KR20240107506A - Multi-layer dental graft material and method for preparing the same - Google Patents

Abstract

One embodiment of the present specification includes the steps of (a) purifying the pericardium or peritoneum of a cow or pig to form a dense layer; (b) forming a porous layer using a collagen solution; (c) salt treatment by adding a salt solution; and (d) drying. It provides a method of manufacturing a dental implant material, including a step of drying.

Description

Multi-layer dental implant material and manufacturing method thereof {MULTI-LAYER DENTAL GRAFT MATERIAL AND METHOD FOR PREPARING THE SAME}

This specification relates to multilayer dental implants and methods for manufacturing the same.

If the bone quality is poor or the amount of bone is insufficient during orthodontic treatment or implant surgery, or if the alveolar bone is destroyed or lost due to periodontitis, a procedure to increase the alveolar bone is necessary, and bone induction is a procedure for bone regeneration. Guided bone regeneration and bone grafting are being performed.

Guided bone regeneration, which is the most commonly used method for bone regeneration, is known to show better results in bone regeneration than simply bone grafting, as it covers the upper part of the bone defect with a shielding membrane, unlike bone grafting. The shielding membrane used in guided bone regeneration maintains space, prevents blood clots from falling out of the defect, and helps bone formation by blocking surrounding connective tissue, which proliferates faster than bone tissue, from entering the space where bone regeneration is needed.

However, when performing guided bone regeneration using a shielding membrane, there is a problem that the shielding membrane is exposed due to insufficient upper gingiva due to the bone graft material, or the upper gingiva is not healed quickly and the shielding membrane is eventually exposed.

On the other hand, when marginal tissue recession occurs, when there is a patient's aesthetic request, and among root coverage techniques performed for root caries or cervical attrition, pedicled soft tissue grafting and free soft tissue grafting use autologous tissue, so the graft It has the advantages of excellent blood supply, fast healing, and natural color mixing with surrounding tissues.

However, there are limitations to autologous tissue collection, and soft tissue regeneration substitutes are being used as an alternative. When soft tissue regeneration substitutes are used, the patient's chair-time at the dentist is reduced because autologous tissue is not collected, and the patient's pain can also be alleviated.

Therefore, when performing guided bone regeneration, the upper gingiva is quickly healed to prevent exposure of the shielding membrane, and the shielding membrane can reduce the risk of infection from the outside, and soft tissue regeneration can replace the autologous tissue used in existing root flap surgery. The development of substitutes is necessary.

The description in this specification is intended to solve the problems of the prior art described above, and one purpose of this specification is to provide a multi-layer dental implant that has the ability to block surrounding soft tissue and induce rapid regeneration at the same time, and can be used as a substitute for shielding membrane and soft tissue regeneration. and providing a manufacturing method thereof.

According to one aspect, (a) purifying the pericardium or peritoneum of a cow or pig to form a dense layer; (b) forming a porous layer using a collagen solution; (c) salt treatment by adding a salt solution; and (d) drying. It provides a method of manufacturing a dental implant material, including a step of drying.

In one embodiment, the purification in step (a) may be performed by one or more of washing, sterilization, base treatment, acid treatment, enzyme treatment, and oxidation treatment.

In one embodiment, the collagen solution may include soluble collagen derived from pork skin, insoluble collagen derived from bovine tendon, or a combination thereof.

In one embodiment, the concentration of the collagen solution may be 0.6 to 1.4% by weight.

In one embodiment, the salt solution may include one selected from the group consisting of phosphate, calcium salt, sodium salt, and combinations thereof.

In one embodiment, the concentration of the salt solution may be 6 to 14 M.

According to another aspect, a dense layer formed by purifying the pericardium or peritoneum of cattle or pigs; and a porous layer containing collagen. It provides a dental implant material comprising a.

In one embodiment, the porosity of the porous layer may be 80 to 90%.

In one embodiment, the average size of pores included in the porous layer may be 260 to 350 μm.

In one embodiment, the thickness ratio of the dense layer and the porous layer may be 1:3 to 1:7.

