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4 protocols using trioctyl trimellitate

1

Phthalate Ester Toxicology Assay

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DEHP (>99.5% pure, CAS 117-81-7), tri-octyl tri-mellitate (TOTM, > 99% pure, CAS 3319-31-1), and peanut oil (CAS 8002-03-7) were purchased from Sigma-Aldrich, (Oakville, ON). Di-octyl succinate (DOS) and di-heptyl succinate (DHPS) were synthesized as previously described [41 (link), 42 (link)].
Nigericin (>98% pure, CAS 28643-80-3) was purchased from Cayman Chemicals (Ann Arbor, MI). Phorbol 12-myristate 13-acetate (PMA, >99% pure, CAS 16561-29-8) was purchased from Abcam, (Toronto, ON). Sulforhodamine B (SRB, (CAS 222-529-8, 85% pure) was purchased from Sigma-Aldrich (Oakville, ON). Antibodies specific for the active p20kD caspase-1 protein were purchased from Adipogen (San Diego, CA) and antibodies specific for IL-1β were purchased from Novus Biologicals (Toronto, ON) or GeneTex (Irvine, CA).
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2

Standardized Phthalate and Plasticizer Analysis

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The eleven phthalates and the two non-phthalates studied plasticizers are listed in Table 1. Benzyl butyl phthalate (BBP; ≥98%), dibutyl phthalate (DBP; ≥99%), di-cyclohexyl phthalate (DCHP; ≥99%), di(2-ethylhexyl) phthalate (DEHP; ≥99.5%), di-(2-ethylhexyl) terephthalate (DEHT; ≥97%), diethyl phthalate (DEP; ≥99.5%), diethyl phthalate-3,4,5,6-d4 (DEP-d4, IS; ≥98%), diisobutyl phthalate (DIBP; ≥99%), diisononyl phthalate (DINP; ≥99%), dimethyl phthalate (DMP; ≥99%), di-n-octyl phthalate (DNOP; ≥98%), diphenyl phthalate (DphP; ≥99%), dipropyl phthalate (DPP; ≥98%), and tri-octyl trimellitate (TOTM; ≥99%) were purchased from Sigma-Aldrich (Saint-Quentin Fallavier, France). HPLC-grade solvents were used. Acetone, dichloromethane, and acetonitrile were purchased from VWR (Fontenay-sous-Bois, France). Ultra-pure water was supplied by a Milli-Q purification system from Millipore (St Quentin en Yvelines, France).
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3

Synthesis and Characterization of Biocompatible Materials

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Sodium nitrite (NaNO2), ethylenediaminetetraacetic acid (EDTA), tetrahydrofuran (THF), trioctyl trimellitate (TOTM), glutaraldehyde, and bis(trimethylsilyl)amine (HMDS) were purchased from Sigma-Aldrich (St. Louis, MO). Glutathione was purchased from GoldBio (St Louis, MO). DI water for all aqueous solutions was obtained using a Milli-Q filter from Millipore Corporation (Billerica, MA). Nitrogen and oxygen gas cylinders were purchased from Airgas (Kennesaw, GA). Hydrochloric acid (HCl), dimethyl sulfoxide (DMSO), and Saint-Gobain Tygon Formula S3 E-3603 poly(vinyl chloride) tubing were purchased from Fisher Scientific (Pittsburgh, PA). Acetone and ethanol were purchased from VWR (Radnor, PA). Drabkin’s reagent was obtained from Ricca Chemical (Arlington, TX). Phosphate-buffered saline (PBS), pH 7.4, supplemented with 138 mM NaCl, 2.7 mM KC, and 10 mM sodium phosphate, and 100 μM EDTA were used. Calcium- and magnesium-free PBS (CMF-PBS) was made with 138 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, and 1.47 mM KH2PO4 with H2O.
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4

Antibacterial Coating Protocol for ETTs

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N-acetyl-d-penicillamine, sodium chloride, potassium chloride, sodium phosphate dibasic, potassium phosphate monobasic, ethylenediaminetetraacetic acid (EDTA), tetrahydrofuran (THF), phosphate-buffered saline (PBS), trioctyl trimellitate (TOTM), and N,N-dimethylacetamide (DMAc) were purchased from Sigma-Aldrich (St. Louis, MO). Methanol, hydrochloric acid, and sulfuric acid were obtained from Thermo Fisher Scientific (Pittsburgh, PA). Acetone was purchased from VWR International (Radnor, PA). Uncuffed, PVC ETTs, 5 mm, were purchased from Dynarex Corporation (Orangeburg, NY). PBS, pH 7.4, containing 138 mM NaCl, 2.7 mM KCl, 10 mM sodium phosphate, 100 μM EDTA was used for all experiments. P. aeruginosa (ATCC 27853) was originally obtained from the American Type Culture Collection (Manassas, VA). Luria broth (LB) and Luria agar were bought from Thermo Fisher BioReagents (Fair Lawn, NJ). PBS was purchased from Sigma-Aldrich.
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