In Vitro Eradication of Planktonic, Saliva and Biofilm Bacteria Using Lingonberry Extract as a Photosensitizer for Visible Light Plus Water-Filtered Infrared-A Irradiation

Abstract

Antimicrobial photodynamic treatment (aPDT) with visible light plus water-filtered infrared-A irradiation (VIS-wIRA) and natural single- or multi-component photosensitizers (PSs) was shown to have potent antimicrobial activity. The aim of this study was to obtain information on the antimicrobial effects of aPDT-VIS-wIRA with lingonberry extract (LE) against bacteria that play a role in oral health. Planktonic bacterial cultures of the Gram-positive E. faecalis T9, S. mutans DSM20523, S. oralis ATCC 35037 and S. sobrinus PSM 203513, the Gram-negative N. oralis 14F2 FG-15-7B, F. nucleatum ATCC 25586, and V. parvula DSM, the anaerobic F. nucleatum ATCC 25586 and V. parvula DSM 2008, and the total mixed bacteria from pooled saliva and supra- and subgingival plaques of volunteers were all treated and compared. aPDT-VIS-wIRA with LE as PS significantly (p < 0.008) reduced the growth of all tested Gram-positive, Gram-negative, as well as aerobic and anaerobic bacterial strains, whereas without irradiation no reductions were seen (p < 0.0001). NaCl, with or without irradiation, was ineffective. After treatment with CHX 0.2%, the highest killing rate (100%) was observed, and no bacteria (0 log10 CFU) were cultivable. The method also significantly reduced all of the bacteria present in saliva and in the gingival biofilms. Three-dimensional visualization of viable and non-viable microorganisms revealed that LE penetrated deeper into the cell wall layers than CHX 0.2%. LE was an appropriate PS for eradicating microorganisms with VIS-wIRA, either in their planktonic form or in saliva and gingival plaque biofilms. These results encourage further investigation in order to determine which LE compounds contribute to the photosensitizing effect and to evaluate the size of the effect on maintaining oral health.

Keywords: Vaccinium vitis-idea; antimicrobial photodynamic treatment; lingonberry; oral pathogenic bacteria; photosensitizer; visible light plus water-filtered infrared-A irradiation.

Figure 1

Absorbance spectrum of the lingonberry extract tested in this study.

Figure 2

The effects of aPDT using VIS+wIRA in combination with LE on bacterial strains. (a) Neisseria oralis; (b) Streptococcus mutans; (c) Streptococcus sobrinus; (d) Enterococcus faecalis; (e) Streptococcus oralis; (f) Veillonella parvula; (g) Fusobacterium nucleatum.

Figure 3

The effects of aPDT using VIS+wIRA in combination with LE on human saliva and oral supra- and subgingival biofilm samples. (a) Saliva aerobic testing; (b) Saliva anaerobic testing; (c) Supragingival plaque aerobic testing; (d) Supragingival plaque anerobic; (e) Subgingival plaque aerobic testing; (f) Subgingival plaque anaerobic testing.

Figure 4

CLSM after Live/Dead Staining, 3D-reconstruction. Viable bacteria were depicted green, while dead bacteria appeared red. (a) Supragingival plaque after aPDT with VIS+wIRA and LE; (b) LE without irradiation; (c) NaCl as negative control; (d) CHX0.2% as positive control.

Figure 4

CLSM after Live/Dead Staining, 3D-reconstruction. Viable bacteria were depicted green, while dead bacteria appeared red. (a) Supragingival plaque after aPDT with VIS+wIRA and LE; (b) LE without irradiation; (c) NaCl as negative control; (d) CHX0.2% as positive control.