In one embodiment, the dense layer and the porous layer may have different decomposition periods.

The method of manufacturing a dental implant material according to one aspect of the present specification has excellent mechanical properties and shielding ability, forming a dense layer that can block surrounding soft tissue and increase the retention period, and absorbing blood and quickly decomposing to promote rapid bone regeneration. It can be applied to the production of multilayer dental implants containing an inducible porous layer.

In addition, the dental implant material according to another aspect of the present specification has the ability to block surrounding soft tissue and induce rapid regeneration at the same time, and can be used as a shield or soft tissue regeneration substitute depending on the direction of use.

The effect of one aspect of the present specification is not limited to the effects described above, and should be understood to include all effects that can be inferred from the configuration described in the detailed description or claims of the present specification.

Figure 1 is an SEM image of a cross-section of a transplant material according to an example and a comparative example of the present specification (Figure 1(a): Example, Figure 1(b): Comparative Example 1, Figure 1(c): Comparison Example 2, Figure 1(d): Comparative Example 3).
Figure 2 shows the results of measuring the suture strength of each of the dense layer, the porous layer, and the two-layer graft material according to an embodiment of the present specification.
Figure 3 shows the results of measuring the decomposition period of the dense layer and porous layer of the transplant material according to an embodiment of the present specification.

Hereinafter, one aspect of the present specification will be described with reference to the attached drawings. However, the description in this specification may be implemented in various different forms, and therefore is not limited to the embodiments described herein. In order to clearly explain one aspect of the specification in the drawings, parts that are not related to the description are omitted, and similar reference numerals are assigned to similar parts throughout the specification.

Throughout the specification, when a part is said to be “connected” to another part, this includes not only cases where it is “directly connected,” but also cases where it is “indirectly connected” with another member in between. . Additionally, when a part is said to “include” a certain component, this does not mean that other components are excluded, but that other components can be added, unless specifically stated to the contrary.

When a range of numerical values is described herein, unless the specific range is stated otherwise, the value has the precision of significant figures given in accordance with the standard rules in chemistry for significant figures. For example, the number 10 includes the range 5.0 to 14.9, and the number 10.0 includes the range 9.50 to 10.49.

Hereinafter, an embodiment of the present specification will be described in detail with reference to the attached drawings.

Manufacturing method of dental implant material

A method for manufacturing a dental implant material according to an aspect of the present specification includes the steps of: (a) purifying the pericardium or peritoneum of a cow or pig to form a dense layer; (b) forming a porous layer using a collagen solution; (c) salt treatment by adding a salt solution; and (d) drying.

Step (a) is a step of forming a dense layer of the transplant material with collagen derived from natural tissues such as the pericardium and peritoneum of bovine or porcine.

The purification in step (a) may be performed by one or more of washing, sterilization, base treatment, acid treatment, enzyme treatment, and oxidation treatment, but is not limited thereto.

Step (a) may include washing the pericardium or peritoneum with water.

Step (a) may include sterilizing the pericardium or peritoneum with an alcohol solution. For example, the alcohol solution may be a 50-90% ethanol solution, but is not limited thereto.

Step (a) may include treating the pericardium or peritoneum with a basic solution. The basic solution may be a sodium hydroxide solution, a calcium hydroxide solution, or a sodium carbonate solution, but is not limited thereto. The pH of the basic solution may be 10.5 to 11.5, and if the pH of the basic solution exceeds the above range, it may cause denaturation of collagen.

Step (a) may include acid treating the pericardium or peritoneum with an acidic solution. The acidic solution may be an acetic acid solution, a hydrochloric acid solution, or a sulfuric acid solution, but is not limited thereto. The pH of the acidic solution may be 2.58 to 2.82.

Step (a) may include enzymatically treating the pericardium or peritoneum with a proteolytic enzyme. The proteolytic enzyme may be trypsin, but is not limited thereto.

Step (a) may include oxidizing the pericardium or peritoneum with an oxidizing agent. The oxidizing agent may be hydrogen peroxide, but is not limited thereto.

