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Published on: July 2, 2013
In Vitro Antimicrobial Photodynamic Therapy for Pseudomonas aeruginosa (P. aeruginosa) and methicillin-resistant
Karen Roa-Tort1, Yael Saavedra2, Angélica Villanueva-Martínez3
1Laboratorio de Optomecatrónica, UPIIH, Instituto Politécnico Nacional, Distrito de Educación, Salud, Ciencia, Tecnología e Innovación, San Agustín Tlaxiaca 42162, Mexico.
Abstract:
Photodynamic therapy (PDT) has been based on using photosensitizers (PS) and applying light of a specific wavelength. When this technique is used for treating infections, it is known as antimicrobial photodynamic therapy (aPDT). Currently, the use of lighting sources for in vitro studies using aPDT is generally applied in multiwell cell culture plates; however, depending on the lighting arrangement, there are usually errors in the application of the technique because the light from a well can affect the neighboring wells or it may be that not all the wells are used in the same experiment. In addition, one must be awarded high irradiance values, which can cause unwanted photothermal problems in the studies. Thus, this manuscript presents an in vitro antimicrobial photodynamic therapy for a Pseudomonas aeruginosa (P. aeruginosa) and methicillin-resistant Staphylococcus aureus (MRSA) inhibition study using an arrangement of thermally isolated and independently illuminated green light source systems for eight tubes in vitro aPDT, determining the effect of the following factors: (i) irradiance level, (ii) exposure time, and (iii) Rose Bengal (RB) concentration (used as a PS), registering the Pseudomonas aeruginosa (P. aeruginosa) and methicillin-resistant Staphylococcus aureus (MRSA) inhibition rates. The results show that in the dark, RB had a poor antimicrobial rate for P. aeruginosa, finding the maximum inhibition (2.7%) at 30 min with an RB concentration of 3 µg/mL. However, by applying light in a correct dosage (time × irradiance) and the adequate RB concentration, the inhibition rate increased by over 37%. In the case of MRSA, there was no significant inhibition with RB in complete darkness and, in contrast, the rate was 100% for those experiments that were irradiated.
Insights
Antimicrobial photodynamic therapy (aPDT) effectively inhibits MRSA and increases Pseudomonas aeruginosa inhibition by over 37% using optimized Rose Bengal concentration and light dosage. This study introduces a novel setup for precise in vitro aPDT experiments.
Area of Science:
- Biochemistry
- Photochemistry
- Microbiology
Background:
- Antimicrobial photodynamic therapy (aPDT) utilizes photosensitizers and light to combat infections.
- Current in vitro aPDT methods face challenges like light leakage between wells and photothermal effects.
- Standardized and precise experimental setups are crucial for reliable aPDT research.
Purpose of the Study:
- To develop and evaluate an in vitro aPDT system for inhibiting *Pseudomonas aeruginosa* and *methicillin-resistant Staphylococcus aureus* (MRSA).
- To investigate the impact of irradiance, exposure time, and Rose Bengal concentration on aPDT efficacy.
- To establish a reliable method for in vitro aPDT studies, minimizing experimental errors.
Main Methods:
- An in vitro aPDT study was conducted using a novel setup with eight independently illuminated, thermally isolated tubes.
- The system allowed precise control over irradiance, exposure time, and Rose Bengal (RB) concentration.
- Inhibition rates for *P. aeruginosa* and MRSA were determined under varying experimental conditions.
Main Results:
- Rose Bengal alone showed minimal inhibition against *P. aeruginosa* in the dark (max 2.7%).
- Optimized aPDT conditions (light dosage and RB concentration) increased *P. aeruginosa* inhibition to over 37%.
- MRSA exhibited 100% inhibition when treated with RB and light, with no significant inhibition in the dark.
Conclusions:
- The developed in vitro aPDT system with controlled light and photosensitizer concentration is highly effective against MRSA and *P. aeruginosa*.
- Precise control over irradiance and exposure time is critical for successful aPDT outcomes.
- This method offers a significant improvement over traditional multiwell plate setups for in vitro aPDT research.
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