3 4 5 dimethylthiazol 2 yl 3 5 diphenyltetrazolium bromide mtt dye test

Phospholipon 90G^® (PL-90G) was kindly provided by Natterman Phospholipid GMBH (Köln, Germany), and contained 93 ± 3% phosphatidylcholine. Absolute ethanol (Ph. Eur. analysis reagent) and poloxamer 407 (P407) were purchased from Sigma-Aldrich (Schnelldorf, Germany). T/C-28a2 and C-28/I2 cells, immortalized chondrocyte cell lines, were obtained from the Research Foundation for the care of cancer “Tommaso Campanella” University Campus of Germaneto–Catanzaro, I-88100, Italy. Dulbecco-modified Eagle medium (D-MEM) culture, fetal bovine serum, penicillin (100 UI/mL)–streptomycin (100 μg/mL) solution (1% v/v), and Trypsin/EDTA (1×) solution were obtained from GIBCO (Invitrogen Corporation, Giuliano Milanese, Milan, Italy). 3-[4,5-dimethylthiazol-2-yl]-3,5-diphenyltetrazolium bromide (MTT) dye test (TLC purity ≥ 97.5%), phosphate buffer (PBS) solution, and sodium dimethyl sulfoxide (DMSO), were purchased from Sigma-Aldrich (Milan, Italy).
[³H]cholesteryl hexadecyl ether ([³H]CHE, 40 Ci/mmoL) was obtained from Perkin Elmer-Italia (Monza, Italy). Double distilled pyrogen-free water was used throughout the experimental investigations. All other materials and solvents used in this study are of analytical grade (Carlo Erba, Milan, Italy).

Phospholipon^® 80H is a Phospholipid GmbH (Cologne) product. Rutin hydrate (MW 610,2, RT), minimum 95%, Tween 80, trehalose from Saccharomyces Cerevisiae ≥ 99%, 3-[4,5-dimethylthiazol-2-yl]-3,5-diphenyltetrazolium bromide (MTT) dye test (TLC purity ≥97.5%), sodium dimethyl sulfoxide (DMSO) and phosphate buffer solution (PBS, pH 7.4) were purchased from Sigma Aldrich (Milan, Italy). Ethanol HPLC grade is a Merck^® product. Acetone HPLC grade was purchased from J.T. Baker^®. Dulbecco’s modified Eagle’s medium (DMEM), heat-inactivated fetal bovine serum (FBS), penicillin (100 UI/mL)/streptomycin (100 µg/mL) solution, amphotericin B (250 μg/mL), glutamine, and trypsin/ethylenediaminetetraacetic acid (EDTA) (1×) solution were obtained from GIBCO (Invitrogen Corporation, Giuliano Milanese, Milano, Italy). Human glioblastoma astrocytoma cells (U373) were purchased from ATCC (American Type Culture Collection) retailer LGC Standards S.r.l. (Sesto San Giovanni, Milano, Italy). The Pierce lactic dehydrogenase (LDH) cytotoxicity assay kit was obtained from Thermo Fisher-Scientific (Waltham, MA, USA). The water used throughout the study was double-distilled, then filtered through 0.2 μm polytetrafluoroethylene (PTFE) filters (Millipore). Dialysis bags were Spectra/Por^® Dialysis Membrane (MWCO: 8000, Spectrum Laboratories, Inc., Rancho Dominguez, CA, USA).