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Staphylococcus aureus is a global challenge to the clinical field and food industry. Therefore, the development of antimicrobial photodynamic therapy (aPDT) has become one of the valuable methods to control this pathogen. The antibacterial activity of photoinactivation by erythrosine (Ery) against S. aureus has been reported, but its modes of action are unclear. This study aimed to employ a proteomic approach to analyze modes of action of Ery-aPDT against S. aureus. We determined the antibacterial effect by Ery-aPDT assays, quantified reactive oxygen species (ROS) and injury to the cell membrane, and determined protein expression using a proteomic approach combined with bioinformatic tools. Ery-aPDT was effective in reducing S. aureus to undetectable levels. In addition, the increment of ROS accompanied the increase in the reduction of cell viability, and damage to cellular membranes was shown by sublethal injury. In proteomic analysis, we found 17 differentially expressed proteins. These proteins revealed changes mainly associated with defense to oxidative stress, energy metabolism, translation, and protein biosynthesis. Thus, these results suggest that the effectiveness of Ery-aPDT is due to multi-targets in the bacterial cell that cause the death of S. aureus.
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BACKGROUND: The efficacy of photodynamic therapy (PDT) depends on the combination of light and a photosensitizer for inactivation of microorganisms. However, finding the ideal conditions for the factors involved in this technique is time and cost-consuming. The rotational composite central design (RCCD) is a tool that can be allied with PDT to achieve precise results within a shorter working time. METHODS: This study used the response surface methodology to optimize the parameters of PDT mediated by Erythrosine (ERY) and green light-emitting diodes (LED) in different Escherichia coli strains by applying RCCD. RESULTS: The RCCD predicted optimum values of ERY and light exposure on PDT. According to the experimental results, the light exposure time showed the most significant influence on the inactivation of the evaluated bacteria. The optimized operating conditions were validated in laboratory tests, and no viable cells were recovered with ERY at 116 µmol L-1 and 30 min of light (33.34 J cm2) for E. coli ATCC 25922, 108 µmol L-1 and 40 min (44.38 J cm2) for E. coli ATCC 35218, and 108 µmol L-1 and 29.3 min (32.5 J cm2) for E. coli O157:H7 EDL 933. CONCLUSION: The adjusted polynomial models provided accurate information on the combined effects of ERY and lighting time with green LED on PDT. The application of the RCCD, in addition to reducing the number of experiments, also allows for increased quantity and quality of the results. Therefore, surface response methodology combined with PDT is a promising approach to inactivate E. coli.
Subject(s)
Escherichia coli , Photochemotherapy , Erythrosine/pharmacology , Photosensitizing Agents/pharmacology , Photochemotherapy/methodsABSTRACT
BACKGROUND: Sporothrix brasiliensis is a pathogenic dimorphic fungus that affects humans and animals causing sporotrichosis. The treatment of this disease with conventional antifungals commonly results in therapeutic failures and resistance. Therefore, this study aimed to evaluate the in vitro effect of curcumin (CUR) mediated by photodynamic therapy (PDT) in its pure state and incorporated into pharmaceutical formulation in gel form, on the filamentous and yeast forms of S. brasiliensis. METHODS: Cells from both forms of the fungus were treated with pure curcumin (PDT-CUR). For this, CUR concentrations ranging from 0.09 to 50 µM were incubated for 15 min and then irradiated with blue LED at 15 J/cm². Similarly, it was performed with PDT-CUR-gel, at lower concentration with fungistatic action. After, a qualitative and quantitative (colony forming units (CFU)) analysis of the results was performed. Additionally, reactive oxygen species (ROS) were detected by flow cytometry. Results PDT with 0.78 µM of CUR caused a significant reduction (p < 0.05) in cells of the filamentous and yeast form, 1.38 log10 and 1.18 log10, respectively, in comparison with the control. From the concentration of 1.56 µM of CUR, there was a total reduction in the number of CFU (≥ 3 log10). The PDT-CUR-gel, in relation to its base without CUR, presented a significant reduction (p < 0.05) of 0.83 log10 for the filamentous form and for the yeast form, 0.72 log10. ROS release was detected after the PDT-CUR assay, showing that this may be an important pathway of death caused by photoinactivation. Conclusion PDT-CUR has an important in vitro antifungal action against S. brasiliensis strains in both morphologies.
