Impact of PVC microplastics in photodynamic inactivation of Staphylococcus aureus and MRSA.

Photodynamic processes have found widespread application in therapies. These processes involve photosensitizers (PSs) that, when excited by specific light wavelengths and in the presence of molecular oxygen, generate reactive oxygen species (ROS), that target cells leading to inactivation. Photodynamic action has gained notable attention in environmental applications, particularly against pathogens and antibiotic-resistant bacteria (ARB) that pose a significant challenge to public health. However, environmental matrices frequently encompass additional contaminants and interferents, including microplastics (MPs), which are pollutants of current concern. Their presence in water and effluents has been extensively documented, highlighting their impact on conventional treatment methods, but this information remains scarce in the context of photodynamic inactivation (PDI) setups. Here, we described the effects of polyvinyl chloride (PVC) microparticles in PDI targeting Staphylococcus aureus and its methicillin-resistant strain (MRSA), using curcumin as a PS under blue light. The presence of PVC microparticles does not hinder ROS formation; however, depending on its concentration, it can impact bacterial inactivation. Our results underscore that PDI remains a potent method for reducing bacterial concentrations in water and wastewater containing ARB, even in highly contaminated scenarios with MPs.

Photodynamic treatment of malignant melanoma with structured light: in silico Monte Carlo modeling.

In this report, we propose a novel strategy for the photodynamic approach to the treatment of melanoma, aiming to mitigate the excessive absorption and consequent thermal effects. The cornerstone of this approach is an innovative structured illumination technique that optimizes light delivery to the tissue. The methodology of this in silico study involves the development of an optical model of human skin with the presence of melanoma and an accurate simulation technique of photon transport within the complex turbid scattering medium. To assess the effectiveness of our proposed strategy, we introduced a cost function reflecting the irradiated volume and optical radiation absorption within the target area/volume occupied by malformation. By utilizing the cost function, we refine the offset illumination parameters for a variety of target system parameters, ensuring increased efficiency of photodynamic therapy. Our computer simulation results introduce a promising new path towards improved photodynamic melanoma treatments, potentially leading to better therapeutic outcomes and reduced side effects. Further experimental validation is needed to confirm these theoretical advancements, which could contribute towards revolutionizing current melanoma photodynamic treatment methodologies.

Femtosecond pulsed laser photodynamic therapy activates melanin and eradicates malignant melanoma.

Photodynamic therapy (PDT) relies on a series of photophysical and photochemical reactions leading to cell death. While effective for various cancers, PDT has been less successful in treating pigmented melanoma due to high light absorption by melanin. Here, this limitation is addressed by 2-photon excitation of the photosensitizer (2p-PDT) using ~100 fs pulses of near-infrared laser light. A critical role of melanin in enabling rather than hindering 2p-PDT is elucidated using pigmented and non-pigmented murine melanoma clonal cell lines in vitro. The photocytotoxicities were compared between a clinical photosensitizer (Visudyne) and a porphyrin dimer (Oxdime) with ~600-fold higher σ2p value. Unexpectedly, while the 1p-PDT responses are similar in both cell lines, 2p activation is much more effective in killing pigmented than non-pigmented cells, suggesting a dominant role of melanin 2p-PDT. The potential for clinical translational is demonstrated in a conjunctival melanoma model in vivo, where complete eradication of small tumors was achieved. This work elucidates the melanin contribution in multi-photon PDT enabling significant advancement of light-based treatments that have previously been considered unsuitable in pigmented tumors.

Influence of temperature combined with photodynamic inactivation on the development of Aedes aegypti.

To reduce the speed of selection of populations resistant to chemical insecticides, photodynamic inactivation (PDI) against Aedes aegypti is a hot-topic and promising alternative technique to vector control. Temperature is an important factor in the survival of Ae. aegypti larvae and mosquitoes as it influences physiology, behavior, and ecology. This work aimed to evaluate parameters of the biological cycle of Ae. aegypti such as: hatching rate, larval development, adult mosquito longevity, sex ratio, weight, and lethal concentration of larval mortality (LC) through the combination of PDI with different temperatures. The number of larvae found after 48 h suggests that temperature affects hatching rate. Additionally, results showed a delay in development of surviving larvae after PDI when compared to control groups, and there was a reduction in the longevity of mosquitoes that undertook photodynamic action. PDI also led to a predominance of male insects, and observed weight indicates that the inactivation method may have also interfered in mosquito size. The results point to a satisfactory performance of PDI at all tested temperatures. Experimental conditions that were not lethal to all larvae implied that PDI impacts the mosquitoes' biological cycle. Though metabolism and development are improved at higher temperatures, so is PDI action, thus maintaining the net benefit. Therefore, it is assumed that the proposed photolarvicide can be useful in reducing arbovirus transmission, and results invite for future research in different abiotic conditions.