The step (b) is a step of forming a porous layer with a pore structure using a collagen solution. The porous layer may be formed by adding a collagen solution on top of the dense layer formed in step (a), or the collagen solution may be poured into a mold to form a porous layer, and then the dense layer may be placed and combined with the dense layer. However, it is not limited to this.

The collagen solution may include one selected from the group consisting of collagen type I, collagen type II, collagen type III, collagen type IV, collagen type VIII, and combinations thereof, for example, collagen I, which is mammalian-derived collagen. type, collagen type III, or a combination thereof, but is not limited thereto.

The collagen solution may include soluble collagen, insoluble collagen, or a combination thereof.

The collagen solution may include soluble collagen derived from porcine dermis, insoluble collagen derived from bovine tendon, or a combination thereof, but is not limited thereto.

The collagen solution may be collagen solubilized by an acidic solution. The acidic solution may be a hydrochloric acid solution, an acetic acid solution, or a sulfuric acid solution. For example, it may be a hydrochloric acid solution with a pH of 2.0 to 3.0, but is not limited thereto.

The concentration of the collagen solution may be 0.6 to 1.4 weight%. For example, it may be 0.6% by weight, 0.7% by weight, 0.8% by weight, 0.9% by weight, 1.0% by weight, 1.1% by weight, 1.2% by weight, 1.3% by weight, 1.4% by weight, or a value between two of these values. If the concentration of the collagen solution is less than the above range, the porosity of the porous layer may be excessively high or the size of the pores may be excessively large, which may result in a decrease in the physical properties of the porous layer or an excessively rapid decomposition rate. If the concentration of the collagen solution exceeds the above range, the porosity of the porous layer may be lowered or the size of the pores may be smaller, resulting in lower absorption and slower decomposition, which may reduce the regeneration rate of soft tissue or bone.

Step (c) is a step of forming collagen fibers by salt-treating the porous layer formed in step (b) or the collagen membrane combining the dense layer and the porous layer.

The salt treatment may be performed for 2 to 24 hours at a temperature range of 10 to 37° C., but is not limited thereto. If the salt treatment temperature exceeds the above range, denaturation of collagen may occur.

The salt solution may include one selected from the group consisting of phosphate, calcium salt, sodium salt, and combinations thereof, but is not limited thereto.

The salt solution may be a PBS (Phosphate Buffer Saline) solution, but is not limited thereto.

The concentration of the salt solution may be 6-14M. For example, 6M, 6.5M, 7M, 7.5M, 8M, 8.5M, 9M, 9.5M, 10M, 10.5M, 11M, 11.5M, 12M, 12.5M, 13M, 13.5M, 14M, or two of these values. It can be a value between . If the concentration of the salt solution is less than the above range, fiberization of the porous layer may not proceed, or may only partially proceed and the desired degree of fiberization may not be achieved, and the formability of the graft material may be reduced accordingly. If the concentration of the salt solution exceeds the above range, the high concentration of salt ions may interfere with cross-linking between collagen molecules, thereby reducing the degree of fibrosis.

The step (d) is a step of drying the salt-treated collagen membrane to obtain a graft material.

The drying may be performed for 24 to 48 hours so that the moisture content in the transplant material does not exceed 10%, but is not limited to this.

Step (d) may be a freeze-drying step, but is not limited thereto. The freeze-drying may be a method of sublimating and removing moisture by reducing atmospheric pressure at a low temperature below room temperature. These freeze-drying conditions are not limited as long as the temperature and pressure allow ice to sublimate.

After step (d), (e) radiation sterilization at 10-40kGy may be further included. For example, step (e) may be a gamma ray sterilization step at 20 kGy, but is not limited thereto.

dental implant material

A dental implant material according to another aspect of the present specification includes a dense layer formed by purifying the pericardium or peritoneum of a cow or pig; and a porous layer containing collagen.

The dental implant material may be manufactured by the method of manufacturing the dental implant material described above, but is not limited thereto.

