Purasorb pdlg 5002a

PLGA 50:50 (Purasorb^® PDLG 5002A) was a gift from Corbion Purac (Amsterdam, The Netherlands) and was used to produce nanoparticles. Dichloromethane and polyvinyl alcohol (PVA, MW 89,000–98,000 Da) were from Sigma-Aldrich (St. Louis, MO, USA). The synthetic XN was from HWI Pharma Services (Rülzheim, Germany). The milli-Q water was prepared in-house. For cell culture studies, B16F10 (ATCC CRL-6475) and RAW 264.7 (ATCC TIB-71) (ATCC, Manassas, VA, USA) mouse cell lines were used. Dulbecco’s modified Eagle’s medium (DMEM), heat-inactivated fetal bovine serum (FBS), penicillin/streptomycin/amphotericin B were from Sigma-Aldrich (St Louis, MO, USA) and used for cell culture media. The CD86 monoclonal rabbit anti-mouse from ABclonal (Massachussets, MA, USA), and the CD206 monoclonal rat anti-mouse from ThermoFisher Scientific (Waltham, MA, USA) were also used. Alexa Fluor 594 conjugated goat anti-rabbit antibody and the Alexa Fluor 555 conjugated donkey anti-rat antibody were purchased from Abcam, UK.

The reagents and solvents used for the synthesis of the ILs were choline hydroxide in methanol [Cho][OH]/MeOH 45% and methanol, both from Sigma-Aldrich (Saint Louis, MO, USA), also acetonitrile from VWR (Fontenay-sous-Bois, France) and the amino acids, L-phenylalanine and glycine, from PanReact AppliChem (Barcelona, Spain).
For the cytotoxicity studies, the following reagents were purchased from Sigma-Aldrich (Saint Louis, MO, USA), phosphate buffered saline (PBS; 0.01 M, pH 7.4), trypsin, penicillin–streptomycin (pen/strep) solution, thiazolyl blue tetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO). Fetal bovine serum (FBS) and Dulbecco’s Modified Eagle’s Medium (DMEM) were from Biowest (Nuaillé, France). The propidium iodide (PI) was purchased from Merck (Darmstadt, Germany). All the rutin solutions had a final concentration of 0.5% (v/v) and were prepared in DMSO, for all assays. The solutions containing ionic liquids were all prepared in sterile water.
For the production of the nanoparticles, the dichloromethane and the polyvinyl alcohol (PVA) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Corbion Purac (Amsterdam, The Netherlands) kindly supplied the Poly(lactic-co-glycolic acid) (PLGA) 50:50 with acidic termination (Purasorb^® PDLG 5002A).
Rutin was obtained from Fagron, São Paulo, Brazil.

Terconazole was provided by Minapharm Pharmaceuticals (Cairo, Egypt). Syloid^®244 FP was received as a gift sample from W.R. Grace & Co. (St. Neots, UK). Poly(D, L-lactic/glycolic acid); PLGA with a lactide to glycolide ratio of 50:50, end-capped with acid terminal and an inherent viscosity of 0.2 dL/g (Purasorb^® PDLG 5002 A); PLGA with a lactide to glycolide ratio of 50:50, end-capped with ester terminal and an inherent viscosity of 0.2 dL/g (Purasorb^® PDLG 5002); PLGA with a lactide to glycolide ratio of 75:25, end-capped with acid terminal and an inherent viscosity of 0.22 dL/g (Purasorb^® PDLG 7502 A) and PLGA with a lactide to glycolide ratio of 75:25, end-capped with ester terminal and an inherent viscosity of 0.22 dL/g (Purasorb^® PDLG 7502) were kindly provided by Corbion (Amsterdam, Netherlands). Acetonitrile (HPLC grade), sodium chloride, sodium dihydrogen phosphate monohydrate and disodium hydrogen phosphate heptahydrate were purchased from El-Nasr Pharmaceutical Chemicals (Cairo, Egypt). Dialysis tubing cellulose membrane (molecular weight cutoff 12000-14000 g/mole) was procured from Sigma Aldrich (Saint louis, MO, USA). The rest of chemicals and solvents were of analytical grade and were utilized as received. Water was used as deionized, bi-distilled water.