WO2013141519A1

WO2013141519A1 - Gingival extension device having periodontal tissue regeneration function

Info

Publication number: WO2013141519A1
Application number: PCT/KR2013/002048
Authority: WO
Inventors: 김민경; 신승화; 최다미; 송향도; 정용일; 송주동; 엄태관; 최규옥
Original assignee: 오스템임플란트주식회사
Priority date: 2012-03-23
Filing date: 2013-03-14
Publication date: 2013-09-26

Abstract

The present invention provides a gingival extension device having a periodontal tissue regeneration function which is grafted into the gingiva before guided bone regeneration-augmentation, wherein a growth factor (GF) and a hydrogel are connected through a peptide linker. According to the present invention, the total implantation period can be reduced by shortening the gingival extension time to be within four weeks, and weakened gingival tissues can be restored to normal gingival tissues within a short time by rapid tissue expansion through the continuous discharge of growth factors.

Description

【Specification】

[Name of invention]

Gingival dilator with periodontal tissue regeneration

Technical Field

The present invention relates to a hydrogel-based gingival expander equipped with periodontal tissue regeneration. More specifically, rapid expansion of gingiva and regeneration of expanded gingiva are simultaneously controlled by enzymes introduced into the hydrogel. Which is about the gingival dilator.

Background Art

<2> An appropriate amount of bone is required around the implant implant as a prerequisite for successful bone adhesion and initial stability during dental implant procedures.

<3> On the other hand, if the bone amount or lack of bone volume is expected before implantation, in the case of exposing the implant according to the positioning of the implant for the manufacture of a normal prosthesis during implant placement, or for various reasons after implant placement Guided bone regeneration-augmentation (GBR) is used when some of the implants are exposed out of the bone due to alveolar bone resorption due to inflammation. The bone guided regeneration is a procedure for forming bone space by using a barrier membrane at the gingival area irrespective of periodontal tissue, and inducing osteoblast regeneration into bone defects in the lower part of the space to promote bone tissue regeneration.

In many cases, absorbent and nonabsorbable dental barriers and various bone materials (autologous bone, allogeneic bone, xenograft bone, synthetic bone) are used in this procedure. Dental barriers are known to prevent fast-growing gingival epithelium from growing into the damaged area first and to help connective tissues with differentiation and bone regeneration due to their adequate space-holding ability. Techniques for dental barrier membranes that can improve the success rate of implant procedures by helping the prognosis of regeneration have been developed.

On the other hand, lack of bone mass or bone volume before implant placement is accompanied by the contraction of the gingiva covering it, there is a problem that it is difficult to secure the space required for alveolar bone regeneration. In order to solve this problem, a technique for implanting a gingival dilator containing a hydrogel prior to a bone guided regeneration procedure to reconstruct the gingival degenerated gingiva using the swelling properties of the hydrogel to its normal state prior to tooth loss Has been developed. When the gingiva retracted in this manner is rebuilt, sufficient space for alveolar bone regeneration may be secured (see FIG. 1).

However, the prior art implants hydrogels into the retracted gingiva and expands the tissue. It takes 6 ~ 8 weeks for a long time, and the expansion effect is also not good, so the Q value (swelling volume / dry volume) is only 5 for 6 ~ 8 weeks, and it is simply concentrated on the gingival expansion technology. There was a problem that the gingival part was weak.

Therefore, it is necessary to develop a technology that can reinforce the thinned gingiva due to the expansion while reducing the gingival expansion time.

[Detailed Description of the Invention]

[Technical problem]

In order to solve the above problems, it is an object of the present invention to reduce the gingival expansion time and to provide an excellent gingival dilator up to the expansion effect.

In addition, the present invention has a maximum gingival expansion effect in a shorter period than the prior art technology to compensate for the problem that the degree of weakening of the gingival relatively large, growth factor (a weakened gingival tissue due to rapid tissue expansion) The goal is to provide a gingival dilator with tissue regeneration that restores normal gingival tissue through sustained release of growth factor (GF).

Technical Solution

<ιο> The present invention, in the gingival dilator implanted into the gingiva before the bone guided regeneration, the hydrogel to secure the gingival space through the body swelling; Gingival regeneration promoting growth factor (GF) mounted in the hydrogel; And a peptide linker having an amino acid sequence cleaved by proteolytic enzymes in the body, wherein the hydration gel and the growth factor are linked through the peptide linker, and provide a gingival dilator loaded with periodontal tissue regeneration. .