Enhanced vegetable production in hydroponic systems using decontamination of closed circulating fluid.

While plant microorganisms can promote plants by producing natural antibiotics, they can also be vectors for disease transmission. Contamination from plant management practices and the surrounding environment can adversely affect plants, leading to infections and hindered growth due to microbial competition for nutrients. The recirculation of nutrient-rich fluids can facilitate the transport of microorganisms between vegetables in the hydroponic production system. This issue can be addressed through the application of the decontamination method in the hydroponic liquid. Ultraviolet light (UV-C) has been employed for microbiology, and its effects on lettuce were evaluated in this study. This study aims to assess the effectiveness of a decontamination system using UV-C in hydroponic solutions during nutrient recirculation in hydroponics. We evaluated the time required for lettuce plants to reach their maximum height, as well as their pigment content, phenolic compounds, antioxidant capacity, and micro and macronutrient levels. The evaluation was conducted under two photoperiods (18 and 20 hours) in lettuce samples exposed to UV-C in the hydroponic fluid, with control groups not exposed to UV-C. The application of the UV-C decontamination system in hydroponic circulation water containing nutrients accelerated plant growth while maintaining nutritional values equal to or higher than those in the control groups without such a system. The results of microorganism control highlight the potential application of this technique for enhancing and expediting vegetable production. This approach reduces production time and enhances nutrient absorption and the content of certain compounds and minerals.

Photodynamic therapy for nodular basal cell carcinoma up to 5mm located on high-risk area: Effectiveness and long-term follow-up results.

SIGNIFICANCE: Response rates evaluation of photodynamic therapy (PDT) for nodular basal cell carcinoma (BCC) treatment located on high-risk and low-risk areas of the face. APPROACH: Two groups of nodular BCC were selected, debulked, and received 20% methyl aminolevulinate (MAL) hydrochloride cream. After 3 h, the first irradiation was performed (20 min, 150 J/cm2). Then, the cream was re-applied, and a second irradiation was performed after 1.5 h (20 min, 150 J/cm2). Clearance at 30 days and recurrence-free survival rate were evaluated. RESULTS: The clearance at 30 days after PDT was 89% for the low-risk area group and 87% for the high-risk group. The recurrence-free survival at 60 months was 82% and 85% for the high-risk and low-risk groups, respectively. CONCLUSIONS: No significant differences were observed between groups nor for clearance at 30 days, nor recurrence-free follow-up. These results make PDT possible option for nodular BCC less than 5 mm located in high-risk areas.

Long-term follow-up results of a pilot study for nodular basal cell carcinoma with PDT using partial home treatment protocol.

SIGNIFICANCE: Evaluate a photodynamic therapy (PDT) protocol for low-risk basal cell carcinoma (BCC) treatment that requires less time spent at the hospital and is less painful. APPROACH: Eight BCCs were selected, debulked, and received 20 % methyl aminolevulinate cream. After 3 h, the first irradiation was performed at the hospital (20 min, 150 J/cm2). Then, the cream was re-applied, and a portable irradiation prototype was fixed to the skin around the lesion. After 1.5 h, the patients turned on the prototype for irradiation at home (for 2 h, totalizing 312 J/cm2). Disease-free survival rate and pain score during irradiations were evaluated. RESULTS: The clearance at 30 days after PDT was 87.5 % by histological analysis. The mean follow-up was 21.5 months and the recurrence-free survival at 22 months was 75 %. The pain score was significantly lower at home. CONCLUSIONS: A potentially less painful and more comfortable PDT treatment protocol with proven long-term efficiency is presented. A randomized clinical trial has been conducted to confirm these results.

Ultrasonic scalpel associated with photodynamic therapy in the treatment of refractory vulvar condyloma acuminatum.