Subject(s)
Curcumin , Photochemotherapy , Humans , Animals , Antifungal Agents/therapeutic use , Curcumin/pharmacology , Curcumin/therapeutic use , Photochemotherapy/methods , Photosensitizing Agents/pharmacology , Photosensitizing Agents/therapeutic use , Saccharomyces cerevisiae , Reactive Oxygen SpeciesABSTRACT
BACKGROUND: Onychomycosis (OM) is a common nail plate disorder caused by dermatophyte molds, yeasts, and non-dermatophyte molds, which use keratin in the nail plate as an energy source. OM is characterized by dyschromia, increased nail thickness, subungual hyperkeratosis, and onychodystrophy, and is typically treated with conventional antifungals despite frequent reports of toxicity, fungal resistance, and OM recurrence. Photodynamic therapy (PDT) with hypericin (Hyp) as a photosensitizer (PS) stands out as a promising therapeutic modality. When excited by a specific wavelength of light and in the presence of oxygen, to lead to photochemical and photobiological reactions on the selected targets. METHODS: OM diagnosis was made in three suspected cases, and the causative agents were identified by classical and molecular methods, and confirmed by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR). Susceptibility of planktonic cells of the clinical isolates to conventional antifungals and PDT-Hyp was evaluated, and photoacoustic spectroscopy (PAS) of Hyp permeation in nail fragments ex vivo was analyzed. Furthermore, the patients opted to undergo PDT-Hyp treatment and were subsequently followed up. The protocol was approved by the human ethics committee (CAAE, number 14107419.4.0000.0104). RESULTS: The etiological agents of OM in patients ID 01 and ID 02 belonged to the Fusarium solani species complex, being identified as Fusarium keratoplasticum (CMRP 5514) and Fusarium solani (CMRP 5515), respectively. For patient ID 03, the OM agent was identified as Trichophyton rubrum (CMRP 5516). PDT-Hyp demonstrated a fungicidal effect in vitro, with reductions of p3 log10 (p < 0.0051 and p < 0.0001), and the PAS analyses indicated that Hyp could completely permeate through both healthy and OM-affected nails. After four sessions of PDT-Hyp, mycological cure was observed in all three cases, and after seven months, clinical cure was confirmed. CONCLUSION: PDT-Hyp showed satisfactory results in terms of efficacy and safety, and thus can be considered a promising therapy for the clinical treatment of OM.
Subject(s)
Nail Diseases , Onychomycosis , Photochemotherapy , Humans , Onychomycosis/drug therapy , Onychomycosis/microbiology , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Photosensitizing Agents/therapeutic use , Photochemotherapy/methods , Nail Diseases/drug therapyABSTRACT
Considering the multifaceted and increasing application of photodynamic therapy (PDT), in recent years the antimicrobial employment of this therapy has been highlighted, because of the antiviral, antibacterial, antiparasitic, and antifungal activities that have already been demonstrated. In this context, research focussed on antimycological action, especially for treatment of superficial infections, presents promising growth due to the characteristics of these infections that facilitate PDT application as new therapeutic options are needed in the field of medical mycology. Among the more than one hundred classes of photosensitizers the antifungal action of hypericin (Hyp) stands out due to its ability to permeate the lipid membrane and accumulate in different cytoplasmic organelles of eukaryotic cells. In this review, we aim to provide a complete overview of the origin, physicochemical characteristics, and optimal alternative drug deliveries that promote the photodynamic action of Hyp (Hyp-PDT) against fungi. Furthermore, considering the lack of a methodological consensus, we intend to compile the best strategies to guide researchers in the antifungal application of Hyp-PDT. Overall, this review provides a future perspective of new studies and clinical possibilities for the advances of such a technique in the treatment of mycoses in humans.
Subject(s)
Anti-Infective Agents , Biological Products , Photochemotherapy , Humans , Photochemotherapy/methods , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Photosensitizing Agents/pharmacology , Anti-Infective Agents/therapeutic useABSTRACT
Chronic wound healing represents an impactful financial burden on healthcare systems. In this context, the use of natural products as an alternative therapy reduces costs and maintains effectiveness. Phytotherapeutic gels applied in photodynamic therapy (PDT) have been developed to act as topical healing medicines and antibiotics. The bioactive system is composed of Spirulina sp. (source of chlorophylls) and Copaifera reticulata oil microdroplets, both incorporated into a polymeric blend constituted by kappa-carrageenan (k-car) and F127 copolymer, constituting a system in which all components are bioactive agents. The flow behavior and viscoelasticity of the formulations were investigated. The photodynamic activity was accessed from studies of the inactivation of Staphylococcus aureus bacteria, the main pathogen of hospital relevance. Furthermore, in vivo studies were conducted using eighteen rabbits with dermatitis (grade III and IV) in both paws. The gels showed significant antibiotic potential in vitro, eliminating up to 100% of S. aureus colonies in the presence or absence of light. The k-car reduced 41% of the viable cells; however, its benefits were enhanced by adding chlorophyll and copaiba oil. The animals treated with the phytotherapeutic medicine showed a reduction in lesion size, with healing and re-epithelialization verified in the histological analyses. The animals submitted to PDT displayed noticeable improvement, indicating this therapy's viability for ulcerative and infected wounds. This behavior was not observed in the iodine control treatment, which worsened the animals' condition. Therefore, gel formulations were a viable alternative for future pharmaceutical applications, aiming at topical healing.