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

Here, the peptide linker connects the side chain group and the growth factor of the hydrogel polymer, and is composed of a peptide sensitive (P P sensitive peptide) to matrix metalloproteinase (匪 P). After the cleavage by the substrate matrix metalloproteinase in the body release the growth factor. Therefore, the present invention, after the peptide linker is cleaved by the substrate metalloproteinase, the hydrogel side cleavage is C00H, S0 ₃ H,

It provides a gingival dilator equipped with periodontal tissue regeneration, characterized in that P0 ₃ H _2l or NH ₃ ⁺ functionalization. On the other hand, the hydrogel serves to secure the gingival space through swelling in the body, and in one embodiment a monomer having a methacrylate group and a monomer having a vinyl group Crosslinked hydrogels can be used.

The growth factor may be epidermal growth factor (EGF), fibroblast growth factor (FGF), convertible enteric factor (TGF-beta), platelet-derived growth factor (PDGF), and vascular endothelial growth factor (VEGF) insulin. Growth factor (IGF-1), thioredoxin (TX), stem cell factor (SCF), HGF (hepatocyte proliferation factor), human growth hormone or angiogenin. Preferably, it is epidermal growth factor (EGF).

<15> The peptide linker is GPQGi IAGQ, GPQG I IWGQ, VPMS1MRGG, IPVSILRSG,

RPFS1MIMG, VPLS LTMG, VPLSlLYSG, IPESiLRAG, SGESPAYlYTA, GPLGiLWAR, PVG UIG, PUULAG, characterized in that it has at least one amino acid sequence selected from the group consisting of. (1: portion where the amino acid sequence is broken by 匪 P)

In addition, various linking compounds may be used to link the peptide linker with the hydrogel and growth factor. In one embodiment, 1-ethyl-3- (3-dimethylaminoprop) carbodiimidehydrochloride may be used. (EDC), N-hydroxysulfosuccinimide (sul fo- NHS), dicyclonucleosilcarbodiimide (DCC), 1-cyclonucleosil-3-3 (imide than 2-morpholinoethylcar) (CMC Can be used or any combination thereof.

In the linking compound, the hydrogel or growth factor and the peptide linker are linked to each other through an acyl substitution reaction, so that a poor leaving group such as _0H may be used as a good leaving group. It helps to react well by converting to leaving group, and is only involved 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 being released through a linker cleavage reaction of the substrate metalloproteinase, and the gingival expansion period including 1 to 2 weeks before the release is 3 to 4 total. Lord.

When the peptide linkage of the peptide linker is interrupted by the substrate metalloproteinase introduced into the hydrogel, release of the growth factor is initiated and at the same time the hydrogel side cleavage portion

C00H S0 ₃ H, P0 ₃ H ₂ , or NH ₃ ⁺ functionalization increases the repulsion between hydrogel polymers. Increased repulsive force widens the hydration space in the hydrogel, which is expressed in the form of a secondary swell followed by a conventional primary swell. In this case, the Q value (swelling volume / drying volume) of the self-loaded hydrogel loaded with the growth factor through the peptide linker is the substrate metalloproteinase. It is further increased during the enzyme action, and the Q value is characterized by 5-10.

Advantageous Effects

The gingival expander of the present invention shortens the expansion time (3-4 weeks), and has an excellent expansion effect.

(Q value is 5 ~ 10), and it has tissue regeneration function that restores weakened gingival tissue to normal gingival tissue due to rapid tissue expansion through sustained release of growth factors regulated through enzyme reaction in the body. Therefore, the present invention has an excellent effect that not only the gingival expansion but also the regeneration of the expanded gingival to normal tissue form at the same time. [Brief Description of Drawings]

Figure 1 shows the gingiva is expanded when a conventional gingival dilator is implanted.

Figure 2 shows the components of the gingival dilator of the present invention.

Figure 3 shows a schematic diagram of the gingival dilator of the present invention before and after the injection of 匪 P.

4 is a graph comparing the swelling characteristics of the present invention and the conventional hydrogel.

5 shows the appearance of the gingival expander of the present invention before swelling (day 0), after primary swelling (13 days) and after secondary swelling (30 days).

