KR102439762B1 - Composition for the prevention and/or treatment of peri-implantitisnear infections - Google Patents

Abstract

The present invention relates to a microbial preparation for preventing peri-implantitis, which uses a culture supernatant of oral lactic acid bacteria Weissella cibaria CMU (oraCMU) and Weissella cibaria CMS1 (oraCMS1), used to inhibit the formation of biofilm of oral bacteria on a titanium disc, which is used as an implant material, thereby having an effect of preventing the occurrence of peri-implantitis.

Description

Composition for preventing and treating peri-implant mucositis

The present invention relates to a microbial preparation for the prevention and treatment of peri-implant mucositis, and implants with the culture supernatant of oral lactic acid bacteria Weissella cibaria CMU (oraCMU) and Weissella cibaria CMS1 ( Weissella cibaria CMS1; oraCMS1) It relates to a composition capable of preventing the occurrence of peri-implant mucositis by inhibiting the formation of a biofilm of oral bacteria on a titanium disk used as a material.

Recently, as the number of dental implant procedures increases, the number of patients suffering from peri-implant disease is increasing. In particular, as a peri-implant disease, peri-implant mucositis, an inflammation of the mucous membrane without loss of support bones, is increasing.

Since the prevention of peri-implant mucositis is very important to avoid alveolar bone loss and to avoid irreversible damage to the tissues around the implant, the microbial preparation of the present invention inhibits the colonization of pathogenic microorganisms, thereby reducing the microflora that can change to a pathological state. It is an oral care solution.

The present inventors found that the culture supernatant of oral lactic acid bacteria Weissella cibaria CMU (oraCMU) and Weissella cibaria CMS1; By reducing the Fusobacterium nucleatum and Prevotella intermedia bacteria, which act as intermediate bridges with other bacteria in the process of completed.

Republic of Korea Patent Publication No. 10-2017-0101890 "The treatment method of peri-implantitis"

The present invention is a microbial preparation for the prevention of peri-implant mucositis, and titanium used as an implant material as a culture supernatant of oral lactic acid bacteria Weissella cibaria CMU (oraCMU) and Weissella cibaria CMS1 (oraCMS1). An object of the present invention is to provide a formulation capable of preventing and treating the occurrence of peri-implant mucositis by inhibiting the formation of a biofilm of oral bacteria on the disc.

The present invention relates to a composition for preventing and treating peri-implant mucositis, comprising the culture supernatant of oral lactic acid bacteria Weissella cibaria CMU (oraCMU) and Weissella cibaria CMS1 ( Weissella cibaria CMS1; oraCMS1) To provide a pharmaceutical composition that can prevent the occurrence of peri-implant mucositis by inhibiting the formation of a biofilm of oral bacteria on a titanium disk used as an implant material.

In addition, the present invention relates to peri-implant mucositis, characterized in that the volume ratio of Weissella cibaria CMU (oraCMU) culture supernatant and Weissella cibaria CMS1; oraCMS1 culture supernatant is 4:1. A pharmaceutical composition for prevention and treatment is provided.

In particular, it reduces the biofilm formation of Porphyromonas gingivalis , a powerful periodontal pathogen, by reducing Fusobacterium nucleatum and Prevotella intermedia bacteria, which act as intermediate bridges with other bacteria in the process of forming a biofilm on the titanium disc by 8 types of bacteria in the oral cavity. This can prevent the occurrence of peri-implant mucositis. Since the prevention of peri-implant mucositis is very important to avoid alveolar bone loss and to avoid irreversible damage to the tissues around the implant, the microbial preparation of the present invention inhibits the colonization of pathogenic microorganisms, thereby reducing the microflora that can change to a pathological state. This provides an oral care solution that has the effect of preventing peri-implant mucositis.

