the present invention refers to a fiber having nano-sized pores through a nanoscale shape realization process for realizing a microsurface in order to improve the structural functionality of the support.

The filament can secure micro-sized pores through which metabolism can be performed smoothly through a three-dimensional internal and external structure, and structurally improve cell activity.

The filament for periodontal tissue regeneration comprising the horse bone nanoceramic and polycaprolactone of the present invention has a mass ratio of horse bone: polycaprolactone of 1:5, 1:7, 1:10, 1:15, or 1:20. can, but is not limited thereto.

The filament may promote proliferation of pulp-derived stem cells, and may promote alveolar bone differentiation.

Another aspect of the present invention provides a periodontal tissue regeneration inducing material for transplantation for inducing regeneration of periodontal tissue comprising the support.

The "regeneration-inducing material" of the present invention is a membranous medical device for Guided Bone Regeneration (GBR), and the most representative medical device using the principle of GBR may be an absorbable periodontal tissue regeneration-inducing material. The regeneration inducing material is a material serving as a shielding film to prevent the growth of surrounding periodontal cells after bone grafting to interfere with alveolar bone regeneration. By applying the membrane-type guide material between the alveolar bone and the periodontal ligament, it can serve to physically separate each tissue.

In the present invention, adhesion with bone tissue can be enhanced by using a horse-derived nanoceramic for the membranous support. It was confirmed that the membrane prevents the soft tissue from penetrating into the bone loss site and promotes bone tissue regeneration (Example 6).

Specifically, in order to increase adhesion with existing bone tissue, a material obtained by mixing horse bone-derived nanoceramics with PCL can be manufactured using a 3D printer. In addition, in order to overcome the disadvantage of poor printing ability due to the non-uniformity of the horse bone-derived nanoceramic particles, a PCL/horse bone mixture dissolved in an organic solvent can be used as a printing material.

The support for periodontal tissue regeneration comprising the horse bone nanoceramic and PCL of the present invention is inserted between the damaged alveolar bone and the periodontal ligament to promote the repair of the alveolar bone tissue and prevent the connective tissue from growing into the periodontal ligament, thereby derived from the tissue surrounding the insertion. Since it plays a role in inducing adhesion of mesenchymal stem cells, proliferation of tooth-derived stem cells, and osteodifferentiation of alveolar bone-derived stem cells, it can be usefully used in related industries such as tooth regeneration.

1 is a view showing a crystal phase after X-ray irradiation of powder obtained after heat treatment of horse bone.
Figure 2 shows the change in horse bone characteristics according to the heat treatment temperature, Figure 2a is a view analyzing the change in mass using DSC-TGA (thermogravimetric analyzer), Figure 2b is a view using FT-IR (infrared spectroscopy) It is a diagram analyzing the functional groups of the powder, and FIG. 2c is a diagram analyzing the mass ratio of inorganic substances using XRF (X-ray fluorescence analyzer).
3 is a diagram illustrating the evaluation of physical properties of PCL/horse bone nanoceramic filaments.
4 is a diagram illustrating the effect of PCL/horse bone nano-ceramic scaffolds on proliferation of dental pulp stem cells (DPSCs) on cell activity, and FIG. 4a is a diagram observed with a fluorescence microscope, and FIG. It is a diagram showing the experimental results.
Figure 5 is a road confirming the effect of PCL / horse bone nano-ceramic support on the bone differentiation of alveolar bone-derived stem cells, Figure 5a is a diagram confirming calcium accumulation according to the horse bone ceramic content, Figure 5b is a diagram showing calcium deposition, 5C is a diagram confirming the expression of bone differentiation-related protein markers.

Hereinafter, the present invention will be described in more detail through examples. These Examples are for explaining the present invention in more detail, and the scope of the present invention is not limited by these Examples.

Example 1. Preparation of PCL scaffolds containing horse bone nanoceramics

Only minerals were extracted from the bones of ponies using a high-temperature heat treatment method, and the extracted horse bones were crushed to a size of 100 to 1000 nanometers, and then the horse bone nanoceramics were dispersed in ethanol using an ultrasonic grinder. After adding polycaprolactone (PCL) to the organic solvent and stirring to prepare a solution, the PCL solution and horse bone nano-ceramics dispersed in ethanol were mixed, and then the solvent was evaporated to prepare a PCL/horse bone nano-ceramic support.

