KR20240057836A - A porous membrane comprising a collagen and a polycarprolacton for regenerating the periodontal complex having improved healing characteristics, and method for preparing the same - Google Patents

Abstract

The present invention relates to a scaffold for porous periodontal tissue regeneration with improved healing properties containing collagen and polycaprolactone and a method for manufacturing the same. More specifically, the periodontal tissue support of the present invention can be injected into the body without side effects, and It relates to a support for porous periodontal tissue regeneration that is decomposed and absorbed in vivo after a certain period of time. It effectively provides pores, which are effective spaces for tissue growth into the existing interior and delivery of new bone, and has high bone regeneration ability, allowing bone regeneration in a short period of time. This is possible.

Description

Shielding membrane for porous periodontal tissue regeneration with improved healing properties containing collagen and polycaprolactone and method for manufacturing the same }

The present invention relates to a shielding membrane for regenerating porous periodontal tissue with improved healing properties containing collagen and polycaprolactone and a method of manufacturing the same.

Bone tissue is the only hard tissue in the living body, and new bone is created by filling bone tissue damaged by trauma, tumor, deformity, or physiological phenomena with bone material. A common method used to recover such bone defects is using bone tissue in other areas. There are autogenous bone transplantation methods in which part of one's own bone is harvested and transplanted, allogeneic bone transplantation methods in which another person's bone is chemically treated and then transplanted, and xenograft bone transplantation methods in which non-human animal bone is chemically treated and then transplanted.

In particular, shielding membranes are used to prevent invasion of bone tissue and skin tissue after bone tissue transplantation, and existing collagen shielding membranes have disadvantages in terms of tensile strength and decomposition period.

Accordingly, there is a need for a shielding material that can achieve sufficient strength, has no possibility of disease transmission, has excellent biocompatibility with performance that can replace existing shielding films, and can be properly absorbed and replaced with regenerated bone when transplanted.

Biodegradable polymer shielding films such as polycaprolactone (PCL) and polylactic acid (PLLA), developed in response to these needs, have excellent physical properties but have the disadvantage of lacking biocompatibility.

Among polymer materials with these characteristics, bioactive polymers include polymer materials extracted from living organisms such as hyaluronic acid and collagen, and natural polymer membranes such as amniotic membrane. Various polymer compounds have been developed as artificial shielding materials, and collagen membranes have been commercialized as shielding membrane materials and are available in various products. The collagen shielding membrane, which has excellent biostability and bone conduction performance, naturally blends with the tissue after transplantation into the bone defect area. It restores the bone shoulder area by biochemically combining with the remaining bone. However, due to its relatively weak tensile strength and short decomposition period, the need for a biodegradable shielding membrane that maintains strength after implantation and induces new bone formation while slowly biodegrading and being absorbed has emerged. Therefore, in response to these demands, the use of a cross-linked collagen membrane, which has high tensile strength, a long decomposition period in the human body, and has a tendency to be completely absorbed by surrounding tissues, and has chemical properties similar to collagen membranes, has been proposed, and is used in the orthopedic and dental fields. It is widely used as a hard tissue replacement material. This cross-linked collagen membrane has the advantage of being decomposed and absorbed within a few weeks after restoration of the bone defect, so it is applied to various products.

Meanwhile, the shielding membrane having the above-mentioned characteristics is a shielding membrane material that can be used safely because it has almost no side effects on surrounding tissues, but due to the nature of the biomaterial, it has a risk of immune response and infection.

In addition, the most important components of the shielding film are strength and decomposition time, which have been improved over the past, but have clear limitations. Therefore, it is necessary to change the material or improve the structure to adjust the tensile strength or decomposition period.

Also, due to its low tensile strength, it has the disadvantage of not being able to be used in areas that experience a lot of shear stress after transplantation or in wide areas, so improvements are needed in this regard.

The present invention continued research to solve the problems of the existing shielding membrane for periodontal tissue regeneration, which was the problem of low tensile strength of the material and the short decomposition period.

Accordingly, by mixing collagen, a previously used material for supporting tissue regeneration, and polycaprolactone, a synthetic polymer material, at a specific content and then performing a freeze-drying process under specific conditions, high porosity and pores of appropriate size are secured, and strong tensile strength is achieved. The present invention was completed after confirming that a periodontal tissue shielding film was formed that can be applied to areas that are subject to a lot of shear stress during bone regeneration or to a wide range of areas.

Therefore, the purpose of the present invention is to provide a periodontal tissue support composition composed of polycaprolactone as a structural material for collagen hardening as a new bone regeneration inducer.

Another object of the present invention is (a) solutionizing the periodontal regeneration support composition; (b) mixing the solutionized support composition at low temperature to form a slurry; And (c) to provide a method of manufacturing periodontal regeneration tissue comprising the step of freeze-drying the slurry composition.