At this time, the bacteria that form the implant biofilm are the oral early colonizers Actinomyces naeslundii, Veillonella sp., Streptococcus gordonii, Streptococcus oralis, Streptococcus sanguinis and It can be characterized by the intermediate colonizer Fusobacterium nucleatum and Prevotella intermedia, and the late colonizer Porphyromonas gingivalis .

According to one embodiment of the present invention, the oral lactic acid bacteria Weissella cibaria CMU ( Weissella cibaria CMU; oraCMU) and Weissella cibaria CMS1 ( Weissella cibaria CMS1; oraCMS1) containing the culture supernatant in a vegetable oil base and It may be provided in a concentrated form or may be characterized in that the concentrate is inhaled into pork potato starch and then freeze-dried and powdered.

According to one embodiment of the present invention, the composition of the present invention may be characterized as a food composition for prevention and recovery of peri-implant mucositis.

The present invention relates to a microbial preparation for preventing peri-implant mucositis, and as an implant material as a culture supernatant of oral lactic acid bacteria Weissella cibaria CMU (oraCMU) and Weissella cibaria CMS1 ( Weissella cibaria CMS1; oraCMS1) By suppressing the biofilm formation of oral bacteria on the titanium disk used, there is an effect to prevent the occurrence of peri-implant mucositis.

1 is a result of measuring the number of viable cells of the oral bacterial biofilm over time.
2 is a real-time polymerase chain reaction result for the biofilm of oral bacteria over time.
3 is a photograph showing the effect of oraCMU and oraCMS1 for inhibiting colony biofilm formation on oral bacteria by phase contrast microscopy.
4 is a graph measuring the colony biofilm formation inhibitory effect of oraCMU and oraCMS1 against oral bacteria by the number of viable cells and real-time polymerase chain reaction.
5 is a photograph and graph of oraCMU and oraCMS1 measured by a confocal scanning laser microscope for the colony biofilm formation inhibitory effect on oral bacteria.
6 is a photograph and graph showing the colony biofilm removal efficacy of oraCMU against oral bacteria with a confocal scanning laser microscope.
7 is a photograph showing the inhibitory effect on implant biofilm formation of oraCMU and oraCMS1 with a phase-contrast microscope.
8 is a graph measuring the absorbance and the number of viable cells for the inhibitory effect on implant biofilm formation of oraCMU and oraCMS1.
FIG. 9 is a graph measuring the inhibitory effect on implant biofilm formation of oraCMU and oraCMS1 by a real-time polymerase reaction.
10 is a graph measuring the absorbance of oraCMU and oraCMS1 to inhibit the formation of an implant biofilm at 595 nm.
11 is a photograph and graph showing the efficacy of oraCMU and oraCMS1 for inhibiting implantation biofilm formation with a confocal scanning laser microscope.
12 is a photograph showing the effect of inhibiting implant biofilm formation of oraCMU and oraCMS1 with a scanning electron microscope.
13 shows a comparison of the effect of inhibiting colony biofilm formation on oral bacteria according to the volume ratio of oraCMU fermented supernatant and oraCMS1 fermented supernatant.
14 shows a comparison of the titanium disc biofilm formation inhibitory effect according to the volume ratio of oraCMU fermented supernatant and oraCMS1 fermented supernatant.
15 is a graph comparing the titanium disk biofilm formation inhibitory effect according to the volume ratio of oraCMU fermented supernatant and oraCMS1 fermented supernatant through qPCR on the machined surface.
16 is a graph comparing the titanium disc biofilm formation inhibitory effect according to the volume ratio of oraCMU fermented supernatant and oraCMS1 fermented supernatant through qPCR on the SLA surface.

Hereinafter, the present invention will be described in more detail through specific examples. The following examples describe a preferred embodiment of the present invention, and the scope of the present invention is not limited and interpreted by the matters described in the following examples.

