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Photodynamic (PDI) and sonodynamic (SDI) inactivation have been successfully employed as antimicrobial treatments. Moreover, sonophotodynamic inactivation (SPDI), which is the simultaneous application of PDI and SDI, has demonstrated greater effects. This study assessed the effects of PDI (PDI group), SDI (SDI group) and SPDI (SPDI group) using curcumin as a sensitizer on the metabolism, adhesion capability, biofilm formation ability and structural effects in a Staphylococcus aureus biofilm. Moreover, the production of reactive oxygen species (ROS) and the degradation spectrum of curcumin under the irradiation sources were measured. SPDI was more effective in inactivating the biofilm than PDI and SDI. All treatments reduced the adhesion ability of the bacteria: 58 ± 2%, 58 ± 1% and 71 ± 1% of the bacterial cells adhered to the polystyrene plate after the SPDI, SDI and PDI, respectively, when compared to 79 ± 1% of the untreated cells (control group). This result is probably related to the metabolism cell reduction after treatments. The metabolism of cells from the PDI group was 89 ± 1% lower than the untreated cells, while the metabolic activity of SDI and SPDI groups were 82 ± 2% and 90 ± 1% lower, respectively. Regarding the biofilm formation ability, all treatments (SPDI, SDI and PDI) reduced the total biomass. The total biomass of the PDI, SDI and SPDI groups were 26 ± 2%, 31 ± 5% and 35 ± 6% lower than the untreated biofilm (control group), respectively. Additionally, all treatments produced ROS and caused significant structural changes, reducing cells and the extracellular matrix. The light caused a greater absorbance decay of the curcumin; however, the US did not expressively alter its spectrum. Finally, SPDI had improved antimicrobial effects, and all treatments exhibited similar effects in the colonization factors evaluated.

The impressive increase in antimicrobial resistance has required the development of alternative treatments that act on multiple non-specific molecular targets and are effective against a broad range of microorganisms. Antimicrobial Photodynamic Therapy (aPDT) is based on microbial inactivation from oxidative stress and represents an important tool for inactivating microorganisms with low risk of resistance selection. Therefore, our research group has been devoted to demonstrating its effectiveness against pathogens that cause pneumonia, one of the most lethal infections worldwide. Previous studies reported the efficiency and safety of an in vitro photoinactivation protocol for Streptococcus pneumoniae and the delivery of infrared light (external illumination) and photosensitizer (PS) in an animal model. However, the in vivo inactivation of microorganisms still poses challenges due to the presence of lung surfactant (LS), which traps PSs, preventing them from reaching the microbial target. This study investigated different approaches such as use of emulsifiers, perfluorocarbon, oxygen nanobubbles, and copolymer towards overcoming LS and optimizing aPDT response. The most promising strategy consisted in combining indocyanine green (ICG) with GantrezTM AN-139 - a Polyvinyl Methyl Ether/Maleic Anhydride copolymer (PVM/MA) – showing high microbial inactivation and safety for human lung epithelial (A549) and fibroblast (MRC-9) cell lines. The in vitro experiments provided an alternative to overcome the limited PS distribution through LS and will serve as the basis for in vivo studies.

BackgroundSporotrichosis is a zoonotic disease caused by the dimorphic fungus Sporothrix spp., leading to skin lesions that can, in some cases, progress and result in the death of infected individuals. Candida albicans is another fungus involved in several skin, oral, and vaginal mucosal infections. Fungal diseases are concerning due to increasing incidence and the limited variety of antifungal classes available for treatment. Furthermore, antifungal medications can cause various side effects, exacerbated by their prolonged use during infection treatment. There is a need to explore alternatives to conventional drugs that are effective, fast, and safe in combating sporotrichosis. This study aimed to achieve in vitro elimination of the fungi Sporothrix brasiliensis and Sporothrix schenckii through Photodynamic Inactivation (PDI), using curcumin as a photosensitizer and in combination with antifungal agents used in the treatment of sporotrichosis.MethodologyYeasts of Candida albicans, Sporothrix brasiliensis, and Sporothrix schenckii were subjected to Photodynamic Inactivation (PDI) using light at a wavelength of 450 ± 10 nm, irradiance of 35 mW/cm2, delivering a fluence of 31.5 J/cm2, with curcumin as the photosensitizer at doses ranging from 0.75 to 150 μg/mL. After determining the Minimum Inhibitory Concentration (MIC) values of the antifungal drugs itraconazole, ketoconazole, and potassium iodide, sub-MIC doses of these antifungals were combined with sub-MIC doses of curcumin in a new PDI session.ConclusionPhotodynamic inactivation is a promising technique in the treatment of sporotrichosis, as well as its combination with antifungals. The combination of curcumin in concentrations ranging from 0.75 g/mL a 7.5 g/mL with sub-MIC concentrations of itraconazole, ketoconazole, and potassium iodide was able to completely inactivate the fungi C. albicans, S. brasiliensis and S. schenckii, indicating that PDI may increase the effectiveness of antifungals. However, further studies are needed to establish protocols for future clinical applications.

