As resistance of bacterial strains to antibiotics is a major problem, there is a need to look for alternative treatments. One option is antimicrobial photodynamic inactivation (aPDI). The pathogenic cells are targeted by a nontoxic photosensitizer while the surrounding healthy tissue is relatively unaffected. The photosensitizer is activated by light of t appropriate wavelength resulting in the generation of reactive oxygen species that are cytotoxic for the pathogens. In this work, the photosensitizer TMPyP and silver nanoparticles (AgNPs) were investigated for their synergistic antibacterial effect. We tested these two substances on two bacterial strains, methicillin-resistant Staphylococcus aureus 4591 (MRSA) and extended-spectrum beta-lactamases-producing Klebsiella pneumoniae 2486 (ESBL-KP), to compare their effectiveness. The bacteria were first incubated with TMPyP for 45 min or 5 h, then irradiated with a LED source with the total fluence of 10 or 20 J/cm2 and then placed in a microbiological growth medium supplemented with AgNPs. To accomplish the synergistic effect, the optimal combination of TMPyP and AgNPs was estimated as 1.56-25 μM for TMPyP and 3.38 mg/l for AgNPs in case of MRSA and 1.56-50 μM for TMPyP and 3.38 mg/l for AgNPs in case of ESBL-KP at 45 min incubation with TMPyP and fluence of 10 J/cm2. Longer incubation and/or longer irradiation led to a decrease in the maximum values of the photosensitizer concentration to produce the synergistic effect. From this work it can be concluded that the combination of antimicrobial photodynamic inactivation with a treatment including silver nanoparticles could be a promising approach to treat bacterial infection.

Department of Medical Biophysics Faculty of Medicine and Dentistry Palacky University in Olomouc Czech Republic

Department of Medical Biophysics Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University in Olomouc Czech Republic

Department of Microbiology Faculty of Medicine and Dentistry Palacky University in Olomouc Czech Republic

Department of Physical Chemistry Regional Centre of Advanced Technologies Faculty of Science Palacky University in Olomouc Czech Republic

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$a Malá, Zuzana, $u Department of Medical Biophysics, Faculty of Medicine and Dentistry, Palacky University in Olomouc, Czech Republic. Electronic address: zuza.mala@seznam.cz $d 1990- $7 xx0267678

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$a The application of antimicrobial photodynamic inactivation on methicillin-resistant S. aureus and ESBL-producing K. pneumoniae using porphyrin photosensitizer in combination with silver nanoparticles / $c Z. Malá, L. Žárská, R. Bajgar, K. Bogdanová, M. Kolář, A. Panáček, S. Binder, H. Kolářová

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$a As resistance of bacterial strains to antibiotics is a major problem, there is a need to look for alternative treatments. One option is antimicrobial photodynamic inactivation (aPDI). The pathogenic cells are targeted by a nontoxic photosensitizer while the surrounding healthy tissue is relatively unaffected. The photosensitizer is activated by light of t appropriate wavelength resulting in the generation of reactive oxygen species that are cytotoxic for the pathogens. In this work, the photosensitizer TMPyP and silver nanoparticles (AgNPs) were investigated for their synergistic antibacterial effect. We tested these two substances on two bacterial strains, methicillin-resistant Staphylococcus aureus 4591 (MRSA) and extended-spectrum beta-lactamases-producing Klebsiella pneumoniae 2486 (ESBL-KP), to compare their effectiveness. The bacteria were first incubated with TMPyP for 45 min or 5 h, then irradiated with a LED source with the total fluence of 10 or 20 J/cm2 and then placed in a microbiological growth medium supplemented with AgNPs. To accomplish the synergistic effect, the optimal combination of TMPyP and AgNPs was estimated as 1.56-25 μM for TMPyP and 3.38 mg/l for AgNPs in case of MRSA and 1.56-50 μM for TMPyP and 3.38 mg/l for AgNPs in case of ESBL-KP at 45 min incubation with TMPyP and fluence of 10 J/cm2. Longer incubation and/or longer irradiation led to a decrease in the maximum values of the photosensitizer concentration to produce the synergistic effect. From this work it can be concluded that the combination of antimicrobial photodynamic inactivation with a treatment including silver nanoparticles could be a promising approach to treat bacterial infection.

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$a Žárská, Ludmila, $u Department of Medical Biophysics, Faculty of Medicine and Dentistry, Palacky University in Olomouc, Czech Republic. Electronic address: ludmila.zarska@centrum.cz $d 1990- $7 xx0267676

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$a Bajgar, Robert $u Department of Medical Biophysics, Faculty of Medicine and Dentistry, Palacky University in Olomouc, Czech Republic. Electronic address: robert.bajgar@tunw.upol.cz

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$a Bogdanová, Kateřina $u Department of Microbiology, Faculty of Medicine and Dentistry, Palacky University in Olomouc, Czech Republic. Electronic address: katerina.bogdanova@upol.cz

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$a Kolář, Milan $u Department of Microbiology, Faculty of Medicine and Dentistry, Palacky University in Olomouc, Czech Republic. Electronic address: milan.kolar@upol.cz

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$a Panáček, Aleš $u Department of Physical Chemistry, Regional Centre of Advanced Technologies, Faculty of Science, Palacky University in Olomouc, Czech Republic. Electronic address: ales.panacek@upol.cz

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$a Binder, Svatopluk $u Department of Medical Biophysics, Faculty of Medicine and Dentistry, Palacky University in Olomouc, Czech Republic. Electronic address: svatopluk.binder@email.cz

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$a Kolářová, Hana $u Department of Medical Biophysics, Faculty of Medicine and Dentistry, Palacky University in Olomouc, Czech Republic; Department of Medical Biophysics, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University in Olomouc, Czech Republic. Electronic address: hana.kolarova@upol.cz

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