The present invention includes a culture supernatant of oral lactic acid bacteria Weissella cibaria CMU (oraCMU) and Weissella cibaria CMS1 (Weissella cibaria CMS1 ; oraCMS1) to provide a composition for preventing and treating peri-implant mucositis To provide a pharmaceutical composition capable of preventing the occurrence of peri-implant mucositis by inhibiting the formation of a biofilm of oral bacteria on a titanium disk used as an implant material.

According to one embodiment of the present invention, Weissella ciberia ( Weissella cibaria CMU) is a Weissella lactic acid bacterium that suppresses anaerobic bacteria that cause bad breath and generates hydrogen peroxide in aerobic and anaerobic conditions, Weissella ciberia CMU KCTC 10650BP make use of

In particular, it reduces the biofilm formation of Porphyromonas gingivalis , a powerful periodontal pathogen, by reducing Fusobacterium nucleatum and Prevotella intermedia bacteria, which act as intermediate bridges with other bacteria in the process of forming a biofilm on the titanium disc by 8 types of bacteria in the oral cavity. This can prevent the occurrence of peri-implant mucositis. Since the prevention of peri-implant mucositis is very important to avoid alveolar bone loss and to avoid irreversible damage to the tissues around the implant, the microbial preparation of the present invention inhibits the colonization of pathogenic microorganisms to reduce the microflora that can change to a pathological state It is an oral care solution that has the effect of preventing peri-implant mucositis.

Hereinafter, the present invention will be described in more detail through specific examples. The following examples describe a preferred embodiment of the present invention, and the scope of the present invention is not limited and interpreted by the matters described in the following examples.

Example 1. in vitro Development of dental implant biofilm model

(One) Induction of oral bacterial biofilm formation

The biofilm formation process of oral bacteria was measured using the colony biofilm method, and the amount of change in the number of viable cells in the biofilm and the amount of change in each of the 8 types of bacteria were measured using real-time PCR. .

(2) Efficacy evaluation of colony biofilm formation inhibition and removal of oral care probiotics

Using the colony biofilm method, we confirmed the evaluation of the inhibition and removal efficacy of oral bacterial biofilm formation of Weissella cibaria CMU ( Weissella cibaria CMU; oraCMU) and Weissella cibaria CMS1 ( Weissella cibaria CMS1; oraCMS1). Efficacy evaluation was confirmed using viable cell count, phase contrast microscopy, confocal scanning laser microscopy, and real-time PCR.

(3) Efficacy evaluation of oral care probiotics in inhibiting dental implant biofilm formation

We confirmed the efficacy evaluation of oral bacterial biofilm formation inhibition of Weissella cibaria CMU ( Weissella cibaria CMU; oraCMU) and Weissella cibaria CMS1 ( Weissella cibaria CMS1; oraCMS1) using a titanium disk, a dental implant material, and the Efficacy evaluation was confirmed using viable cell count, phase contrast microscopy, confocal scanning laser microscopy, and real-time PCR.

Preparation Example 1

Weissella cibaria CMU used in the present invention suppresses anaerobic bacteria that cause bad breath and generates hydrogen peroxide in aerobic and anaerobic conditions Weissella cibaria CMU (accession number: KCTC 10650BP) ) was cultured in MRS medium at 37° C. for 24 hours, followed by centrifugation (5,000 g, 10 min, 4° C.) to prepare a supernatant.

Preparation 2

It is the same as in Preparation Example 1 except for using Weissella cibaria CMS1 instead of Weissella cibaria CMU.

<Test Example 1> Induction of oral bacterial biofilm in vitro

To confirm the biofilm formation process of oral bacteria, Actinomyces naeslundii, Veillonella sp., Streptococcus gordonii, Streptococcus oralis, Streptococcus sanguinis, which are early colonizers, and Fusobacterium nucleatum and Prevotella intermedia, which are intermediate colonizers, and Porphyromonas gingivalis , which are late colonizers, were cultured anaerobically and aerobically, respectively.

