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Updated: May 13, 2026

LED-Based In Vitro Screening for Assessing Photoactivable Molecules in Bacterial Photodynamic Inactivation
Published on: January 24, 2025
Study of photodynamic therapy in the control of isolated microorganisms from infected wounds--an in vitro study
Denise Pereira de Lima Carvalho1, Juliana Guerra Pinto, Camila Di Paula Costa Sorge
1Laboratory of Photodynamic Therapy, LEVB-Institute of Research & Development-Univap, University of Vale do Paraíba/Faculty of Health Sciences/IP&D, Av. Shishima Hifumi, 2911, Urbanova São José dos Campos, 12244, São Paulo, Brazil.
Abstract:
The effective treatment of infected wounds continues to be a serious challenge, mainly due to the rise of antibiotic-resistant bacteria. Photodynamic therapy (PDT) refers to the topical or systemic administration of a non-toxic, photosensitizing agent (PS), followed by irradiation with visible light of a suitable wavelength. The possibility of applying the PDT locally is what makes it so favorable to the treatment of infected wounds. The goal of this study was to evaluate the action of the PDT in the inactivation in vitro of microorganisms coming from infected wounds, using methylene blue (MB) and photodithazine (PDZ) as the PS and comparing the efficacy of these two compounds for PDT on bacteria. For the application of PDT, isolated microorganisms identified from material collected from wounds were suspended in a saline solution containing 10(6) viable cells/ml. Each isolated microorganism was submitted to PDT with MB and with PDZ in accordance with the following treatment groups: N/T--no treatment; T1--PDT with PDZ; T2--PDT with MB; T3--irradiation without PS; T4--treatment with PDZ without light; and T5--treatment with MB without light. As a light source, an LED-based device was used (Biopdi/Irrad-Lead 660), composed of 54 LEDs, each with 70 mW of power in the 660 nm region of the electromagnetic spectrum. Each tray of 96 wells was irradiated with an intensity of 25 mW/cm(2) and a dose of light of 50 J/cm(3) for 33 min. All the tests were made in duplicate. It was then concluded that the PDT with PDZ was capable of inhibiting the growth of gram-positive bacteria samples, however it did not have the same effect on gram-negative bacteria, which showed growth greater than 100,000 CFU; the PDT with MB showed an effectiveness on gram-positive as well as gram-negative bacteria, for it was able to inhibit bacterial growth in both cases.
Insights
Photodynamic therapy (PDT) using methylene blue (MB) effectively inactivated both gram-positive and gram-negative bacteria from infected wounds. Photodithazine (PDZ) inhibited gram-positive bacteria but not gram-negative bacteria.
Area of Science:
- Microbiology
- Biochemistry
- Photomedicine
Background:
- Antibiotic resistance poses a significant challenge in treating infected wounds.
- Photodynamic therapy (PDT) offers a localized treatment approach for wound infections.
- Photosensitizing agents (PS) combined with light are key to PDT efficacy.
Purpose of the Study:
- To evaluate the in vitro efficacy of PDT in inactivating microorganisms from infected wounds.
- To compare the effectiveness of methylene blue (MB) and photodithazine (PDZ) as photosensitizing agents in PDT.
- To assess PDT's impact on both gram-positive and gram-negative bacteria.
Main Methods:
- Microorganisms isolated from infected wounds were suspended in saline (10^6 cells/ml).
- Treatments included PDT with MB, PDT with PDZ, irradiation alone, and PS alone (no light).
- An LED device (660 nm, 25 mW/cm^2, 50 J/cm^3 for 33 min) was used for irradiation.
Main Results:
- PDT with photodithazine (PDZ) inhibited gram-positive bacteria but not gram-negative bacteria.
- PDT with methylene blue (MB) demonstrated effectiveness against both gram-positive and gram-negative bacteria.
- Control groups (no treatment, light only, PS only) showed no significant bacterial inhibition.
Conclusions:
- Methylene blue (MB) is a more effective photosensitizing agent than photodithazine (PDZ) for broad-spectrum bacterial inactivation in PDT for infected wounds.
- PDT holds promise as an alternative therapeutic strategy against antibiotic-resistant bacteria in wound infections.
- Further research is warranted to optimize PDT protocols for clinical application in wound care.

