Performance of urinary and gene expression biomarkers in detecting the nephrotoxic effects of melamine and cyanuric acid following diverse scenarios of co-exposure

Although standard nephrotoxicity assessments primarily detect impaired renal function, KIM-1, clusterin, NGAL, osteopontin and TIMP-1 were recently identified biomarkers proposed to indicate earlier perturbations in renal integrity. The recent adulteration of infant and pet food with melamine (MEL) and structurally-related compounds revealed that co-ingestion of MEL and cyanuric acid (CYA) could form melamine-cyanurate crystals which obstruct renal tubules and induce acute renal failure. This study concurrently evaluated the ability of multiplexed urinary biomarker immunoassays and biomarker gene expression analysis to detect nephrotoxicity in F344 rats co-administered 60 ppm each of MEL and CYA in feed or via gavage for 28 days. The biomarkers were also evaluated for the ability to differentiate the effects of the compounds when co-administered using diverse dosing schedules (i.e., consecutive vs. staggered gavage) and dosing matrixes (i.e., feed vs. gavage). Our results illustrate the ability of both methods to detect and differentiate the severity of adverse effects in the staggered and consecutive gavage groups at much lower doses than previously observed in animals co-exposed to the compounds in feed. We also demonstrate that these urinary biomarkers outperform traditional diagnostic methods and represent a powerful, non-invasive indicator of chemical-induced nephrotoxicity prior to the onset of renal dysfunction.

► Urinary biomarkers have been identified that are proposed to detect renal damage prior to the onset of kidney dysfunction. ► Multiplexed ELISA and gene transcript analyses were used to assess the effects of MEL and CYA via diverse exposure scenarios. ► The biomarkers were able to detect and differentiate the severity of adverse effects induced by dosing schedule and matrix. ► Compared to feed, each method indicated that co-exposure via gavage induced more severe adverse effects at a much lower dose. ► Multiplexed ELISA is a powerful, non-invasive method to assess chemical exposure to prior to the onset of renal dysfunction.