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6 protocols using triethyl phosphate tep

1

Synthesis of Hybrid Silica-Calcium Phosphate Biomaterials

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ε-Caprolactone (CL) monomer (purity 99%), Tin(II) 2-ethylhexanoate (TEH) catalyst (analytical grade), poly(ethylene glycol) (PEG) (average Mn 10,000 g/mol), tetraethyl orthosilicate (TEOS) (reagent grade 98%), triethyl phosphate (TEP) (purum >99.8%), calcium nitrate tetrahydrate Ca(NO3)2·4H2O) (ACS reagent 99%), cetyltrimethylammonium bromide (CTAB) and ethanol were purchased from Sigma-Aldrich chemical company (Saint Louis, MO, USA). Sodium hydrate pellets were supplied from Mallinckrodt Company (Staines-upon-Thames, UK). All the other reagents used were of analytical grade and purchased by Sigma-Aldrich.
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2

Synthesis of Calcium-Silicate-Phosphate Bioceramic Nanoparticles

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MBN were synthesized by modified sol-gel method [2 (link)]. Briefly, in 150 mL of distilled water, 10 mL 2-ethoxyethanol (Sigma-Aldrich, St. Louis, MO, USA), 2 mL aqueous ammonia (Samchun, Seoul, Korea), 1.4 g calcium nitrate tetrahydrate (Ca (NO3) 2.4H2O) (Sigma-Aldrich, St. Louis, MO, USA), 20 mL ethanol (Samchun, Seoul, Korea), and then 1 g hexadecyltrimethylammonium bromide (CTAB, Sigma-Aldrich, St. Louis, MO, USA) were added. After 30 min stirring, 5 mL and 0.25 mL of tetradecyl acetate (TEOS, Sigma-Aldrich, St. Louis, MO, USA), Triethyl phosphate (TEP, Sigma-Aldrich, St. Louis, MO, USA) were added sequentially and stirring for 4 h vigorously. The mole (Mol%) ratio of CaO: SiO2: P2O5 was 36:60:4. The white precipitates were obtained, dried at 60 °C for 24 h, and calcinated at 600 °C for 6 h with a heating rate of 1 °C min−1 and then furnace cooled naturally.
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3

Synthesis and Characterization of Phosphate Esters

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All chemicals for the syntheses were purchased from commercial sources and were used without further purification: trimethyl phosphate (TMP) (97%, Sigma-Aldrich, Burlington, MA, USA), triethyl phosphate (TEP) (99%, Sigma-Aldrich), tri-n-propyl phosphate (TnPP) (99%, Sigma-Aldrich), triphenyl phosphate (TPhP) (99%, Merck Schuchardt OHG, Hohenbrunn, Germany), di-n-butyl phosphate (DBP) (97%, Sigma-Aldrich), bis(2-ethylhexyl) phosphate (BEHP) (97%, Sigma-Aldrich), zinc oxide (≥99.0%, Merck KGaA, Darmstadt, Germany), zinc acetate dihydrate (ZnOAc·2H2O) (99%, Merck KGaA), calcium chloride ≥ 97.0%, Sigma-Aldrich), sodium bicarbonate (pure, POCh S.A., Gliwice, Poland), Bisphenol A diglycidyl ether (BADGE) (Brookfield viscosity of 5279 mPa·s at 25 °C, Sigma-Aldrich).
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4

Mesoporous Silica-Based Biomaterials

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This study used the following reagents: tetraethylorthosilicate (TEOS) (98%, Sigma-Aldrich, Darmstadt Germany), triethylphosphate (TEP) (99% Sigma-Aldrich, Darmstadt, Germany), calcium nitrate tetrahydrated (Ca(NO3)2·4H2O) (99% Sigma-Aldrich, Darmstadt, Germany) and cerium(III) nitrate hexahydrate (99% Sigma-Aldrich, Darmstadt, Germany) as silica, phosphate-, calcium- and cerium-oxide precursors, respectively, hydrochloric acid (HCl) (Sigma-Aldrich, Darmstadt, Germany) as a catalyst, PEG-PPG-PEG, called Pluronic® P123 (Sigma-Aldrich, Darmstadt, Germany) as structure directing agent and poly methyl methacrylate (Alfa Aesar, Ward Hill, MA, USA).
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5

Synthesis of Mesoporous Bioactive Nanoparticles

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MBN was synthesized using a modified sol-gel process. Briefly, 3.12 g calcium nitrate tetrahydrate (Ca(NO3)2·4H2O) (Sigma-Aldrich, St. Louis, MO, USA), 2 mL aqueous ammonia (Samchun, Seoul, Korea), 10 mL 2-ethoxyethanol (Sigma-Aldrich, St. Louis, MO, USA), 20 mL ethanol (Samchun, Seoul, Korea), and 1 g hexadecyltrimethylammonium bromide (CTAB) (Sigma-Aldrich, St. Louis, MO, USA) were mixed in 150 mL distilled water. The mixture was stirred at room temperature for 30 min. Then, 5 mL tetraethyl orthosilicate (TEOS; Sigma-Aldrich, St. Louis, MO, USA) was added and stirred at room temperature for 30 min. Subsequently, 0.25 mL triethyl phosphate (TEP; Sigma-Aldrich, St. Louis, MO, USA) was added and the mixture was vigorously stirred for 4 h at room temperature. A white precipitate was formed and dried in a vacuum oven at 60 °C for 24 h. The dried gel powder was calcined at 600 °C for 5 h. The molar ratio of SiO2:CaO:P2O4 was calculated to be 60:36:4.
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6

Binding Assay for Thyroid Hormone Disruption

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Tris-EDTA, non-radiolabelled thyroxine (T4; ≥ 98% purity), and lyophilized transthyretin isolated from human plasma (hTTR; ≥ 95% purity) were obtained from Sigma-Aldrich (Oakville, ON, Canada). The radioligand 125 I-labelled T4 (50 μCi) was obtained from MP Biomedicals (Solon, OH). The polychlorinated biphenyl (PCB) metabolite 4-hydroxy-2,2′,3,4′,5,5′,6-heptachloro-biphenyl (4-OH-CB-187; > 98% purity) was obtained from Wellington Laboratories (Guelph, ON, Canada). The OP esters, tris(butoxyethyl) phosphate (TBOEP; 94% purity), triethyl phosphate (TEP; 99% purity), triphenyl phosphate (TPHP; 99% purity), and diphenyl phosphate (DPHP; 99% purity) were obtained from Sigma-Aldrich (St. Louis, MO). Tris(1,3dichloro-2-propyl) phosphate (TDCIPP; > 95% purity) was obtained from TCI America (Portland, OR), and para-hydroxy triphenyl phosphate (p-OH-TPHP; 98% purity) was generously donated by Dr. Heather Stapleton (Duke University, DC) (Fig. 1).
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