The periodontal tissue support of the present invention relates to a support for porous periodontal tissue regeneration that can be injected into the body without side effects and is decomposed and absorbed in the body after a certain period of time. It provides an effective space for tissue growth into the existing interior and delivery of new bone. It effectively provides pores and has a high bone regeneration ability, enabling bone regeneration in a short period of time.

Figure 1 is an SEM photograph of Example 1-1 of the present invention.
Figure 2 is an SEM photograph of Example 2-1 of the present invention.
Figure 3 is an SEM photograph of Example 3-1 of the present invention.
Figure 4 is an SEM photograph of Example 4-1 of the present invention.
Figure 5 is an SEM photograph of Example 5-1 of the present invention.
Figure 6 is an SEM photograph of Example 6-1 of the present invention.
Figure 7 is an FT-IR graph of Example 2-1 of the present invention.
Figure 8 is a photograph showing the cell proliferation test results of Examples 2-1, 3-1, 4-1, 5-1, and 6-1 of the present invention.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention is,

A periodontal regeneration support consisting of a structural material and a new bone regeneration inducing material, wherein the structural material includes polycaprolactone, and the new bone regeneration inducing material includes collagen.

In particular, the weight ratio of polycaprolactone:collagen = 1:0.09 ~ 0.11, more preferably the weight ratio of polycaprolactone:collagen = 1:0.1, can exhibit excellent cell proliferation, and is a porous periodontal structure with improved healing properties. A retardant for regeneration can be provided.

The collagen has a molecular weight within the range of 5 to 15 kDa, more preferably within a molecular weight of about 10 kDa, and not only has a desirable decomposition period, but also exhibits high tensile strength suitable for the role of a support and has excellent cell proliferation properties.

The porous structure of the scaffold for periodontal tissue regeneration of the present invention is preferably in the form of a sponge.

Another aspect of the present invention is,

A method of manufacturing a periodontal regeneration support consisting of a structural material and a new bone regeneration inducing material,

(a) solutionizing the periodontal regeneration support composition;

(b) mixing the solutionized support composition at low temperature to form a slurry; and

(c) including the step of freeze-drying the slurry composition,

The step (a) is characterized by mixing polycaprolactone and collagen in an organic solvent and forming a solution at a weight ratio of polycaprolactone:collagen = 1:0.9 to 1.1.

The organic solvent may be one or more selected from acetic acid, chloroform, dichloromethane, tetrahydrofuran, dimethylformamide, and dimethylsulfoxide. may be selected, and acetic acid may be preferably selected.

The freeze-drying step (c) above may affect the final physical properties of the support. Unlike general drying, freeze-drying does not change the volume of the material even when the solvent is volatilized and dried, and the space where the solvent was left remains as a void, creating uniform and high-quality pores. It may be advantageous to secure .

In the present invention, the freeze-drying conditions are not particularly limited, and a person skilled in the art can set appropriate freeze-drying conditions, but it is preferable to proceed under vacuum conditions of -40°C and 5 mTorr, and 24 to 24 mTorr. It may be conducted for 122 hours, more preferably 48 to 96 hours, and even more preferably 72 to 96 hours.

The present invention will be described in detail below through examples. However, it should be made clear that the following examples are only for detailed description of the invention and are not intended to limit the scope of rights.

Example

Selection of polycarprolactone (PCL: Polycarprolactone) molecular weight

It was confirmed that the decomposition period of polycaprolactone (PCL) is different depending on the molecular weight. Accordingly, in order to select polycaprolactone suitable for the purpose of the support for porous periodontal tissue regeneration of the present invention, polycaprolactone sponge was manufactured and the decomposition period was It was measured using a common method in the technical field.

The results were as shown in Table 1.

Molecular weight (Da) Decomposition period PCL sponge 10,000 6 months 20,000 8 months 40,000 1 year 80,000 1 year and 6 months

Considering various conditions such as the implant surgery period and cell regeneration time, the decomposition period of PCL forming the support skeletal structure can be considered to be approximately 6 months to 1 year, and is preferably around 8 months. Polycaprolactone (PCL) of approximately 20,000 was selected and used.

Manufacturing of collagen and polycarprolactone (PCL) solution

In the present invention, the components were mixed according to the ratios shown in Table 2 below and homogenized by stirring in an acetic acid solvent at 25°C for one day.

(1) Structural material: Polycaprolactone (PCL) was used as a material that prevents loss of new bone regeneration material and provides self-hardening.

(2) Organic solvent: Acetic acid was used to prepare a solution for preparing a support for periodontal regeneration.

(3) New bone regeneration material: Collagen was used as a material for the alveolar bone regeneration effect, and five types of materials with different molecular weights were used.