Figure 6 is a graph of the emission pattern of the growth factor of the present invention.

For reference, FIGS. 4 and 6 are graphs showing results of the same sample.

[Best form for implementation of the invention]

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

The gingiva dilator of the present invention is a gingival dilator which is implanted into the gingiva prior to guided bone regeneration (GBR), a hydrogel that secures the gingival space through swelling of the body, and is mounted in the hydrogel A gingival regrowth growth factor (GF), and a peptide linker having an amino acid sequence cleaved by proteolytic enzymes in the body, wherein the hydrogel and the growth factor are linked through the peptide linker It is characterized by.

<30>

<3i> The conventional gingival dilator takes a long time for 6-8 weeks to expand the tissue, and the expansion effect is also not good, and the Q value (swelling volume / drying volume) is only 5 for a period of 6-8 weeks. The problem is that the gingival area of the gingiva is weakened by focusing only on the gingival expansion technique.

Complementing this problem of the prior art, the present invention provides the maximum value in a short period of time. Is characterized by having a tissue regeneration function to restore and strengthen the gingival tissue through the expansion effect and the continuous release of growth factors.

In order to achieve this dual effect, the gingiva dilator of the present invention is a hydrogel that secures the gingival space through swelling in the body, a growth factor (GF) that promotes gingival regeneration, and a proteolytic enzyme in the body. A peptide linker having an amino acid sequence to be cleaved, wherein the growth factor and the hydrogel are linked via the peptide linker.

Subsequently, when the matrix metalloproteinase is introduced into the hydrogel, the peptide linker is cleaved to release the growth factor, and at the same time, the secondary hydrogel increases in addition to the primary hydrogel swelling before the enzyme inflow. Swelling begins. The secondary swelling is due to the cleavage of the peptide linker by the substrate metalloproteinase, followed by hydrogel side cleavage addition functionalization, thereby increasing the repulsive force between the hydrogel polymers.

In addition, growth factors released by cleavage of the peptide linker may promote cell proliferation of epithelial cells and endothelial cells, promote cell proliferation of fibroblasts that synthesize collagen, which is a component of the dermis, and blood vessels at the site of skin damage. Fibronectin, a substance that induces the release of angiogenesis and other regeneration promoters, and is a substance that orients skin tissue in order

It plays a key role in gingival expansion and regeneration, including the promotion of fibronectin. The growth factor has a release period of about 14 days.

On the other hand, the hydrogel used in the present invention serves to secure the gingival space through swelling in the body, various hydrogels that can cause primary swelling in the body may be used, preferably methacrylic Hydrogels crosslinked with monomers having methacrylate and monomers having vinyl groups can be used.

On the other hand, the hydrogel in which the monomer having methacrylate and the monomer having a vinyl group is crosslinked is methyl methacrylate (MMA) and vinyl-pyridone ( VP) can be synthesized by crosslinking at a mass ratio of 6 to 7: 3 to 4, in addition to hydroxyethyl methacrylate (HEMA) and vinyl-pyrrolidone (VP), or methacrylate F carbon number, 4 , 8,12) and vinyl-pyridone (VP) can be synthesized by crosslinking. (= 4 butyl methacrylate, n = 8 octyl methacrylate, n = 12 lauryl methacrylate)

In the past, hydrogel was mainly used as a support, but now, it may be used for drug release by adding drugs or cells to the hydrogel. This is because it is excellent in biocompatibility and easy to maintain activity of cells and drugs, especially protein drugs. In addition, growth factors used in the present invention may be epithelial growth factor (EGF), fibroblast growth factor (FGF), conversion growth factor (TGF-beta), platelet-derived growth factor (PDGF), vascular endothelial cell growth Factor (VEGF), insulin-like growth factor (IGF-1), thioredoxin (TX), stem cell factor (SCF), HGF (hepatocyte growth factor), human growth hormone or angiogenin ) to be. Preferably, it is epidermal growth factor (EGF).

