The effect of PDT on H. influenzae biofilm in vivo

Biofilm formation has been demonstrated for many mucosal pathogens such as Haemophilus influenzae. The presence of mucosal biofilms with chronic otitis media with effusion (COME) suggests that bacteria do not clear by antibiotics. Aim: To test the effect of photodynamic therapy (PDT) on H. influenzae induced biofilm in vivo. Methods: Gerbils were divided into control (C), HI group, Laser (L), PS, PDT A, and PDT B groups. The C group received no treatment. HI group was injected with 20μl (108CFU/ml) of H. influenzae into the bullae and formation of biofilms in the bullae was obtained by 5 days. For L group, 120 J/cm2 (100 mw × 20 min) of 632 nm LD laser was irradiated by a fiber inserted into the bullae 5 days after the H. influenzaeinjection. For PS group, photofrin 40μl (1mg/ml) were injected into the bullae 5 days after the H. influenzae injection. PDT A group received photofrin 1 mg/ml and LD laser 120 J/cm2 that were administered into the bullae 5 days after the H. influenzae injection. PDT B group received photofrin 2 mg/ml and laser 150 J/cm2 5 days after the H. influenzae injection. The mucosal tissues in bullae were examined by H/E staining, and SEM. Results: The C group showed normal mucosa of bullae. The HI, L, and PS groups have shown well formed biofilm. Twenty five percent of the PDT A group and 50 % of the PDT B group have shown completely or partially resolved biofilm. Conclusion: The results of this study demonstrated that PDT appears to be effective to treat experimental H. influenzae induced biofilms in vivo. Clinical implication: PDT may be an alternative to antibiotic treatment on otitis media with biofilm formation.

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