The dental implant material may be a multilayer dental implant material comprising one or more dense layers and one or more porous layers. For example, it may be a double-layer, triple-layer, or quadruple-layer dental implant material, but is not limited thereto.

The porosity of the porous layer may be 80-90%. For example, it may be 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, or a value between two of these values. If the porosity of the porous layer is less than the above range, the absorbency of the porous layer may decrease and the decomposition rate may slow down, thereby reducing the regeneration rate of soft tissue or bone. If it exceeds the above range, the physical properties of the porous layer may decrease or the decomposition rate may become too fast. The ability to induce soft tissue or bone regeneration may be reduced.

The average size of pores included in the porous layer may be 260 to 350 μm. For example, it may be 260 μm, 270 μm, 280 μm, 290 μm, 300 μm, 310 μm, 320 μm, 330 μm, 340 μm, 350 μm, or a value between two of these values. If the average pore size of the porous layer is less than the above range, the absorbency of the porous layer decreases and the decomposition rate slows, which may reduce the regeneration rate of soft tissue or bone. If it exceeds the above range, the physical properties of the porous layer may decrease or the decomposition rate may decrease. If it becomes too fast, the ability to induce soft tissue or bone regeneration may be reduced.

The thickness ratio of the dense layer and the porous layer may be 1:3 to 1:7. For example, it may be 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, or a ratio between two of these. . If the thickness ratio of the porous layer to the dense layer is less than the above range, the decomposition speed of the porous layer may become too fast, which may reduce the ability to induce regeneration of soft tissue or bone. If it exceeds the above range, the decomposition period of the porous layer may be prolonged, resulting in regeneration of soft tissue or bone. Speed may decrease.

The thickness of the dense layer may be 0.1 to 1 mm. For example, it may be 0.1mm, 0.2mm, 0.3mm, 0.4mm, 0.5mm, 0.6mm, 0.7mm, 0.8mm, 0.9mm, 1mm, or a value between two of these values, but is not limited thereto. It can be determined depending on the thickness of the pericardium or peritoneum of the cow or pig that underwent the procedure.

The thickness of the porous layer may be 0.3 to 7 mm. For example, it could be 0.3mm, 0.5mm, 1mm, 1.5mm, 2mm, 2.5mm, 3mm, 3.5mm, 4mm, 4.5mm, 5mm, 5.5mm, 6mm, 6.5mm, 7mm, or any value between these two values. However, it is not limited to this.

The dense layer and the porous layer may have different decomposition periods. The dense layer has a longer decomposition period than the porous layer, which can lead to a long maintenance period when applied to the body, and the porous layer has a shorter decomposition period than the dense layer, which can induce rapid regeneration of soft tissue or bone.

The dense layer has excellent mechanical properties and shielding ability, and gradually decomposes while maintaining sufficient volume during the desired function maintenance period, thereby blocking the invasion of surrounding soft tissue or bacteria, securing space for bone regeneration, and increasing the maintenance period. can be performed.

The porous layer can absorb blood and decompose quickly, leading to rapid regeneration of soft tissue or bone.

When the dense layer of the dental implant material is used in contact with the bone, it can be applied as a dental shield capable of open surgery. In this case, the dense layer is maintained for a long period of time to serve as a barrier and is porous. The layer can speed up healing of the upper soft tissue and prevent exposure of the shield, thereby lowering the risk of infection from the outside.

When the porous layer of the dental implant material is used in contact with bone, it can be applied as a soft tissue regeneration substitute for gingival augmentation or root flap surgery, and by not performing autologous tissue collection, the time the patient stays at the dentist (Chair-time) can reduce and alleviate the patient's pain.

Hereinafter, embodiments of the present specification will be described in more detail. However, the following experimental results describe only representative experimental results among the above examples, and the scope and content of the present specification cannot be interpreted as being reduced or limited by the examples. Each effect of various implementations of the present specification that are not explicitly presented below will be described in detail in the corresponding section.

