CN103830115A - Application of epigallocatechin gallate in protecting dentin - Google Patents

Abstract

The invention discloses an application of epigallocatechin gallate in protecting dentin. Tea polyphenol has been added into tooth health-care products, but the specific mechanism of action has been not clear. Tea polyphenol has various functional components in different structures and effects, so that application of tea polyphenol is limited. According to the application of the EGCG (epigallocatechin gallate), which is disclosed by the invention, the activity of MMPs (Matrix Metalloproteinases) in dentin can be inhibited, the dentin is protected by maintaining the structure of a dentin collagen stent, and no related dental health-care products which act in the mechanism have been seen. The EGCG is the unique but most efficient MMPs-inhibiting component among all tea polyphenol extracts. The EGCG selected as the effective component of the dental health-care product can realize the best curative effect at the lowest drug concentration, and possible side effects are avoided.

Description

The application of EGCG fat aspect protection dentin

Technical field

The present invention relates to the application of EGCG fat aspect protection dentin.

Background technology

Oral cavity microenvironment complexity, antibacterial, enzyme and food etc. are repeatedly, long-term stimulation can cause the damage of hard tooth tissue structure.Dentin is the organizational structure containing blood vessel and cell not, can not self-regeneration as other sclerous tissueses of human body, because be rich in organic matter and collagen in structure, and can quick, progressive formation large defect time under attack.Be mainly various containing fluoride preparation for the protection of Dentinal health care prevention product at present, improve Dentinal resistance to demineralization ability by induction remineralization and play a role.Research is in recent years found, Dentinal collagen fiber web frame is the Dentinal resistance to demineralization ability of raising, maintains the natural cover for defense that Dentin Structure is complete.In dentin, comprise a large amount of matrix metalloproteinases (Matrix Metalloproteinases; MMPs); the activation of MMPs is the necessary process of dentin collagen degraded; adopt effective MMPs inhibitor; protecting dentin collagen by the activity that suppresses MMPs, is the new way that can effectively protect Dentin Structure.

Epigallocatechingallate (Epigallocatechingallate, EGCG) be the main active of the tea polyphenols that extracts from Chinese green tea, EGCG is a kind of effectively MMPs inhibitor, therefore EGCG has the effect that suppresses collagen degradation and improve the resistance to demineralization ability of dentin, at present domestic its research that is applied to gear division health product or commodity that there is not yet.

Summary of the invention

The object of the invention is to utilize the biological characteristics of EGCG; suppress targetedly the activity of MMPs in dentin by targeting; play the resistance to demineralization ability of raising dentin, protect Dentinal object, EGCG to can be applicable to develop the health-related product of the gear division such as toothpaste, collutory.

Its structural formula of EGCG fat is as follows:

Molecular formula is C22H18011, and molecular weight is 458.4 grams/mol.

The application of described EGCG fat aspect protection dentin.

EGCG fat is by share as the Dentinal gear division health product of protection containing fluoride preparation separately or with some.

The invention has the beneficial effects as follows: in dental care product, add tea polyphenols long-standing, but because its concrete mechanism of action is still not clear, and the tea polyphenols functional component that has various structures difference, effect to differ, this has all limited its application.In the present invention, the application of EGCG can suppress the activity of MMPs in dentin, reaches the Dentinal effect of protection by maintaining the structure of dentin collagen support, has not yet to see the relevant gear division health product of this machining function; EGCG is in all tea polyphenols extract, and composition unique and that the most effectively suppress MMPs, selects EGCG as the effective ingredient in gear division health product in invention, can reach best therapeutic effect with minimum drug level, and avoid possible side effect.

Accompanying drawing explanation

Fig. 1 is the inhibitory action of variable concentrations EGCG to tooth source property MMPS

Fig. 2 is the pH circulation process figure of one embodiment of the present of invention

Fig. 3 is the Surface Ultrastructure (SEM, × 1000, × 5000, × 20000 times) of the rear each processed group of PH circulation of one embodiment of the present of invention

The specific embodiment

In order to make objects and advantages of the present invention clearer, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.

This concrete enforcement comprises the steps:

1) determine that EGCG suppresses the valid density of MMPs in dentin

Each sample takes 0.2g Dentin, with the demineralization 6 hours at 4 ℃ of 1ml10% phosphoric acid, 3000rpm/2min4 ℃ discards supernatant after centrifugal, Dentin rinses with 1ml100mMTris-HCl, 3000rpm/2min4 ℃ discards supernatant after centrifugal, repeat 3 times, then add 250 μ l sodium lauryl sulphate (SDS) solution [100mMTris-HCl (pH6.8), 4%SDS, 20% glycerol] from the Dentin of demineralization, extract MMPs, 14000rpm/10min4 ℃ of centrifugal rear extraction supernatant is for subsequent use.First add the extracting solution of 100 μ l to every hole in 96 orifice plates, be divided at random 6 groups (n=36), add respectively EGCG solution (the 100 μ g/ml of 80 μ l variable concentrations, 200 μ g/ml, 300 μ g/ml, 400 μ g/ml), deionized water (negative control group), and 0.2%CHX (positive controls), then in every hole, add 20 μ l fluorescence collagen substrates (100 μ g/ml).Shaking after 30 seconds, is within 0,2,4,8,12,24,36,48 hour, to detect its fluorescent value intensity by microplate reader at time point, and experimental result is shown in Fig. 1.