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.

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.

Resumo Fundamento A formação de células espumosas ocorre devido ao aumento em lipoproteína plasmática de baixa densidade (LDL) e desregulação da inflamação, sendo importante para o desenvolvimento da aterosclerose. Objetivo Avaliar o perfil do fator de necrose tumoral alfa (TNF-α) e da interleucina-6 (IL-6) no método de formação da célula espumosa existente, otimizando esse protocolo. Métodos A LDL foi isolada, oxidada e marcada com sonda de isotiocianato de fluoresceína (FITC). As células espumosas foram geradas de célula derivada de monócitos humanos THP-1 e incubadas na ausência (controle) ou presença de FITC-ox-LDL (10, 50, 100, 150 ou 200 μg/mL), por 12, 24, 48 ou 72 horas. A FITC-ox-LDL na célula foi quantificada por microscopia. O ensaio de imunoabsorção enzimática foi avaliado para quantificar a IL-6 e o TNF-α, com um p

Considerable efforts have been spent on the development of biodefensives based on the encapsulation of essential oils for controlling of urban pests from their larval stage, especially as anopheline controlling agents. The larval source management of Anopheles aquasalis is important for malaria prevention. For this reason, this research proposes larvicidal biodefensives based on polymeric particles loaded with Piper nigrum essential oil, considering the influence of temperature (35 °C) and preservatives on the formulation stability. The biodefensive containing the preservative phenoxyethanol/methylisothiazolinone (PNE) resulted in 5 months of shelf-life storage with an Encapsulation Efficiency (EE%) of essential oil of 70%. The biodefensive PNE (containing 500 µg.mL−1 of encapsulated essential oil) presented a polydisperse particle size distribution, ranging from D10 = (127 ± 10) nm to D90 = (472 ± 78) nm and a particle mean size of (236 ± 34) nm. The AFM images revealed a spherical morphology with an external surface almost regular and smooth. The controlled release of the essential oil was evaluated up to 72 h according to the Korsmeyer-Peppas mathematical model, confirming the anomalous transport (n = 0.64 in pH = 3 and pH = 10, and n = 0.65 in pH = 7). The total larvae mortality on the in loco bioassays was almost reached (92%) after 24 h. However, according to the in vitro bioassays applying the in natura essential oil alone, the concentration of 454 μg.mL−1 resulted on the mortality of 70% of the larvae after 24 h. For this reason, the highest efficiency of the biodefensive PNE may be related to the encapsulation of essential oil, delivering the loaded particles more efficiently inside the larvae. From this perspective, the present study shows that a formulation based on P. nigrum essential oil may be taken into account in the integrated management of disease vector mosquitoes.

One of the most widely used molecules used for photodynamic therapy (PDT) is 5-aminolevulinic acid (5-ALA), a precursor in the synthesis of tetrapyrroles such as chlorophyll and heme. The 5-ALA skin permeation is considerably reduced due to its hydrophilic characteristics, decreasing its local bioavailability and therapeutic effect. For this reason, five different systems containing polymeric particles of poly [D, L–lactic–co–glycolic acid (PLGA)] were developed to encapsulate 5-ALA based on single and double emulsions methodology. All systems were standardized (according to the volume of reagents and mass of pharmaceutical ingredients) and compared in terms of laboratory scaling up, particle formation and stability over time. UV-VIS spectroscopy revealed that particle absorption/adsorption of 5-ALA was dependent on the method of synthesis. Different size distribution was observed by DLS and NTA techniques, revealing that 5-ALA increased the particle size. The contact angle evaluation showed that the system hydrophobicity was dependent on the surfactant and the 5-ALA contribution. The FTIR results indicated that the type of emulsion influenced the particle formation, as well as allowing PEG functionalization and interaction with 5-ALA. According to the 1H-NMR results, the 5-ALA reduced the T1 values of polyvinyl alcohol (PVA) and PLGA in the double emulsion systems due to the decrease in molecular packing in the hydrophobic region. The results indicated that the system formed by single emulsion containing the combination PVA–PEG presented greater stability with less influence from 5-ALA. This system is a promising candidate to successfully encapsulate 5-ALA and achieve good performance and specificity for in vitro skin cancer treatment.