Add 2.5 mM DL-dithiothreitol to 10 mL of saliva, stir for 10 minutes, and then centrifuge (9,000 g, 10 min, 4℃) to mix the supernatant in the same amount with phosphate buffered saline (PBS), 0.22 μm pore size filter It was prepared by filtration using

Put a polycarbonate membrane (0.22 μm, 25 mm) on sheep blood agar containing hemin and menadione, place it in an anaerobic incubator at 37°C so that the medium is adsorbed overnight, and then centrifuge ( 5,000 g, 10 min, 4° C.) and the remaining pellet was suspended in saliva. After taking an equal amount of each suspended bacterial solution and mixing, 90 μL each was taken and dropped on a membrane for anaerobic culture.

On days 0, 1, 3, and 6, the membrane was recovered, put into 10 mL of peptone water previously reduced in an anaerobic incubator, and homogenized on ice for 30 seconds using a homogenizer. Stair-diluted with peptone water was inoculated on sheep blood agar, and the number of viable cells was measured by anaerobic culture for 5 days. For real-time PCR measurement, the homogenate was centrifuged again (5000 g, 10 min, 4℃) Genomic DNA was isolated from the obtained pellet.

1 is a result of measuring the number of viable cells of the oral bacterial biofilm over time. As shown here, it was confirmed that the number of viable cells increased with the lapse of time.

2 is a real-time polymerase chain reaction result for the biofilm of oral bacteria over time. (A) is the absolute amount of each microbe over time, and (B) is the change amount of each microbe over time. As shown here, as time passes, A. naeslundii and Veillonella sp., which are early colonizers, decreased, while F. nucleatum, P. intermedia, which are intermediate colonizers, increased significantly, and P. gingivalis , acting as a late colonizer, was the most an increase was observed.

<Test Example 2> Efficacy evaluation of colony biofilm formation inhibition and removal of in vitro oral care probiotics

To confirm the biofilm formation process of oral bacteria, the early colonizers Actinomyces naeslundii, Veillonella sp., Streptococcus gordonii, Streptococcus oralis, Streptococcus sanguinis and the intermediate colonizer Fusobacterium nucleatum, Prevotella intermedia, and the anaerobic and late colonizer Porphyromonas , respectively, were used. cultured.

oraCMU and oraCMS1 were prepared by filtration using a 0.22 μm pore size filter from the supernatant obtained by centrifugation (5,000 g, 10 min, 4° C.) after culturing in MRS broth for 24 hours. 0.8% soft agar was prepared by mixing 10 mL of sheep blood agar (agar 1.6%) containing hemin and menadione and 10 mL each of oraCMU or oraCMS1 filtered supernatant, 0.8% soft agar mixed with 10 mL of MRS broth was used as a control medium.

Add 2.5 mM DL-dithiothreitol to the saliva, stir for 10 minutes, and then centrifuge (9,000 g, 10 min, 4℃) to mix the supernatant in the same amount with phosphate buffered saline (PBS), and use a 0.22 μm pore size filter. It was prepared by filtration.

Put a polycarbonate membrane (0.22 μm, 25 mm) on the soft sheep blood agar prepared above, place it in an anaerobic incubator at 37°C so that the medium is adsorbed overnight, then adjust the absorbance of the oral bacterial culture to 0.5 at 600 nm and centrifuge (5,000 g) , 10 min, 4°C), and the remaining pellet was suspended in saliva. After taking an equal amount of each suspended bacterial solution and mixing, 90 μL each was taken and dropped on a membrane for anaerobic culture.

On days 0, 1, 3, and 6, the membrane was recovered, put into 10 mL of peptone water previously reduced in an anaerobic incubator, and homogenized on ice for 30 seconds using a homogenizer. 10 μL of this solution was analyzed with a phase-contrast microscope, and for measuring the number of viable cells, step-diluted with peptone water was inoculated into sheep blood agar and cultured anaerobically for 5 days. In addition, for real-time PCR measurement, the homogenate was centrifuged again (5000 g, 10 min, 4℃) to separate genomic DNA from the pellet obtained. For analysis with a confocal scanning laser microscope, the membrane was not homogenized and stained using the Live/Dead BacLight Bacterial Viability Kit to measure and compare live and dead cells.

