Measurement of Osteoblast Cytotoxicity Induced by S. aureus Secreted Toxins

Abstract

Staphylococcus aureus is a Gram-positive bacterium that is capable of infecting and inducing tissue pathology in nearly every organ system. The pathogenesis of staphylococcal infection is dictated, in part, through the production of toxins that induce cellular death through receptor-dependent and -independent mechanisms, thereby contributing to tissue injury. One common manifestation of invasive staphylococcal infection is osteomyelitis, or infection of bone. Osteomyelitis triggers extreme bone loss, in part, through production of secreted toxins. Cytotoxicity assays, therefore, can be instrumental in elucidating how S. aureus triggers bone loss, and such assays are rapidly adaptable to study of tissue damage across multiple cell types and organ systems. Additionally, in conjunction with proteomic approaches, cytotoxicity studies may help identify toxins capable of inducing host cell death. Here, a protocol is described for the isolation and stimulation of primary osteoblasts with S. aureus supernatants for rapid detection of cytotoxicity. This assay provides an excellent in vitro system to better understand how staphylococcal secreted toxins impact skeletal cell biology to induce changes in bone homeostasis.