Application of miR-296 and simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation

A technology for bone marrow mesenchymal and osteogenic differentiation, applied in the field of medicine, can solve problems such as many complications, and achieve the effect of increasing bone formation and promoting the osteogenic differentiation of bone marrow mesenchymal stem cells

Active 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-296 and simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation
  • Application of miR-296 and simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation
  • Application of miR-296 and simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: miR-296 promotes BMSCs bone formation in vivo

[0038] 1 Experimental method

[0039] 1.1 The BMSCs transfected with miR-296 in vitro were subcutaneously transplanted by the new biomaterial hydroxyapatite for 8 weeks, and the effect of miR-296 transfection on bone formation in graft tissue was observed by Masson staining

[0040] 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-296 mimic, mimic-NC, miR-296 AMO, AMO-NC

[0041] 1.3 Experimental testing items: Masson staining

[0042] 2 Observations

[0043] like figure 1 As shown, compared with the mimic-NC group, the bone formation at the transplantation site was significantly increased in the miR-296 mimic group, whereas in the miR-296 AMO group, the bone formation at the transplantation site was significantly decreased.

Embodiment 2

[0044] Example 2: miR-296 regulates the osteogenic differentiation of bone marrow mesenchymal stem cells

[0045] 1 Experimental method

[0046]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

[0047] 1.2 Experiment grouping design: The experiment was divided into the following four groups according to the transfection control group and treatment group: miR-296 mimic, mimic-NC, miR-296 AMO, AMO-NC.

[0048] 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 3

[0062] Example 3: miR-296 promotes the expression level of osteogenesis-related proteins

[0063] 1 Experimental method

[0064] 1.1 Western blot was used to detect the expression levels of osteogenesis-related proteins in BMSCs after 7 days of transfection and osteogenesis induction

[0065] 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-296 mimic, mimic-NC, miR-296 AMO, AMO-NC

[0066] 1.3 Transfection method: same as Example 1

[0067] 1.4 Osteogenic induction method: same as Example 1

[0068] 1.5 Experimental testing items: Western blot (Western blot) experiment

[0069] 2 Observations

[0070] Seven days after osteogenesis induction, Western blot experiments were performed to detect the expression levels of osteogenesis-related proteins Runx2 and Osterix. like Figure 4 As shown, compared with mimic-NC, the ...

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Abstract

The invention discloses an application of miR-296 and a simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation. The regulation effect of themiR-296 on osteogenic differentiation of the bone marrow mesenchymal stem cells and bone formation is explored through in-vitro experiments. In-vivo experiments prove that the formation of new bones can be promoted through the transfection of the miR-296, and the in-vitro experiments prove that the miR-296 can improve osteogenic differentiation ability of the bone marrow mesenchymal stem cells andpromote 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 Osterix and Runx2 which are 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-296, in particular to the application of miR-296 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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Patent Type & Authority Applications(China)
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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