In the step of preparing the solution, polycaprolactone was added to the organic solvent and stirred at a speed of 300 to 500 rpm for 2 to 3 hours.

The organic solvent added to the PCL may be chloroform, dichloromethane, tetrahydrofuran, dimethylformamide, or dimethylsulfoxide.

The horse bone nanoceramics of the present invention were obtained by completely removing organic matter from the bones of ponies using a high temperature heat treatment method to obtain only bone minerals (horse bone nanoceramics), and a temperature of 800° C. or higher was used to eliminate the risk of further infection.

In the step of preparing the ethanol in which the horse bone nano-ceramic is dispersed, 1 to 2 g/ml of horse bone nano-ceramic is mixed with 99% ethanol, and then treated with an ultrasonic grinder at an output of 60 watts (w) for 3 to 5 minutes. did.

Thereafter, the PCL solution was mixed with 99% ethanol in which horse bones were dispersed and stirred at 300 to 500 rpm for 1 to 2 hours. When preparing the PCL/horse bone nanoceramic solution, the amount of the solution was adjusted so that the mass ratio of PCL to horse bone was 20:1 to 5:1 (PCL: horse bone).

After mixing the PCL/horse bone nanoceramic solution, the solution was poured on a square plate and the solvent was completely evaporated for 5 to 10 hours in a fume hood.

The PCL/horse bone nanoceramic solution was put into a hot melt syringe and then heat-treated at a temperature of 80 to 110° C. to sufficiently heat the material for 1 to 2 hours.

The heat-treated PCL/horse bone nanoceramics were printed at a head speed of 0.1 to 10 mm/s through an 18 g nozzle at a pressure of 100 to 500 kPa to prepare a PCL scaffold containing horse bone nanoceramics.

Example 2. Confirmation of Effect of High-Temperature Heat Treatment on Crystallinity of Horse Bone Powder

Horse bones from which the flesh was removed were heat-treated at a temperature of 600° C. for 2 hours to prepare a powder having a particle size of 200 μm or less.

The horse bone ceramic powder having a particle size of 200 μm or less was put into an electric furnace and heat-treated at a temperature of 600, 900, 1100, and 1200 °C for 4 hours, respectively, and an electric furnace was fabricated to have a rising temperature of 5 °C/min, respectively.

After heat treatment, it was confirmed that all organic matter was removed from the horse bone ceramic powder, and the powder was analyzed using an X-ray diffraction analyzer to confirm the crystal structure of each powder. The crystal structure was analyzed through data published in JCPDS, and the crystal structure of hydroxyapatite prepared through synthesis was also analyzed at the same time. In addition, a commercially available bone graft material, Bio-OSS, was used as a control material.

As shown in FIG. 1 , the heat-treated horse bone powder showed a characteristic band in which constructive interference occurred at the same angle as that of hydroxyapatite, a comparative material, confirming that it was a material having a similar crystal structure. It was confirmed that the peak obtained at a specific angle narrows as the heat treatment temperature increases, indicating that this is due to a phenomenon in which the size of the crystal increases.

Through this, it was found that the crystal size of horse bone powder was increased by heat treatment.

Example 3. Confirmation of Effect of High-Temperature Heat Treatment on Chemical Composition of Horse Bone Powder

To investigate the effect of high-temperature heat treatment on horse bone powder in more detail, the characteristics of horse bone fragments were observed using a thermogravimetric analysis (DSC-TGA) apparatus.

Through thermogravimetric analysis, bounded water is present near 110℃, organic substances remaining at 340~360℃, carbonic acid (CO ₃ ^2- ) group at 800℃, and phosphoric acid group (PO ₄ ^{3 ) at 1100℃ or higher. It} was investigated that the hydroxyapatite structure was decomposed due to the decomposition of - ), and the results are shown in FIG. 2a.

Analysis of the FT-IR results shown in FIG. 2b showed that carbonic acid groups were decomposed after 900°C and phosphoric acid groups were decomposed at 1300°C. It can be seen that the result is maintained at about 1.9 until ℃, and then exceeds 2.0 after 1300℃.

Through this, it was confirmed that the structure of hydroxyapatite was decomposed and the molecular ratio of Ca/P increased when high-temperature heat treatment was performed.

Example 4. Confirmation of effect on tensile strength and surface of PCL/horse bone support

In order to measure the strength of a scaffold including horse bone ceramic, a filament in which the two materials were uniformly blended was prepared.