This is a case report of a patient with vulvar condyloma acuminatum associated with hyperkeratosis and folliculitis, which had been refractory to two previous therapies. The patient had been successfully treated using a combination of promising techniques: the ultrasonic scalpel and MAL photodynamic therapy.

Hydrogen peroxide preoxidation as a strategy for enhanced antimicrobial photodynamic action against methicillin-resistant Staphylococcus aureus.

Antimicrobial photodynamic treatment (aPDT) is a photooxidative process based on the excitation of a photosensitizer (PS) in the presence of molecular oxygen, under specific wavelengths of light. It is a promising method for advanced treatment of water and wastewater, particularly targeting disinfection challenges, such as antibiotic-resistant bacteria (ARB). Research in improved aPDT has been exploring new PS materials, and additives in general. Hydrogen peroxide (H2O2) a widely applied disinfectant, mostly in the food industry and clinical settings, present environmentally negligible residuals at the usually applied concentrations, making it friendly for the water and wastewater sectors. Here, we explored the effects of preoxidation with H2O2 followed by blue light-mediated (450 nm) aPDT using curcumin (a natural-based PS) against methicillin-resistant Staphylococcus aureus (MRSA). Results of the sequential treatment pointed to a slight hampering in aPDT efficiency at very low H2O2 concentrations, followed by an increasing cooperative effect up to a deleterious point (≥7 log10 inactivation in CFU mL-1), suggesting a synergistic interaction of preoxidation and aPDT. The increased performance in H2O2-pretreated aPDT encourages studies of optimal operational conditions for the assisted technology and describes potentials for using the described strategy to tackle the issue of ARB spread.

Spectroscopic Identification of Bacteria Resistance to Antibiotics by Means of Absorption of Specific Biochemical Groups and Special Machine Learning Algorithm.

FTIR (Fourier transform infrared spectroscopy) is one analytical technique of the absorption of infrared radiation. FTIR can also be used as a tool to characterize profiles of biomolecules in bacterial cells, which can be useful in differentiating different bacteria. Considering that different bacterial species have different molecular compositions, it will then result in unique FTIR spectra for each species and even bacterial strains. Having this important tool, here, we have developed a methodology aimed at refining the analysis and classification of the FTIR absorption spectra obtained from samples of Staphylococcus aureus, with the implementation of machine learning algorithms. In the first stage, the system conforming to four specified species groups, Control, Amoxicillin induced (AMO), Gentamicin induced (GEN), and Erythromycin induced (ERY), was analyzed. Then, in the second stage, five hidden samples were identified and correctly classified as with/without resistance to induced antibiotics. The total analyses were performed in three windows, Carbohydrates, Fatty Acids, and Proteins, of five hundred spectra. The protocol for acquiring the spectral data from the antibiotic-resistant bacteria via FTIR spectroscopy developed by Soares et al. was implemented here due to demonstrating high accuracy and sensitivity. The present study focuses on the prediction of antibiotic-induced samples through the implementation of the hierarchical cluster analysis (HCA), principal component analysis (PCA) algorithm, and calculation of confusion matrices (CMs) applied to the FTIR absorption spectra data. The data analysis process developed here has the main objective of obtaining knowledge about the intrinsic behavior of S. aureus samples within the analysis regions of the FTIR absorption spectra. The results yielded values with 0.7 to 1 accuracy and high values of sensitivity and specificity for the species identification in the CM calculations. Such results provide important information on antibiotic resistance in samples of S. aureus bacteria for potential application in the detection of antibiotic resistance in clinical use.

Photoantimicrobial chitosan-gelatin-pomegranate peel extract films for strawberries preservation: From microbiological analysis to in vivo safety assessment.

