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A strain of Lactobacillus fermentum capable of preventing and/or treating periodontitis and its application

A technology for fermenting lactobacillus and toothpaste, applied in the field of microorganisms, can solve the problems of high cost, increase production cost, cannot inhibit the growth of Porphyromonas gingivalis, etc., and achieve the effect of relieving periodontitis and having a huge application prospect.

Active Publication Date: 2021-08-24
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Publication number is disclosed in the Chinese patent application of CN101190239B a kind of product that contains Lactobacillus casei, and this type Lactobacillus casei is to periodontal disease pathogenic bacteria Porphyromonas gingivalis (Porphyromonas gingivalis), intermediate Prevotella (Prevotella intermedia), periodontal centipede bacteria (Centipeda periodontii) have inhibitory effect, but, as can be seen from the application text, this type of Lactobacillus casei only has inhibitory effect on a single bacterial species, and there are multiple pathogenic bacteria in the periodontitis environment Bacterial strains, and there will also be interactions between bacterial strains, so that Lactobacillus casei that inhibits a single strain in vitro does not necessarily have an inhibitory effect on the mixed bacterial film in the periodontitis environment. Therefore, it is still necessary to find a method that can inhibit the mixed bacteria probiotic preparation
[0008] Another example is that the patent document CN111642746 A discloses a product that is used in combination with Lactobacillus paracasei and Lactobacillus fermentum to inhibit periodontitis pathogenic bacteria Porphyromonas gingivalis. When Lactobacillus casei and Lactobacillus fermentum LBF128 were co-cultured with Porphyromonas gingivalis separately, the number of Porphyromonas gingivalis at 6 hours was more than that when cultured alone, which proved that when the two were used alone, there was no There is no inhibitory effect on Porphyromonas gingivalis, especially when Lactobacillus fermentum LBF128 and Porphyromonas gingivalis are co-cultured, not only can it not inhibit the growth of Porphyromonas gingivalis, but it has a certain effect on promoting its growth ; and it is disclosed in the application text that only when the two are used together, will there be an effect of inhibiting the growth of Porphyromonas gingivalis
However, in order to exert the effect of inhibiting Porphyromonas gingivalis, this product needs to have two kinds of probiotics at the same time, which virtually increases the production cost. Secondly, this product can only inhibit the growth of Porphyromonas gingivalis strains, while There are many kinds of bacteria in the environment of periodontitis, so only inhibiting the growth of one kind of bacteria can not achieve the purpose of treating periodontitis, and the mixed bacteria in the environment of periodontitis will also interact with each other, so how can we Obtaining a probiotic preparation that can inhibit the growth of mixed bacteria has become a research hotspot and difficulty
[0009] As can be seen from the above, according to the existing methods, it is necessary to add a variety of lactobacilli to exert beneficial effects, and the cost is relatively high.
In addition, the existing methods only aim at reducing the biomass of oral pathogenic bacteria, but do not involve the amount of biofilm formed by various pathogenic bacteria in the oral cavity and the inflammatory response caused by periodontitis, and the therapeutic effect is limited

Method used

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  • A strain of Lactobacillus fermentum capable of preventing and/or treating periodontitis and its application
  • A strain of Lactobacillus fermentum capable of preventing and/or treating periodontitis and its application
  • A strain of Lactobacillus fermentum capable of preventing and/or treating periodontitis and its application

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Experimental program
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Effect test

preparation example Construction

[0064] The method of preparing the bacterial suspension involved in the following examples is as follows:

[0065] With the suspension of the nucleus, the Nucleus Bacillus is inoculated into the BHI medium in the inoculation amount of 2% (v / v) in the total volume of BHI medium, and then at 37 ° C for anaerobic culture for 48 h, with BHI Medium-based modifier concentration is 1 × 10 9 CFU / ml.

[0066] Gubprinocensis Suspension: Inoculation of the gingival porphty cells inocatalyst in the BHI medium in the amount of 2% (v / v) of the total volume of BHI medium, 48 h at anaerobic culture at 37 ° C After using BHI medium, the hydrochloride concentration is 1 × 10 9 CFU / ml.

[0067] Intermediate Placeonius suspension: Intermediate P. aeruginus is inoculated into BHI medium in a inoculation amount of 2% (V / V) in BHI medium, and in anaerobic culture at 37 ° C for 48 hours. Based sized hydrochloride concentration is 1 × 10 9 CFU / ml.