Prepare a solution in which 20 g of polycarprolacton (PCL) was added to 100 ml of dichloro-methane (DCM), and stirred at a speed of 300 rpm for 4 hours so that PCL was completely dissolved in DCM. The solution was prepared.

In addition, in order to evenly disperse the horse bone ceramic, 2 g of horse bone ceramic pulverized to a nanometer size of 100 to 1000 was put in 40 ml of 99% ethanol, and then treated with an output of 60 W for 5 minutes using an ultrasonic grinder. .

After mixing the two solutions, the mixture was stirred at a speed of 300 rpm for 1 hour so that the two solutions were sufficiently mixed, poured into a square plate, and dried in a fume hood until the solvent was completely evaporated. The dried PCL/horse bone ceramics had a film shape and were cut to have a size of 2 cm or less to be used as a material for 3D printing.

PCL/horse bone ceramics were placed in a syringe for loading materials of a hot melt extrusion (HME) type 3D printer and heated at 100° C. for 1 hour. A 14 gauge needle was installed in an extruder to produce a filament, and the material was injected at a pressure of 150 kPa.

As shown in FIG. 3 , the thickness of the prepared PCL/horse bone ceramic filaments was found to be about 1.6 in all treatment groups. Tensile strength was observed to decrease when 10% of horse bone ceramic was contained.

Example 5. Confirmation of the effect of PCL/equine bone 3D scaffold on the proliferation of tooth-derived stem cells

For in vitro examination, pulp-derived stem cell passage 5 (human dental pulp stem cell 2; DPSC cells) collected from human extracted tooth tissue was used. After thawing the frozen DPSC cells, they were cultured in a medium prepared by mixing alpha MEM, 10% FBS (Fatal Bovine Serum), and 1% penicillin in a 37°C, 5% CO2 incubator for 72 hours. After confirming that the culture density of DPSC cells became confluent, they were separated from the culture plate using trypsin-EDTA.

The experimental group used PCL containing horse bone ceramic prepared in Example 1, and as shown in FIG. 4, a support mixed with horse bone ceramic content of 1, 5, and 10% and a support composed of only PCL were used. In vitro tests were performed.

Thereafter, the four types of patches were prepared in a circle having a diameter of 6 mm in a 96-well plate and fixed to each plate. Thereafter, 1 × 103 DPSC cells per well were seeded in a 96-well plate, respectively.

Then, cell activity was measured by the WST-1 test method at intervals of 1, 3, and 7 days.

In order to investigate the role of horse bone ceramics on cell adhesion, the adhesion shape of DPSC cells on the PCL/equine bone ceramic scaffold according to Example 1 was analyzed using a fluorescence microscope.

As shown in Fig. 4a, the higher the content of horse bone ceramic, the better the cells were attached, and it was confirmed that it was remarkably displayed at 7 days. It was confirmed that the quantitative experimental result of FIG. 4b also showed the same aspect as the result of observation under a fluorescence microscope. That is, it was observed that cell proliferation was most active in the PCL/equine bone ceramic scaffold having a horse bone content of 10% prepared according to Example 1.

This indicates that cells adhere well when PCL having a high content of horse bone ceramic is used.

Example 6. Confirmation of the effect of PCL/equine bone 3D scaffold on osteogenic differentiation of alveolar bone-derived stem cells

For in vitro testing, DPSC cell passage 5 obtained from human tooth extractions was used. After thawing the frozen DPSC cells, they were cultured in a medium prepared by mixing alpha MEM, 10% FBS (Fatal Bovine Serum), and 1% penicillin in a 37°C, 5% CO2 incubator for 72 hours. After confirming that the culture density of DPSC cells became confluent, they were separated from the culture plate using trypsin-EDTA.

The experimental group used PCL containing horse bone ceramic prepared in Example 1, and as shown in FIG. 4, a support mixed with horse bone ceramic content of 1, 5, and 10% and a support composed of only PCL were used. The degree of bone differentiation of DPSC was tested.

Then, for observation under a fluorescence microscope, the four types of patches were prepared in a circle having a diameter of 6 mm in a 96-well plate and fixed to each plate. ^{Then, 1 × 10 3} DPSC cells per well were seeded in 96-well plates, respectively.

In addition, in order to measure the generation of calcium minerals using Alizarin Red staining technique, the four types of patches were prepared in a circular shape having a diameter of 10 mm in a 24-well plate and fixed to each plate. Thereafter, 5 × 10 ³ DPSC cells per well were seeded in 48 well plates, respectively.

