KR101446964B1 - Gingiva expander with the regenerating function of periodental tissue - Google Patents

Abstract

The present invention relates to a gingival expander which is implanted into the gingiva before bone induction regeneration, wherein the gingival dilator is a structure in which a growth factor (GF) and a hydrogel are connected via a peptide linker. Provides gingival expanders with tissue regeneration. According to the present invention, by shortening the gingival expansion time to 4 weeks or less, the total implantation period can be shortened and the gingival tissue weakened by rapid tissue expansion due to sustained release of growth factors can be restored to normal gingival tissues in a short period of time have.

Description

[0001] GINGIVA EXPANDER WITH THE REGENERATING FUNCTION OF PERIODENTAL TISSUE [0002]

The present invention relates to a gingival expander based on a hydrated gel loaded with a periodontal regeneration function, and more particularly, to a gingival expander based on a hydrogel-based regeneration function, .

As a precondition for successful osseointegration and initial stabilization in implant dentistry, an appropriate amount of bone is required around the implant.

On the other hand, when the shortage of bone mass or bone volume before implantation is expected, it is considered that, when the implant is implanted, the implant is exposed to the implant due to the positioning of the implant for preparing the normal prosthesis, (GBR) is used when a part of the implants is exposed to the outside of the bone. Regardless of the periodontal tissue, the bony induction regeneration regenerates the bone tissue by forming a space using a blocking membrane in the edentulous region and inducing osteoblast flow into the bone defect portion under the space.

Currently, absorbent, nonabsorbable dental shields and various bone materials (autograft, allogeneic bone, xenograft, synthetic bone) are used in many cases in this procedure. It is known that the dental barrier prevents the gingival epithelium that is early to regenerate from reaching the injured part and prevents it from entering, and has a proper space maintenance ability, which helps to differentiate connective tissue and regenerate bone. In this regard, techniques for dental restoration membranes that can improve the success rate of implant treatment by helping the prognosis of bone induction regeneration have been developed.

On the other hand, the shortage of bone mass or bone volume before implantation is accompanied by the degeneration of the gingiva covering the tooth, which makes it difficult to secure space necessary for alveolar bone regeneration. In order to solve this problem, there has been developed a technique of implanting a gingival expander including a hydrogel before the bone induction regeneration procedure and reconstructing the gingival recession, which has been regressed by using the swelling property of the hydrogel, to a normal state before tooth loss Has come. When the restored gingiva in this manner is reconstructed, a space necessary for alveolar bone regeneration can be sufficiently secured (see FIG. 1).

However, the prior art has shown that the Q value (swelling volume / dry volume) of the hydrogel is transferred to the gingival recession and extended for 6 to 8 weeks, 5, and there was also a problem in that the extended portion of the gingiva could be weakened by merely focusing on the gingival expansion technique.

Therefore, it is necessary to develop a technique for shortening the gingival expansion time and reinforcing the gingival thinning due to expansion.

In order to solve the above-mentioned problems, it is an object of the present invention to provide a gingival expander that shortens the gingival expansion time and is superior in expanding effect.

In order to solve the problem that the degree of gingival weakness is relatively increased due to the maximum gingival expansion effect in a shorter time than the conventional technique, the gingival tissue weakened by rapid tissue expansion is called a growth factor (growth factor) GF), which is capable of regenerating normal gingival tissue through continuous release of the gingival expander.

The present invention relates to a gingival expander which is implanted into a gingiva before bone induction regeneration, comprising: a hydrogel for securing a space in the gingiva through swelling in the body; A gingival regeneration promoting growth factor (GF) in the hydrogel; And a peptide linker having an amino acid sequence cleaved by an internal protease, wherein the hydrogel and the growth factor are linked via the peptide linker. .

In addition, the gingival expander of the present invention includes the structure of the polymer-peptide linker-growth factor in which the growth factor and the hydrated gel polymer are linked through the peptide linker.

Here, the peptide linker is composed of a peptide (MMP-sensitive peptide) that binds to the side chain of the hydrated gel polymer and the growth factor and is sensitive to matrix metalloproteinase (MMP) The growth factor is released. Accordingly, the present invention, the periodontal tissue, characterized in that the peptide linker is then cut by a matrix metalloproteinase protease is Hydrogels side cut vaporized COOH, SO ₃ H, PO ₃ H _2, or NH ₃ ⁺ function Featured gingival expander.

