Intracellular reactive oxygen species in monocytes generated by photosensitive chromophores activated with blue light

Dent Mater. 2008 Aug;24(8):1070-6. doi: 10.1016/j.dental.2007.12.004. Epub 2008 Feb 19.

Abstract

Objectives: Disinfection of the tooth pulp-canal system is imperative to successful endodontic therapy. Yet, studies suggest that 30-50% of current endodontic treatments fail from residual bacterial infection. Photodynamic therapy using red-light chromophores (630 nm) to induce antimicrobial death mediated by generated reactive oxygen species (ROS) has been reported, but red-light also may thermally damage resident tissues. In the current study, we tested the hypothesis that several blue light chromophores (380-500 nm) generate intracellular reactive oxygen species but are not cytotoxic to mammalian cells.

Methods: THP1 monocytes were exposed to 10 microM of four chromophores (chlorin e6, pheophorbide-a, pheophorbide-a-PLL, and riboflavin) for 30 min before activation with blue light (27J/cm(2), 60s). After activation, intracellular ROS were measured using a dihydrofluorescein diacetate technique, and cytotoxicity was determined by measuring mitochondrial activity with the MTT method.

Results: All photosensitizers produced intracellular ROS levels that were dependent on both the presence of the photosensitizer and blue light exposure. Riboflavin and pheophorbide-a-PLL produced the highest levels of ROS. Photosensitizers except riboflavin exhibited cytotoxicity above 10 microM, and all except pheophorbide-a-PLL were more cytotoxic after blue light irradiation.

Significance: The current study demonstrated the possible utility of blue light chromophores as producers of ROS that would be useful for endodontic disinfection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Chlorophyll / administration & dosage
  • Chlorophyll / analogs & derivatives
  • Chlorophyll / pharmacology
  • Chlorophyllides
  • Coloring Agents
  • Fluoresceins
  • Fluorescent Dyes
  • Humans
  • Light
  • Mitochondria / drug effects
  • Monocytes / drug effects*
  • Photosensitizing Agents / administration & dosage
  • Photosensitizing Agents / pharmacology
  • Polylysine / administration & dosage
  • Polylysine / pharmacology
  • Porphyrins / administration & dosage
  • Porphyrins / pharmacology
  • Radiation-Sensitizing Agents / administration & dosage
  • Radiation-Sensitizing Agents / pharmacology*
  • Reactive Oxygen Species / metabolism*
  • Riboflavin / administration & dosage
  • Riboflavin / pharmacology
  • Succinate Dehydrogenase / drug effects
  • Tetrazolium Salts
  • Thiazoles

Substances

  • Chlorophyllides
  • Coloring Agents
  • Fluoresceins
  • Fluorescent Dyes
  • Photosensitizing Agents
  • Porphyrins
  • Radiation-Sensitizing Agents
  • Reactive Oxygen Species
  • Tetrazolium Salts
  • Thiazoles
  • dihydrofluorescein
  • Chlorophyll
  • Polylysine
  • phytochlorin
  • Succinate Dehydrogenase
  • thiazolyl blue
  • pheophorbide a
  • Riboflavin