2) determine that EGCG has the effect that improves the resistance to demineralization ability of dentin

Intercept the dentin that people is preced with middle part, 20 of the dentin test specimens of preparation 2mm × 2mm × 4mm size, self curing plastic embedding test specimen, and make dentin piece be exposed to outer not embedding near adamantine surface.Smooth with the speed of the 120 revs/min dentin face of polishing with 600,1200,1500,2000,2500,3000 order carborundum papers successively under flowing water cooling, guarantee that test specimen upper and lower surface is parallel, and with the antiscuffing paste polished surface of 0.3 μ m granularity.In the district of windowing of the reserved 2mm × 2mm size of each test specimen dentin surface.Good sanding and polishing test specimen is divided into two groups at random, every group 10, one group of negative matched group, use deionized water processing, experimental group selects 400 μ g/ml alcoholic solution as surface treatment according to definite valid density, each group test specimen carries out as shown in Figure 2 pH circulation (demineralization processing) under 37 ℃ of conditions, circulates every day 2 times, and totally 6 days.After loop ends, collect total acid demineralization liquid of each test specimen, adopt inductively coupled plasma atomic emission spectrometer (inductively-coupled plasma atomic emission spectrometer, ICP-AES) measure calcium ion concentration in acidic buffer, and disengage rate (calcium depletion rate, CDR) according to the calcium that following formula calculates unit are in the unit interval of each test specimen in the acid demineralization liquid of unit: calcium content in CDR=demineralization liquid (μ g)/acid treatment time (h) × demineralization area (mm ²), it is higher that calcium disengages rate, illustrates that the resistance to demineralization ability of test specimen is lower, and experimental result shows that (calcium disengages rate 2.93 ± 0.18 μ g/h mm compared with matched group ²), EGCG processing can significantly improve Dentinal resistance to demineralization ability, and (calcium disengages rate 1.52 ± 0.24 μ g/h mm ²).

Example one

EGCG contains the comparison of fluoride preparation to the effect of the resistance to demineralization ability of dentin with tradition

Intercept the dentin that people is preced with middle part, 50 of the dentin test specimens of preparation 2mm × 2mm × 4mm size, self curing plastic embedding test specimen, and make dentin piece be exposed to outer not embedding near adamantine surface.Smooth with the speed of the 120 revs/min dentin face of polishing with 600,1200,1500,2000,2500,3000 order carborundum papers successively under flowing water cooling, guarantee that test specimen upper and lower surface is parallel, and with the antiscuffing paste polished surface of 0.3 μ m granularity.In the district of windowing of the reserved 2mm × 2mm size of each test specimen dentin surface.Adopt dimension formula durometer application 50g power to maintain 15s and measure the district's surface microhardness (Surface Microhardness, SMH) of windowing, each test specimen is measured 5 points, and every at a distance of 100 μ m.Calculate the average SMH of each test specimen, and it is for subsequent use to select 45 test specimens of SMH in (60-80) Vickers hardness number (Vickers Hardness Numbers, VHN) scope.45 test specimens are divided into 3 groups (n=15) at random.Negative control group is deionized water processing, and positive controls adopts 0.1% sodium fluoride (NaF) solution, and experimental group adopts 400 μ g/ml alcoholic solution.Each group test specimen carries out as shown in Figure 2 pH circulation, every day 2 times, totally 6 days under 37 ℃ of conditions.After PH loop ends, every group is selected the ultrastructure pattern (Fig. 3) of 5 test specimens field emission microscopy observation dentin surface at random.After every group of remaining 10 test specimen PH circulation, carry out again micro-hardness testing, and calculate the microhardness difference △ SMH=SMH before and after demineralization ₁-SMH ₂, SMH ₁and SMH ₂the microhardness value that is respectively demineralization front and back, experimental result is in table 1.

Table 1: the variation of microhardness before and after each processed group PH circulation (Mpa, n=15,

Note: have significant difference (P < 0.05) between two groups of the different subscript alphabetic flags of every string.

In sum, use EGCG to process the reduction that can significantly suppress Dentinal microhardness amount, dentinal tubule shows as locking, in dentin surface and tubule, all can be observed a large amount of mineral deposits, illustrate that EGCG processing can significantly improve Dentinal resistance to demineralization ability, its action effect can be with traditional NaF solution phase when (△ SMH there was no significant difference, under SEM, ultrastructure is similar).NaF is the generally acknowledged dentin remineralization liquid of a kind of effect; to reach dentin remineralization by calcium binding free in surperficial fluorion and dentin to protect Dentinal effect; EGCG can reach the protection Dentinal effect suitable with NaF by suppressing the activity of MMPs in dentin, points out it separately or share as the Dentinal gear division health product of protection and have reliable effect and wide application prospect containing fluoride preparation with some.

The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.

Claims (1)

1. the application of EGCG fat aspect protection dentin.

Images