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A kind of cultivation method of dental plaque biofilm model and optimized biofilm live bacteria counting method and application

A culture method and technology for counting viable bacteria, which are used in the cultivation of dental plaque biofilm models, evaluating the bactericidal efficacy and bacteriostatic efficacy of oral care products, and in the field of biofilm viable bacteria counting methods, which can solve the problem of decreased sensitivity to antibacterial agents, The problems of large differences in the tested individuals and unfavorable screening work have achieved obvious effects, strong repeatability, and improved viable count results.

Active Publication Date: 2020-12-01
GUANGZHOU LIBY
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] (3) Strong tolerance and decreased sensitivity to antibacterial agents;
Clinical trials are a more realistic and objective way to evaluate the antibacterial efficacy of products against biofilms. The results are relatively reliable and representative, but there are also some obvious disadvantages: (1) It takes a long time, about 1 week; (2) ) The cost is high; (3) The tested individuals have great differences; (4) The tested individuals need a high degree of cooperation
Preliminary formulation research of oral care products requires the screening of a large number of antibacterial raw materials with different concentrations, and clinical trials are not conducive to the screening of a large number of formulations of oral care products in the early stage

Method used

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  • A kind of cultivation method of dental plaque biofilm model and optimized biofilm live bacteria counting method and application
  • A kind of cultivation method of dental plaque biofilm model and optimized biofilm live bacteria counting method and application
  • A kind of cultivation method of dental plaque biofilm model and optimized biofilm live bacteria counting method and application

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preparation example Construction

[0089] Preparation method of dental plaque biofilm

[0090] The preparation mode of dental plaque biofilm involved in the present invention is as follows:

[0091] S101. In an oxygen-free atmosphere and a constant temperature environment, respectively streak and culture various oral pathogenic bacteria on a TSA-supplemented medium plate for 2 to 6 days until a large number of bacterial lawns appear on the plate;

[0092]S102. Inoculate the various oral pathogenic bacteria cultivated in step S101 into the BHI-enriched medium, and culture them in an oxygen-free atmosphere and a constant temperature environment for 1 to 3 days until the BHI-enriched medium becomes turbid to obtain various Mature bacterial suspension of oral pathogenic bacteria;

[0093] S103. Add various mature bacterial suspensions obtained in step S102 to the BHI enrichment medium containing an adhesion promoter, and the volume percentage of each mature bacterial suspension is 2% to 8%; then in an oxygen-fre...

Embodiment 1

[0214] Specifically include the following steps:

[0215] 1) Culture of dental plaque biofilm

[0216] Dental plaque biofilms were cultured in TC-treated 24-well cell culture plates according to the steps in the preparation example of the dental plaque biofilm model. Calcium carbonate suspension was added as an adhesion promoter, and the incubation time was 16 hours.

[0217] 2) Antibacterial testing of oral care products using dental plaque biofilm

[0218] Draw TC to handle the bacterial suspension in the two small holes of the cell culture plate, and discard it; add 0.2mL~1mL oral care product samples of a certain concentration along the hole wall (the test sample solution in this embodiment is the test sample solution of the toothpaste sample) , keep for 3 minutes, draw the sample, and discard it; immediately add an equal volume of BHI-enriched medium to the small well along the wall of the hole, absorb the BHI-enriched medium, and discard it; then add it twice along the...

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Abstract

The invention relates to the technical field of antibacterial effect evaluation of oral care products, in particular to a dental plaque biofilm model cultivation method, an optimized biofilm viable counting method and a method for rapidly evaluating antibacterial effects of the oral care products. According to the dental plaque biofilm model cultivation method, a dental plaque biofilm model capable of improving evaluation accuracy of the antibacterial effects of the oral care products can be cultivated by optimizing cultivation conditions. According to the optimized biofilm viable counting method, viable counting results can be improved and are closer to the number of viable bacteria on a real biofilm. According to the method for rapidly evaluating the antibacterial effects of the oral care products, detection results of the method are closer to evaluation results after practical use of consumers as compared with a free bacteria method, the effect evaluation time of the oral care products can be greatly reduced, data support can be provided for rapidly distinguishing the effects of the oral care products, repeatability is strong, result stability is good, the cost is low, and the cultivation conditions are easily met.

Description

technical field [0001] The invention relates to the technical field of antibacterial efficacy evaluation of oral care products, in particular to a method for cultivating a dental plaque biofilm model and an optimized method for counting viable biofilm bacteria, and evaluating the bactericidal efficacy of oral care products using the dental plaque biofilm model and antibacterial methods. Background technique [0002] There are a variety of microorganisms living in the oral cavity, including bacteria, fungi, viruses, mycoplasma, protozoa, etc., and their existence forms include free state and aggregated state. Free state microorganisms are called planktonic microorganisms, and aggregated state microorganisms are mainly dental plaque biofilms. present in the human oral cavity. Compared with the free state, the microbial population in the form of biofilm has the following characteristics: [0003] (1) Microorganisms accumulate and colonize each other, with spatial and environm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12Q1/06C12Q1/18C12R1/46C12R1/225C12R1/04C12R1/01
CPCC12N1/20C12Q1/06C12Q1/18G01N2333/195G01N2333/315G01N2333/335G01N2333/36
Inventor 李淑钰江丹钟美邱振名黄亮张利萍
Owner GUANGZHOU LIBY