Inflammation parameters were determined only at the end of experi

Inflammation parameters were determined only at the end of experiment in each setting at 90 min, 150 min or 270 min. For each blood sampling animals were substituted with a 0.5 ml bolus of 0.9% NaCl via the femoral artery (with the additional effect to keep the catheter functional). In order to obtain plasma, blood was centrifuged at 4000g for 15 min at room temperature. The gained plasma was stored at 4 °C until use (maximal within 4 h). Arterial blood pH, oxygen and carbon dioxide partial pressures (pO2, pCO2), base excess (BE), bicarbonate and lactate were assessed with a blood gas analyzer (ABL 715, Radiometer,

Copenhagen, p38 MAPK cancer Denmark). The plasma activity of lactate dehydrogenase (LDH) as a general marker of cell injury, creatine kinase (CK) as a marker for muscle cell injury, aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) as markers for hepatocyte injury as well as the plasma creatinine concentration as a marker of renal function were determined using a fully automated clinical chemistry analyzer (Vitalab Selectra E, VWR International, Darmstadt, Germany) using commercially available reagent kits (DiaSys, Holzheim, Germany). The plasma concentration of the cytokines Interferon-gamma (IFN-γ), Interleukin-1alpha (IL-1α), IL-1β, IL-4, IL-5, IL-6, IL-10 and tumor necrosis factor alpha (TNF-α) were

determined by using a rat-specific multiplex bead suspension Histone demethylase array (Bio-Plex Cytokine Assay) in conjunction with a Bio-Plex Array Reader (Bio-Rad, check details Muenchen, Germany). The plasma concentration of complement factors 3 (C3) and 4a (C4a) were assessed with rat-specific ELISA kits (Rat complement fragment 4a ELISA kit, Cusabio, Wuhan, China, and

Rat Complement Factor 3, GenWay Biotech Inc, San Diego, CA, USA) according to the manufacturer’s instructions. In vivo microscopy analysis (IVM) was performed with a Leica DMLM epifluorescence microscope (Leica Microsystems, Wetzlar, Wetzlar, Germany, ×160 magnification). Anesthesia Analgesia, and surgical procedures were the same as in the setting without IVM. The left lateral liver lobe was exteriorized on a specially designed stage 40 min after catheterization of the femoral vessels. The abdominal cavity was kept moist. One milliliter/kilogram body weight of fluorescein isothiocyanate-dextran 150,000 (FITC-dextran, 5% solution in 0.9% NaCl) was injected intravenously for fluorescent staining of liver microcapillaries [ 17]. Fifteen min after application of FITC-dextran, PFD-filled PLGA microcapsules (either 1.5 or 1 µm), PLGA microspheres (1.5 µm) or 0.9% NaCl solution were infused continuously for 30 min into the right femoral vein using a syringe pump (20 ml/kg body weight × h). During the measurement, animals were biomonitored continuously as described.

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