Repeated hyperbaric oxygen (HBO2) treatments at 2.2 ATA for 90 minutes each are used to treat chronically ill patients with problem wounds, but there are concerns about the cytotoxicity of oxygen to blood cells and platelet function during prolonged HBO2 therapy. We recruited 31 consenting patients scheduled for multiple HBO2 treatments to evaluate oxidative metabolism in platelets, platelet aggregation, and hematology (mean age +/- standard error, 61 +/- 2.6 years, 20 males, 11 females). Venous blood was collected before and after the 1st and 20th HBO2 treatments. No effect of HBO2 was observed on red cell counts, hematocrit, hemoglobin, mean red cell volume (MCV), platelet counts, basal levels of lactate production by platelets, ferric reducing ability of plasma (FRAP), or plasma protein. The capacity for oxidative metabolism (lactate ratio) in platelets was not affected by HBO2, except in smokers where it increased by the 20th HBO2 treatment. Mean lymphocyte count was increased by 38% after the 20th treatment. There was also a 23% increase in platelet protein content, and a 24% increase in arachidonic acid-dependent platelet activation. Collagen-dependent platelet aggregation was unaffected. Blood glucose showed HBO2-dependent variability, but remained in the normal range. Plasma lactate levels decreased significantly from 3.2 to 2.5 mmol/l by the end of the study. Overall, we found no evidence that 20 HBO2 sessions caused adverse effects on platelet aggregation or oxidative metabolism in platelets, red or white cell counts, or total antioxidant status of the plasma.