Sophorolipid-toluidine blue conjugates for improved antibacterial photodynamic therapy through high accumulation

Abstract

Anti-bacterial photodynamic therapy is the most promising treatment protocol for bacterial infection, but low accumulation of photosensitizers has seriously hindered their development in clinical application. Here, with inherent outstanding affinity to bacterial cell envelope, sophorolipid produced from Candida bombicola has been conjugated to toluidine blue (SL-TB) through amidation reaction. The structure of SL-TB conjugates was identified by ¹H-NMR, FT-IR and ESI-HRMS. The interfacial assembly and photophysical properties of SL-TB conjugates have been disclosed through surface tension, micro-polarity, electronic and fluorescence spectra. After light irradiation, the log₁₀ (reduced CFU) of free toluidine blue to P. aeruginosa and S. aureus were 4.5 and 7.9, respectively. In contrast, SL-TB conjugates showed a higher bactericidal activity, with a reduction of 6.3 and 9.7 log₁₀ units of CFU against P. aeruginosa and S. aureus, respectively. The fluorescence quantitative results showed that SL-TB could accumulate 2850 nmol/10¹¹ cells and 4360 nmol/10¹¹ cells by P. aeruginosa and S. aureus, which was much higher than the accumulation of 462 nmol/10¹¹ cells and 827 nmol/10¹¹ cells of free toluidine blue. Through the cooperation of triple factors, including sophorose affinity to bacterial cells, hydrophobic association with plasma membrane, and electrostatic attraction, higher SL-TB accumulation was acquired, which has enhanced antibacterial photodynamic efficiencies.