A new systematic structural study was performed using the Atomic Force Microscopy (AFM) reporting statistical parameters of polymeric particles based on gelatin and poly-ε-caprolactone (PCL) containing essential oil from Lippia origanoides. The developed biocides are efficient alternative controlling agents of Conotrachelus humeropictus and Moniliophtora perniciosa, the main pests of Theobroma grandiflorum. Our results showed that the particles morphology can be successfully controlled by advanced stereometric parameters, pointing to an appropriate concentration of encapsulated essential oil according to the particle surface characteristics. For this reason, the absolute concentration of 1000 µg·mL−1 (P1000 system) was encapsulated, resulting in the most suitable surface microtexture, allowing a faster and more efficient essential oil release. Loaded particles presented zeta potential around (–54.3 ± 2.3) mV at pH = 8, and particle size distribution ranging from 113 to 442 nm. The hydrodynamic diameter of 90% of the particle population was found to be up to (405 ± 31) nm in the P1000 system. The essential oil release was evaluated up to 80 h, with maximum release concentrations of 63% and 95% for P500 and P1000, respectively. The best fit for the release profiles was obtained using the Korsmeyer–Peppas mathematical model. Loaded particles resulted in 100% mortality of C. humeropictus up to 48 h. The antifungal tests against M. perniciosa resulted in a minimum inhibitory concentration of 250 µg·mL−1, and the P1000 system produced growth inhibition up to 7 days. The developed system has potential as alternative controlling agent, due to its physical stability, particle surface microtexture, as well as pronounced bioactivity of the encapsulated essential oil.

The development of conjugated polymer-based nanocomposites by adding metallic particles into the polymerization medium allows the proposition of novel materials presenting improved electrical and optical properties. Polyaniline Emeraldine-salt form (ES–PANI) has been extensively studied due to its controllable electrical conductivity and oxidation states. On the other hand, tungsten oxide (WO3) and its di-hydrated phases, such as WO3·2H2O, have been reported as important materials in photocatalysis and sensors. Herein, the WO3·2H2O phase was directly obtained during the in-situ polymerization of aniline hydrochloride from metallic tungsten (W), allowing the formation of hybrid nanocomposites based on its full oxidation into WO3·2H2O. The developed ES–PANI–WO3·2H2O nanocomposites were successfully characterized using experimental techniques combined with Density Functional Theory (DFT). The formation of WO3·2H2O was clearly verified after two hours of synthesis (PW2 nanocomposite), allowing the confirmation of purely physical interaction between matrix and reinforcement. As a result, increased electrical conductivity was verified in the PW2 nanocomposite: the DFT calculations revealed a charge transfer from the p-orbitals of the polymeric phase to the d-orbitals of the oxide phase, resulting in higher conductivity when compared to the pure ES–PANI.

Carbon-based photosensitizers are more attractive than the other ones based on their low cost, high stability, broadband of light absorption, tunable emission spectra, high quantum yield, water solubility, high resistance to metabolic degradation, and selective delivery. These properties allow multiple applications in the field of biology and medicine. The present study evaluated in vitro and in vivo the antimicrobial photodynamic effect of a one-pot microwave produced C-DOTS based on citric acid. The in vitro assays assessed the effectiveness of illuminated C-DOTS (C-DOTS + light) against Staphylococcus aureus suspension and biofilm. The concentrations of 6.9 and 13.8 mg/mL of C-DOTS and light doses of 20 and 40 J/cm2 were able to reduce significantly the microorganisms. Based on these parameters and results, the in vivo experiments were conducted in mice, evaluating this treatment on wounds contaminated with S. aureus. The viability test showed that C-DOTS–mediated photodynamic inactivation reduced 104 log of the bacteria present on the skin lesions. These results, altogether, showed that antibacterial photodynamic therapy using C-DOTS is a promising and viable treatment for Gram-positive bacteria-infected wounds.