Meanwhile, the hydrated gel plays a role of securing a space in the gingiva through swelling in the body. In one embodiment, a monomer having a methacrylate group and a monomer having a vinyl group are crosslinked A hydrogel may be used.

The growth factor may be selected from the group consisting of epithelial growth factor (EGF), fibroblast growth factor (FGF), transforming growth factor (TGF-beta), platelet-derived growth factor (PDGF), vascular endothelial growth factor IGF-1), thioredoxin (TRX), stem cell factor (SCF), HGF (hepatocyte growth factor), human growth hormone or angiogenin. Preferably, it is an epithelial growth factor (EGF).

(SEQ ID NO: 1), GPQG ↓ IWGQ (SEQ ID NO: 2), VPMS ↓ MRGG (SEQ ID NO: 3), IPVS ↓ LRSG (SEQ ID NO: 4), RPFS ↓ MIMG LIGG (SEQ ID NO: 6), VPLS LYSG (SEQ ID NO: 7), IPES LRAG (SEQ ID NO: 8), SGESPAY YTA (SEQ ID NO: 9), GPLG LWAR 11) and PLG ↓ LAG (SEQ ID NO: 12). (↓: part of amino acid sequence is broken by MMP)

In addition, various linking compounds can be used to link the peptide linker with the hydrated gel and the growth factor. In one embodiment, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) N-hydroxysuccinimide One of the sulfo-NHS, dicyclohexylcarbodiimide (DCC) and 1-cyclohexyl-3- (2-morpholinoethylcarbodiimide) (CMC) May be used.

The linking compound may be a good leaving group, such as a poor leaving group, such as -OH, in which the hydrated gel or growth factor and the peptide linker are linked through an acyl substitution reaction. To help the reaction occur well. It is involved only in the reaction process and does not contribute to the structure of the final product.

The growth factor has a release period of about 14 days after releasing through a linker cleavage reaction of the substrate metalloproteinase, and the period of gingival expansion including the period of gingival spreading 1 to 2 weeks before release is 3 to 4 weeks in total.

When the peptide linkage of the peptide linker is cleaved by the substrate metalloproteinase introduced into the hydrogel, the release of the growth factor is initiated and at the same time the cleavage side of the hydrogel is reacted with COOH SO ₃ H, PO ₃ H ₂ , or NH ₃ ⁺ And the repulsive force between the hydrogel polymer is increased. The increased repulsive force expands the hydration space in the hydrated gel, which is expressed in the form of a secondary swelling following the existing primary swelling. In this case, the Q value (swelling volume / dry volume) of the hydrated gel on which the growth factor is loaded through the peptide linker is further increased when the substrate metalloproteinases are operated, and the Q value is characterized by 5 to 10 .

The gingival expander of the present invention has the advantages of shortened expansion time (3 to 4 weeks), excellent expansion effect (Q value of 5 to 10), rapid tissue expansion through sustained release of growth factors Has a tissue regeneration function that restores weakened gingival tissue to normal gingival tissue. Therefore, the present invention has an excellent effect that not only the gingival expansion effect but also the regenerating action of the extended gingiva into the normal tissue form progress together.

1 shows a state in which the gingiva is expanded when a conventional gingival expander is implanted.
Figure 2 shows the components of the gingival expander of the present invention.
Fig. 3 shows a schematic diagram of the gingival expander of the present invention before and after the MMP injection.
4 is a graph comparing the swelling characteristics of the present invention and a conventional hydrogel.
Figure 5 shows the appearance of the gingival expander of the present invention after swelling (0 day), primary swelling (13 days) and secondary swelling (30 days).
Figure 6 is a graph of the release pattern of the growth factor of the present invention.
4 and 6 are graphs showing the results of the same sample.

Hereinafter, a gingival expander equipped with the periodontal tissue regeneration function of the present invention will be described in detail with reference to the accompanying drawings.

The gingival dilator of the present invention is a gingival expander which is implanted into the gingiva prior to the guided bone regeneration-augmentation (GBR). The gingival dilator includes a hydrating gel securing a space in the gingiva through swelling in the body, A growth factor (GF), and a peptide linker having an amino acid sequence cleaved by an internal protease, wherein the hydrated gel and the growth factor are linked through the peptide linker .

Conventional gingival expanders require 6 to 8 weeks of prolonged time to expand the tissue, and the expansion effect is not excellent. Therefore, the Q value (swelling volume / dry volume) is only 5 for 6 to 8 weeks, There is a problem that the gingival portion of the extended portion is weakened by concentrating only on the technique.

