Application of mangiferin in preparing drug for promoting bone defect repair

A mangiferin and bone defect technology, applied in the field of biomedicine, can solve the problems of cell apoptosis, seed cell apoptosis repair effect, unsatisfactory, etc., to achieve the effect of enhancing survival ability, enhancing bone formation effect, and good application prospects

Inactive Publication Date: 2017-12-22
ARMY MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem of repairing a large bone defect based on a large endochondral osteogenesis system, when its seed cells enter the bone defect area, it is easy to generate a large amount of reactive oxygen species (ROS) due to hypoxia, resulting in cell apoptosis. The present invention provides a A method of using mangiferin to protect seed cells from apoptosis while promoting endochondral bone repair, so as to avoid the problems of premature apoptosis of seed cells and unsatisfactory repair effect when repairing large segmental bone defects

Method used

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  • Application of mangiferin in preparing drug for promoting bone defect repair
  • Application of mangiferin in preparing drug for promoting bone defect repair
  • Application of mangiferin in preparing drug for promoting bone defect repair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1, the cultivation of bone marrow mesenchymal stem cells (BMSCs)

[0027] Use a heparin-treated syringe to extract 2-3 mL of human bone marrow under sterile conditions, immediately add it to a centrifuge tube filled with an equal amount of DMEM low-sugar medium, and quickly and gently blow it to make a cell suspension. Gently add to the centrifuge tube with an equal volume of lymphocyte separation medium, centrifuge at 2000r / min for 20min, collect the mononuclear cells in the buffy coat layer in another centrifuge tube, wash with DMEM (centrifuge at 1000r / min for 5min) twice, The supernatant was discarded, and the cells were resuspended in DMEM-L complete medium containing 15% fetal bovine serum, and diluted with 1×10 6 / mL cell density inoculated in a culture flask, recorded as the primary generation, placed at 37°C, 5% CO 2 Culture in a constant temperature incubator, change the medium every 3 days, observe the cell growth and morphological characteristics...

Embodiment 2

[0029] Embodiment 2, mangiferin can reduce the BMSCs cell viability that hypoxia causes to reduce

[0030] The above BMSCs were divided into 5×10 3 piece / cm 2 Inoculate 96-well plates and 6-well plates at different densities. After 24 hours, the medium was discarded and replaced with CoCl 2 solution, treated for 12h. Add 200 μM of mangiferin concentration of 1 μM, 5 μM, 10 μM and 20 μM solutions respectively. After 24 hours, CCK-8 was used to detect cell viability, and flow cytometry was used to detect cell apoptosis. The results were as follows: figure 1 shown. The results showed that when the concentration of mangiferin was greater than 20 μM, it could reduce the decrease of cell viability caused by hypoxia in vitro (p<0.05), and reduce the apoptosis of BMSCs (p<0.05).

Embodiment 3

[0031] Embodiment 3, mangiferin promotes BMSCs to promote cartilage differentiation

[0032] When the bone marrow BMSCs cultured above were fused to about 90%, they were digested with trypsin with a mass fraction of 0.25%, and then incomplete cartilage induction medium (i.e. 1% ITS, 100mg / mL streptomycin added to DMEM high-glucose medium) , 100 U / mL penicillin, 100 μg / mL vitamin C, 40 μg / mL proline and 100 nM dexamethasone) and adjust the cell density to 5.0×10 5 / mL, the resulting cell suspension was divided into 0.5mL cell suspension (2.5×10 5 cells) for aliquoting, and then centrifuged at 500g for 10min to agglomerate the cells. There is no need to discard the supernatant or resuspend the cells after the centrifugation. Induction medium (i.e. DMEM low-glucose medium supplemented with 10ng / mL TGF-β3, 1%ITS, 100mg / mL streptomycin, 100U / mL penicillin, 50μg / mL vitamin C, 40μg / mL proline and 100nM dextrose Measone), 500 μL of complete chondrogenic induction medium was added to...

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Abstract

The invention relates to an application of mangiferin in preparing a drug for promoting bone defect repair and solves the problems that a host micro-environment in a conventional bone repair mode is short of blood supply, apoptosis of seed cells in an anaerobic environment is easily caused, the ossifying effect is poor and the like. By means of a policy of combining mangiferin with an endochondral ossification system for bone repair as the mangiferin not only plays a protecting role, but also promotes the osteogenic capability of seed cells, mesenchymal stem cells are planted on a porous bone scaffold material to construct a tissue engineering composite, then chondrogenic differentiation inductive culture is performed in vitro for two weeks, and the composite is implanted into a defect region to repair after hypertrophic chondrogenic differentiation inductive culture is performed for two weeks; the mangiferin can be loaded on the porous bone scaffold material and can be integrally systematically administrated; an in vivo research result shows that the repair method provided by the invention can repair bone defects successfully, and has a good application prospect in tissue engineering bone construction and bone defect repair.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of mangiferin in the preparation of medicines for promoting bone defect repair. Background technique [0002] Bone defect and nonunion are difficult clinical problems in orthopedics. Due to the congenital defects of autologous bone and allograft bone in terms of origin, disease transmission, and shape and size, tissue engineered bone, as an excellent alternative strategy, has been widely used clinically in recent years. [0003] However, due to the characteristics of traditional tissue engineered bone based on intramembranous osteogenesis, its central area cannot be effectively vascularized, which limits the size of traditional tissue engineered bone, making it unable to effectively repair large bone defects. In order to overcome the above shortcomings, in recent years, some scholars have proposed tissue engineered bone based on endochondral osteogenesis system. When cons...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P19/08A61L27/54A61L27/56
Inventor 白赟董世武
Owner ARMY MEDICAL UNIV
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