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Application of mitochondrial transplantation in treatment of periodontitis

A technology for mitochondria and periodontitis, applied in the application field of mitochondrial transplantation in the treatment of periodontitis, can solve the problems of loss of periodontal support tissue and ineffective repair, and achieve the solution of ineffective recovery, enhance osteogenic differentiation potential, and promote Bone effect

Pending Publication Date: 2021-11-12
HOSPITAL OF STOMATOLOGY SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are various treatment methods for periodontitis, and new technologies and drugs are constantly being developed and updated, the existing treatment methods are still unable to effectively repair the loss of periodontal support tissue that has occurred
There is no report on the use of mitochondrial transplantation in the treatment of periodontitis

Method used

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  • Application of mitochondrial transplantation in treatment of periodontitis
  • Application of mitochondrial transplantation in treatment of periodontitis
  • Application of mitochondrial transplantation in treatment of periodontitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Mitochondria Isolation and Electron Microscopic Observation and Activity Test

[0032] 1. Mitochondria Isolation

[0033] The mitochondria of healthy human periodontal ligament stem cells (hPDLSCs) were extracted for subsequent experiments. The specific method is as follows:

[0034] (1) Primary culture of hPDLSCs, inoculation of hPDLSCs in the logarithmic phase of growth in 150mm culture dishes, when the cell density reached 90%, the mitochondria were extracted according to the instructions of the mitochondrial extraction kit.

[0035] (2) Conventionally digest and centrifuge the above hPDLSCs cells, collect the cell pellet, resuspend the cell pellet in 2ml PBS, and transfer the cell suspension to a new 2ml EP tube;

[0036] (3) After centrifuging at 1200 rpm for 5 min, discard the supernatant carefully, and add 800 μl of mitochondrial extraction reagent A. Place the EP tube on the vortexer and vortex at the maximum speed for 5s, then immediately place it ...

Embodiment 2

[0059] Example 2 Effect of mitochondria on the osteogenic differentiation ability of hPDLSCs in the inflammatory microenvironment

[0060] Experimental group:

[0061] ① hPDLSCs: routinely cultured, and the medium was changed synchronously with the rest of the groups.

[0062] ② hPDLSCs+OS (osteoinduction, osteogenesis induction): the osteogenesis induction medium was replaced synchronously with the rest of the groups.

[0063] ③hPDLSCs+P.g-LPS (Porphyromonas gingivalis lipopolysaccharide): stimulated with α-MEM medium containing a final concentration of 1 μg / ml P.g-LPS for 24 hours, and then replaced with fresh medium.

[0064] ④hPDLSCs+P.g-LPS+OS: After being stimulated for 24 hours with α-MEM medium containing P.g-LPS at a final concentration of 1 μg / ml, the osteogenic induction medium was replaced synchronously.

[0065] ⑤ hPDLSCs+P.g-LPS+MT (Mitochondria, mitochondria): After being stimulated with α-MEM medium containing a final concentration of 1 μg / ml P.g-LPS for 24 h...

Embodiment 3

[0090] Example 3 Effect of Mitochondria on the Immunomodulatory Function of Inflammation-Injured hPDLSCs

[0091] (1) Method

[0092] 1) Preparation of hPDLSCs supernatant: hPDLSCs in the logarithmic growth phase were inoculated into 10 cm culture dishes. The experimental groups were as follows: Control group, Pg-LPS group, Pg-LPS+MT group. On the second day after inoculating the cells, when the cell density reached 40%, the LPS group and the Pg-LPS+MT group were stimulated with 1 μg / ml LPS-P.G for 24 hours and then replaced with fresh medium, and the Pg-LPS+MT group was added with the extracted mitochondria ( According to the ratio of the number of recipient cells: the number of donor cells = 1:3, that is, the number of recipient cells is 1.5×10 6 , the number of donor cells is 4.5×10 6 hPDLSCs) were treated for 24 h, and 10 ml of fresh complete medium was replaced after the treatment. The control group and the Pg-LPS group were changed synchronously, and the supernatant ...

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Abstract

The invention discloses an application of mitochondrial transplantation in treatment of periodontitis. The mitochondria is extracted from healthy human periodontal ligament stem cells through a kit and a differential centrifugation method, and the obtained mitochondria has a mitochondrial structure and function integrity. In an in-vitro experiment, the mitochondria can be captured by the human periodontal ligament stem cells damaged by inflammation, the osteogenic differentiation capability of the human periodontal ligament stem cells can be enhanced, the immunoregulation function of the periodontal ligament stem cells in an inflammation microenvironment can be adjusted, macrophages can be promoted to be differentiated to am M2 type, and inflammation can be relieved, so that the mitochondria can be used for treating periodontitis and other inflammatory diseases. A periodontitis animal model research shows that mitochondrial transplantation can relieve inflammatory periodontal tissue damage and promote periodontal tissue osteogenesis, and therefore, the problem that lost periodontal tissues cannot be effectively recovered by existing treatment means is solved.

Description

technical field [0001] The invention relates to the technical field of medicines for treating periodontitis, and more specifically, relates to the application of mitochondria transplantation in treating periodontitis. Background technique [0002] Periodontitis is a chronic inflammatory disease that occurs in the periodontal supporting tissue. It is initiated by plaque microorganisms and the host immune response is involved. It is characterized by gingival inflammation, periodontal pocket formation, alveolar bone resorption and tooth loosening. cause tooth loss. Periodontal disease is not only the main cause of tooth loss in adults, but also closely related to systemic diseases, such as atherosclerosis, diabetes, chronic kidney disease, senile pneumonia, rheumatoid arthritis, Alzheimer's disease, etc. This makes the treatment of periodontitis more difficult. The treatment of periodontitis is mainly through non-surgical or surgical treatment of periodontal tissue regenerati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/32A61P1/00A61P29/00
CPCA61K35/32A61P1/00A61P29/00
Inventor 王彦于乐李逸曹泽源欧乾民
Owner HOSPITAL OF STOMATOLOGY SUN YAT SEN UNIV
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