Example

To purify bovine pericardium, it was washed with water, sterilized with a 70% ethanol solution, treated with a base of sodium hydroxide solution at pH 11, washed with water, treated with an acetic acid solution at pH 2.7, and then pH Washed with water until 6.5-7.0 was obtained. Afterwards, it was treated with trypsin enzyme at 37°C, oxidized with 30% aqueous hydrogen peroxide, and washed with water to obtain a dense layer.

A collagen solution was prepared by adding insoluble collagen derived from bovine tendon to a hydrochloric acid solution of pH 2.0 at a concentration of 1.0 wt%, and then hydrated for about 1 hour. It was ground for about 5 minutes using a blender, placed in a centrifuge, and centrifuged at 4000 rpm for 20 minutes to remove air bubbles. Afterwards, the collagen swelling liquid from which air bubbles were removed was poured into the mold to form a porous layer, and a dense layer was placed on top of it and then frozen.

The frozen collagen swelling liquid was treated with salt by adding a 10M Phosphate Buffer Saline (PBS) solution at a volume ratio of 1/2 that of the swelling liquid.

After washing the salt-treated collagen membrane, it was pre-frozen at -60°C for 1 hour and freeze-dried at -86°C and 5 torr for 24 hours. The dried collagen graft material was sterilized by gamma rays at 20 kGy to obtain the graft material.

Comparative Example 1

A graft material was manufactured in the same manner as in the above example, except that insoluble collagen was added to the hydrochloric acid solution at a concentration of 0.5 wt%.

Comparative Example 2

A graft material was manufactured in the same manner as in the above example, except that insoluble collagen was added to the hydrochloric acid solution at a concentration of 1.5 wt%.

Comparative Example 3

A graft material was manufactured in the same manner as in the above example, except that insoluble collagen was added to the hydrochloric acid solution at a concentration of 2.0 wt%.

Experimental Example 1

In order to confirm the change in pores of the porous layer depending on the concentration of the collagen solution, the graft materials prepared in Examples and Comparative Examples 1 to 3 were frozen using liquid nitrogen, then cut, and the cross-section of the graft material was examined with a scanning electron microscope ( This was confirmed through Scanning Electron Microscope (SEM). SEM images of cross-sections of each of the graft materials prepared in Examples and Comparative Examples 1 to 3 are shown in Figure 1 (Figure 1(a): Example, Figure 1(b): Comparative Example 1, Figure 1 ( c): Comparative Example 2, Figure 1(d): Comparative Example 3).

Referring to Figure 1, it was confirmed that the porous layer of the transplant material prepared in the example had an appropriate pore size and that the pore size was uniform. On the other hand, it was confirmed that the pores of the porous layer of the transplant material prepared in Comparative Example 1 were somewhat large in size, and the pores of the porous layer of the transplant material prepared in Comparative Examples 2 and 3 were confirmed to be small in size. In addition, the pores of the porous layer of the graft materials prepared in Comparative Examples 1 and 2 were irregular in size and structurally unstable.

The thickness ratio of the dense layer and the porous layer of the transplant material prepared in Examples was 1:5.85, and the thickness ratios of the dense layer and porous layer of the transplant material prepared in Comparative Examples 1 to 3 were 1:6.36 and 1:3.51, respectively. , 1:3.52.

Experimental Example 2

In order to confirm the change in pores of the porous layer according to the concentration of the collagen solution, mercury was injected into the graft materials prepared in Examples and Comparative Examples 1 to 3 by changing pressure, and the porosimeter was injected into the porous layer using a mercury intrusion method (Porosimeter). The porosity and average pore diameter were measured. The results are shown in Table 1 below.

division Example Comparative Example 1 Comparative Example 2 Comparative Example 3 Porosity (%) 88 92 78 72 Average pore size (μm) 280 362 256 252

Referring to Table 1, it was confirmed that the porous layer of the graft material prepared in the Examples satisfied a porosity of 80 to 90% and an average pore size of 260 to 350 μm, and had the most appropriate porosity and pore size. The porous layer of the graft material prepared in Comparative Example 1 using a low-concentration collagen solution was confirmed to have a higher porosity and larger pore size than the Example, and the graft material prepared in Comparative Examples 2 and 3 using a high-concentration collagen solution It was confirmed that the porous layer had a low porosity and small pore size compared to the example.

