A composition for inhibiting pathogenic bacteria on the surface of oral accessories and a preparation method thereof

Abstract

The application discloses a composition for inhibiting pathogenic bacteria on the surface of oral accessories and a preparation method thereof. The composition for inhibiting pathogenic bacteria on the surface of oral accessories comprises composition 1 and composition 2 which are stored separately. The composition 1 comprises, in mass fraction, 1-2% of punicalagin, 9-11% of malic acid, 2-3% of cellobiase, 2-3% of endocellulase, 3-5% of sodium lauryl sulfate, 15-20% of disodium ethylenediaminetetraacetate, 1-3% of glycerol, and the rest of deionized water. The composition 2 comprises, in mass fraction, 0.1-0.5% of myrtenol, 1-3% of glycerol, 3-5% of ethanol, 4-7% of propylene glycol, 1-2% of borax, and the rest of deionized water. The composition can specifically inhibit porphyromonas, streptococcus and aggregatibacter actinomycetemcomitans, can destroy the collagen scaffold on the surface of oral accessories, and can specifically disintegrate the nutrition transport channel outside the bacterial body.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of stomatology, in particular to a composition for inhibiting pathogenic bacteria on the surface of oral accessories and a preparation method thereof. BACKGROUND

[0002] In oral treatment, various accessories for treatment are often worn in the oral cavity, such as orthodontic accessory brackets, artificial tooth crowns, inlays and veneers and other artificial objects for oral repair. There are various bacteria in daily life, and it is impossible to completely eliminate all bacteria in the living environment. The three specific oral pathogenic bacteria that are more harmful to the oral cavity include porphyromonas, streptococcus and aggregatibacter actinomycetemcomitans, which are specific bacteria causing various oral diseases and easily leading to tooth loosening and shedding. Therefore, the first step in treating accessories worn in the oral cavity is to inhibit these three specific bacteria.

[0003] In the living environment, various bacteria and dirt will contaminate the accessories needed to be worn in the oral cavity; another situation is that the surface of the accessory in the oral cavity has a collagen matrix, which is not a direct component of bacteria, but helps bacteria form a stable external scaffold structure and assist in forming a three-dimensional community structure, which is not easy to fall off from the surface of the oral accessory.

[0004] In view of the above situation, the stomatology field uses various methods to remove them, such as 75% alcohol disinfection, mouthwash soaking, potassium permanganate liquid soaking, or hydrogen peroxide flushing, but many methods have inherent defects. For example, 75% alcohol soaking often accelerates the aging of the accessory, the disinfection efficiency of mouthwash is low, and there are problems such as pigment residue after potassium permanganate treatment. More importantly, the ordinary bacteriostatic and bactericidal effect mainly refers to killing the bacteria itself, but cannot cut off the external nutrient structure (mycelium) of the bacteria, and cannot also break down the collagen scaffold on the surface of the dirt.

[0005] In order to solve the above technical problems, it is necessary to develop a composition for inhibiting pathogenic bacteria on the surface of oral accessories. SUMMARY

[0006] The purpose of the present application is to provide a composition for inhibiting pathogenic bacteria on the surface of oral accessories, which can specifically inhibit the three oral pathogenic bacteria of porphyromonas, streptococcus and aggregatibacter actinomycetemcomitans, and can also break down the collagen scaffold on the surface of the oral accessory and specifically break down the nutrient transport channel (mycelium) outside the bacteria.

[0007] In order to achieve the above purpose, the present application adopts the following technical scheme:

[0008] In the first aspect of the present application, a composition for inhibiting pathogenic bacteria on the surface of oral accessories is provided, which comprises composition 1 and composition 2 stored separately,

[0009] The composition 1 is composed of 1-2% maslinic acid, 9-11% malic acid, 2-3% cellobiase, 2-3% endofibroglycan, 3-5% sodium lauryl sulfate, 15-20% disodium ethylenediaminetetraacetate, 1-3% glycerol, and the rest is deionized water, with the total amount being 100% by mass fraction.

[0010] The composition 2 is composed of 0.1-0.5% myrtenol, 1-3% glycerol, 3-5% ethanol, 4-7% propylene glycol, 1-2% borax, and the rest is deionized water, with the total amount being 100% by mass fraction.