[0068] The method of equipped with the PBS b...

Embodiment 1

[0070] Example 1: Screening and strain identification of Lactobacillus Fermentum CCFM1139

[0071] 1, screening

[0072] Since the feces sample from the healthy person from Yangzhou area, after pretreatment, the sample is preserved in the -80 ° C refrigerator, remove the thaw, mix the sample, and absorb 0.5ml sample to 4.5ml 9g / L Physiological saline was diluted, selecting a suitable gradient diluent coated on MRS medium containing 20 g / L agar 48 h at 37 ° C for 48 h, and a typical colonies of fermentation Lactobacillus were selected to a MRS medium containing 20 g / L agar. Undiren purification, picking up a single colony transfer to MRS medium, 30% glycerol deposit, resulting in strain CCFM1139 and synchronous strains 1 to 19; wherein the typical colonies of the fermented Lactobacillus are small white and translucent. Round.

[0073] 2, identification

[0074] The genome of strain CCFM1139 and strains 1 to 19 was extracted, and 16S RDNA of strain CCFM1139 and strain 1 to 19 ...

Embodiment 2

[0075] Example 2: The effect of Lactobacillus Fermentum CCFM1139 on the amount of mixed biofilm is divided into two groups, which are fermented Lactobacillus and blank control groups;

[0076] Among them, the fermentation Lactobacillus system is handled in accordance with the following method:

[0077] (1) The fermented Lactobacillus CCFM1139, a bacterium, fermented Lactobacillus 1 to 19, respectively, inoculated into the MRS medium in the MRS medium total volume of 2%, at 37 ° C for 24 h, obtained culture solution, will be cultured The fluids were centrifuged at 8000 r / min, 4 ° C for 5 min, and the supernatant was obtained, and the supernatant was filtered at 0.22 μm sterile filmers, respectively, respectively. 9 CFU / mL of the fermentation of Lactobacillus CCFM1139, the supernatant of the fermentation of Lactobacillus 1 to 19;

[0078] (2) In the 96-well plate, 40 μl of the gingival porphty cells and the nucleus of the nucleus, and the mixed bacteria suspension is obtained, a...

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Abstract

The invention discloses a strain of Lactobacillus fermentum capable of preventing and / or treating periodontitis and its application, belonging to the technical field of microorganisms. According to the invention, a strain of Lactobacillus fermentum CCFM1139 is obtained by screening, and the Lactobacillus fermentum CCFM1139 has the effect of relieving periodontitis, which is embodied in that it can inhibit the mixed organisms of Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum. The formation of film reduces the amount of biofilm by 52.45%; the content of TNF-α in the periodontitis cell model can be reduced from 131.37pg / mL to 83.31pg / mL, and the content of IL-8 can be reduced from 147.70pg / mL to 121.12 pg / mL; reduced the colonization of Porphyromonas gingivalis and Fusobacterium nucleatum in the oral cavity by 1-2 orders of magnitude; reduced alveolar bone resorption from 1838.0 μm to 805.7 μm.

Description

Technical field [0001] The present invention relates to a fermented Lactobacillus capable of preventing and / or treating teeth and its applications, belonging to the microbial technology. Background technique [0002] Periodontitis is a common oral disease that can cause loss of periodontal tissue, eventually leading to the teeth. The main pathogenic bacteria of periodontitis include gingival porphyridomycetes, derogatorium, and intermediate P., the infection caused by microorganisms can not only lead to the damage of gums and periodontal tissues. It will stimulate the body to produce an immune response, so that the host continuously releases various inflammatory factors, so that the body is in an inflammatory state for a long time. [0003] Under normal circumstances, there is a balance between the various microorganisms in the gingival area and the environment of the immune inflammatory medium, which can realize the steady state in the host periodontal tissue. Once the balance...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/747A61K8/99A61Q11/00A61P1/02A61P31/04A23L33/135A23C9/123A23G4/12C12R1/225
CPCA61K35/747A61K8/99A61Q11/00A61P1/02A61P31/04A23L33/135A23C9/1234A23G4/123A23V2002/00A23V2400/143A23V2200/312C12N1/20C12N1/205C12R2001/225A23V2400/11
Inventor 陈卫张秋香徐晚晴陆文伟张灏赵建新
Owner JIANGNAN UNIV
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