In order to solve the problem of the prior art as described above, the present invention is characterized in that it has a maximum gingival expansion effect in a short period of time and has a tissue regeneration function for restoring and strengthening gingival tissue through continuous release of growth factors.

In order to achieve such a dual effect, the gingival dilator of the present invention comprises a hydrogel for securing a space in the gingiva through swelling in the body, a growth factor (GF) for promoting gingival regeneration, and an amino acid Sequence, wherein the growth factor and the hydrated gel are linked via the peptide linker.

When the substrate metalloproteinase is introduced into the hydrated gel, the peptide linker is cleaved and the growth factor is released. At the same time, the swelling of the first hydrated gel before the entry of the enzyme increases, and the swelling of the second hydrated gel . The secondary swelling is attributed to the increase in the repulsive force between the hydrated gel polymers due to the functionalization of the hydrolyzate side cleavage site after cleavage of the peptide linker by the matrix metalloproteinases.

In addition, the growth factor released by the cleavage of the peptide linker can be used for promoting cell proliferation of epithelial cells and endothelial cells, promoting cell proliferation of fibroblasts synthesizing collagen as a component of the dermis, Inducing the secretion of regeneration promoting factors, and promoting the synthesis of fibronectin, which is a substance that causes the skin tissue to form an orientation and form a net, thereby playing a key role in gingival expansion and regeneration. The growth factor has a release period of about 14 days.

Meanwhile, the hydrogel used in the present invention plays a role of securing a space in the gingiva through swelling in the body, and various hydrogels capable of causing primary swelling in the body may be used, but methacrylate ) And a monomer having a vinyl group are cross-linked with each other may be used.

Meanwhile, the hydrated gel in which the monomer having methacrylate and the monomer having vinyl group are cross-linked is prepared by mixing methyl methacrylate (MMA) and vinyl-pyrrolidone (VP) with 6 (HEMA) and vinyl-pyrrolidone (VP), or n -methacrylate ( n = number of carbon atoms, 4) ( N = 4: butyl methacrylate, n = 8: octyl methacrylate, n = 12: lauryl methacrylate) can be synthesized by crosslinking vinyl pyrrolidone (VP)

In the past, hydrated gel was mainly used as a support, but nowadays, a drug or cell is added to a hydrogel to be used as a drug release. This is because the biocompatibility is excellent and the activity of cells and drugs, particularly protein drugs, is easily maintained.

In addition, the growth factor used in the present invention may be selected from the group consisting of epithelial growth factor (EGF), fibroblast growth factor (FGF), transforming growth factor (TGF-beta), platelet-derived growth factor (PDGF), vascular endothelial growth factor (IGF-1), thioredoxin (TRX), stem cell factor (SCF), HGF (hepatocyte growth factor), human growth hormone or angiogenin. Preferably, it is an epithelial growth factor (EGF).

Meanwhile, the peptide linker used in the present invention serves to link GF with the side chain of the hydrated gel polymer. The peptide linker is composed of a peptide sensitive to a substrate metalloproteinase. In one embodiment, GPQG ↓ IAGQ (SEQ ID No. 1), GPQG ↓ IWGQ (SEQ ID No. 2), VPMS ↓ MRGG (SEQ ID No. 3), IPVS ↓ LPSG (Sequence Number 4), RPFS ↓ MIMG (Sequence Number 5), VPLS ↓ LTMG (Sequence No. 6), VPLS ↓ LYSG (Sequence No. 7), IPES ↓ LRAG (Sequence No. 8), SGESPAY ↓ YTA ), GPLG ↓ LWAR (SEQ ID NO: 10), PVG ↓ LIG (SEQ ID NO: 11) and PLG ↓ LAG (SEQ ID NO: 12). (↓: part of amino acid sequence is broken by MMP)

Meanwhile, various linking compounds can be used to link the peptide linker with the hydrated gel and the growth factor. In one embodiment, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC ), N-hydroxysulfosuccinimide (sulfo-NHS), dicyclohexylcarbodiimide (DCC) or 1-cyclohexyl-3- (2-morpholinoethylcarbodiimide) Or a mixture thereof. Preferably, a combination of EDC and sulfo-NHS is used.

The linking compound may be a good leaving group, such as a poor leaving group, such as -OH, in which the hydrated gel or growth factor and the peptide linker are linked through an acyl substitution reaction. To help the reaction occur well. It is involved only in the reaction process and does not contribute to the structure of the final product.

