Effect of N-acetyl-L-cysteine on ROS production and cell death caused by HEMA in human primary gingival fibroblasts

Gianrico Spagnuolo, Vincenzo D'Antò, Claudia Cosentino, Gottfried Schmalz, Helmut Schweikl, Sandro Rengo

Research output: Contribution to journalArticlepeer-review


Previous investigations have shown that 2-hydroxyethyl methacrylate (HEMA) causes reactive oxygen species (ROS) production, which in turn affects cell survival and cell death. The purpose of this study was to evaluate the effects of the antioxidant N-acetyl-l-cysteine (NAC) on HEMA-induced toxicity in human primary gingival fibroblasts (HGF). HGF were treated with various concentrations of HEMA (0-12 mm) in the absence and presence of NAC (1, 5, and 10 mm). The 3-(4,5 dimethyiazol-2-1)-2-5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate the mitochondrial dehydrogenase activity after HEMA exposure. Viability and cell death were determined by flow cytometry using Annexin V and PI staining. ROS production was detected by the increasing fluorescence of the oxidation-sensitive dye 2′,7′-dichlorofluorescein diacetate (DCFH-DA) after HEMA treatment. After a 24 h incubation period, HEMA concentrations higher then 10 mm caused a decrease of cell viability, mitochondrial activity, and an increase of cell death. HEMA concentrations of 4-12 mm markedly increased ROS levels in a dose-dependent manner. High NAC concentrations (5 and 10 mm) significantly reduced cell death, and restored the mitochondrial activity after a 24 h co-treatment, but 1 mm NAC increased HEMA toxicity (p

Original languageEnglish
Pages (from-to)1803-1809
Number of pages7
Issue number9
Publication statusPublished - Mar 2006


  • Dental resin
  • HEMA
  • Human gingival fibroblasts
  • NAC
  • Reactive oxygen species

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering


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