This study aimed to investigate the application of biopolymeric materials (chitosan, gelatin, and pomegranate peel extract as photosensitizer) and antimicrobial photodynamic therapy (aPDT) on the physicochemical and microbial safety of strawberries. The photosensitizer potential of the materials was confirmed by a light-dose-dependent photobleaching profile. The application of light (525 nm; 50 J cm-2) decreased by >2 log CFU mL-1 the survival of Staphylococcus aureus on the surface of the photoactive-biopolymeric films. Moreover, the materials did not present in vivo cytotoxicity using Danio rerio (Zebrafish) as well as cytophytotoxic, genotoxic, or mutagenic potentials against Allium cepa plant model, which points out their safety to be used as films without posing a risk to the humans and the environment. The photoactive-polymeric coatings were able to maintain the strawberries weight, and the association with green light was 100 % effective in delaying fungal contamination. These coated-strawberries presented a significant reduction in S. aureus survival after light application (5.47-4.34 log CFU mL-1). The molecular level analysis of the photoactive compound cyanidin-3-glucoside indicates absorption on UV-Vis consistent with aPDT action. Therefore, this study showed that the antimicrobial effects of aPDT combined with photoactive-biopolymeric coatings were enhanced, while the quality of the strawberries was maintained.

Recovering the susceptibility of antibiotic-resistant bacteria using photooxidative damage.

Multidrug-resistant bacteria are one of the most serious threats to infection control. Few new antibiotics have been developed; however, the lack of an effective new mechanism of their action has worsened the situation. Photodynamic inactivation (PDI) can break antimicrobial resistance, since it potentiates the effect of antibiotics, and induces oxidative stress in microorganisms through the interaction of light with a photosensitizer. This paper addresses the application of PDI for increasing bacterial susceptibility to antibiotics and helping in bacterial persistence and virulence. The effect of photodynamic action on resistant bacteria collected from patients and bacteria cells with induced resistance in the laboratory was investigated. Staphylococcus aureus resistance breakdown levels for each antibiotic (amoxicillin, erythromycin, and gentamicin) from the photodynamic effect (10 µM curcumin, 10 J/cm2) and its maintenance in descendant microorganisms were demonstrated within five cycles after PDI application. PDI showed an innovative feature for modifying the degree of bacterial sensitivity to antibiotics according to dosages, thus reducing resistance and persistence of microorganisms from standard and clinical strains. We hypothesize a reduction in the degree of antimicrobial resistance through photooxidative action combats antibiotic failures.

Photon spectra effects tested on the vegetal model Allium cepa.

The use of artificial light sources in plants is considered a type of photobiomodulation (PBM), a trend in agriculture and food industries, aiming at decontamination, pest control, and increased production yield. However, literature lacks a broader assessment to address the effects of photon light spectra on plant characteristics. Here, we aimed to describe the effects of visible light, infrared, and ultraviolet light upon Allium cepa, a known bioindicator, under various light doses. Samples irradiated under visible and infrared light did not show cytotoxicity, genotoxicity, or mutagenicity in any of the evaluated doses. Light induction at 460 and 635 nm significantly stimulated root development of the test organism. In contrast, 254 nm irradiation proved to be cytotoxic, genotoxic, and mutagenic. This work reveals and quantifies the spectral response of A. cepa seeds, suggesting that it can be proposed as a model for future research on mechanisms of PBM in plants.

Using Antimicrobial Photodynamic Therapy with Ultrasound Devices and Bioactive Glasses as a Combined Approach for Treating Dentin Caries Lesions.

Novel approaches for caries lesion removal and treatment have been proposed. This study evaluates the combined use of an experimental ultrasound, aPDT (antimicrobial photodynamic therapy) and bioactive glasses on the removal, decontamination and remineralization of dentin caries lesions. A biological model created with a duo species biofilm (Streptococcus mutans and Lactobacillus acidophilus) was used for the development of a caries-like lesion over the dentin for 7 days. Bovine dentin specimens (4 × 4 × 2 mm) were randomized according to the following caries removal techniques: bur (BUR) or ultrasound (ULT), decontamination (with or without aPDT) and remineralization materials (45S5 or F18 bioactive glasses). The following different groups were investigated: caries lesion (control); sound dentin (control); BUR; BUR + aPDT; ULT; ULT + aPDT; BUR + 45S5, BUR + F18; ULT + 45S5; ULT + F18; BUR + aPDT + 45S5; BUR + aPDT + F18; ULT + aPDT + 45S5; and ULT + aPDT + F18. Transverse microradiography (TMR), cross-sectional microhardness (CSH), FT-Raman spectroscopy and confocal microscopy (CLSM) were performed. A two-way ANOVA and Tukey's test were used (α = 0.05). (3) Results: The TMR revealed a lesion depth of 213.9 ± 49.5 µm and a mineral loss of 4929.3% vol.µm. The CSH increases as a function of depth, regardless of the group (p < 0.05). Removal with BUR (24.40-63.03 KHN) has a greater CSH than ULT (20.01-47.53 KHN; p < 0.05). aPDT did not affect the CSH (p > 0.05). No difference was observed between 45S5 or F18 (p > 0.05), but a change was observed for ULT (p > 0.05). The FT-Raman shows no differences for the phosphate (p > 0.05), but a difference is observed for the carbonate and C-H bonds. The CLSM images show that aPDT effectively inactivates residual bacteria. A combination of ULT, aPDT and bioactive glasses can be a promising minimally invasive treatment.