In addition, the substrate metalloproteinases may be collagenolytic enzymes such as MMP 1, MMP 18 or MMP 13 or gelatinase, for example MMP 2 or MMP 9.

When the body fluid is introduced into the hydrogel contained in the gingival expander of the present invention, the first swelling of the hydrogel begins and the gingiva is expanded. The primary swelling itself is a passive swelling due to the fundamental characteristics of the hydrated gel polymer, and is an important process that is the beginning of the growth factor release.

That is, as the primary swelling occurs, the MMP in the body enters the hydrogel, and the peptide linker is degraded by MMP to initiate the release of growth factors. The cleaved part of the hydrogel containing the growth factor releases COOH SO ₃ H, PO ₃ H ₂ , or NH ₃ ⁺ functionalization, and the second swelling proceeds due to an increase in intermolecular repulsive force of the hydrogel.

In this way, the growth factor release and the secondary swelling are performed in a series of processes. Through this series of processes, rapid expansion of the gingiva and regeneration of the expanded gingiva into the normal tissue are simultaneously carried out.

The growth factor is continuously released for up to two weeks to promote regeneration of the gingiva thinned due to rapid swelling and to restore the normal thickness. The gingival expansion period including the second swelling period is 3 to 4 weeks in total, and the Q value (swelling volume / dry volume) of the hydrated gel-linker-growth factor in the gingival dilator is 5 to 10. The final Q value due to secondary swelling is 8.

Hereinafter, the present invention will be described in more detail with reference to specific examples. However, these embodiments are only for describing the present invention more specifically, and the scope of the present invention is not limited by these embodiments.

Example  One: Hydrating Gel  Synthesis of polymer

Methyl methacrylate (MMA), vinyl-pyrrolidone (VP), 2-vinyl-pyrrolidone (VP) and the like are mixed so that the crosslinked methyl methacrylate (MMA) , 0.1 wt% of 2'-azobisisobutylonitrile (polymerization initiator), 0.2 wt% of hydroxypropyl methacrylate (crosslinking agent), and methylpyrrolidone (solvent) After such blending, the blend was thermally polymerized at a temperature of 65 DEG C for 24 hours. Then, the residue of the solvent and the like were removed and hydrolyzed to synthesize a hydrogel in which methyl methacrylate (MMA) and vinyl-pyrrolidone (VP) were crosslinked.

Example  2: "Polymer- Peptides  Linker-growth factor " Hydrated gel  synthesis

(EDC) in a proportion corresponding to 2/5 to 1/4 of the methyl methacrylate synthesized in Example 1 was added to 100 to 300 mg of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride To synthesize the hydrated gel and the peptide linker in an amount corresponding to 3 equivalents of the EDC.

Wherein the peptide linker uses a general structural protein having the amino acid sequence of Sequence Listing 1. Then, 1 g to 1 mg of FGF (growth factor) was ligated to the peptide linker connected to the hydrated gel through a condensation reaction using the base material after separation to obtain a pure hydrated gel-peptide linker, 1 g of "hydrated gel-peptide linker-FGF (growth factor)" was synthesized.

Example  3: "Polymer- Peptides  Linker-Growth Factor "swelling experiment

The polymer-peptide linker-growth factor of the present invention synthesized in Example 2 and the conventionally used polymer in which the methyl methacrylate (MMA) and the vinyl-pyrrolidone (VP) were crosslinked and polymerized was immersed in 0.9% NaCl solution , And the swelling test (in vitro test) was performed at 37 캜 for 30 days. At this time, 14 days after the impregnation, 1 ug to 1 mg of MMP 1, MMP 18, MMP 13, or a mixture thereof was added as a collagenase.

In the case of the conventional hydrogel, the volume gradually increased. After about 2 weeks, the volume increased slightly and the final Q value was measured to be 5. On the other hand, in the case of the "polymer-peptide linker-epithelial cell growth factor" hydrated gel of the present invention, the final Q value was measured as 8. As shown in FIG. 4, the volume of the pre-swelling hydrogel of the present invention was 0.13 ml, but the volume of the final hydrogel after the second swelling was 1.04 ml. On the other hand, the dental extender of the present invention is shown in FIG. 5 before swelling and after 13 days and 30 days of swelling in 37%, 0.9% NaCl solution.

