Evaluation of Metallothionein formation as a proxy for zinc absorption in an <i>in vitro</i> digestion/Caco‐2 Cell culture Model

TL;DRAbstract

Caco‐2 cell metallothionein (MT) formation was studied to determine if MT could be used as a proxy for zinc (Zn) bioavailability in an in vitro /cell culture model. The intracellular concentration of MT was determined using a cadmium/hemoglobin affinity assay. Total cellular Zn uptake was determined on acid treatment (5% HNO 3 ) using inductively‐coupled argon‐plasma emission spectroscopy (ICPES). The effect of phytic acid (PA) on cellular MT formation was also studied. Caco‐2 cells were treated with media containing 0, 2, 5, 10, 25, 50, and 75 μmol/L Zn (as ZnCl 2 ). Some Zn absorption studies employed radio‐Zn ( 65 ZnCl 2 ). The effect of varying Zn:PA molar ratios (i.e., 1:0, 1:1, 1:5, 1:10, and 1:20) on Zn uptake was determined. A positive correlation between Zn‐media concentrations and cellular 65 Zn uptake and cellular MT formation was observed. The concentrations of Zn and MT in cells treated with increasing levels of non‐radioactive Zn (i.e., 25, 50 or 75 μmol/L Zn) increased a

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