Absence of NF-kappaB activation by a new polystyrene-type adsorbent designed for hemoperfusion
| Autor: | Morteza Mansouri, Luigi Massimino Sena, Annalisa Davit, James F. Winchester, Antonella Dragonetti, Giuseppina Loduca, Elisa Menegatti, Debora Di Simone, Giulio Mengozzi, Dario Roccatello, Gian Piero Sancipriano, Claudio Ronco, Giorgio Marietti |
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| Jazyk: | angličtina |
| Rok vydání: | 2005 |
| Předmět: |
Biocompatibility
medicine.medical_treatment Gene Expression Nitric Oxide Hemoglobins chemistry.chemical_compound Adsorption Reference Values Materials Testing medicine Humans Chromatography Reverse Transcriptase Polymerase Chain Reaction Tumor Necrosis Factor-alpha NF-kappa B Hematology General Medicine Elisa assay Hemoperfusion Interleukin 1β Blood donor chemistry Biochemistry Nephrology Luminescent Measurements Cytokines Polystyrenes Polystyrene Nf κb activation Interleukin-1 |
| Popis: | Aim: The aim of the study was to evaluate biocompatibility of anew polystyrene-type adsorbent (BetaSorbTM) designed for hemoperfusion, using second-level biomolecular analyses. The device has recently been developed to enhance β2-microglobulin removal during hemodialysis. Molecular structure and chemical modifications of the surface beads of this cartridge should prevent exposure of dense hydrophobic surface sites to proteins, and avoid the major drawbacks of previous polystyrene-type adsorbent materials. Methods: Whole blood of healthy donors was incubated in sterile minicolumns packed with BetaSorb Cuprophan, Hemophan, polysulfone and cellulose acetate. In parallel experiments, whole blood was recirculated for 180 min in a sham dialysis circuit equipped with the study sorbent or Hemophan or polysulfone. Biocompatibility was assessed by means of new biomolecular approaches focused on nuclear factor ĸB (NF-ĸB) activation (assessed by electrophoretic mobility shift assay), TNF-α and IL-1β gene expression (evaluated by real-time PCR), TNF-α and IL-1β production (measured by Western blot assay and ELISA), nitric oxide (NO) generation (detected by electron paramagnetic resonance), free oxygen radical production (by chemiluminescence in a biological assay) and the generation of the complement breakdown product C3d. Results:In coincubation experiments, 5-min contact with any dialysis device, but BetaSorb, was enough to induce activation of NF-ĸB. The amount of TNF-α precursor form was found to increase after 5 min of exposure to each tested polymer, but no traces of mature forms of TNF-α or IL-1β were detected in in vitro experimental conditions using healthy blood. NO and free oxygen radical generation were significantly lower in blood samples exposed to BetaSorb than in control dialysis devices. C3d levels were found to be increased with Hemophan, unaffected by polysulfone, and remarkably decreased with the BetaSorb device. In the sham hemodialysis experiments, NF-ĸB activation and C3d and NO profiles were similar to direct incubation experiments. Compared to basal levels, quantitation of TNF-α and IL-1β mRNA revealed a 15- and 9-fold increase, respectively, in samples exposed to Hemophan for 180 min. Conclusions: The new BetaSorb device not only appears to be highly biocompatible, but shares properties that make it probably able to interfere with the activation of the inflammatory state. |
| Databáze: | OpenAIRE |
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