Anti‑inflammatory effects of ozonated water in an experimental mouse model

Background

Acute inflammation drives a wide range of illness and injury, and much of the tissue damage it causes comes from an excess of pro-inflammatory signalling molecules such as tumour necrosis factor-alpha (TNF-alpha) combined with a build-up of reactive oxygen species that outpaces the body's own antioxidant defences, including the enzyme superoxide dismutase (SOD). Ozone and ozonated water have previously been reported to influence oxidative stress pathways, leading researchers to ask whether ozonated water could measurably dampen an acute inflammatory response in a living animal, rather than in a test tube alone.

Study design

Researchers used a mouse model to test this question. Nine female BALB/c mice, seven weeks old, were split into three groups of three: an untreated control group, a saline-treated control group, and an ozonated-water group. The ozonated-water group received daily intraperitoneal injections of 10 ppm ozonated water for five days, while the saline group received an equivalent volume of saline. Twenty-four hours after the final injection, all mice were given an intraperitoneal dose of lipopolysaccharide (LPS), a bacterial component widely used in research to trigger a rapid, acute inflammatory response. Blood samples were taken one hour after the LPS dose. TNF-alpha levels were measured using an ELISA assay, and SOD activity was assessed with the nitroblue tetrazolium dye test.

Findings

Mice pre-treated with ozonated water showed significantly lower serum TNF-alpha levels than both control groups (P<0.01 versus the untreated group, P<0.05 versus the saline group). SOD activity was also higher in the ozonated-water group, at 38±2%, compared with 26±1% in the untreated group and 28±1% in the saline group.

What the authors concluded

The authors concluded that ozonated water exerted a critical anti-inflammatory effect in this model, reducing a key inflammatory marker while boosting the body's own antioxidant enzyme activity. On this basis, they suggested ozonated water could represent a therapeutic option worth exploring for acute inflammation.

Limitations of this study

This is preclinical animal evidence from a small mouse study, with only three animals per group, and it has not yet been replicated at larger scale or in other species. Ozonated water was delivered by intraperitoneal injection rather than by any route used in everyday human contexts, so these findings cannot be directly extended to oral, topical or inhaled use in people. The model also used a single, acute LPS challenge rather than a chronic or naturally occurring inflammatory disease, and blood was sampled at only one time point shortly afterwards, so longer-term effects and safety were not assessed. Human clinical trials would be needed before any conclusions could be drawn about ozonated water as a treatment for inflammatory conditions in people.