3 is a photograph showing the effect of oraCMU and oraCMS1 for inhibiting colony biofilm formation on oral bacteria by phase contrast microscopy. As shown here, it was confirmed that the colony biofilm formation hardly occurred in the place treated with oraCMU and oraCMS1 compared to the control on the 6th day, so that almost no bacterial observation was observed.

4 is a graph showing the effect of colony biofilm formation inhibition on oral bacteria of oraCMU and oraCMS1 by measuring the number of viable cells and real-time polymerase chain reaction. (A) is a measurement of the number of viable cells, (B) is a real-time polymerase It measures the chain reaction. As shown here, a significant decrease in the number of viable cells was observed from the 1st day in the place treated with oraCMU and oraCMS1, and the number of viable cells could not be measured on the 3rd day of culture. could

5 is a photograph and graph of oraCMU and oraCMS1 measured by a confocal scanning laser microscope for the colony biofilm formation inhibitory effect on oral bacteria. As shown here, it was confirmed that live cells (green) decreased and dead cells (red) increased in the place treated with oraCMU and oraCMS1 over time.

6 is a photograph and graph showing the colony biofilm removal efficacy of oraCMU against oral bacteria with a confocal scanning laser microscope. As shown here, the colony biofilm formed on the 6th day was transferred to a soft agar containing oraCMU and observed after 1 or 3 days. As a result, the number of dead bacteria started to increase from the 1st day, and on the 3rd day, the amount of dead bacteria compared to the 1st day Since it was confirmed that more of these were observed, the biofilm removal efficacy of oraCMU was also confirmed.

7 is a photograph showing the inhibitory effect on implant biofilm formation of oraCMU and oraCMS1 with a phase-contrast microscope. As shown here, it was confirmed that the number of bacteria was significantly reduced on the surface of the titanium disk, which is the implant material, and in the oraCMU and oraCMS1 treated groups, F. was not observed. This is because F. nucleatum is known to increase biofilm by accelerating the biofilm of anaerobic bacteria that cause periodontal disease through the bridge role between aerobic bacteria, which act as initial settlers, and absolute anaerobes, such as P. gingivalis . The absence of F. nucleatum is a result confirming the biofilm reduction effect.

8 is a graph measuring the absorbance and the number of viable cells for the inhibitory effect on implant biofilm formation of oraCMU and oraCMS1. (A) showed that the biofilm formation was relatively decreased on both the machined surface and the SLA surface titanium disk treated with oraCMU and oraCMS1 at an absorbance of 600 nm. A decrease in bacteria was observed.

FIG. 9 is a graph measuring the inhibitory effect on implant biofilm formation of oraCMU and oraCMS1 by a real-time polymerase reaction. As shown here, Veillonella sp., F. nucleatum, P. gingivalis, P. intermedia, A. naeslundii, S. gordonii, S. oralis in biofilm on both machined surface and SLA surface titanium disks treated with oraCMU and oraCMS1. , S. sanguinis were all decreased, and in particular, it was confirmed that the reduction of P. intermedia , an obligate anaerobic bacterium involved in periodontitis and acute lumpy ulcerative gingivitis, was remarkable.

10 is a graph measuring the absorbance of oraCMU and oraCMS1 to inhibit the formation of an implant biofilm at 595 nm. As shown here, it was confirmed that there was a decrease in the biofilm concentration on both the machined surface and the SLA surface titanium disk treated with oraCMU and oraCMS1.

11 is a photograph and graph showing the efficacy of oraCMU and oraCMS1 for inhibiting implantation biofilm formation with a confocal scanning laser microscope. As shown here, it was confirmed that dead cells (red) were increased in both the machined surface and the SLA surface titanium disk treated with oraCMU and oraCMS1.