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BACKGROUND: Outbreaks related to food contamination by resistant microorganisms is a worldwide concern that, motivates industries and research institutions to search for affordable solutions. Among the solutions that have been proposed, Photodynamic Inactivation (PDI) of microorganisms has gained prominence, among other aspects, because it is easy to apply and does not generate microbial resistance. METHODS: In this study, we used the association between curcumin solubilized with Tween and light in the photodynamic inactivation process, using light-emitting diodes with a wavelength of 430 nm for decontamination S. Typhimurium and K. pneumoniae from bovine casings used as wrappers for meat products. The result was verified by counting and comparing the number of colony-forming units of the treatment concerning the negative control. RESULTS: The solubilizer, Tween 80, used does not change the optical absorption of curcumin. An optical fluence of 150J/cm2 induces a microbial log reduction of 3.8±0.2 and 2.7±0.1 for S. Typhimurium, and K. pneumoniae contaminated guts, respectively. For the 200µM concentration of curcumin, the PDI provided a microbial log reduction of 3.16±0.03 for S. Typhimurium. For K. pneumoniae, the minimal inhibitory concentration of curcumin occurs up to 12.5µM, causing an microbial log reduction of 2.08±0.03. CONCLUSION: Both curcumin and tween are already used as additives in food production and do not pose health risks at the concentrations used. Furthermore, in the case of the material studied, the addition of curcumin favors the organoleptic quality associated with the color of the food, unlike the green or blue photossensitizers. The results pave the way for possible application of curcumin in finished meat products.
Subject(s)
Anti-Infective Agents , Curcumin , Photochemotherapy , Cattle , Animals , Curcumin/pharmacology , Photosensitizing Agents/pharmacology , Photochemotherapy/methods , BacteriaABSTRACT
The antifungal application of photodynamic therapy (PDT) has been widely explored. According to superficial nature of tinea capitis and the facility of application of light sources, the use of nanoencapsulated hypericin in P-123 associated with PDT (P123-Hy-PDT) has been a poweful tool to treat this pathology. Thus, the aim of this study was to evaluate the efficiency of P123-Hy-PDT against planktonic cells and in a murine model of dermatophytosis caused by Microsporum canis. In vitro antifungal susceptibility and in vivo efficiency tests were performed, including a skin toxicity assay, analysis of clinical signs by evaluating score, and photoacoustic spectroscopy. In addition, tissue analyses by histopathology and levels of pro-inflammatory cytokines, such as quantitative and qualitative antifungal assays, were employed. The in vitro assays demonstrated antifungal susceptibility with 6.25 and 12.5 µmol/L P123-Hy-PDI; these experiments are the first that have used this treatment of animals. P123-Hyp-mediated PDT showed neither skin nor biochemical alteration in vivo; it was safe for dermatophytosis treatment. Additionally, the treatment revealed rapid improvement in clinical signs at the site of infection after only three treatment sessions, with a clinical score confirmed by photoacoustic spectroscopy. The mycological reduction occurred after six treatment sessions, with a statistically significant decrease compared with untreated infected animals. These findings showed that P123-Hy-PDT restored tissue damage caused by infection, a phenomenon confirmed by histopathological analysis and proinflammatory cytokine levels. Our results reveal for the first time that P123-Hy-PDT is a promising treatment for tinea capitis and tinea corporis caused by M. canis, because it showed rapid clinical improvement and mycological reduction without causing toxicity.
