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Cytotoxicity of Simulated Gastric Leachates of 3D Printer Metal-Containing Filaments in Rat and Human Intestinal Models
Citation:
Hughes, M., H. Clapper, G. Tedla, AND K. Rogers. Cytotoxicity of Simulated Gastric Leachates of 3D Printer Metal-Containing Filaments in Rat and Human Intestinal Models. SOT, Nashville, TN, March 19 - 23, 2023. https://doi.org/10.23645/epacomptox.22105208
Impact/Purpose:
N/A
Description:
Printer filaments that contain metal particles (metal-fill) are commonly used to manufacture a variety of 3D-printed objects. Exposure to these objects may pose health risks to individuals, possibly due to the release of toxic metals. This study investigated the cytotoxicity of print object leachates in rat (IEC-6) and human intestinal (EpiIntestinal) cells. Copper-, bronze-, and steel-fill filament print objects were incubated for 2 hr in serum-free cell culture media acidified to pH 2. Leachates were then adjusted to pH 7, serum (10% final concentration) was added, and delivered to cells for 4 or 24 hr. Concentration- and time-dependent decreases in viability of rat and human cells were observed using a colorimetric assay (MTS assay, rats; MTT assay, human) and confirmed using microscopic imaging. Copper- and bronze-fill leachates were more toxic to rat and human cells than the steel-fill leachate. ICP-MS data showed the highest copper concentrations in copper-fill leachates followed by bronze-fill leachates. Steel-fill leachates consisted primarily of iron. Subsequent studies focused on the copper-fill print object. A 4-hr exposure to CuSO4 (25-1000 µg/ml), resulted in concentration-dependent decreases in rat cell viability. Addition of the copper chelator, bathocuproine disulphonate (1mM), to CuSO4 (250 µg/ml) and copper-fill leachate restored rat cell viability to levels comparable to untreated cells, suggesting that decreased viability was a result of copper ion exposure. Glutathione levels in rat cells decreased following a 4-hr exposure to copper-fill leachate, suggesting the formation of reactive oxygen species. Hydrogen peroxide levels increased in rat cells exposed for 4 hr to copper-fill leachate. Overall, our data indicate that metals released from the simulated gastric acidic exposure of print objects using metal-fill filaments, especially copper, are toxic to rat and human intestinal cells in a concentration- and time-dependent manner. Metals ions, particularly copper, released from metal-fill printer objects by this exposure may form reactive oxygen species leading to cytotoxicity. This abstract does not represent EPA policy.
URLs/Downloads:
DOI: Cytotoxicity of Simulated Gastric Leachates of 3D Printer Metal-Containing Filaments in Rat and Human Intestinal Models
HUGHES ET AL SOT 2023.PDF (PDF, NA pp, 952.947 KB, about PDF)