On the other hand, the peptide linker used in the present invention,

It connects GF. The peptide linker is composed of a peptide sensitive to substrate metalloproteinase, and in one embodiment, GPQGi lAGQ, GPQG 1 IWGQ, VPMS4MRGG, IPVSILRSG, RPFS1MIMG, VPLSILTMG, VPLSlLYSG, IPESlLRAG, SGESPAYlYTA, GPLG1LWAR, PVG1LLI Have one or more amino acid sequences selected from the group consisting of amino acid sequences (1: amino acid sequence is cleaved by MMP)

Meanwhile, various linking compounds may be used to link the peptide linker with the hydrogel and growth factor. In one embodiment, 1-ethyl-3 ′ (3-dimethylaminopropyl) carbodiimide Hydrochloride (EDO, N-hydroxysulfosuccinimide

(sulfo-NHS), dicyclonucleosilcarbodiimide (DCC), 1-cyclonucleosil-3- (2-morpholinoethylcarbodiimide) (CMC), or a combination thereof. . Preferably, a combination of the above EDC and sulfo-NHS is used.

In the linking compound, the hydrogel or growth factor and the peptide linker are linked through an acyl substitution reaction, and thus, —a poor leaving group such as 0H may be used as a good leaving group ( It converts into a good leaving group and helps the reaction occur well. It only participates in the reaction process and does not contribute to the structure of the final product.

In addition, the substrate metalloproteinase is a collagenase, for example, 謹 P 1, MMP

18 or “P 13” or gelatinase, for example “P 2 or” P 9.

<44>

When the body fluid flows into the hydrogel included in the gingival expander of the present invention, the primary swelling of the hydrogel begins to expand. The primary swelling itself is a passive swelling due to the fundamental characteristics of the hydrogel polymer, and is an important process that initiates growth factor release.

That is, as the primary swelling occurs, the P in the body is introduced into the hydrogel, and the peptide linker is decomposed by the P to initiate the release of the growth factor. Number of Growth Factors Released The cleavage portion of the gel is functionalized with C00H S0 ₃ H, PO3H2, or NH ₃ , and the repulsive force between the hydrogel polymers is increased so that secondary swelling proceeds.

As such, growth factor release and secondary swelling are a series of processes that, in turn, rapidly expand gingival and regenerate gingival normal tissues.

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

<49>

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

<51>

Example 1: Synthesis of Hydrogel Polymer

Methyl methacrylate (MMA), vinylpipyrrolidone (MMA) and vinylpipyrrolidone (VP) have a mass ratio of 6-7: 3-4. VP), 2,2'-azobisisobutylonitrile (polymerization initiator) 0.1 wt%, hydroxypropyl methacrylate (crosslinker) 0.2 wt%, and methylpyridone (solvent) to combine the concentration of the total solution After blending to 0.8M, the blend was thermally polymerized at a temperature of 65 ^° C. for 24 hours. Thereafter, residues such as a solvent were removed and hydrolyzed to synthesize a hydrogel cross-linked with methyl methacrylate (XA) and vinyl-pyrrolidone (VP).

<54>

Example 2: Synthesis of "polymer-peptide linker-growth factor" hydration gem

1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) at a rate of 100 to 2 / 5-1 / 4 of the methyl methacrylate synthesized in Example 1 above -300 mg was added to connect (synthesize) the hydrogel with the peptide linker in an amount equivalent to 3 equivalents of the EDC.

In this case, the peptide linker used a general structural protein having an amino acid sequence of SEQ ID NO: 1. Then, after the separation operation to obtain a pure hydrogel-peptide linker, by condensation reaction using the base material (FGF) lug ~ lmg is connected to the peptide linker linked to the hydrogel ， "Hydraulic gel-peptide Linker-FGF (growth factor) lg was synthesized.

<58>

Example 3: Swelling test of "polymer-peptide linker-growth"

0.9% NaCl, respectively, of polymers cross-polymerized with methyl methacrylate (MMA) and vinylpyrrolidone (VP), and polymer-peptide linker growth factor of the present invention synthesized in Example 2, respectively. The solution was impregnated and subjected to swelling experiment (in vitro experiment) for 30 days at 37 ^° C. At this time, after 14 days of impregnation, 1 ug to 1 mg of 匪 P 1, 匪 P 18 or fflP 13, or a mixture thereof was added as collagenase.

