Effect of increased partial pressure of oxygen on growth and metabolism of fibroblasts - results of pilot study
Abstract
Normobaric and hyperbaric oxygen therapy is commonly used treatment method, a positive or negative effects and events at the cellular level are not sufficiently described, it is not clear how the cells and systems respond to nonphysiological levels of dissolved oxygen in the blood. At the Faculty of Medicine in Plzen have been carried out pilot experiments in order to get an idea of the areas.The experiments were focused to investigate the effect of the environment with different pO2 on cultivation media in order to find conditions under which there will be no changes in pH of the medium, to compare the growth curves of cell lines (HFL1) grown in atmosphere with increased pO2 (95kPa pO2 pCO2 + 5 kPa), and optimization of the cultivation to achieve sufficient amount of cells for mitochondrial respiration measurement at the Oroboros device (method HRR) and simultaneously build and optimize a suitable protocol.Among the significant outputs of the first part of experiments belong finding the necessary presence of pCO2 in the atmosphere (5 kPa), otherwise there is a significant change in pH of the medium and decrease the antioxidant capacity (e.g.
DMEM), at increased concentrations of O2 and 5 kPa CO2, there is no significant difference in the growth of influenced and control cells, and a sufficient quantity of cells to reliable measurement of oxygen consumption by HRR is at least 400,000 / ml. It is possible to determine the mitochondrial respiration of intact cells and focus on the contribution of individual mitochondrial respiratory complexes on total consumption of oxygen.
Preliminary results indicate that cells exposed to higher concentrations of O2 have up to twice the consumption of O2 versus controls.Lack of publications and the results of the pilot experiments lead us to the conclusion that this area of basic research is scientifically interesting and it makes sense to continue.