Toxicity measurement of orthopedic implant alloy degradation products using a bioluminescent bacterial assay.

The toxicity of aqueous metal solutions representative of ionic degradation products from orthopedic implant alloys was determined using a bacterial bioluminescence assay, Microtox. The toxicity of forms of the individual elements released from ASTM F75 Co-Cr-Mo (Co-Cr-Mo), F138 316L stainless steel (316L), and F136 Ti-6Al-4V (Ti-6Al-4V) was first determined, and a mathematical model was developed to predict the toxicity of mixtures of these ions. Aqueous metal solutions were then mixed according to the proportions of the ions found in these alloys, and their toxicity was measured with Microtox. Mixture behavior was classified as synergistic, antagonistic, or additive by comparing measured toxicity to predicted toxicity. Since relating these tests to actual implant corrosion processes can be confounded by selective leaching, the predicted and measured toxicity of aqueous metal solutions mixed according to proportions representative of selective leaching were next determined, and the mixture behaviors were classified as before. The most toxic individual alloying elements were found to be hexavalent Cr, Ni, and Co, in that order: a finding in accord with prior biocompatibility research. Co-Cr-Mo was found to be the most toxic alloy mixture of both those combined according to alloy composition and those combined to reflect selective leaching. The Ti-6Al-4V mixtures were found to behave synergistically, while the Co-Cr-Mo and 316L mixtures behaved antagonistically. By providing insight into degradation product toxicity and elemental interaction, these experiments demonstrate the utility of employing bioluminescent bacterial assays to investigate biocompatibility of implant materials. Further studies to more closely simulate in vivo conditions, though, are required to fully gauge their potential in this regard.