Subject(s)
Nanostructures/chemistry , Perylene/analogs & derivatives , Photochemotherapy/methods , Poloxamer/analogs & derivatives , Tinea/drug therapy , Animals , Anthracenes , Capsules , Mice , Perylene/chemistry , Perylene/pharmacology , Photosensitizing Agents/chemistry , Photosensitizing Agents/pharmacology , Poloxamer/chemistry , PolymerizationABSTRACT
In this study we report a novel theranostic lipid-polymer liposome, obtained from DPPC and the triblock copolymer F127 covalently modified with 5(6)-carboxyfluorescein (CF) for photodynamic applications. Due to the presence of F127, small unilamellar vesicle (SUV) liposomes were synthesized by a simple and fast thin-film hydration method without the need for an extrusion process. The vesicles have around 100 nm, low polydispersity and superb solution stability. The clinically used photosensitizer verteporfin (VP) was entrapped into the vesicles, mostly in monomeric form, with 90% loading efficiency. Stern-Volmer and fluorescence lifetime assays showed heterogeneous distribution of the VP and CF into the vesicles, ensuring the integrity of their individual photophysical properties. The theranostic properties were entirely photoactivatable and can be trigged by a unique wavelength (470 nm). The feasibility of the system was tested against the Glioblastoma multiforme cell line T98G. Cellular uptake by time-resolved fluorescence microscopy showed monomerized VP (monoexponential decay, 6.0 ns) at nucleus level, while CF was detected at the membrane by fluorescence microscopy. The strategy's success was supported by the reduction of 98% in the viability of T98G cells by the photoactivated lipid-polymer liposome with [VP] = 1.0 µmol L-1. Therefore, the novel theranostic liposome is a potential system for use in cancer and ocular disease therapies.
Subject(s)
Photochemotherapy/methods , Verteporfin/administration & dosage , Verteporfin/pharmacology , Cell Line, Tumor , Drug Stability , Humans , Kinetics , Liposomes , Verteporfin/therapeutic useABSTRACT
We induce nanometer-scale surface deformation by exploiting momentum conservation of the interaction between laser light and dielectric liquids. The effect of radiation force at the air-liquid interface is quantitatively assessed for fluids with different density, viscosity and surface tension. The imparted pressure on the liquids by continuous or pulsed laser light excitation is fully described by the Helmholtz electromagnetic force density.
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Strong light absorption and high levels of singlet oxygen production indicate erythrosin B as a viable candidate as a photosensitizer in photodynamic therapy or photodynamic inactivation of microorganisms. Under light irradiation, erythrosin B undergoes a photobleaching process that can decrease the production of singlet oxygen. In this paper, we use thermal lens spectroscopy to investigate photobleaching in micellar solutions of erythrosin ester derivatives: methyl, butyl, and decyl esters in low concentrations of non-ionic micellar aqueous solutions. Using a previously developed thermal lens model, it was possible to determine the photobleaching rate and fluorescence quantum efficiency for dye-micelle solutions. The results suggest that photobleaching is related to the intensity of the dye-micelle interaction and demonstrate that the thermal lens technique can be used as a sensitive tool for quantitative measurement of photochemical properties in very diluted solutions.
Subject(s)
Erythrosine/chemistry , Fluorescent Dyes/chemistry , Photosensitizing Agents/chemistry , Esters/chemistry , Micelles , Photobleaching , Photolysis , Water/chemistryABSTRACT
An adaptive spatial filter is used as an optical novelty filter to detect photothermal mirror (PM) signals in high absorbing materials using continuous wave laser excitation. The optical novelty filter uses an optical beam-fanning limiter based on single domain barium titanate (BaTiO3), cut and poled 45° relative to the c-axis. The optical novelty filter approach relaxes the requirement for high sample surface smoothness because the effect aperture adapts to the surface, reducing the stationary background from the optical signal and provides a means of developing the photothermal mirror signal. Time-dependent probe laser phase shifts due to photothermal surface deformation pass through the optical novelty filter and are detected as an intensity increase over the stationary or "mundane" signal. Experimental studies are performed using four well-characterized metals using both the conventional photothermal mirror and optical novelty filter apparatuses in order to understand the complicated signal behavior. Signal behavior is analyzed in different excitation intervals using pseudo-chopped sample excitation with different duty cycles. Optical novelty filter signals show fast response for changes in the spatial beam profile followed by long relaxation time. Reasons for the optical novelty filter response are described.
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We investigate the dynamics of the 2009 influenza A (H1N1/S-OIV) pandemic by analyzing data obtained from World Health Organization containing the total number of laboratory-confirmed cases of infections--by country--in a period of 69 days, from 26 April to 3 July, 2009. Specifically, we find evidence of exponential growth in the total number of confirmed cases and linear growth in the number of countries with confirmed cases. We also find that, i) at early stages, the cumulative distribution of cases among countries exhibits linear behavior on log-log scale, being well approximated by a power law decay; ii) for larger times, the cumulative distribution presents a systematic curvature on log-log scale, indicating a gradual change to lognormal behavior. Finally, we compare these empirical findings with the predictions of a simple stochastic model. Our results could help to select more realistic models of the dynamics of influenza-type pandemics.