In addition, in order to measure the expression level of bone differentiation-related protein markers using a Western blot assay technique, the four types of patches were prepared in a circle having a diameter of 130 mm in a 150 pi TCPS dish, and each fixed to the dish. Thereafter, 1 × 10 ⁶ cells per plate were each seeded in 150 pi dishes.

Then, in a medium prepared by mixing alpha MEM, 10% FBS (Fatal Bovine Serum), and 1% penicillin, it was cultured at 37° C. and 5% CO2 in an incubator for 72 hours. Then, in a medium made by mixing alpha MEM, 10% FBS (Fatal Bovine Serum), Dexamethasone (11mmol), and 1% penicillin, at 37°C, 5% CO2 incubator was incubated for 1, 2, 3, 4 weeks.

To investigate the role of horse bone ceramics on the differentiation of DPSC cells into osteoblasts, the expression of osteodifferentiation-related proteins (Osteopontin, OPN, green) of DPSC cells on the PCL/equine bone ceramic scaffold according to Example 1 was examined under a fluorescence microscope. was analyzed through

As shown in FIG. 5A , it was confirmed that the higher the content of horse bone ceramic, the higher the expression level of bone differentiation-related proteins. This trend was also confirmed through the ARS experiment of FIG. 5b, and it was confirmed that the higher the content of horse bone ceramic, the more influence it had on the increase of calcium mineral accumulation.

In addition, as shown in FIG. 5c , as a result of confirming the expression levels of bone differentiation-related protein markers at 1, 2, 3, and 4 weeks, the higher the content of horse bone ceramic, the higher the expression of the initial bone differentiation markers at 1 and 2 weeks. After 3 weeks of activity, late markers were expressed, confirming that the differentiation of stem cells into osteoblasts was promoted.

Through this, it was confirmed that the cell differentiation was most active in the PCL/equine bone ceramic scaffold having a horse bone content of 10% prepared according to Example 1, suggesting that differentiation into osteoblasts is promoted as the horse bone content is higher.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the following claims and their equivalents.

Claims (11)

A support for periodontal tissue regeneration, which promotes proliferation of pulp-derived stem cells, including horse bone nano-ceramics and polycaprolactone (PCL).
The support for periodontal tissue regeneration according to claim 1, wherein the support has a mass ratio of horse bone:polycaprolactone (PCL) of 1:5 to 1:20.
According to claim 1, wherein the support will further promote alveolar bone differentiation, periodontal tissue regeneration support.
(a) extracting minerals from horse bones through high-temperature heat treatment;
(b) pulverizing the extracted horse bone to a size of 100 to 1000 nanometers;
(c) adding ethanol to the pulverized horse bone nanoceramics and dispersing them with an ultrasonic grinder; and
(d) mixing the organic solvent containing the horse bone nano-ceramic and PCL and evaporating the solvent, a method for producing a support for periodontal tissue regeneration, the support for periodontal tissue regeneration comprising the horse bone nano-ceramic and PCL is to promote the proliferation of pulp-derived stem cells, a method for producing a scaffold for periodontal tissue regeneration.
5. The method of claim 4, wherein the organic solvent is any one selected from the group consisting of chloroform, dichloromethane, tetrahydrofuran, dimethylformamide, and dimethylsulfoxide. That will, a method of manufacturing a support for periodontal tissue regeneration.
5. The method of claim 4, wherein after step (d)
(e) heat-treating the PCL/horse bone nanoceramic at 80 to 150°C; and
(f) 3D printing the heat-treated PCL/horse bone nanoceramic at a pressure of 100 to 500 kPa using a nozzle of 10 to 20 g, the method of manufacturing a scaffold for periodontal tissue regeneration.
The method of claim 6, wherein the 3D printing is made at a head speed of 0.01 to 20 mm/s.
Horse bone nano-ceramic and polycaprolactone (polycarprolacton; PCL), which promotes the proliferation of pulp-derived stem cells, periodontal tissue regeneration filament.
The filament for periodontal tissue regeneration according to claim 8, wherein the filament has a mass ratio of horse bone: polycaprolactone (PCL) of 1:5 to 1:20.
The filament for periodontal tissue regeneration according to claim 8, wherein the filament further promotes alveolar bone differentiation.
Claims 1 to 3, comprising the support of any one of claims, periodontal tissue regeneration inducing material for transplantation.

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