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Application of melatonin in preparation of drugs for promoting mesenchymal stem cell chondrogenic differentiation

A technology of stem cells and melatonin, applied in the field of medicine, can solve the problems of potential protective effect and lack of mechanism

Inactive Publication Date: 2014-01-08
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few reports on the potential protective effect and mechanism of melatonin in promoting chondrogenic differentiation of mesenchymal stem cells in the inflammatory microenvironment of joint diseases

Method used

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  • Application of melatonin in preparation of drugs for promoting mesenchymal stem cell chondrogenic differentiation
  • Application of melatonin in preparation of drugs for promoting mesenchymal stem cell chondrogenic differentiation
  • Application of melatonin in preparation of drugs for promoting mesenchymal stem cell chondrogenic differentiation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Effect of melatonin on intracellular reactive oxygen species, SOD activity and cell proliferation of synovial mesenchymal stem cells in IL-1β or TNF-α inflammatory microenvironment

[0037] P4 synovial mesenchymal stem cells at 3000cell / cm 2 Inoculate 12-well plates and culture in 5% CO 2Replace the basal medium (α-MEM, 10% FBS, 100 U / mL penicillin, 100 μg / mL streptomycin, 0.25 μg / mL amphotericin B) every other day. After the cell density reaches 95%, replace with cartilage medium (DMEM / HIGH GLUCOSE, 40 μg / mL proline, 100 nM dexamethasone, 100 U / mL penicillin, 100 μg / mL streptomycin, 0.25 μg / mL Amphotericin B, 0.1 mM L-ascorbic acid, ITS® Premix, 20 ng / mL TGF-β1) for chondrogenic induction, adding 10ng / ml IL-1β or 10ng / ml TNF-α to simulate the microenvironment in vivo, plus or No melatonin was used as the experimental group, and the cartilage medium was the control group, marked as: CTRL (control group), IL (IL-1β without melatonin), MT+IL (IL-1β plus mela...

Embodiment 2

[0048] Example 2 Effect of melatonin on chondrogenic differentiation of synovial mesenchymal stem cells in vitro in IL-1β or TNF-α inflammatory microenvironment

[0049] The cell culture method is the same as that in Example 1, and the P4 generation of synovial mesenchymal stem cells is 3000 cell / cm 2 Inoculate 12-well plates and culture in 5% CO 2 After the cell density reaches 95%, replace with cartilage medium, add 10ng / ml IL-1β or 10ng / ml TNF-α to simulate the microenvironment in vivo, the experimental group is marked as: CTRL (control group), IL (IL-1β without melatonin), MT+IL (IL-1β plus melatonin), TNF (TNF-α without melatonin), MT+TNF (TNF-α plus melatonin), every Change medium every day.

[0050] 1. Chondrogenesis was induced for 14 days. The cells in the orifice plate were fixed overnight with 4% paraformaldehyde, washed with PBS and then washed with 1% paraformaldehyde.

[0051] Alcian blue (dissolved in 3% acetic acid) at pH = 2.5 was used to stain cartilage...

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Abstract

The invention discloses new application of melatonin, fills the technical blanks in the prior art, and discloses the application of the melatonin in the preparation of drugs for promoting mesenchymal stem cell chondrogenic differentiation, the application of the melatonin in the preparation of drugs for treatment of joint diseases, and the application of the melatonin in the preparation of drugs for treatment of cartilage diseases. The application introduces two representative proinflammatory cytokines including IL-1 beta (Interleukin-1 beta) and TNF-alpha (tumor necrosis factor-alpha), creates two in-vitro arthritis cell models for studying to determine mesenchymal stem cell in-vitro chondrogenic differentiation promoted by the melatonin and a mechanism thereof, and establishes a foundation for further application of the melatonin in the preparation of the drugs for promoting the mesenchymal stem cell chondrogenic differentiation, in the preparation of the drugs for treatment of the joint diseases, and in the preparation of the drugs for treatment of the cartilage diseases.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to the application of melatonin in the preparation of drugs for promoting chondrogenic differentiation of mesenchymal stem cells. Background technique [0002] Stem cells are a class of cells with self-renewal and differentiation potential, including embryonic stem cells, mesenchymal stem cells and pluripotent induced stem cells, which can be further differentiated into various tissues and constitute various complex body organs, and have been widely used in the field of regenerative medicine and tissue engineering. Mesenchymal stem cells (MSCs) play an important role in development, growth, self-regeneration of tissues and organs, and maintenance of microenvironment balance in the body. Due to their easy extraction, strong self-renewal ability and multi-directional differentiation It has been widely used in regenerative medicine, gene therapy, tissue engineering, cytokine replac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4045A61P19/02A61P29/00A61P19/08
Inventor 龚逸鸿何帆刘小珍蒋庆
Owner SUN YAT SEN UNIV
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