12 is a photograph showing the effect of inhibiting implant biofilm formation of oraCMU and oraCMS1 with a scanning electron microscope. A) untreated disc, B) positive control disc, C) oraCMU treated group, and D) oraCMS1 treated group. As shown here, the amount of bacteria was remarkably low on both the machined surface and the SLA surface titanium disk treated with oraCMU and oraCMS1, and in particular, bacteria of F. nucleatum , a long rod-shaped bacteria, could not be observed.

<Test Example 3> In vitro evaluation of the efficacy of oral care probiotics for inhibiting the formation of dental implant biofilms

A titanium disk, which is mainly used as an implant material, was manufactured, and the surface treatment on one side was a machined surface and a surface using spray treatment and acid corrosion (SLA; sand-blasted large-grit, acid-etched). The disk was made of 8 x 2 mm in size.

After putting the disk surface-treated side up in a 24-well plate, 0.2 mL of the prepared saliva was dropped on the disk, wrapped with a wrap to prevent drying, and incubated with shaking at 37°C for 4 hours.

Early colonizers Actinomyces naeslundii, Veillonella sp. , Streptococcus gordonii, Streptococcus oralis, Streptococcus sanguinis , the intermediate colonizers Fusobacterium nucleatum and Prevotella intermedia, and the late colonizer Porphyromonas gingivalis were cultured by smear or aerobic culture on sheep blood agar containing hemin and menadione, respectively.

oraCMU and oraCMS1 were prepared by filtration using a 0.22 μm pore size filter from the supernatant obtained by centrifugation (5,000 g, 10 min, 4° C.) after culturing in MRS broth for 24 hours.

Weissella cibaria CMU ( Weissella cibaria CMU; oraCMU) and Weissella cibaria CMS1 ( Weissella cibaria CMS1; oraCMS1) to confirm the efficacy evaluation of the inhibition of titanium disc biofilm formation, colonies of oral bacteria grown on sheep blood agar were scraped and suspended in tryptic soy broth containing hemin and menadione, and the absorbance was adjusted to 0.05 at 600 nm and mixed in equal amounts. 0.5 mL of the suspension was added to the prepared 24-well plate containing the disk, and 0.5 mL of the oraCMU or oraCMS1 supernatant was added and cultured in an anaerobic incubator at 37°C for 3 days.

Remove the culture solution, wash 3 times with PBS, put the disk in a 5 mL tube containing 2 mL of peptone water previously reduced in an anaerobic incubator, vortex vigorously for 2 minutes to drop the bacteria attached to the disk surface, and then take a portion It was measured with a phase contrast microscope, the absorbance was 600 nm and the number of viable cells was measured by inoculating sheep blood agar after stepwise dilution with peptone water.

In addition, in order to measure the number of bacteria for each oral bacteria, genomic DNA was isolated from the pellet obtained by transferring all of the bacterial solution and centrifuging (12,000 rpm, 3 min). A real-time polymerase chain reaction was performed using primers for each bacteria, and the reaction conditions were repeated 40 times: 2 minutes at 50°C, 2 minutes at 95°C, 15 seconds at 95°C, and 60 seconds at 60°C.

To measure the density of the disc biofilm stained with crystal violet, open the lid of the disc washed 3 times with sterile water, leave it at room temperature for about 5 to 10 minutes, dry the disc, and then use 0.1% crystal violet at room temperature for 15 minutes. After staining, washing with sterile water 3 times, and then dissolving in 0.5 mL of 99% ethanol, the biofilm concentration was measured at absorbance 595 nm.

In order to check the anti-biofilm effect of oraCMU and oraCMS1 using confocal scanning laser microscopy, the disk washed 3 times with sterile water was dropped with Live/Dead BacLight Bacterial Viability reagent and light was blocked for 20 minutes After the reaction, the mixture was washed three times with sterile water, and the mounting oil was dropped on the confocal dish, and the disk was placed on it and measured under a microscope.

