Milligram scale

Uncoated MWCNTs (U-MWCNT) and aluminum oxide coated (A-MWCNT) were weighed using a milligram scale (Mettler, Toledo, OH) suspended in a sterile, 0.1% pluronic F-68 (Sigma–Aldrich, St. Louis, MO) in phosphate buffer solution to achieve the final concentration of 10 mg/mL. Vials containing the suspended nanomaterials were dispersed using a cuphorn sonicator (Qsonica, Newton CT) at room temperature for 1 min prior to dosing. A limulus amebocyte lysate (LAL) chromogenic assay (Lonza Inc., Walkersville, MD) was used to test the nanomaterials for endotoxin contamination. All MWCNTs, both U-MWCNT and A-MWCNT, tested negative (<0.3 EU/mL) for endotoxin.

Uncoated MWCNTs (U-MWCNT), aluminum oxide coated (A-MWCNT), and zinc oxide coated (Z-MWCNT) were weighed using a milligram scale (Mettler, Toledo OH) suspended in a sterile 0.1% pluronic F-68 (Sigma-Aldrich, St. Louis MO) phosphate buffer solution to achieve the final concentration of 10 mg/mL. Vials containing the suspended nanomaterials were dispersed using a cuphorn sonicator (Qsonica, Newton CT) at room temperature for 1 minute prior to dosing. The A-MWCNT and Z-MWCNT concentrations were normalized to the U-MWCNT nanoparticle number in order to account for the mass increase caused by the surface modification of the CNT. The A-MWCNT were dosed at 2.5 times the U-MWCNT dose and the Z-MWCNT were dosed at 2.85 times the U-MWCNT dose. A limulus amebocyte lysate chromogenic assay (Lonza Inc., Walkersville MD) was used to test the nanomaterials for endotoxin contamination. All MWCNTs tested negative (< 0.3 EU/mL) for endotoxin.