The effect of blood ozonation on mitochondrial function and apoptosis of peripheral blood mononuclear cells in the presence and absence of plasma antioxidants

Abstract

Ozone-autohemotherapy (O 3 -AHT) has recently gained interest as a form of alternative and complementary medicine. There is, however, some concern regarding its toxicity and effectiveness. Ozone is a powerful oxidant and when introduced into biological fluids react with most cellular components including proteins, lipids and DNA. We assessed the effect of O 3 -AHT on peripheral blood mononuclear cells (PBMC) viability, apoptosis and mitochondrial function in the presence and absence of plasma antioxidants. Exposure to ozone increased lactate dehydrogenase (LDH) release and caspase 3/7 activity in PBMC. A decrease in mitochondrial function was measured as a decrease in ATP levels and an increase in NADH/ NAD + ratio. Complex IV (cytochrome c oxidase) of the respiratory chain was almost completely inhibited by ozone. These results indicated that the death of PBMC was probably through apoptosis. These effects were more evident in the absence of plasma antioxidants. Therefore, high concentrations of ozone were damaging to the cells, but this effect was diminished by antioxidants present in plasma. It is not certain if the in vitro damage will be propagated when ozonated blood is injected back into individuals. One must bear in mind that only a fraction of the total blood volume is ozonated.

Mitochondrial Features of Mouse Myoblasts Are Finely Tuned by Low Doses of Ozone: The Evidence In Vitro

“…It is known that impaired mitochondrial function and decreased ATP production play a crucial role in cell death [26]. In C2C12 cells, the fine morphology of mitochondria was affected by both O 2 and O 3 treatments.…”

“…Conversely, 20 µg O 3 induced a significant decrease in cell viability at both 24 h and 48 h after treatment. Accordingly, after treatment with 20 µg O 3 a decreased cell viability characterized by a high level of lactic dehydrogenase (as necrosis marker) and increased caspase activity was described in peripheral blood mononuclear cells [26]. Twenty µg O 3 was found to increase the death rate also in human adipose-derived adult stem cells [30].…”

“…Mitochondria, essential cell organelles involved in more than just adenosine triphosphate (ATP) synthesis [23,24], are known to be sensitive to even mild oxidative stress [25], thus representing a susceptible target for O 3 . However, studies focused on the effects of low O 3 concentrations on mitochondria are still scarce [26,27].…”