Effect of Hyperbaric Oxygen on Differentiation of Osteoblasts in Vitro
Abstract
Objective To investigate the effect of hyperbaric oxygen (HBO) on the differentiation of osteoblasts in vitro. Methods Osteoblasts were seeded in 96 well plates at a cell density of 10 000 cells per well. After 3 days normal culture, medium was changed to osteogenic medium. Subsequently, cultures were exposed daily to HBO of 2.4 ATA for 90 minutes and 1.5 ATA for 90 minutes up to 19 days. Mineralization was evaluated by calcium deposition assay, alkaline phosphatase (ALP) activity and von Kossa staining. Results Significant calcium deposition was observed after 3 days of HBO treatment and the calcium deposited was consistently higher in HBO treatments period, there was no difference in the amount of calcium deposition between the treatment of hyperbaric air and untreated control group. In addition, ALP activity and the biomarker for osteogenic differentiation were also significantly increased within the HBO-treated cultures. von Kossa staining showed that bone nodule formation of HBO group was more than the control group. Conclusions HBO treatment could significantly stimulate osteogenic differentiation, which implies a potential application of HBO in bone tissue engineering.
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