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Application of miRNA in preparation of medicine for preventing and treating osteoarthritis, exosome with high expression of miRNA, and application of exosome

A technology for osteoarthritis and exosomes, which is applied in the field of miRNA and exosomes, can solve problems such as unimproved joint damage, achieve the effects of promoting chondrocyte proliferation, promoting cartilage regeneration, and reducing osteoarthritis damage

Active Publication Date: 2021-01-08
SUZHOU MUNICIPAL HOSPITAL (NORTH DISTRICT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are still many difficulties to be overcome in the treatment of osteoarthritis, because most of the drugs for osteoarthritis can only relieve joint pain, and the joint damage has not been improved

Method used

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  • Application of miRNA in preparation of medicine for preventing and treating osteoarthritis, exosome with high expression of miRNA, and application of exosome
  • Application of miRNA in preparation of medicine for preventing and treating osteoarthritis, exosome with high expression of miRNA, and application of exosome
  • Application of miRNA in preparation of medicine for preventing and treating osteoarthritis, exosome with high expression of miRNA, and application of exosome

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preparation example Construction

[0028] In the present invention, the preparation method of the SMSC-155-5p-Exos is a conventional preparation method, preferably comprising: extracting synovial tissue from the knee articular cartilage of a patient undergoing total knee arthroplasty for osteoarthritis (Osteoarthritis, OA), Remove fat and some connective tissues, cut them into pieces, add DMEM medium containing collagenase Ⅱ and 20% fetal bovine serum (FBS), and digest overnight in a CO2 incubator; suspend and inoculate the cell pellet on a 60mm culture dish, and change the medium after 24 hours Remove non-adherent cells. Change the medium every 3 days. When the cells grew to 80% confluence, the cells were placed in a medium containing 10% fetal bovine serum (FBS), 25 μg / ml ascorbic acid 2-phosphate and 1% penicillin streptomycin at 37 °C and 5% carbon dioxide. cultured in DMEM medium to obtain synovial mesenchymal stem cells (SMSCs). use 2000 transfection reagent cultured SMSCs at a concentration of 100nM ...

Embodiment 1

[0033] The cartilage tissue was extracted from the knee articular cartilage of patients undergoing total knee arthroplasty for osteoarthritis, and the cartilage tissue was extracted with collagenase II to obtain primary chondrocytes. OA chondrocytes were obtained by culturing in DMEM medium containing 10% fetal bovine serum (FBS), 25 μg / ml ascorbic acid 2-phosphate and 1% penicillin-streptomycin. use 2000 transfection reagent was used to culture OA chondrocytes at a concentration of 100 nM and transfect miR-155-5p mimics to obtain transfected chondrocytes. SMSCs were transfected with miR-155-5p mimics, and then separated and concentrated to obtain SMSC-155-5p-Exos.

[0034] The method of separation and concentration is as follows: the transfected chondrocytes are cultivated with serum-free DMEM medium to obtain a culture supernatant; the first supernatant is obtained after the culture supernatant is subjected to the first centrifugation (centrifugal force is 1000r / min). Sup...

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Abstract

The invention relates to the field of miRNAs and exosomes, in particular to an application of miRNA in the preparation of a medicine for preventing and treating osteoarthritis, an exosome with the high expression of the miRNA, and an application of the exosome. The invention provides an application of miR-155-5p in the preparation of a medicine for preventing and / or treating osteoarthritis, and anucleotide sequence of the miR-155-5p is as shown in SEQ ID NO: 1. The application provided by the invention can effectively improve the expression of the miR-155-5p, and the miR-155-5p can promote cartilage cell proliferation, migration and extracellular matrix secretion and inhibit cell apoptosis by targeting Runx2, and thereby achieves the effect of preventing or treating osteoarthritis.

Description

technical field [0001] The present invention relates to the field of miRNA and exosomes, in particular to the application of miR-155-5p in the prevention and treatment of osteoarthritis, exosomes derived from synovial mesenchymal stem cells with high miR-155-5p expression, and applications thereof. Background technique [0002] Osteoarthritis (OA) is one of the most common joint diseases and has become a leading cause of disability in the elderly. Studies have found that various factors such as trauma, abnormal mechanical load, lack of nutrition, and genetic predisposition can lead to the occurrence of osteoarthritis. At present, there are still many difficulties to be overcome in the treatment of osteoarthritis, because most of the drugs for osteoarthritis can only relieve joint pain, and the joint damage has not been improved. [0003] The occurrence of osteoarthritis is mainly related to the decrease of the number of chondrocytes in the joint tissue, the increase of apop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7088A61K35/28A61P19/02A61P29/00
CPCA61K31/7088A61K35/28A61P19/02A61P29/00C12N2310/141C12N15/113C12N2320/32C12N5/0668C12N5/0662
Inventor 郝跃峰杨兴周静许檬磊耿德春
Owner SUZHOU MUNICIPAL HOSPITAL (NORTH DISTRICT)
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