[0011] In the second aspect of the present application, a preparation method of a composition for inhibiting pathogenic bacteria on the surface of oral appendages is provided, and the method comprises:

[0012] First, the components of the composition 1 are weighed, and the disodium ethylenediaminetetraacetate is dissolved in deionized water; then, the sodium lauryl sulfate is added and dissolved, and then the maslinic acid, the malic acid, the cellobiase and the endofibroglycan are added and dissolved, to obtain the composition 1.

[0013] First, the components of the composition 2 are weighed, and the glycerol is dissolved in deionized water; then, the ethanol, the propylene glycol, the borax and the myrtenol are added and dissolved in sequence, to obtain the composition 2.

[0014] In the above technical solution, the components are added in the above order to prevent interaction between them and facilitate dissolution.

[0015] The one or more technical solutions in the embodiments of the present application have at least the following technical effects or advantages:

[0016] 1. The composition for inhibiting pathogenic bacteria on the surface of oral appendages provided by the present application has the following ratio: 1-2% maslinic acid and 0.1-0.5% myrtenol. It is found through experiments that the combined use of the two components has 17 times better effect than the use of maslinic acid or myrtenol alone, and the two components have a synergistic effect and can specifically inhibit three specific oral bacteria.

[0017] 2. The 0.1-0.5% myrtenol in the composition for inhibiting pathogenic bacteria on the surface of oral appendages provided by the present application has a destructive effect on the collagen scaffold on the surface of oral appendages, which is not found in other documents and inventions. 3. The existing oral bactericidal method kills bacteria and reduces the amount of bacteria, but the nutrient transport (mycelium) outside the bacteria is not broken down. The composition for inhibiting pathogenic bacteria on the surface of oral appendages provided by the present application can also specifically break down the nutrient transport channel (mycelium) outside the bacteria. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed in the embodiment description will be briefly introduced. Obviously, the drawings in the following description are some embodiments of the present application, and other drawings can be obtained by those skilled in the art without any creative effort.

[0019] Figure 1 Resulting graph of the inhibition of bacteria by different gradient concentration combinations.

[0020] Figure 2 Resulting graph of the obvious synergistic effect of punicalagin and myrtenol on the inhibition of bacteria.

[0021] Figure 3 Resulting graph of the obvious degradation of collagen scaffold by myrtenol.

[0022] Figure 4 Resulting graph of the whole plaque thickness shown by atomic force microscopy. Comparison of the whole plaque thickness before treatment (left) and the whole plaque thickness after treatment according to the present application (right).

[0023] Figure 5 Resulting graph of the topographic map of the data reconstruction of the bacterial colony morphology shown by the surface morphology microscope before and after treatment.

[0024] Figure 6 Inhibition of the nutrition transport pathway (hyphae) outside the bacterial body.

[0025] Figure 7 Comparison of the inhibition of 3 kinds of oral cavity specific pathogenic bacteria detected by PCR before and after treatment; PCR detects 3 kinds of oral cavity main pathogenic bacteria of the whole colony species, including porphyrin mononuclear bacteria, streptococcus and accompanying actinomyces. Comparison of 4 kinds of main pathogenic bacteria before treatment (left) and after treatment according to the present application (right).

[0026] Figure 8 Comparison of the changes of harmful products of plaque detected by Western blot; comparison of the total products LPS, IL-1β and TNF-α of the whole colony before treatment and the whole colony products LPS, IL-1β and TNF-α after treatment according to the present application.

[0027] Figure 9 Comparison of the collagen morphology and content in the bacterial colony scaffold. DETAILED DESCRIPTION

[0028] The advantages and various effects of the present application will be more clearly presented by the following specific embodiments and examples. Those skilled in the art should understand that these specific embodiments and examples are used to illustrate the present application, rather than limit the present application.

[0029] Throughout the specification, unless otherwise specifically indicated, the terms used are as generally used in the art. Thus, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. If there is a conflict between the present specification and the definitions of terms, the present specification controls.

[0030] Unless otherwise specifically indicated, various raw materials, reagents, instruments and equipment used in the present application can be purchased on the market or obtained by existing methods.

