Application of microRNA‑29b as a drug target in articular cartilage repair

A technology of chondrocytes and bone marrow mesenchyme, applied in the field of cartilage tissue engineering, to prevent cartilage fibrosis and heterotopic ossification, realize tissue engineered cartilage repair, and promote maintenance and stability

Active Publication Date: 2017-07-18
ARMY MEDICAL UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on repairing defective articular cartilage from the perspective of inhibiting hypertrophy of MSCs-derived chondrocytes in the prior art

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of microRNA‑29b as a drug target in articular cartilage repair
  • Application of microRNA‑29b as a drug target in articular cartilage repair
  • Application of microRNA‑29b as a drug target in articular cartilage repair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Mesenchymal stem cell culture and hypertrophic chondrogenic differentiation induction experiment

[0036] (1) Acquisition and culture of mouse bone marrow mesenchymal stem cells

[0037] Under sterile conditions, wash the mouse femoral bone marrow with a 1mL syringe, transfer it into a centrifuge tube filled with 5mL DMEM medium, and blow it gently to make a cell suspension; transfer the cell suspension into a tube containing 5mL lymphocyte In the centrifuge tube of separation liquid, centrifuge 20min under the condition of 2000r / min; After centrifugation, collect the mononuclear cells of cloudy buffy coat and place in another centrifuge tube, wash twice with DMEM low-sugar medium, centrifuge (1000r / min) min, 5min), the supernatant was discarded, and the cells were resuspended with DMEM basal medium containing 15% fetal bovine serum, and finally the primary cells were diluted with 1×10 6 Cells / mL were inoculated into culture flasks and placed at 37°C, 5% CO ...

Embodiment 2

[0040] Example 2 Increased expression of microRNA-29b promotes hypertrophy of chondrocytes

[0041] (1) Collect the cell clusters of 14d, 21d and 28d of chondrogenic induction, extract total RNA and reverse transcribe it into cDNA. The specific operation of reverse transcription is as follows: put the PCR tube on ice, and add 5 μL 5×RT Reaction Buffer, 1 μL 25mMdNTP mixture, 1 μL 25U / μL RNase inhibitor, 1 μL 200U / μL M-MLV reverse transcriptase and 4 μL of DEPC water, then add the mixture of RNA and primers to make the total reaction volume 25 μL, mix the above components and bathe in 42°C water for 60 minutes, and finally heat to 85°C for 5 minutes to get the cDNA solution, put the solution in- Store at 70°C for later use.

[0042] The mRNA sequences of type II collagen (Col II), type X collagen (Col X), Sox9, and internal reference gene GAPDH from mice were obtained from the Nucleotide database, and the corresponding PCR primers were designed with Primer Premier 5.0 software...

Embodiment 3

[0065] Example 3 Transfection of microRNA-29b inhibitor can significantly inhibit hypertrophic chondrocyte-related gene expression

[0066] Transfection experiment: using Lipofectamin TM 2000 kit, take normal cultured mesenchymal stem cells and culture them in cell micromass, use medium B (complete chondrogenic induction medium containing TGF-β3) for chondrogenic induction culture in the first 14 days, and replace with medium from the 15th day C (hypertrophic cartilage induction complete medium) continued to be cultured, and samples were collected on the 14th day, 21st day, and 28th day of culture for subsequent experimental detection. The experiment was divided into two groups, the control group and the experimental group. The experimental group was transfected with microRNA-29b inhibitor, and the control group was transfected with microRNA-29b inhibitor negative control (bioinformatics analysis showed that it was compatible with human, mouse, rat genomes, and All miRNAs in...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses an application of microRNA-29b in articular cartilage repair as a drug target. According to the application disclosed by the invention, the research indicates that microRNA-29b is a key miRNA for regulating the hypertrophy change of MSCs after chondrogenic differentiation, therefore, the hypertrophy of MSCs-derived cartilage cells is inhibited by virtue of an microRNA-29b inhibitor, the phenotypes of the MSCs-derived cartilage cells are stabilized, and the fibrosis and the heterotopic ossification of tissue engineering cartilages are prevented, thus further realizing tissue engineering cartilage repair, and providing a new thought for tissue engineering cartilage construction, cartilage framework material design and cartilage repair.

Description

technical field [0001] The invention belongs to the technical field of cartilage tissue engineering, and in particular relates to the application of MicroRNA-29b as a drug target in articular cartilage repair. Background technique [0002] Damage to articular cartilage on the surface of bone joints can be caused by various causes such as trauma, osteoarthritis, tumors, etc., and often presents progressive damage, which seriously affects the quality of life of patients. Since articular cartilage lacks self-repair ability, how to realize the functional repair of articular cartilage defect is a major problem in the medical field. Tissue engineering technology combines cell biology and biomaterials to repair damaged tissue physiologically, which provides an ideal repair method for articular cartilage defect repair. Among them, selecting and optimizing suitable seed cells is an important proposition in this field. Mesenchymal stem cells (MSCs) are considered as promising seed c...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61P19/04
Inventor 董世武康菲苗莹杨波李建美曹震窦策赵春容
Owner ARMY MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap