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5 protocols using Gantrez™-AN119

1

Characterization of Biodegradable Polymers for Vaginal Applications

MTZ was purchased from Tokyo Chemical Industry, Tokyo, Japan. PCL 6506 (Mw = 50,000 Da) henceforth referred to as H-PCL and PCL 2054 (Mw = 550 Da) henceforth referred to as L-PCL were kindly provided by Perstop (Malmö, Sweden). Gantrez™-AN119, a copolymer of methyl vinyl ether and maleic anhydride, was a gift from Ashland, (Kidderminster, UK). Porcine mucin was obtained from Sigma Aldrich (Dorset, UK). Microcrystalline cellulose (MCC), Avicel PH 102, was acquired by IMCD UK Limited (Sutton, UK). Gardnerella vaginalis NCTC 10287 was incubated anaerobically on brain heart infusion (BHI) broth at 37 °C in for 48–72 h to perform the disk diffusion test. All the chemicals used were of pharmaceutical grade.
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2

Synthesis and Characterization of Gantrez-based Biomaterials

Gantrez®® AN 119 was supplied by Ashland Inc. (Barcelona, Spain). Mannosamine hydrochloride, zein, mannitol, lysine, tween 20, bromoethylamine-hydrobromide and Bovine Serum Albumin (BSA) were purchased from Sigma-Aldrich (Spain). Acetone was obtained from VWR-Prolabo (Spain) and O- phtalaldehide was provided by Invitrogen (Thermo Fisher, Waltham, Ma. USA). Ethanol, formaldehyde, NaOH and DMSO was supplied by Panreac (Spain). TSB was obtained from bioMérieux (Marcy l’Etoile, France). RPMI was obtained from Gibco-BRL (UK). Coomassie brilliant blue and sample buffer was purchased from Bio-Rad (Spain). All other reagents and chemicals used were of analytical grade.
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The copolymer of methylvinylether and maleic anhydride or poly(anhydride) (Gantrez® AN 119) was supplied by Ashland Inc. (Barcelona, Spain). Thiamine hydrochloride, zein, mannitol, lysine, agarose, glutaraldehyde, propylene oxide, sodium cacodylate and EPON™ were purchased from Sigma-Aldrich (Madrid, Spain). Ethanol were provided by Panreac (Barcelona, Spain). Acetone was obtained from VWR-Prolabo, (Linars del Vallès, Spain). Perylene-Red (BASF Lumogen® F Red 305) was from Kremer Pigmente GmbH & Co. (Aichstetten, Germany) and OCT™ Compound Tissue-Tek from Sakura Finetek Europe (Alphen aan Der Rijn, The Netherlands). 4′,6-diamidino-2-phenylindole (DAPI) was obtained from Biotium Inc. (Madrid, Spain). Glass bottom imaging dishes (35 mm diameter dish with a glass coverslip at 1.5 mm thick and 10 mm diameter) were from MatTek Corporation (Ashland, USA). PLGA nanoparticles (PDLG-5002 containing lactic:glycolic at 50%:50%, MW 17 KDa.) with a mean size of 161 ± 0.03 nm and a zeta potential of −29.2 ± 2.11, were supplied by Nanomi B.V. (Oldenzaal, The Netherlands).
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Pluronic F127 (PF127) and bovine serum albumin (BSA) were obtained from Sigma-Aldrich (USA). Hybrane S1200 (HS1200) was acquired from Polymer Factory (Sweden). Gantrez AN119 (AN119) was kindly provided by Ashland (Spain). Trehalose 6,6´dibehenate (TDB) and dimethyldioctadecylammonium bromide (DDAB) were purchased from Avanti Polar Lipids Inc. (USA). Cell Strainers presenting a pore size of 40 μm and 100 μm were purchased from Corning Incorporated (USA). Purified rat anti-mouse IL-17A, biotin rat anti-mouse IL-17A, purified rat anti-mouse IFN-𝛾, biotin rat anti-mouse IFN-𝛾 and streptavidin HRP were purchased from BD Pharmingen (USA). Rabbit anti-mouse IgG (H+L)-HRP was obtained from Invitrogen (USA). Goat anti-mouse IgA-HRP was purchased from Southern Biotech (USA). Carbonate buffer pH 9.6 was supplied by SSI Diagnostics (Denmark). TMB PLUS2 was obtained from Kem-En-Tec Diagnostics (Denmark). Chlamydia trachomatis vaccine CTH522 was supplied by Statens Serum Institut (SSI) (Denmark).
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Poly(methyl vinyl ether-co-maleic anhydride) or poly(anhydride) (Gantrez® AN 119) was supplied by Ashland, (Ashland, USA). Peanut extract was kindly provided by Diater® Laboratories SA (Madrid, Spain). Ethanol and rose Bengal were provided by Panreac (Barcelona, Spain). Acetone was obtained from (VWR-Prolabo). Cholera toxin, Dmannitol, D-mannosamine and Tween 20 were from Sigma-Aldrich (Germany).
Lumogen® F red 305 was from Kremer (Aichstetten, Germany). Tissue-Tek® OCT compound was obtained from Sakura (Alphen, Netherlands) 4´, 6-diamidino-2phenylindole (DAPI) was obtained from Biotium Inc. (Hayward, CA). Micro-BCA™ Protein Assay Reagent Kit was from Pierce® (Rockford, USA). Veratox Peanut Kit was from Neogen (Lansing, MI, USA) and CAPITAN MANI® soft peanut butter was from Alimentación Varma, S.L. (Alcobendas, Madrid). Sodium hydroxide (NaOH) was from Scharlau (Spain).
PLGA nanoparticles (PLGA-NP) were kindly supplied by Nanomi B.V. (The Netherlands).
These nanoparticles displayed a mean size of 161 nm with a polydispersity index of 0.03 and negative zeta potential of -29 mV.
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