Span 80

Fluvastatin sodium was kindly donated by Eipico (Cairo, Egypt), Span 60 (sorbitan monostearate), Span 80 (sorbitan monooleate), Tween 80 (polyoxyethylene sorbitan monooleate), Brij 35 (polyoxyethylene lauryl ether), acetonitrile (HPLC grade), methanol (HPLC grade), diethyl ether (HPLC grade), complete Freund's adjuvant (CFA), and methotrexate were acquired from Sigma-Aldrich (St. Louis, MO). Dialysis bags with a molecular weight cut off of 12,000 Da were obtained from SERVA Electrophoresis GmbH (Heidelberg, Germany). Serum rheumatoid factor (RF), cartilage oligomeric matrix protein (COMP), and antinuclear antibody (ANA) ELISA kits were purchased from CliniLab Company (Cairo, Egypt). Serum tumor necrosis factor-alpha (TNF-α) ELISA kit was obtained from Glory Science company (St. Del Rio, TX). Serum interleukin-10 (IL-10) ELISA kit was obtained from Cusabio Biotech Co. (Wuhan, China). Serum malondialdehyde (MDA) and glutathione (GSH) reduced colorimetric kits were purchased from Biodiagnostic Company (Giza, Egypt). Acrylamide Kit (SDS-PAGE) (for sodium dodecyl sulfate polyacrylamide gel electrophoresis) was provided by Bio-Rad Laboratories (TNC, Hercules, CA) catalog no. 161-0181. All other ingredients used were of analytical grade.

Thyme essential oil (ρ = 0.917 g/cm³ at 25 °C) from Sigma-Aldrich^® (St. Louis, MO, USA) (Lot: MKCF4333) is used to form the oil nucleus. Chitosan (MW = 50–190 kDa, μ = 20–300 cP in CH₃COOH solution (1 g/L) at 25 °C) (Lot: STBH6262) as the encapsulating polymer, Tween^® 80 and Span^® 80 (Lot: MKBV4425V) were purchased from Sigma Aldrich^®. The distilled water was Milli-Q^® grade (Millipore Corporation, Bedford, MA, USA). ABTS (2,2′-azino-bis- (3-ethylbenzthiazoline)-6-sulfonic acid) (Lot: SLBD8908V), DPPH (2,2-diphenyl-1-picrylhydracil) (Lot: STBG9431), FRAP (2,4,6-tri (2-pyridyl)-s-triazine) (Lot: BCBW6989) were purchased from Sigma Aldrich^® (St. Louis, MO, USA). Ascorbic acid (C₆H₈O₆) (Lot: A0616289), potassium persulfate (K₂S₂O₈) (Lot: M1010511), potassium dihydrogen phosphate (KH₂PO₄) (Lot: M0918575), dipotassium hydrogen phosphate (KH₂PO₄) (Lot: M1014575), and glacial acetic acid (CH_3-COOH) (Lot: A0414235) were purchased from Meyer^® (Reactivos Química Meyer, Ciudad de México, México). Cyclohexane (C₆H₁₂) and methanol (CH₃OH) (Lot: 824333) were purchased from Fermont^® (Productos Químicos Monterrey, México).

All solvents were purchased from Fisher and used as received. Phosphate-buffered saline (PBS) solution, Dulbecco’s modified Eagle’s medium (DMEM), fungizone antimycotic (Fz), penicillin (Pen), streptomycin (Strep) and trypsin-EDTA were supplied by Gibco Life Technologies. Poly(ε-caprolactone) oligomers, hexamethylene diisocyanate, Span^® 80, dibutyltin dilaurate, collagenase, paraformaldehyde (PFA), 2-phospho-l-ascorbic acid, β-glycerophosphate, DAPI, alizarin red, Triton X100 and cetylpyridinium chloride were purchased from Sigma-Aldrich. Fetal bovine serum (FBS) was obtained from PANBiotech GmbH.
Human dermal fibroblasts were isolated, using standard procedures, from foreskins of 4-year-old children with informed consent of the parents. Fibroblasts were used for the experiments between passages 4 and 6. hMSC were obtained by plastic adhesion from bone marrow samples collected from hematologically healthy patients undergoing routine total hip replacement surgery. All samples were obtained after informed consent from donors from the Hôpital d’Instruction des Armées Percy (Clamart, France).

The commercial oils screened were olive oil (Gallo, Alges, Portugal), sunflower oil (Fula, Alges, Portugal) and corn oil (Fula, Alges, Portugal). Surfactants used for emulsification were PEG 300 (Clariant, Muttenz, Switzerland), Span^® 80, Tween^® 80, Triton™ X-100 Detergent and SDS (≥99% (GC), dust-free pellets), which were procured from Sigma Aldrich, Portugal (made in Lyon, France). Potassium carbonate (>99.5% pure, Scharlab S.L., Sentmenat, Spain) was used to prepare a solution to adjust the pH to 11. Carbonic anhydrase from bovine erythrocytes (≥95% (SDS-PAGE), specific activity ≥3500 W-A units/mg protein, lyophilised powder) was acquired from Sigma Aldrich, Portugal (made in St. Louis, MI, USA). The gases used, carbon dioxide (98% purity) and methane (99% purity) were purchased from Praxair (Huelva, Spain). The commercial membranes used in this study are detailed in Table 1.

PLGA (LA:GA = 50:50) and PDLLA with molecular weight of 100 kDa (as indicated by their inherent viscosity 0.6–0.8 dL/g) were purchased from Lakeshore Biomaterials, USA. Chloroform was supplied by Uni Chem Co., Korea. The human β-NGF with Enzyme Linked Immunosorbent Assay (ELISA) Kit, human GDNF with ELISA Kit were purchased from Peprotech Inc. and R&D Systems, Inc., respectively. Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin (P/S), and trypsin/EDTA were purchased from Invitrogen, Inc., USA. Mouse anti-neurofilament (M + H) primary antibody, goat anti-mouse IgG secondary antibody (Alexa Fluor^® 594), phalloidin (Alexa Fluor^® 488), and DAPI (4’, 6-diamidino-2-phenylindole) were purchased from Life Technologies. Triton X-100, Span-80, phosphate buffered saline (PBS) tablets, heparin, bovine serum albumin (BSA), paraformaldehyde, glutaraldehyde, sodium cacodylate, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), and sucrose were Sigma-Aldrich products. Other chemicals were used as received.

Retinol, retinaldehyde and retinoic acid were obtained from Sigma-Aldrich (St. Louis, MO) and were prepared as 1 mM stock in DMSO and stored in the vapor phase of nitrogen liquid. Corexit-EC9500A and Corexit-EC9527A (Nalco Energy Services, L.P., Sugar Land, TX) were kindly provided by Dr. Paddy Wiesenfeld at the U.S. FDA [41 (link)]. SPAN®80 (CAS#1338-43-8), TWEEN®80 (CAS#9005-65-6), TWEEN®85 (CAS#9005-70-3), Dioctyl sulfosuccinate sodium salt (DOSS, CAS#577-11-7), Di-(propylene glycol) butyl ether (CAS#29911-28-2), 2-Butoxyethanol (CAS#111-76-2), and 1,2-Propanediol (CAS#57-55-6) were purchased from Sigma-Aldrich. Chemical concentration of 1ppm is defined as 0.0001% v/v except for DOSS, which is 0.0001% w/v.