Graphene oxide (GO) with their interesting properties including thermal and electrical conductivity and antibacterial characteristics have many promising applications in medicine. The prevalence of resistant bacteria is considered a public health problem worldwide, herein, GO has been used as a broad spectrum selective antibacterial agent based on the photothermal therapy (PTT)/photodynamic therapy (PDT) effect. The preparation, characterization, determination of photophysical properties of two different sizes of GO is described. In vitro light dose and concentration-dependent studies were performed using Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus bacteria based on the PTT/PDT effect used ultra-low doses (65 mW cm–2) of 630 nm light, to achieve efficient bacterial decontamination. The results show that GO and nanographene oxide (nGO) can sensitize the formation of 1O2 and allow a temperature rise of 55°C to 60°C together nGO and GO to exert combined PTT/PDT effect in the disinfection of gram-positive S. aureus and gram-negative E. coli bacteria. A complete elimination of S. aureus and E. coli bacteria based on GO and nGO is obtained by using a dose of 43–47 J cm–2 for high concentration used in this study, and a dose of around 70 J cm–2 for low dose of GO and nGO. The presence of high concentrations of GO allows the bacterial population of S. aureus and E. coli to be more sensitive to the use of PDT/PTT and the efficiency of S. aureus and E. coli bacteria disinfection in the presence of GO is similar to that of nGO. In human neonatal dermal fibroblast, HDFs, no significant alteration to cell viability was promoted by GO, but in nGO is observed a mild damage in the HDFs cells independent of nGO concentration and light exposure. The unique properties of GO and nGO may be useful for the clinical treatment of disinfection of broad-spectrum antimicrobials. The antibacterial results of PTT and PDT using GO in gram-positive and gram-negative bacteria, using low dose light, allow us to conclude that GO and nGO can be used in dermatologic infections, since the effect on human dermal fibroblasts of this treatment is low compared to the antibacterial effect.

(1) Background: Cervical cancer is the third most commonly diagnosed cancer and the fourth leading cause of cancer death in women worldwide. The highest incidence rates are in Africa, followed by South-Central Asia and South America. According to the Brazilian National Institute of Cancer (INCA), 16,370 new cases of cervical cancer were estimated for each year of the biennium of 2018−2019. About 90% of cervical cancers originate from the malignant progression of cervical intraepithelial neoplasia (CIN) which is classified based on cytohistological characteristics (low- and high-grade lesions). The present study reports the long-term effectiveness of topical photodynamic therapy (PDT) for CIN grades 1 and 2/3 with up to two years of follow up. (2) Methods: A total of 56 patients with CIN 1, ten with CIN 2, and 14 patients for the placebo group were enrolled in this study. (3) Results: 75% (n = 42) of CIN 1 patients presented a complete response to PDT and only 23.2% (n = 13) of recurrence, progression, and/or lesions remaining two years after PDT. For CIN 2/3 patients, 90% were observed to be cured after one and two years of follow up. (4) Conclusions: PDT presented best results two years after a non-invasive, fast, and low-cost procedure and in comparison with the placebo group, preventing the progression of cervical intraepithelial neoplasia and preserving the cervix.

The success of the onychomycosis treatment is directly associated with factors such as the choice of the medication, the administration route, and the pharmaceutical formulation. Photodynamic therapy (PDT) is an emerging and promising technique indicated for onychomycosis treatment. For this application, the main challenge is the efficient delivery of the photosensitizer (PS). Curcumin is widely used as a PS, however it is an unstable molecule and it is a challenge to develop a formulation with good penetration into the nail plate, maintaining the stability of curcumin. In this study, the molecular mechanisms underlying the efficacy of two topical formulations containing curcumin used in a clinical trial for onychomycosis treatment were analyzed by Raman microspectroscopy. It is shown that curcumin is present in both formulations in aggregated and non-aggregated states, and in aggregates it is present in different conformations, depending on the interaction with the solvent. This proves to be critical for efficient and uniform PS delivery to the nail and its complete use during the treatment. These analyses are showing how promising Raman microspectroscopy is in understanding the molecular mechanisms of the efficiency of photosensitizers and are helping to improve the development of pharmaceutical formulations.

The photodynamic therapy (PDT) is an alternative technique that can be indicated for superficial basal cell carcinoma (sBCC), Bowen’s disease and actinic keratosis with high efficiency. The objective of this study is to present the importance of fluorescence imaging for PDT guidance and monitoring in real time. Confirming that the lesion is well prepared and the photosensitizer shows a homogenous distribution, the outcome after few PDT sessions will be positive and the recurrence should be lower. Our proposition in this study is use the widefield fluorescence imaging to evaluate the PDT protocol in situ and in real time for each lesion. This evaluation procedure is performed in two steps: first with the monitoring of the production of protoporphyrin IX (PpIX) induced by methyl aminolevulinate (MAL), an derivative of 5-aminolevulinic acid (ALA) and second with the detection of PpIX photobleaching after illumination. The fluorescence images provide information correlated with distinct clinical features and with the treatment outcome. Eight BCC lesions are presented and discussed in this study. Different fluorescence patterns of PpIX production and photobleaching could be correlated with the treatment response. The presented results show the potential of using widefield fluorescence imaging as a guidance tool to customized PDT.