[0031] The technical solution of the embodiments of the present application is to solve the above technical problems, and the general idea is as follows:

[0032] According to a typical embodiment of the present application, a composition for inhibiting pathogenic bacteria on the surface of oral accessories is provided, which comprises composition 1 and composition 2 stored separately,

[0033] The composition 1 comprises, by mass fraction: 1-2% of punicalagin, 9-11% of malic acid, 2-3% of cellobiase, 2-3% of endofibroglycanase, 3-5% of sodium lauryl sulfate, 15-20% of disodium ethylenediaminetetraacetate, 1-3% of glycerol, and the balance is deionized water;

[0034] The composition 2 comprises, by mass fraction: 0.1-0.5% of myrtenol, 1-3% of glycerol, 3-5% of ethanol, 4-7% of propylene glycol, 1-2% of borax, and the balance is deionized water.

[0035] In the above technical solution,

[0036] The combination of 1-2% of punicalagin and 0.1-0.5% of myrtenol can specifically inhibit the three oral bacteria: porphyromonas, streptococcus and paraburkholderia aggregata; in addition, 0.1-0.5% of myrtenol can destroy the collagen scaffold on the surface of oral accessories.

[0037] The function of malic acid is to inhibit the formation of soft dirt on the surface of oral accessories.

[0038] The functions of cellobiase and endofibroglycanase are to degrade food residues in dirt

[0039] According to another typical embodiment of the present application, a method for using the composition for inhibiting pathogenic bacteria on the surface of oral accessories is also provided, which comprises the following steps:

[0040] Step 1: The accessories of orthodontic or repair, dirt, bloodstains, saliva, soft dirt, etc. are rinsed with deionized water.

[0041] Step 2: Put the accessory into composition 1 for treatment, and rinse it with deionized water.

[0042] Step 3: Put the accessory into composition 2 for treatment, and rinse it with deionized water, and use dental triple syringe for rinsing.

[0043] Step 4: Put the treated accessory into ultrasonic cleaner for final shock rinsing.

[0044] Preferably, the treatment time of step 2 is 3-5 minutes.

[0045] Preferably, the treatment time of step 3 is 5-10 minutes.

[0046] Preferably, the rinsing time of step 3 using triple syringe is 10-30 seconds.

[0047] Preferably, the rinsing liquid of step 3 using triple syringe is deionized water.

[0048] Preferably, the ultrasonic cleaning time of step 4 is 2-5 minutes.

[0049] A composition for inhibiting pathogenic bacteria on the surface of oral accessories will be described in detail below in combination with examples, comparative examples and experimental data.

[0050] Example 1

[0051] 1. The present example provides a composition for inhibiting pathogenic bacteria on the surface of oral accessories, which comprises composition 1 and composition 2 stored separately,

[0052] The composition 1 comprises, by mass fraction: 1.5% of punicalagin, 10% of malic acid, 2.5% of cellobiase, 2.5% of endofibroglycanase, 4% of sodium lauryl sulfate, 17.5% of disodium ethylenediaminetetraacetate, 2% of glycerol, and the rest is deionized water.

[0053] The composition 2 comprises, by mass fraction: 0.25% of myrtenol, 2% of glycerol, 4% of ethanol, 5% of propylene glycol, 1.5% of borax, and the rest is deionized water.

[0054] 2. Method for use:

[0055] (1) Under the premise of obtaining the consent of the patient, remove the fallen tooth inlay.

[0056] (2) Use deionized water to rinse the dirt, bloodstains, saliva, soft dirt, etc. on the inlay.

[0057] (3) Put the accessory into composition 1 for treatment, and rinse it with deionized water.

[0058] (4) Put the accessory into composition 2 for treatment, and the treatment time is 10 minutes.

[0059] (5) Rinse with deionized water, and use dental three-purpose air gun for rinsing. The rinsing time is 30 seconds.

[0060] (6) Put the treated accessory into ultrasonic cleaning machine for final shock rinsing. The cleaning time is 5 minutes.

[0061] Example 2

[0062] The example provides a composition for inhibiting pathogenic bacteria on the surface of oral accessories, which comprises composition 1 and composition 2 stored separately.

