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Application of miR-92b and simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation

A 1.mir-92b, bone marrow mesenchymal technology, applied in the field of medicine, can solve problems such as many complications, and achieve the effect of promoting the osteogenic differentiation of bone marrow mesenchymal stem cells

Inactive Publication Date: 2019-03-29
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These traditional treatment methods have certain limitations and trauma in clinical application, and there are many complications. Therefore, it is particularly important to explore and discover a new treatment method.

Method used

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  • Application of miR-92b and simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation
  • Application of miR-92b and simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation
  • Application of miR-92b and simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Example 1: miR-92b regulates the osteogenic differentiation of bone marrow mesenchymal stem cells

[0034] 1 Experimental method

[0035] 1.1 Alizarin red staining to detect the number of calcium nodules formed by osteoblasts and alkaline phosphatase staining to indirectly reflect the activity of the marker enzyme ALP of osteoblasts

[0036] 1.2 Experiment grouping design: The experiment was divided into the following four groups according to the transfection control group and treatment group: miR-92bmimic, mimic-NC, miR-92b AMO, AMO-NC.

[0037] 1.3 Transfection method: seed bone marrow mesenchymal stem cells in a well plate, and perform transfection when the confluence of the cells reaches about 40%. Discard the culture medium in the well plate, replace with fresh serum-free OPTI-DMEM medium, starve the cells for 2 hours, and start transfection after 2 hours. Mix microRNA mimic and AMO with transfection reagent X-treme well and add to the well plate, so that the fin...

Embodiment 2

[0051]Example 2: miR-92b promotes the expression of Runx2, an important regulatory transcription factor in the process of osteogenic differentiation

[0052] 1 Experimental method

[0053] 1.1 Immunofluorescence staining to detect the expression of Runx2, an important regulatory transcription factor in the process of osteogenic differentiation

[0054] 1.2 Experimental grouping design: the experiment was divided into 4 groups: the experiment was divided into the following four groups according to the transfection control group and the treatment group: miR-92b mimic, mimic-NC, miR-92b AMO, AMO-NC

[0055] 1.3 Transfection method: same as Example 1

[0056] 1.4 Osteogenic induction method: same as Example 1

[0057] 1.5 Experimental detection items: Runx2 immunofluorescence staining

[0058] 2. Data processing

[0059] The results of the present invention are represented by standard deviation±standard error. Graphpad prism 5.0 was used for graphing, and the correlation betw...

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Abstract

The invention discloses an application of miR-92b and a simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation. The regulation effect of themiR-92b on osteogenic differentiation of the bone marrow mesenchymal stem cells and bone formation is explored through in vitro experiments. The miR-92b can be proved to improve osteogenic differentiation ability of the bone marrow mesenchymal stem cells and promote in-vivo bone formation by promoting the formation of osteoblast calcium nodules and the activity of a marker enzyme ALP, as well as promoting the expression of Runx2 which is important regulatory transcription factors in the process of osteogenic differentiation. The invention provides a new technical means for the treatment of osteoporosis.

Description

technical field [0001] The present invention relates to a new therapeutic application of miR-92b, in particular to the application of miR-92b and its mimics in promoting osteogenic differentiation and bone formation of bone marrow mesenchymal stem cells. The invention belongs to the technical field of medicine. Background technique [0002] Osteoporosis, or osteoporosis, is a metabolic bone lesion characterized by loss of bone mass per unit volume caused by various reasons. This disease is seen in postmenopausal women and the elderly. It often manifests as bone pain, increased bone fragility, and prone to fractures, causing serious damage to the human body. At present, mild symptoms can be improved through diet control, exercise, and appropriate supplementation of calcium and vitamin D. For those with signs of osteoporosis or fractures, drugs that resist bone resorption or promote bone formation are often used to improve symptoms , Severe cases need to be supplemented by s...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61P19/10A61P19/08A61P19/00
CPCA61K31/7105A61P19/00A61P19/08A61P19/10
Inventor 杨磊蔡本志袁野杨帆杜伟杰
Owner HARBIN MEDICAL UNIVERSITY
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