In the case of the conventional hydrogel, the volume gradually increased. After about two weeks, the hydrogel increased slightly and the final Q value was 5. On the other hand, in the case of the "polymer-peptide-linked epithelial cell growth factor ¹ " hydrogel of the present invention, the final Q value was measured to 8. As shown in Figure 4, the volume of the hydrogel before swelling of the present invention is It was 0.13 ml but the volume of the final hydrogel after the second swelling was 1.04 ml, while the gingival dilators of the present invention before swelling and after 13 and 30 days of swelling in 37 ^° C., 0.9% NaCl solution were shown in FIG. Presented.

<62>

Example 4 Emission Pattern of One Growth of the Present Invention

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

<65>

On the other hand, the above embodiments are intended to help explain the present invention, and the present invention is not limited by the above embodiments, and those skilled in the art should understand that the present invention is within the spirit scope of the present invention. Various modifications and equivalent other embodiments are possible based on the claims and the content of the invention.

【Sequence List Free Text】

<67> Amino acid sequence cleaved by proteolytic enzymes in the peptide linker (sequence of 1 inker peptide)

<68> 1.Gly Pro Gin Gly lie Ala Gly Gin

2.Gly Pro Gin Gly lie Trp Gly Gin

3.Val Pro Met Ser Met Arg Gly Gly

<7i> 4.He Pro Val Ser Leu Arg Ser Gly _// : _/ O ₈ ss _o 2 ₀ 2Ml _> d 6 _{Ϊ £ Ϊ} 320ΖAV

1s¾ _ll J _Ι Λ9 ¾ρ _J n¾ n¾ P _J ΛνGHVs _I sH∞ _/ JE § ^ S _J n ₉ Λν

Claims

[Range of request]

[Claim 1]

In the gingival dilator implanted into the gingiva prior to guided bone regener at ion—augment at ion (GBR),

Hydrogel to secure the space in the gingiva through swelling in the body;

Gingival regeneration promoting growth factor (GF) mounted in the hydrogel; And a peptide linker having an amino acid sequence cleaved by a proteolytic enzyme in the body,

The hydrogel and the growth factor gingival dilator equipped with a periodontal tissue regeneration function that is connected through the peptide linker ᅳ

[Claim 2]

The method of claim 1,

The growth factor and the hydrogel are connected through the peptide linker gingival dilator with a periodontal tissue regeneration function to form a structure of the hydrogel-peptide linker-growth factor.

[Claim 3]

The method according to claim 1 or 2,

The peptide linker is cleaved by Matrix Metal loproteinase (謹 P), and the hydrogel side cleavage is C00H, S0 ₃ H, P0 ₃ H ₂ , or N¾ ⁺ functionalized to cause secondary swelling. Periodontal tissue regeneration function is characterized in that the gingival expansion.

[Claim 4]

Tax in Clause 1 or 2

The hydrogel is a gingival dilator equipped with a periodontal tissue regeneration function, characterized in that the hydrogel is a cross-linked hydrogel with a monomer having a methacrylate and a monomer having a vinyl (vinyl) group.

[Claim 5]

The method according to claim 1 or 2,

The growth factor is epithelial growth factor (EGF), fibroblast growth factor (FGF), convertible enteric factor (TGF-beta), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), insulin-like growth Factor (IGF-1), thioredoxin (TRX), stem cell factor (SCF), HGF (hepatocyte growth factor), human growth hormone or angiogenin Gum dilator equipped with a periodontal tissue regeneration function, characterized in that.

[Claim 6]

The method according to claim 1 or 2,

The peptide linker has one or more amino acid sequences selected from the group consisting of amino acid sequences of GPQGUAGQ, GPQGI I GQ, VPMS4MRGG, IPVSILRSG, RPFS MIMG, VPLSiLTMG, VPLSILYSG, IPESILRAG, SGESPAYiYTA, GPLGILWAR, PVG ILIG, PLGiLAG. Periodontal tissue regeneration function with gingival expansion. (@ ： Part where amino acid sequence is broken by MMP)

[Claim 7]

The method of claim 6,

As a linking compound for linking the peptide linker with the hydrogel and growth factor,

1-ethyl-3- (3'dimethylaminopropyl) carbodiimidehydrochloride (EDC), N-hydroxysulfosuccinimide (sulfo-NHS), dicyclonucleosilcarbodiimide (DCC), 1-cyclo A gingival dilator with a periodontal tissue regeneration function, characterized in that any one or a combination thereof is used as nuclear silo3- (2-morpholinoethylcarbodiimide) (CMC).