Synergic vascular photodynamic activity by methylene blue-curcumin supramolecular assembly.

A supramolecular assembly was obtained by combining methylene blue (MB) with a natural plant extract, curcumin (Curc), in a stoichiometric ratio of 1:4 in aqueous solution (90% PBS + 10% ethanol) at room temperature. The MB-Curc supramolecular assembly was evidenced by absorption and fluorescence spectroscopies, and the stoichiometry and bonding constant were obtained using Cielens model. Its stability and photostability were evaluated by chromatographic analysis and UV-Vis absorption. The MB-Curc avoids the aggregation of both isolated compounds and efficiently produces singlet oxygen (ΦΔ= 0.52 ± 0.03). Its potential for photodynamic antiangiogenic treatments was evaluated through the vascular effect observed in chicken chorioallantoic membrane (CAM) assay. The results showed intense damage in CAM vascular network by MB-Curc after irradiation, which is higher than the effect of isolated compounds, indicating a synergistic vascular effect. This combination can be essential to prevent cancer revascularization after photodynamic application and improve the efficacy of this approach. The characteristics exhibited by MB-Curc make it a potential candidate for use in cancer treatments through photodynamic antiangiogenic therapy.

Sonophotodynamic inactivation of Pseudomonas aeruginosa biofilm mediated by curcumin.

The inactivation of Pseudomonas aeruginosa biofilm is a major challenge, as biofilms are less responsive to conventional treatments and responsible for persistent infections. This has led to the investigation of alternative approaches for biofilm control such as photodynamic (PDI) and sonodynamic (SDI) inactivation. The combination of them, known as Sonophotodynamic Inactivation (SPDI), has improved the effectiveness of the process. Curcumin, a well-established photosensitizer, has been identified as a potential sonosensitizer. This study evaluated the most effective combination for SPDI against P. aeruginosa biofilms in vitro, varying curcumin concentrations and ultrasound intensities. The results indicated that the inactivation was directly proportional to the curcumin concentration. Using curcumin 120 µM and 3.0 W.cm-2 of ultrasound intensity, SPDI demonstrated the highest and the best synergistic results, equivalent to 6.9 ± 2.1 logs of reduction. PDI reduced 0.7 ± 0.9 log and SDI had no effect. In conclusion, SPDI with curcumin is a promising approach for biofilm inactivation.

Photobleaching Kinetics and Effect of Solvent in the Photophysical Properties of Indocyanine Green for Photodynamic Therapy.

Indocyanine green is an attractive molecule for photodynamic therapy due to its near infrared absorption, resulting in a higher tissue penetration. However, its quantum yields of the triplet and singlet state have been reported to be low and then, reactive oxygen species are unlikely to be formed. Aiming to understand the ICG role in photodynamic response, its photobleaching behavior in solution has been studied under distinct conditions of CW laser irradiation at 780 and 808 nm, oxygen saturations and solvents. Sensitizer bleaching and photoproduct formation were measured by absorption spectroscopy and analyzed using the PDT bleaching macroscopic model to extract physical parameters. ICG photobleaching occurs even at lower oxygen concentrations, indicating that the molecule presents more than one way of degradation. Photoproducts were produced even in solution of less than 4 % oxygen saturation for both solvents and excitation wavelengths. Also, the amplitude of absorption related to J-dimers was increased during irradiation, but only in 50 % PBS solution. The formation of photoproducts was enhanced in the presence of J-type dimers under low oxygen concentration, and the quantum yields of triplet and singlet states were one order of magnitude and two times higher, respectively, when compared to ICG in distilled H2 O.

Structural basis of antibacterial photodynamic action of curcumin against S. aureus.

