Photodynamic effects of toluidine blue on human oral keratinocytes and fibroblasts and Streptococcus sanguis evaluated in vitro

Abstract

Background and objective: Some oral bacteria are susceptible to killing by red light after their sensitization with toluidine blue O (TBO). The photochemotherapy of periodontal disease in vivo would require a therapeutic window where bacteria could be killed without adjacent normal tissue damage.

Study design/materials and methods: The laser-induced effects of TBO on normal human gingival keratinocytes and fibroblasts have been studied in vitro. For the assessment of viability, the CellTiter 96TM AQueous Non-Radioactive Cell Proliferation Assay was used.

Results: TBO was cytotoxic at low concentrations (5.0 micrograms/ml). Sensitization of keratinocytes and fibroblasts with 2 and 5.0 micrograms/ml TBO, respectively, for 5 min and exposure to light from a 7.3 mW Helium/Neon (HeNe) laser for up to 2 min (0.876J) did not reduce cell viability. However, killing of Streptococcus sanguis was achieved following exposure to HeNe light for 75 sec (0.547J) in the presence of TBO at a concentration of 2.5 micrograms/ml.

Conclusion: The development of a system for the lethal photosensitization of bacteria responsible for periodontal disease may be possible.