Effects of hyperbaric oxygen on the osteogenic differentiation of adipose-derived stem cells
10.3760/cma.j.issn.1009-6906.2017.05.007
- VernacularTitle:高压氧对脂肪干细胞成骨分化的影响
- Author:
Yanhua ZHOU
1
;
Haifeng ZHENG
;
Qingxin ZENG
;
Shaojing FU
;
Shaobin WANG
Author Information
1. 内蒙古医科大学第三附属医院神经内科, 内蒙古自治州包头,014010
- Publication Type:Journal Article
- Keywords:
Hyperbaric oxygen;
Stem Cell;
Adipose-derived stem cell;
Differentiation
- From:
Chinese journal of nautical medicine and hyperbaric medicine
2017;24(5):365-369
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of hyperbaric oxygen (HBO) on the osteogenic differentiation of adipose-derived stem cells.Methods Adipose-derived stem cells were isolated,cultured and passed to next generations.The fourth adipose-derived stem cells were collected and seeded to the 96-well plates (2 × 105 cells/well) for culture in osteogenic induction medium,and were randomly divided into the control group and the hyperbaric oxygen (HBO) group.In the entire experiment,the control group was only kept in the 5％ CO2 incubator at a temperature of 37 ℃,while the HBO group was treated with HBO for 24 hours a day at the same culture conditions as the control group.(The HBO treatment cells were placed in the HBO chamber,which was washed with pure oxygen at a ventilation rate of 10 L/min for 15min.When oxygen flow rate was down-regulated to 2-3 L/min,the chamber was compressed for 20-25 min to a pressure of 2.5 ATA,which was maintained for 60 minutes.Then,uniform decompression was implemented for 20-25 minutes.At days 7,14 and 21,alkaline phosphatase activity of the cells was detected by alkaline phosphatase kit.At days14 and 21,osteocalcin concentration was detected by using the osteocalcin quantitative test kit,and at day 21,the calcified nodules were stained with Alizarin red.The data obtained were analyzed by 2 independent samples t test,paired sample t test and repeated test ANOVA.Results The cultured cells could express such mesenchymal stem cell markers as CD90 and CD105,but could not express hematopoietic markers such as CD34.After 7,14 and 21 days of culture,the activity of alkaline phosphatase in the HBO group was significantly increased,as compared with that of the control group A (P ＜ 0.01).Following culture for 14 and 21 days,the expression level of osteocalcin in the HBO group was significantly elevated,as compared with that of the control group (P ＜ 0.05).At day 21,the detection results by alizarin red staining showed that the number of calcified nodules for the HBO group was increased markedly as compared with that of the control group,and the calcified size became larger.Conclusions Treatment of HBO could increase osteogenic differentiation of adipose-derived stem cells.
