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Application of aloin in improvement of osteogenic capability of bone mesenchymal stem cell

A technology of bone marrow mesenchyme and aloin, applied in the field of biomedicine, can solve the problems of unclear influence of aloin, and achieve the effect of great application prospect.

Inactive Publication Date: 2018-02-06
AFFILIATED HOSPITAL OF GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effect of aloin on the osteogenic differentiation of BMSCs is unclear

Method used

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  • Application of aloin in improvement of osteogenic capability of bone mesenchymal stem cell
  • Application of aloin in improvement of osteogenic capability of bone mesenchymal stem cell
  • Application of aloin in improvement of osteogenic capability of bone mesenchymal stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Primary Isolation, Culture, Identification of Human and Rat Bone Marrow Mesenchymal Stem Cells

[0033] 1. With the approval of the Medical Ethics Committee of the Affiliated Hospital of Guangdong Medical University, and with the informed consent of the patient, 10ml of bone marrow was aseptically collected from the proximal femur of a patient undergoing hip replacement, and separated by Ficoll-Histopaque (d=1.077g / ml) For mononuclear cells, resuspend the cells in low-glucose DMEM medium (containing 10% fetal bovine serum and 1% PSN), at 37°C, 5% CO 2 Subculture in a cell incubator; Femur and tibia of healthy 4-week-old rats were taken, and the bone marrow cavity was repeatedly washed with sterile saline using a syringe, and mesenchymal stem cells were separated from the washed bone marrow. The separation and subculture methods were the same as those of human bone marrow Mesenchymal stem cells.

[0034] 2. Phenotype identification of mesenchymal stem cells: t...

Embodiment 2

[0036] Example 2 Effect of Different Concentrations of Aloe Vera on the Viability of Mesenchymal Stem Cells

[0037] Mesenchymal stem cells (2000 cells per well) were inoculated in a 96-well cell culture plate. After the cells were completely attached to the wall, the cells were treated with 0, 10, 20, and 50 μM aloin for 24, 48, and 72 hours, respectively. The CellTiter 96AQueous OneSolution Cell Proliferation Assay kit was used to identify cell viability, and the absorbance value in the 570nm wavelength range was detected by a spectrophotometer. The experiment was repeated three times, and statistical analysis was performed using GraphPad software. The results showed that compared with the control group, 10, 20, and 50 μM aloin had no significant effect on cell viability, suggesting that low concentrations of aloin had no obvious toxic effects on human and rat bone marrow mesenchymal stem cells (see figure 2 ).

Embodiment 3

[0038] Example 3 Aloe vera promotes the differentiation of human and rat bone marrow mesenchymal stem cells into osteoblasts

[0039] 1. In order to evaluate the effect of aloin on the differentiation of bone marrow mesenchymal stem cells into osteoblasts, different concentrations of aloin (0, 10, 50 μM ) were cultured for 14 days and stained with alizarin red. The results showed that cells treated with aloin produced more mineralized nodules than the control group (see image 3 A). In order to further identify the effect of aloin on the osteogenic differentiation of bone marrow mesenchymal stem cells, different concentrations of aloin (0, 10, 50 μM) were added for 3 days under osteogenic induction culture conditions, and the cells were collected and rinsed with pre-cooled PBS 3 times, permeabilize the cells with 10mM Tris–HCl (pH 7.4) buffer (containing 0.1% Triton X-100), centrifuge at 1,000g, 4°C for 10min, use the ALP detection kit to detect the ALP activity of the cells...

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Abstract

The invention discloses application of aloin in improvement of osteogenic capability of a bone mesenchymal stem cell (BMSC). According to the application of the invention, the BMSC is treated by aloins with different concentrations, and cell viability of the BMSC is detected; influence of the aloin on osteoblast differentiation of the BMSC is evaluated by adopting alkaline phosphatase activity andmineralization experiments; real-time fluorescent quantitative PCR detection is carried out for detecting expression situation of osteogenesis-related genes such as Runx2, ALP, OCN and OPN; variationof an ERK1 / 2-Runx2 signal channel is detected by virtue of a Western blotting process and an immumofluorescence process; and results show that the aloin promotes the osteoblast differentiation of theBMSCs by activating the ERK1 / 2-Runx2 signal channel, the osteogenic capability of the BMSCs is improved, and the research results provide data support for research and development of a new drug usedfor preventing and treating osteoporosis, so that the aloin has a greater application prospect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. More specifically, it relates to the application of aloin in improving the osteogenic ability of bone marrow mesenchymal stem cells. Background technique [0002] Osteoporosis is a systemic skeletal disease characterized by low bone mass and microarchitectural disruption of bone tissue, leading to increased bone fragility and increased fracture risk. The main cause of osteoporosis is the disruption of the balance between bone formation and bone resorption. Generally, there are two methods for clinical prevention and treatment of osteoporosis. Antiresorptive drugs, such as bisphosphonates, are widely used in the treatment of osteoporosis, mainly because of their ability to slow down bone resorption; another approach is to use steroid drugs, including human parathyroid hormone (hPTH1 -34), used to stimulate bone formation. However, long-term use of these drugs has certain limitations and si...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/351A61P19/10
CPCA61K31/351
Inventor 楚佳奇李鹏全娟花赵名艳王家丰荆凯鹏魏波魏劲松黄瑞高翔闫守泉罗远明牛艳茹孔俊超陈祝明黄帅吕贵何宗治贤
Owner AFFILIATED HOSPITAL OF GUANGDONG MEDICAL UNIV
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