Example  4: Release pattern of the growth factor of the present invention

As shown in FIG. 6, the growth factor of the present invention was hardly released until 14 days after injection of collagenase, but was rapidly released after 14 days after the addition of collagenase. Thus, it was confirmed that the maximum release period of the growth factor was 14 days.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, And various other modifications and equivalent embodiments are possible based on the contents of the invention.

<110> OSSTEMIMPLANT CO., LTD. <120> GINGIVA EXPANDER WITH THE REGENERATING FUNCTION OF PERIODENTAL          TISSUE <130> P2013-097 <150> 12 / 029,844 <151> 2012-03-23 <160> 12 <170> Kopatentin 2.0 <210> 1 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 1 Gly Pro Gln Gly Ile Ala Gly Gln   1 5 <210> 2 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 2 Gly Pro Gln Gly Ile Trp Gly Gln   1 5 <210> 3 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 3 Val Pro Met Ser Met Arg Gly Gly   1 5 <210> 4 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 4 Ile Pro Val Ser Leu Arg Ser Gly   1 5 <210> 5 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 5 Arg Pro Phe Ser Met Ile Met Gly   1 5 <210> 6 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 6 Val Pro Leu Ser Leu Thr Met Gly   1 5 <210> 7 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 7 Val Pro Leu Ser Leu Tyr Ser Gly   1 5 <210> 8 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 8 Ile Pro Glu Ser Leu Arg Ala Gly   1 5 <210> 9 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 9 Ser Gly Glu Ser Pro Ala Tyr Tyr Thr Ala   1 5 10 <210> 10 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 10 Gly Pro Leu Gly Leu Trp Ala Arg   1 5 <210> 11 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 11 Pro Val Gly Leu Ile Gly   1 5 <210> 12 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> sequence of linker peptide <400> 12 Pro Leu Gly Leu   1 5

Claims (7)

Prior to guided bone regeneration-augmentation (GBR), in a gingival expander implanted into the gingiva,
Hydrating gel securing space in the gingiva through swelling in the body; A gingival regeneration promoting growth factor (GF) in the hydrogel; And a peptide linker having an amino acid sequence cleaved by an internal protease,
The hydrated gel and the growth factor are connected via the peptide linker,
(SEQ ID NO: 1), GPQG ↓ IWGQ (SEQ ID NO: 2), VPMS ↓ MRGG (SEQ ID NO: 3), IPVS ↓ LRSG (SEQ ID NO: 4), RPFS ↓ MIMG LIGG (SEQ ID NO: 6), VPLS LYSG (SEQ ID NO: 7), IPES LRAG (SEQ ID NO: 8), SGESPAY YTA (SEQ ID NO: 9), GPLG LWAR 11) and an amino acid sequence of PLG ↓ LAG (SEQ ID NO: 12). (↓) A gingival expander equipped with a periodontal regeneration function. (↓: An amino acid sequence is deleted by MMP Losing part) The method according to claim 1,
Wherein the growth factor and the hydrated gel are connected through the peptide linker to form a structure of a hydrated gel-peptide linker-growth factor. 3. The method according to claim 1 or 2,
The peptide linker is the substrate metal protease: vaporizes (Matrix Metalloproteinase MMP) The after Hydrogel side cut is _{COOH, SO 3 H, PO 3} H 2, or NH ₃ ⁺ function cut by said taking place the secondary swelling A gingival expander with periodontal regeneration function. 3. The method according to claim 1 or 2,
Wherein the hydrated gel is a hydrated gel in which a monomer having methacrylate and a monomer having vinyl group are crosslinked. 3. The method according to claim 1 or 2,
The growth factor may be selected from the group consisting of epithelial growth factor (EGF), fibroblast growth factor (FGF), transforming growth factor (TGF-beta), platelet-derived growth factor (PDGF), vascular endothelial growth factor Wherein the periodontal regeneration function is characterized by being characterized by a periodontal regeneration function (IGF-1), thioredoxin (TRX), stem cell factor (SCF), HGF (hepatocyte growth factor), human growth hormone or angiogenin This mounted gingival expander. delete The method according to claim 1,
As a linking compound for linking the peptide linker with the hydrated gel and the growth factor,
(EDC), N-hydroxysulfosu NHS, dicyclohexylcarbodiimide (DCC), 1-cyclohexyl (1-cyclohexyl) ethylcarbodiimide 3- (2-morpholinoethylcarbodiimide) (CMC), or a mixture thereof is used as the gum expander.

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