[0063] The composition 1 comprises, by mass fraction, 1% of maslinic acid, 9% of malic acid, 2% of cellobiase, 2% of endofibroglycane, 3% of sodium lauryl sulfate, 15% of disodium ethylenediaminetetraacetate, 1% of glycerol, and the balance of deionized water.

[0064] The composition 2 comprises, by mass fraction, 0.1% of myrtenol, 1% of glycerol, 3% of ethanol, 4% of propylene glycol, 1% of borax, and the balance of deionized water.

[0065] Example 3

[0066] The example provides a composition for inhibiting pathogenic bacteria on the surface of oral accessories, which comprises composition 1 and composition 2 stored separately.

[0067] The composition 1 comprises, by mass fraction, 2% of maslinic acid, 11% of malic acid, 3% of cellobiase, 3% of endofibroglycane, 5% of sodium lauryl sulfate, 20% of disodium ethylenediaminetetraacetate, 3% of glycerol, and the balance of deionized water.

[0068] The composition 2 comprises, by mass fraction, 0.5% of myrtenol, 3% of glycerol, 5% of ethanol, 7% of propylene glycol, 2% of borax, and the balance of deionized water.

[0069] Comparative Example 1

[0070] In the comparative example, the simple composition 1 comprises maslinic acid. The composition is configured as follows: 1.5% of maslinic acid, 10% of malic acid, 2.5% of cellobiase, 2.5% of endofibroglycane, 4% of sodium lauryl sulfate, 17.5% of disodium ethylenediaminetetraacetate, 2% of glycerol, and the balance of deionized water.

[0071] Comparative Example 2

[0072] The comparative example 2 is a simple composition 2: myrtenol. The configuration is as follows: myrtenol 0.25%, glycerol 2%, ethanol 4%, propylene glycol 5%, borax 1.5%, and the rest is deionized water.

[0073] Comparative example 3

[0074] The comparative example 3 does not contain myrtenol, and the other components are the same as those in the example 1. The configuration is as follows: glycerol 2%, ethanol 4%, propylene glycol 5%, borax 1.5%, and the rest is deionized water.

[0075] Example 4

[0076] The composition 1 is composed of the following compounds: punicalagin 2%, malic acid 10%, cellobiase 3%, endocelloglucanase 3%, sodium lauryl sulfate 5%, disodium ethylenediaminetetraacetate 20%, glycerol 3%, and the rest is deionized water.

[0077] The composition 2 is composed of the following compounds: myrtenol 0.5%, glycerol 3%, ethanol 5%, propylene glycol 7%, borax 2%, and the rest is deionized water.

[0078] Examples 5-10

[0079] The component settings of the examples 5-7 are shown in Table 1, and the component settings of the examples 8-10 are shown in Table 2.

[0080] Comparative examples 4-15

[0081] The component settings of the comparative examples 4-8 are shown in Table 1, and the component settings of the comparative examples 9-15 are shown in Table 2.

[0082] Experimental example 1

[0083] 1. Gradient concentration group comparison of punicalagin (examples 5-7 and comparative examples 4-8): 8 gradient content groups are set, containing punicalagin at a content of 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 5.0%, and 10.0%, respectively, and the rest of the components are: malic acid 10%, cellobiase 2.5%, endocelloglucanase 2.5%, sodium lauryl sulfate 4%, disodium ethylenediaminetetraacetate 17.5%, glycerol 2%, and the rest is deionized water. These are divided into 8 groups, which are respectively applied to the surface of the bracket, and then the content of the three specific bacteria is measured by real-time PCR method. The groups are shown in Table 1.

[0084] Table 1

[0085]

[0086] From the results of Table 1, it can be seen that the efficiency of inhibiting specific bacteria is the greatest when the specific content of punicalagin is between 1.0-2.0%.

[0087] 2. Ten gradient content groups (Examples 5-7 and Comparative Examples 4-8) were set up, each containing myrtenol at a content of 0.01%, 0.05%, 0.1%, 0.25%, 0.5%, 0.6%, 1.0%, 2.0%, 5.0%, and 10.0%, and the rest of the components were: glycerol 2%, ethanol 4%, propylene glycol 5%, borax 1.5%, and the rest was deionized water. The 10 groups were divided, and each was applied to the surface of the bracket, and then the content of the three specific bacteria was measured by real-time PCR. The groups are shown in Table 2.