Antimicrobial photodynamic therapy (aPDT) is an alternative tool to commercial antibiotics for the inactivation of pathogenic bacteria (e.g., S. aureus). However, there is still a lack of understanding of the molecular modeling of the photosensitizers and their mechanism of action through oxidative pathways. Herein, a combined experimental and computational evaluation of curcumin as a photosensitizer against S. aureus was performed. The radical forms of keto-enol tautomers and the energies of curcumin's frontier molecular orbitals were evaluated by density functional theory (DFT) to point out the photodynamic action as well as the photobleaching process. Furthermore, the electronic transitions of curcumin keto-enol tautomers were undertaken to predict the transitions as a photosensitizer during the antibacterial photodynamic process. Moreover, molecular docking was used to evaluate the binding affinity with the S. aureus tyrosyl-tRNA synthetase as the proposed a target for curcumin. In this regard, the molecular orbital energies show that the curcumin enol form has a character of 4.5% more basic than the keto form - the enol form is a more promising electron donor than its tautomer. Curcumin is a strong electrophile, with the enol form being 4.6% more electrophilic than its keto form. In addition, the regions susceptible to nucleophilic attack and photobleaching were evaluated by the Fukui function. Regarding the docking analysis, the model suggested that four hydrogen bonds contribute to the binding energy of curcumin's interaction with the ligand binding site of S. aureus tyrosyl-tRNA synthetase. Finally, residues Tyr36, Asp40, and Asp177 contact curcumin and may contribute to orienting the curcumin in the active area. Moreover, curcumin presented a photoinactivation of 4.5 log unit corroborating the necessity of the combined action of curcumin, light, and O2 to promote the photooxidation damage of S. aureus. These computational and experimental data suggest insights regarding the mechanism of action of curcumin as a photosensitizer to inactivate S. aureus bacteria.

Bacterial Adhesion Strength on Titanium Surfaces Quantified by Atomic Force Microscopy: A Systematic Review.

Few studies have been able to elucidate the correlation of factors determining the strength of interaction between bacterial cells and substrate at the molecular level. The aim was to answer the following question: What biophysical factors should be considered when analyzing the bacterial adhesion strength on titanium surfaces and its alloys for implants quantified by atomic force microscopy? This review followed PRISMA. The search strategy was applied in four databases. The selection process was carried out in two stages. The risk of bias was analyzed. One thousand four hundred sixty-three articles were found. After removing the duplicates, 1126 were screened by title and abstract, of which 57 were selected for full reading and 5 were included; 3 had a low risk of bias and 2 moderated risks of bias. (1) The current literature shows the preference of bacteria to adhere to surfaces of the same hydrophilicity. However, this fact was contradicted by this systematic review, which demonstrated that hydrophobic bacteria developed hydrogen bonds and adhered to hydrophilic surfaces; (2) the application of surface treatments that induce the reduction of areas favorable for bacterial adhesion interfere more in the formation of biofilm than surface roughness; and (3) bacterial colonization should be evaluated in time-dependent studies as they develop adaptation mechanisms, related to time, which are obscure in this review.

Nanostructure-Driven Indocyanine Green Dimerization Generates Ultra-Stable Phototheranostics Nanoparticles.

Indocyanine green (ICG) is the only near-infrared (NIR) dye approved for clinical use. Despite its versatility in photonic applications and potential for photothermal therapy, its photobleaching hinders its application. Here we discovered a nanostructure of dimeric ICG (Nano-dICG) generated by using ICG to stabilize nanoemulsions, after which ICG enabled complete dimerization on the nanoemulsion shell, followed by J-aggregation of ICG-dimer, resulting in a narrow, red-shifted (780 nm→894 nm) and intense (≈2-fold) absorbance. Compared to ICG, Nano-dICG demonstrated superior photothermal conversion (2-fold higher), significantly reduced photodegradation (-9.6 % vs. -46.3 %), and undiminished photothermal effect (7 vs. 2 cycles) under repeated irradiations, in addition to excellent colloidal and structural stabilities. Following intravenous injection, Nano-dICG enabled real-time tracking of its delivery to mouse tumors within 24 h by photoacoustic imaging at NIR wavelength (890 nm) distinct from the endogenous signal to guide effective photothermal therapy. The unprecedented finding of nanostructure-driven ICG dimerization leads to an ultra-stable phototheranostic platform.