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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  • Summary
  • 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.

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

Example Embodiment

[0026] Example 1. Cultivation of bone marrow mesenchymal stem cells (BMSCs)

[0027] Use a heparin-treated syringe to extract 2 to 3 mL of human bone marrow under aseptic conditions, immediately add it to a centrifuge tube containing the same amount of DMEM low-sugar medium, and quickly and gently pipette to make a cell suspension. The resulting cell suspension is light along the tube wall. Gently add it to a centrifuge tube with an equal volume of lymphocyte separation solution, centrifuge at 2000r / min for 20min, collect the mononuclear cells in the albuginea 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 6 Cell density per mL is inoculated in the culture flask, recorded as the primary generation, placed at 37℃, 5% CO 2 Culture in a constant temperature incubator, change the medium every 3 days, observe the cell...

Example Embodiment

[0029] Example 2. Mangiferin can reduce the decrease of BMSCs cell viability caused by hypoxia

[0030] Put the above BMSCs according to 5×10 3 Pcs / cm 2 Inoculate 96-well plates and 6-well plates at the same density. Discard the medium after 24h and change to CoCl 2 Solution, treat for 12h. The mangiferin concentration of 1μM, 5μM, 10μM and 20μM solution 200μM were added respectively. After 24 hours, CCK-8 was used to detect cell viability and flow cytometry was used to detect cell apoptosis. The results are as follows figure 1 Shown. The results show that mangiferin concentrations greater than 20 μM can reduce the decrease in cell viability caused by hypoxia in vitro (p <0.05), and reduce BMSCs apoptosis (p <0.05).

Example Embodiment

[0031] Example 3. Mangiferin promotes BMSCs to promote cartilage differentiation

[0032] When the above-mentioned cultured bone marrow BMSCs are fused to about 90%, they are digested with 0.25% trypsin, and then used incomplete cartilage induction medium (that is, DMEM high glucose medium with 1% ITS, 100mg / mL streptomycin , 100U / mL penicillin, 100μg / mL vitamin C, 40μg / mL proline and 100nM dexamethasone) resuspend and adjust the cell density to 5.0×10 5 / mL, the resulting cell suspension is calculated according to 0.5mL cell suspension (2.5×10 5 Cells) were aliquoted, and then centrifuged at 500g for 10 minutes to make the cells agglomerate. After centrifugation, there is no need to discard the supernatant or resuspend the cells. Place the centrifuge tube gently in a constant temperature incubator, let it stand for 24 hours, and then replace it with complete cartilage Induction medium (ie DMEM low-sugar medium supplemented with 10ng / mL TGF-β3, 1% ITS, 100mg / mL streptomycin, 100U / ...

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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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