[0088] Table 2

[0089]

[0090] From Table 2, it can be seen that the efficiency of inhibiting specific bacteria is the greatest when the specific content of myrtenol is between 0.1-0.5%.

[0091] Experimental Example 2

[0092] The compositions of Example 1, Comparative Example 1, and Comparative Example 2 were applied to the surface of the bracket, and then the content of the three specific bacteria was measured. The results are shown in Table 3.

[0093] Table 3

[0094]

[0095] From Table 3, it can be seen that:

[0096] The specific bacteriostatic effect (P. cinerea, S. mutans, and A. actinomycetemcomitans) was determined by real-time PCR, and the effect of using Composition 1 and Composition 2 together was nearly 10.53-17.74 times that of using Composition 1 or Composition 2 alone, which was a statistically significant difference, indicating that the specific concentration and specific combination of the composition are the outstanding features of the present application.

[0097] Experimental Example 3, Anti-collagen scaffold active ingredient detection

[0098] Example 1 and Comparative Example 3 were applied to the surface of the bracket, and then the content of collagen before and after treatment was measured.

[0099] The results are shown in Table 4. Figure 3As shown, there is no obvious difference in the collagen content before and after treatment for the group without myrtenol. However, for the group containing myrtenol in the composition of the present patent, the collagen content is significantly reduced after treatment compared with before treatment. This shows that myrtenol plays a role in inhibiting and destroying the collagen of oral accessories.

[0100] Experiment Example 4: Specifically disintegrating the nutrition transport channel outside the bacterial body

[0101] I. Preoperative detection of the composition of Example 4 on the oral bracket accessory. The detection content and method are as follows:

[0102] 1. Atomic force microscope shows the thickness of plaque: using a microscope, the bracket surface is observed, the thickness of the soft dirt in the vertical distance direction of the side is recorded, and 3D reconstruction is completed. Organize into a data graph.

[0103] 2. Surface morphology microscope shows the shape of the bacterial population: mainly including two aspects, the shape of the bacterial population and the maturity of the bacterial colony. The growth form of the bacterial colony is detected, and the surface layout is calculated by the topographic map calculation formula of the atomic force microscope. The difference in the shape of the bacterial population is calculated to comprehensively evaluate the maturity of the bacterial population form.

[0104] 3. PCR result measurement: PCR mainly detects the presence or absence of three kinds of specific pathogenic bacteria in the oral cavity, including porphyromonas, streptococcus and paraplastomyces. All bacteria are scraped off with a scalpel, and a bacterial genomic DNA extraction kit is used to extract standard strain genomic DNA. The reaction of different strains and strain-specific primers in the PCR detection process is observed, the product is cut and recovered, the DNA segment is extracted, and PCR amplification is performed to measure the amount of pathogenic bacteria on the bracket.

[0105] 4. Western blot detection of changes in harmful products of plaque: mainly detecting the harmful products of the whole bacterial population, LPS, IL-1β and TNF-α. The lysate is subjected to SDS-PAGE gel electrophoresis. The primary antibody is incubated in TBST at 4°C overnight. The membrane is washed with TBST and incubated with HRP secondary antibody at room temperature, and then washed with TBST. Then visual analysis is performed, and the blot is analyzed using an immunoblot analysis detection system (Biorad).

[0106] II. Use the composition of the present application to treat the bracket accessory.

[0107] The method is as follows:

[0108] Step 1: Use deionized water to rinse the dirt, bloodstains, saliva, plaque, etc. on the accessory.

[0109] Step 2: Put the accessory into composition 1 for treatment, the treatment time is 3 minutes. And rinse it with deionized water.

[0110] Step 3: Place the attachment into composition 2 for treatment for 5 minutes.

[0111] Step 4: Rinse thoroughly with deionized water, then use a dental air gun for 10 seconds.

[0112] Step 5: Place the processed attachments into an ultrasonic cleaner for a final agitation rinse. The cleaning time is 2 minutes.