Experimental Example 3

The difference in physical properties between the dense layer and the porous layer of the graft material prepared in the above example was evaluated through a suture pull-out test. The suture pulling method is a method of measuring the strength of the suture to tear the sample when a suture (3-0 nylon) is hung on the top of a sample of a certain size and pulled to both sides, and the dense layer and porosity of the graft material prepared in the above example are measured. The suture strength of each layer and graft material containing two layers was measured. The results are shown in Figure 2.

Referring to Figure 2, it was confirmed that the dense layer showed the highest sealing strength, and the porous layer had a very low sealing strength. In addition, it was confirmed that the overall physical properties of the implant material (D+P layer) including the dense layer and the porous layer depend on the dense layer.

Experimental Example 4

In order to confirm the difference in the decomposition period between the dense layer and the porous layer of the transplant material prepared in the above example, the decomposition resistance of each layer was evaluated using collagenase, a collagen decomposing enzyme.

After separating the dense layer and the porous layer of the transplant material prepared in the above example and preparing a sample by cutting each layer into about 10 mg, 2 ml of 0.05 M calcium chloride solution (pH 7.4) dissolved in Tris buffer solution was added and 37 It was pretreated for 1 hour in a constant temperature incubator at ℃. Afterwards, a collagenase solution containing collagenase added to 0.1 M Tris-HCl at a concentration of 50 Unit/ml was added at a rate of 1 ml per 5 mg of sample, and then reacted in a constant temperature incubator at 37°C. Weight changes over time were measured. The weight of the samples was measured every hour by washing each sample 2-3 times with purified water and then lyophilizing it. Collagenase derived from Clostridium histolyticum, which has a high decomposition rate, was used. The results are shown in Figure 3.

Referring to Figure 3, the porous layer was completely decomposed within 1 to 3 hours, and the dense layer was completely decomposed within 8 to 10 hours, confirming that the decomposition period of the two layers was different. did.

The description of the present specification described above is for illustrative purposes, and a person skilled in the art to which an aspect of the present specification pertains can easily transform it into another specific form without changing the technical idea or essential features described in the present specification. You will be able to understand it. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive. For example, each component described as single may be implemented in a distributed manner, and similarly, components described as distributed may also be implemented in a combined form.

The scope of the present specification is indicated by the claims described below, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be construed as being included in the scope of the present specification.

Claims (11)

(a) Purifying the pericardium or peritoneum of cattle or pigs to form a dense layer;
(b) forming a porous layer using a collagen solution;
(c) salt treatment by adding a salt solution; and
(d) drying; a method of manufacturing a dental implant material, including the step. According to paragraph 1,
The purification in step (a) is performed by one or more of washing, sterilization, base treatment, acid treatment, enzyme treatment, and oxidation treatment. According to paragraph 1,
The collagen solution is a method of manufacturing a dental implant material comprising soluble collagen derived from pork skin, insoluble collagen derived from bovine tendon, or a combination thereof. According to paragraph 1,
Method for producing a dental implant material, wherein the concentration of the collagen solution is 0.6 to 1.4% by weight. According to paragraph 1,
The salt solution includes one selected from the group consisting of phosphate, calcium salt, sodium salt, and combinations thereof. According to paragraph 1,
A method of manufacturing a dental implant material, wherein the concentration of the salt solution is 6 to 14M. A dense layer formed by purifying the pericardium or peritoneum of cattle or pigs; and
A dental implant material comprising a porous layer containing collagen. In clause 7,
A dental implant material wherein the porous layer has a porosity of 80 to 90%. In clause 7,
A dental implant material, wherein the average size of the pores included in the porous layer is 260 to 350 μm. In clause 7,
A dental implant material, wherein the thickness ratio of the dense layer and the porous layer is 1:3 to 1:7. In clause 7,
A dental implant material, wherein the dense layer and the porous layer have different decomposition periods.