Table 5.

A list of commonly used parameters for nephrotoxicity testing

Assessment	Measurements
Cell viability assays	Degree of cell death or damage in response to a drug or other compound, which can indicate potential NT assessed CCK-8, MTT and live/death kits
Biomarker analysis	Expression of biomarkers associated with kidney function (e.g., albumin, nephrin) for the effects of a drug or compound on kidney cells
Histological analysis	3D kidney tissue constructs under a microscope can reveal structural changes indicative of kidney damage
OCR/ECAR	Metabolic parameters can provide information on cellular respiration and glycolysis, which can be affected by nephrotoxic compounds, seahorse analysis
ROS	ROS levels can provide insight into this aspect of nephrotoxicity
Inflammatory markers	Cytokines genes and proteins: TNF-α, IL-1β, IL-6, IL-8, MCP-1, HMGB1, CRP, PGE2 and NO, assessed by q-PCR and western-blot
Kidney-specific protein markers	Expression of proteins: e.g., aquaporins (AOP1, AQP3), transporters (i.e., OAT1, OAT3, OCT2, MRP2, BCRP, SGLT2)
Mitotoxicity	MMP, ATP production, complexes I-V expression and activity, mitochondrial morphology, mtDNA content and OCR/ECAR

OCR oxygen consumption rate, ECAR extracellular acidification rate, ROS reactive oxygen species assays, MMP Mitochondrial membrane potential, ATP adenosine triphosphate, mtDNA content-mitochondrial DNA content