Effect of Ca+ on the photobactericidal efficacy of methylene blue and toluidine blue against gram-negative bacteria and the dye affinity for lipopolysaccharides

• Published in: Lasers in surgery and medicine 2006-12, Vol.38 (10), p.946-954
• Main Authors: Usacheva, Marina N., Teichert, Matthew C., Sievert, Chet E., Biel, Merrill A.
• Format: Article
• Language: English
• Subjects: bacterial damage; CaCl2 inhibition; Calcium Chloride - metabolism; Calcium Chloride - pharmacology; Coloring Agents - metabolism; Coloring Agents - pharmacology; Gram-Negative Bacteria - drug effects; Lasers; Lipopolysaccharides - metabolism; Methylene Blue - metabolism; Methylene Blue - pharmacology; phenothiazine dyes; photobleaching; proteins of bacterial outer membrane; red laser light; Spectrophotometry; Tolonium Chloride - metabolism; Tolonium Chloride - pharmacology
• ISSN: 0196-8092; 1096-9101
• DOI: 10.1002/lsm.20400

Background and Objectives Methylene blue (MB) and toluidine blue (TB) form metachromatic complexes with lipopolysaccharides (LPS). The greater photobactericidal efficacy of TB may be explained by its affinity for LPS. This study aims to elucidate the difference in photobactericidal efficacies between the dyes using Ca2+ as a competitor for dye‐binding sites on the bacterial outer membrane. Study Design/Materials and Methods Fixed dye concentration solutions with gram‐negative bacteria and increasing concentrations of CaCl2 were exposed to red laser light. Bacterial survival and spectrophotometry were used to describe the effect of Ca2+ on dye interaction with bacteria and LPS. Results MB‐mediated photokilling was inhibited more significantly than that of TB. CaCl2 inhibited dye photobleaching and suppressed the metachromatic reaction between the dyes and LPS, in particular TB. Conclusions CaCl2 inhibits bacterial photokilling by binding with LPS, as well as other anionic polymers including outer membrane proteins. LPS is chiefly involved in TB‐mediated photokilling, whereas outer membrane proteins probably are more involved in MB‐mediated photokilling. Lasers Surg. Med. 38:946–954, 2006. © 2006 Wiley‐Liss, Inc.