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Medicine for treating periodontal ligament cell dysfunction and alveolar bone loss caused by periodontal pathogen infection

A periodontal disease and drug technology, applied in the direction of gene therapy, drug combination, in vivo test preparations, etc., can solve the problems of insignificant therapeutic effect, inability to achieve a radical cure, and large side effects, and achieve the purpose of inhibiting the loss of periodontal ligament tissue , prevent the occurrence and development, and reduce the effect of the disease

Pending Publication Date: 2019-11-12
HOSPITAL OF STOMATOLOGY SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Antibiotics or traditional Chinese medicines are used in the treatment of periodontitis in the prior art, which have complex components, large side effects, insignificant therapeutic effect, and cannot achieve the purpose of radical cure

Method used

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  • Medicine for treating periodontal ligament cell dysfunction and alveolar bone loss caused by periodontal pathogen infection
  • Medicine for treating periodontal ligament cell dysfunction and alveolar bone loss caused by periodontal pathogen infection
  • Medicine for treating periodontal ligament cell dysfunction and alveolar bone loss caused by periodontal pathogen infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Silk thread ligation combined with Porphyromonas gingivalis periodontitis modeling

[0064] (1) Bacterial culture

[0065] Mix 0.5% hemin vitamin solution and bovine heart brain infusion juice medium (BHI medium) according to 1:100, inoculate the P.gingivalis strain in the medium, in 85% nitrogen, 10% hydrogen, 5% Cultivate under strict anaerobic conditions of carbon dioxide and 37°C;

[0066] After two generations of culture, the bacterial concentration reached 1×10 8 For CFU / mL, resuspend the bacteria in PBS buffer containing 2% sodium carboxymethylcellulose for use;

[0067] The bacteria cultured for 48 hours were identified according to the colony morphology, Gram staining, cell morphology and biochemical reaction results;

[0068] (2) modeling

[0069] Choose C57BL / 6 mice with complete dentition, no caries and periodontal disease for 8 weeks, and give them in an environment with good lighting conditions, clean, quiet, well-ventilated, room temperature...

Embodiment 2I

[0073] Example 2 Preparation of IL-18 receptor siRNA exosomes

[0074] (1) Design and synthesis of IL-18Rα siRNA sequence:

[0075] The nucleic acid sequence of IL-18Rα siRNA is shown in SEQ ID NO: 1-2:

[0076] SEQ ID NO: 1 (sense strand): 5'-AAACUCGGCAUCCUUCAGGUU-3';

[0077] SEQ ID NO: 2 (antisense strand): 5'-AACCUGAAGGAUGCCGAGUUUTT-3';

[0078] (2) Separation and purification of exosomes derived from mouse serum

[0079] ① Take the mouse serum and centrifuge it at 300g for 10min at 4°C, discard the precipitate and remove the cells;

[0080] ②Take the supernatant, centrifuge at 2000g for 10min, discard the precipitate, and remove dead cells;

[0081] ③Take the supernatant, centrifuge at 10000g for 30min, discard the precipitate, and remove the cell debris;

[0082] ④Take the supernatant and centrifuge at 100,000g for 70min, the resulting precipitate is exosomes;

[0083] ⑤Discard the supernatant, resuspend the pellet with PBS, and centrifuge at 100,000g for 70min;

...

Embodiment 3IL-18

[0088] Example 3 Effect of IL-18 on the secretion of pro-inflammatory cytokines by periodontal ligament cells

[0089] In this example, periodontal ligament cells were stimulated and cultured with IL-18 for 96 hours, and the cell supernatant was collected, and the expression levels of IFN-γ, GM-CSF, IL-2 and TNF-α were detected by ELISA.

[0090] The results are shown in Figure 1(A), Figure 1(B), Figure 1(C), Figure 1(D) and Table 1, the expressions of the above cytokines in the culture supernatant after IL-18 stimulation were all significantly increased ( *P<0.05).

[0091] Table 1 Levels of cytokines secreted by periodontal ligament cells after IL-18 stimulation (pg / mL)

[0092] Cytokines Control group (pg / mL) IL-18 stimulation (pg / mL) IFN-γ 82.11±17.24 687.1±109.3* GM-CSF 32.90±7.29 89.27±19.47* IL-2 114.2±28.39 197.6±54.28* TNF-α 881.7±263.2 2269±746.1*

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Abstract

The invention provides a medicine for treating periodontal ligament cell dysfunction and alveolar bone loss caused by periodontal pathogen infection. The medicine includes an IL-18 antibody and / or anIL-18R receptor siRNA exosomes. The medicine of the present invention uses the IL-18 antibody and / or the IL-18R receptor siRNA exosomes as a functional component, by combining with IL-18 in periodontal tissue, the secretion of proinflammatory cytokines by periodontal ligament cells can be inhibited, the periodontal ligament cell loss and inflammatory cell infiltration caused by periodontal ligament cell dysfunction under inflammation can be improved, the occurrence and development of periodontal disease can be prevented, and the medicine has potential application value for the clinical prevention and treatment of periodontal disease.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a medicine for treating periodontal ligament cell dysfunction and alveolar bone loss caused by periodontal pathogenic bacteria infection. Background technique [0002] Periodontitis is a common oral disease, mainly caused by pathogenic microorganisms in the plaque biofilm to initiate innate immunity, adaptive immunity and inflammatory response, manifested as a series of complex inflammatory syndromes, mainly affecting the periodontal ligament and surrounding and the tissues supporting the teeth, leading to progressive loss of periodontal alveolar bone, which in turn leads to tooth loss. [0003] The pathogenic microorganisms in the dental plaque biofilm are the main cause of periodontal disease, and the gingival microbial community is transplanted in the periodontal pocket, so that the dominant microbial community in the periodontal pocket changes from Gram-positive aerobic bacter...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61K31/7105A61K48/00A61P1/02A01K67/027A61K49/00
CPCA01K67/027A01K2207/30A01K2227/105A01K2267/03A61K31/7105A61K39/395A61K49/0008A61P1/02A61K2300/00
Inventor 谭家莉李丹枫匡威
Owner HOSPITAL OF STOMATOLOGY SUN YAT SEN UNIV
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