Example Acetic acid (mL) Polycaprolactone (mg) Collagen (mg)* 1-1 40 200 40 (1kDa) 1-2 40 (3kDa) 1-3 40 (5kDa) 1-4 40 (10kDa) 1-5 40 (30kDa) 2-1 400 40 (1kDa) 2-2 40 (3kDa) 2-3 40 (5kDa) 2-4 40 (10kDa) 2-5 40 (30kDa) 3-1 800 40 (1kDa) 3-2 40 (3kDa) 3-3 40 (5kDa) 3-4 40 (10kDa) 3-5 40 (30kDa) 4-1 1200 40 (1kDa) 4-2 40 (3kDa) 4-3 40 (5kDa) 4-4 40 (10kDa) 4-5 40 (30kDa) 5-1 1600 40 (1kDa) 5-2 40 (3kDa) 5-3 40 (5kDa) 5-4 40 (10kDa) 5-5 40 (30kDa) 6-1 2000 40 (1kDa) 6-2 40 (3kDa) 6-3 40 (5kDa) 6-4 40 (10kDa) 6-5 40 (30kDa) * Five types of collagen with different molecular weights were used, and the number of collagen with different molecular weights was the branch number of each example (1: 1kDa, 2: 3kDa, 3: 5kDa, 4: 10kDa, 5: 30kDa).

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 12 to 72 hours. The organic solvent added to the polycaprolactone was acetic acid and chloroform ( chloroform, dichloromethane, tetrahydrofuran, dimethylformamide, or dimethylsulfoxide can be used, but acetic acid is preferred.

Low-temperature mixing of polycarprolacton (PCL) and collagen

In order to slurry the periodontal regeneration support prepared above, each of the examples was added to a 100 ml glass bottle and then stirred at a speed of 300 to 500 rpm for 12 to 72 hours at 4°C.

Freeze drying

The frozen periodontal regeneration support solution prepared above was freeze-dried under vacuum conditions of -40°C and 5 mTorr for 72 to 96 hours using a freeze dryer to give sponge-shaped pores.

Surface confirmation via SEM and FT-IR

The porosity of the manufactured periodontal regeneration scaffold was confirmed through SEM.

When the ratio of collagen to polycaprolactone in Examples 1-1 to 1-5 was less than 50%, collagen could not be bound, and a powder form, not a sponge form, appeared (see Figure 1).

On the contrary, in Examples 2-1 to 2-5, 3-1 to 5, 4-1 to 4-5, 5-1 to 5-5, and 6-1 to 6-5, a sponge-type pore structure appeared ( 2 to 6). However, there were some differences between the examples in terms of average pore size, porosity, and pore shape, which is likely to affect cell proliferation, which will be described later.

Meanwhile, in order to confirm that the manufactured periodontal regeneration support was a well-mixed support, the components were confirmed through FT-IR analysis (Figure 7).

The peaks seen at 2900 and 1700 show the presence of PCL, and the peaks seen at 1500 were confirmed, confirming the presence of collagen. This shows that the mixing was good.

Confirmation of cell proliferation

In order to confirm the cell proliferation of the manufactured periodontal regeneration scaffold, a cell experiment was performed. However, in Examples 1-1 to 1-5, it was in powder form and was excluded from cell experiments.

The results were as shown in FIG. 8, and it was confirmed that cell proliferation was the best in Example 2-1 and tissue differentiation occurred only after the 3rd day. As a result, it was confirmed that the ratio of polycaprolactone:collagen = 1:0.1 was the most desirable in terms of cell proliferation.

Check tensile strength and decomposition period

Material properties such as tensile strength and decomposition period, which are important properties for commercialization, were measured for Examples 2-1 to 2-5, which were the most effective in terms of cell proliferation.

The measurement results were as shown in Table 3 below.

Example Molecular weight [kDa] Tensile strength [MPa] Decomposition period [weeks] cell proliferation [%] 2-1 One One One 120 2-2 3 2 2 130 2-3 5 3 4 140 2-4 10 6 12 160 2-5 30 6 22 130

As shown in Table 3, the tensile strength of collagen tends to increase with molecular weight, but is not directly proportional to it. It was confirmed that as the molecular weight increases, the effect of increasing the tensile strength due to the increase in molecular weight gradually decreases. In addition, the molecular weight is about 10 kDa. It is suitable for use as a cell support both internally and externally, and it was confirmed that 10 kDa with a decomposition period of 12 weeks is most suitable for use as a bone graft material.

Claims (1)

In the periodontal regeneration support composed of structural material and new bone regeneration inducing material,
The structural material includes polycaprolactone,
The new bone regeneration inducer includes collagen,
Polycaprolactone: Collagen = 1: 0.09 ~ 0.11 weight ratio,
The support is a shielding membrane for periodontal tissue regeneration, characterized in that it has a porous structure.