[0113] The processed bracket accessories were re-inspected using the aforementioned detection method. The main detection items, as described in Part II of this example, include: atomic force microscopy to show plaque thickness, surface morphology microscopy to detect the surface morphology of the bacterial community, PCR to detect plaque quantity, and Western blot to detect changes in harmful products from the plaque.

[0114] A comparison of the overall plaque thickness before treatment (left image) and the overall plaque thickness after treatment with the composition for inhibiting pathogenic bacteria on the surface of oral appendages in Example 1 of this invention (right image) is shown below. Figure 4 As shown, the plaque thickness in the treated group of the present invention is significantly reduced. This indicates that the composition of the present invention thoroughly cleans the bacterial colony thickness.

[0115] Figure 5 This indicates that the composition of the present invention can effectively eliminate the amount of Porphyromonas colonies; Figure 6 This indicates that the composition of the present invention disrupts extrabacterial nutrient transport (hyphae), reducing the amount of three specific oral bacteria (see appendix). Figure 7 The harmful bacterial byproducts, including LPS, IL-1β, and TNF-α, are significantly inhibited and eliminated (see appendix). Figure 8 ), breaking down the collagen fiber scaffold on the surface of the bracket (see attached document) Figure 9 ).

[0116] In summary, one or more technical solutions of the embodiments of the present invention have at least the following advantages:

[0117] 1. Punicin and myrtol only exhibit significant bactericidal effects within the specific concentration ranges specified in this patent.

[0118] 2. When punicalin and myrtol are used together, the effect is 10.53-17.7 times greater than when punicalin or myrtol are used alone. It has been found that their specific combination is more effective.

[0119] 3. Myrtol disrupts the collagen scaffold on the surface of oral appendages, thereby damaging the external survival environment of bacteria.

[0120] 4. Specifically targets bacteria on three types of oral appendages.

[0121] 5. Other fungicides are killing the bacteria itself, reducing the amount of bacteria. But the outside of the bacteria nutrient transport (hyphae) is not collapsed. The present invention can also specifically collapse the nutrient transport channel (hyphae) outside the bacteria body.

[0122] Finally, it should be noted that the terms "comprises", "comprising", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but can also include other elements not expressly listed or inherent to such process, method, article, or apparatus.

[0123] While the preferred embodiments of the application have been described, it should be understood that various modifications and changes can be made by those skilled in the art which follow in the spirit of the application and the scope of the appended claims. Accordingly, it is intended that all such modifications and changes be included within the scope of the application as claimed.

[0124] Obviously, numerous modifications and variations of the present application are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims and their equivalents, the application can be practiced otherwise than as specifically described.

Claims

1. A composition for inhibiting pathogenic bacteria on the surface of oral appendages, characterized in that, The composition for inhibiting pathogenic bacteria on the surface of oral appendages comprises composition 1 and composition 2, which are stored separately. The composition 1, by mass fraction, comprises: 1-2% punicin, 9-11% malic acid, 2-3% cellobiase, 2-3% celloglucan endopeptidase, 3-5% sodium lauryl sulfate, 15-20% disodium EDTA, 1-3% glycerol, and the balance being deionized water; The composition 2, by mass fraction, comprises: 0.1-0.5% myrtle alcohol, 1-3% glycerol, 3-5% ethanol, 4-7% propylene glycol, 1-2% borax, and the balance being deionized water.

2. The composition for inhibiting pathogenic bacteria on the surface of oral appendages according to claim 1, characterized in that, The composition 1, by mass fraction, comprises: 1.5% punicin, 10% malic acid, 2.5% cellobiase, 2.5% celloglucan endopeptidase, 4% sodium lauryl sulfate, 17.5% disodium EDTA, 2% glycerol, and the balance being deionized water; The composition 2, by mass fraction, consists of: 0.25% myrtle alcohol, 2% glycerol, 4% ethanol, 5% propylene glycol, 1.5% borax, and the balance being deionized water.

3. A method for processing an accessory inserted into the oral cavity, characterized in that, The method includes: The accessory to be inserted into the oral cavity is sequentially cleaned with deionized water, soaked in composition 1 according to any one of claims 1-2, rinsed with deionized water, soaked in composition 2 according to any one of claims 1-2, and rinsed.

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