Lornoxicam

Celecoxib, diclofenac, lornoxicam and diazepam were a kind gift of Dr. Maierhofer (AGES, PharmMed, Austria), whereas ibuprofen (I1892, SigmaAldrich, Austria), meloxicam (M3935, SigmaAldrich, Austria), piroxicam (P5654, SigmaAldrich, Austria), tenoxicam (T0909, SigmaAldrich, Austria), carboxyfluorescein (21877, Fluka, Switzerland), probenecid (P8761, SigmaAldrich, Austria) and verapamil (94837, Fluka, Switzerland) were purchased from commercial sources. Iscove’s modified Dulbecco’s medium (IMDM), Ham reg. nutrient mixture F12 (Ham’sF-12), newborn calf serum (NCS), l-glutamine and penicillin/streptomycin were obtained from Invitrogen Life technologies (GibcoTM, Carlsbad, CA). Heparin and collagen solution (predominantly collagen I, 150703) were purchased from MP Biomedicals (Irvine, CA). Amphotericin B, transferrin, 8-aminopyrene-1,3,6-trisulfonate (APTS) and gelatine were from SigmaAldrich (Austria), whereas dextran (av. MW 6000) was from Fluka (Switzerland). Fibronectin was obtained from BD Biosciences (Bedford, MA) as well as Transwell inserts (FalconTM) and six-well plates (FalconTM). Basal endothelial and astrocyte media and components for primary rat endothelial cells and astrocytes were provided from Biopredic Int. (France). Inorganic salts and all other reagents were of analytical grade.

The stock solution of 0.1 mol L⁻¹ KCl was prepared by dissolution of the appropriate amount of powder (Ing. Petr Švec-PENTA s.r.o., Prague, Czech Republic) in the distilled water and the standard solutions of 6.25 × 10⁻⁴ and 2.5 × 10⁻³ mol L⁻¹ K₃[Fe(CN)₆] and [Ru(NH₃)₆]Cl₃ (both of purity ≤99%, Sigma-Aldrich, Merck, Prague, Czech Republic) were prepared in the solution of 0.1 mol L⁻¹ KCl. Britton–Robinson buffer (BRB) consisting of the mixture of an acidic (H₃PO₄, H₃BO₃, and CH₃COOH (0.4 mol L⁻¹), Penta-Švec, Czech Republic) and alkaline component (NaOH (0.2 mol L⁻¹), Penta-Švec, Czech Republic) was used as a supporting electrolyte. The stock solution of 0.001 mol L⁻¹ lornoxicam (≥98%, Sigma-Aldrich) was prepared by dissolution of its powder in methanol (Penta-Švec, Czech Republic) and stored in a refrigerator (+4 °C) without light access. Solutions of lower concentrations were prepared daily fresh by dilution with a supporting electrolyte.