Metal-on-metal hip arthroplasty is associated with elevated levels of cobalt and chromium ions. The effects of cobalt and chromium ions on cell number, activity, expression of osteoprotegerin (OPG) and receptor activator of nuclear factor kappa B ligand (RANKL) and oxidative stress on human osteoblast-like cells were addressed. Saos-2 cells were supplemented with Co 2þ , Cr 3þ , or Co 2þ þ Cr 3þ (1:2) at 0, 1, 10, and 100 mg/L and incubated for 24, 48, 72, and 96 h. Cell activity was assessed by MTT-assay and cell number by Crystal Violet staining. RNA levels of OPG and RANKL were evaluated using real-time quantitative polymerase chain reaction (qPCR). Compared to controls Co 2þ reduced cell numbers: at 10 mg/L by 17 AE 8% after 48 h and at 100 mg/L after 24 h by 35 AE 8%. Cr 3þ decreased cell numbers at 10 mg/L after 48 and 72 h. Co 2þ þ Cr 3þ combined at 1 mg/L lowered cell numbers after 24 and 96 h (17 AE 13, resp. 13 AE 4%). The 10 and 100 mg/L concentrations reduced cell numbers significantly after 24, 48, and 96 h. Cr 3þ reduced osteoblast activity at 1, 10, and 100 mg/L at all incubation times. The strongest reduction (11 AE 1%) was seen at 100 mg/L after 96 h. The OPG/RANKL ratio was reduced after 72 h with almost all Co 2þ and Cr 3þ concentrations. After 96 h, glutathione, superoxide dismutase, and catalase levels were indicative for an oxidative stress response in all samples. In conclusion, cobalt and chromium ions reduce human osteoblast activity, reduce OPG/RANKL ratio and lead to oxidative stress. Keywords: metal-on metal; osteoblasts; oxidative stress; OPG; RANKL Total hip arthroplasty (THA) is a well-accepted therapy for hip osteoarthritis, with long lasting good clinical results. Survival rates of most THA's are documented at more than 95% over 10-15 years. Wear, osteolysis, and loosening have emerged as the dominant concerns among orthopedic hip surgeons. Polyethylene wear particles trigger a foreign body granulomatous reaction, inflammation, and immune reactions leading to the secretion of various bone resorbing agents (cytokines, prostaglandins). This results in peri-prosthetic bone loss, contributing to loosening and failure of the prosthesis.1 One of the key factors in peri-prosthetic osteolysis is receptor activator of nuclear factor kappaB ligand (RANKL) and strong correlations have been found between its expression and the amount of polyethylene wear debris and the degree of osteolysis. 3 also related the presence of polyethylene to RANKL expression. In general, osteoclastogenesis is largely regulated by receptor activator of nuclear factor kappaB (RANK), RANKL, and osteoprotegerin (OPG). RANKL stimulates differentiation of osteoclast precursor cells into mature osteoclasts and is required for osteoclast activity; in fact, RANKLtreated mice show increased bone resorbtion. On the other hand, OPG decreases the number of osteoclasts and mice treated with OPG exhibit lowered osteoclast activity and increased bone volume. 4 In order to avoid polyethylene wear-related loosening and t...