Cytotoxic and genotoxic potential of the type I photoinitiators BAPO and TPO on human oral keratinocytes and V79 fibroblasts

Dent Mater. 2018 Dec;34(12):1783-1796. doi: 10.1016/j.dental.2018.09.015. Epub 2018 Oct 17.

Abstract

Objectives: Phenylbis(acyl) phosphine oxide (BAPO) and diphenyl(acyl) phosphine oxide (TPO) are alternative photoinitiators to camphorquinone (CQ) in dental resinous materials. Aim of this study was to investigate their cytotoxic/genotoxic potential in human oral keratinocytes (OKF6/Tert2) and Chinese hamster lung fibroblasts (V79) in comparison to CQ.

Methods: Cells were exposed to different concentrations of BAPO and TPO (1-50μM). Cytotoxicity was evaluated using H33342 and MTT assay, cell proliferation by BrdU proliferation assay and microscopy. Effects on cellular redox homeostasis were assessed by detecting intracellular levels of reactive oxygen/nitrogen species (ROS/RNS) using the DCFH2 assay and by quantification of mRNA expression of oxidatively regulated, cyto-protective enzymes. Genotoxic potential was determined by use of micronucleus (MN) assay.

Results: BAPO and TPO induced a concentration-dependent decrease of cell number. BAPO and TPO showed 50- to 250-fold higher cytotoxicity than CQ. In contrast to CQ, both photoinitiators revealed no increase of intracellular ROS/RNS. However, BAPO (10μM) at least significantly induced mRNA-expression of redox-regulated proteins after 24h similar to 2.5mM CQ. Additionally, BAPO significantly raised the number of micronuclei, but only in V79 cells (10μM: 12±1, 2.5mM CQ: 15±1, medium control: 6±3). However, it also significantly decreased proliferation of these cells (10μM BAPO: 19.8%±7.3% compared to controls).

Significance: BAPO and TPO revealed concentration-dependent cytotoxic effects in human oral keratinocytes and V79 cells. However, in contrast to CQ, no generation of intracellular ROS/RNS was found. Only BAPO induced genotoxicity in V79 cells.

Keywords: Camphorquinone; Cytotoxicity; Diphenyl(acyl) phosphine oxides; Genotoxicity; Phenylbis(acyl) phosphine oxides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Camphor / analogs & derivatives
  • Camphor / toxicity
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Cricetulus
  • Cyclic N-Oxides / toxicity*
  • Fibroblasts / drug effects*
  • Humans
  • In Vitro Techniques
  • Keratinocytes / drug effects*
  • Lung / cytology
  • Materials Testing
  • Micronucleus Tests
  • Mutagenicity Tests
  • Oxidative Stress
  • Photoinitiators, Dental / toxicity*
  • Reactive Oxygen Species

Substances

  • Cyclic N-Oxides
  • Photoinitiators, Dental
  • Reactive Oxygen Species
  • Camphor
  • 4-benzylideneamino-2,2,6,6-tetramethylpiperidine-1-oxyl
  • camphorquinone