In order to check the anti-biofilm effect by scanning electron microscopy, the discs washed three times with sterile water were pre-fixed with 2.5% glutaraldehyde, post-fixed with 1% osmium tetroxide, and then dehydrated with 50% ~ 100% ethanol. After the process, it was dried by substitution with hexamethyldisilazane, pre-treated, and then analyzed by magnification up to 10,000 times with a scanning electron microscope.

Example 2. Formulation process

When the whey solution obtained in Preparation Examples 1 and 2 is inhaled into pork potato starch rich in inulin component and dietary fiber component, vacuum freeze-dried and powdered to form powder, it spreads evenly in the mouth and has a long-lasting effect. It has been shown to have a maintenance effect.

In addition, during oral administration of the present invention, in addition to formulations such as powders, tablets, capsules, etc., it may be formulated as a film, toothpaste, mouthwash solution, spray, dressing solution, coating agent, and the like.

Example 3. Effect according to the volume ratio of fermented supernatant of oraCMU and oraCMS1

Example 3-1. Colony biofilm formation inhibitory effect on oral bacteria

Table 1 and FIG. 13 show the effect of inhibiting colony biofilm formation on oral bacteria according to the volume ratio of oraCMU fermented supernatant and oraCMS1 fermented supernatant.

Specifically, according to Table 1 and FIG. 13, when the volume ratio of the fermented supernatant of oraCMU and oraCMS1 is 4:1, the remaining 1:1, 2:1, 8:1, 16:1, 1:2, 1:4 , 1:8, 1:16, it can be seen that the colony biofilm formation inhibitory effect on oral bacteria is significantly superior.

Example 3-2. Titanium disc biofilm formation inhibitory effect

Table 2 and FIG. 14 show the titanium disc biofilm formation inhibitory effect according to the volume ratio of the oraCMU fermented supernatant and the oraCMS1 fermented supernatant.

Specifically, according to Table 2 and FIG. 14, when the volume ratio of the fermented supernatant of oraCMU and oraCMS1 is 4:1, the remaining 1:1, 2:1, 8:1, 16:1, 1:2, 1:4 , 1:8, 1:16, it can be seen that the titanium disk biofilm formation inhibitory effect is significantly superior to that of the case.

Furthermore, the effect of inhibiting the formation of machined surface and SLA surface titanium disk biofilm according to the volume ratio of oraCMU fermented supernatant and oraCMS1 fermented supernatant was confirmed through qPCR, and the results are shown in Tables 3, 4 and 15 and 16 below.

According to Tables 3, 4 and 15 and 16, the remaining 1:1, 2:1, 8:1, 16: when the volume of the fermented supernatant of oraCMU and oraCMS1 is 4:1 on both the machined surface and the SLA surface: 1, 1:2, 1:4, 1:8, it can be seen that compared to the case of 1:16, the titanium disk biofilm formation inhibitory effect is significantly superior.

Claims (4)

Prevention and treatment of peri-implant mucositis, comprising a mixture of oral lactic acid bacteria Weissella cibaria CMU (oraCMU) culture supernatant and Weissella cibaria CMS1; oraCMS1 culture supernatant as an active ingredient A pharmaceutical composition for
The composition is peri-implant mucositis, characterized in that the Weissella cibaria CMU (orCMU) culture supernatant and the Weissella cibaria CMS1; oraCMS1 culture supernatant are mixed in a volume ratio of 4:1. Pharmaceutical compositions for prophylaxis and treatment. delete The pharmaceutical composition for preventing and treating peri-implant mucositis according to claim 1, wherein the culture supernatant is inhaled into porcine potato starch and then freeze-dried to powder. A food composition for the prevention and recovery of peri-implant mucositis comprising the composition of claim 1.

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