Thermoresponsive Hydrogel-Loading Aluminum Chloride Phthalocyanine as a Drug Release Platform for Topical Administration in Photodynamic Therapy.

Phthalocyanine aluminum chloride (Pc) is a clinically viable photosensitizer (PS) to treat skin lesions worsened by microbial infections. However, this molecule presents a high self-aggregation tendency in the biological fluid, which is an in vivo direct administration obstacle. This study proposed the use of bioadhesive and thermoresponsive hydrogels comprising triblock-type Pluronic F127 and Carbopol 934P (FCarb) as drug delivery platforms of Pc (FCarbPc)-targeting topical administration. Carbopol 934P was used to increase the F127 hydrogel adhesion on the skin. Rheological analyses showed that the Pc presented a low effect on the hydrogel matrix, changing the gelation temperature from 27.2 ± 0.1 to 28.5 ± 0.9 °C once the Pc concentration increases from zero to 1 mmol L-1. The dermatological platform showed matrix erosion effects with the release of loaded Pc micelles. The permeation studies showed the excellent potential of the FCarb platform, which allowed the partition of the PS into deeper layers of the skin. The applicability of this dermatological platform in photodynamic therapy was evaluated by the generation of reactive species which was demonstrated by chemical photodynamic efficiency assays. The low effect on cell viability and proliferation in the dark was demonstrated by in vitro assays using L929 fibroblasts. The FCarbPc fostered the inhibition of Staphylococcus aureus strain, therefore demonstrating the platform's potential in the treatment of dermatological infections of microbial nature.

Hypericin photodynamic activity in DPPC liposomes - part II: stability and application in melanoma B16-F10 cancer cells.

Hypericin (Hyp) is considered a promising photosensitizer for Photodynamic Therapy (PDT), due to its high hydrophobicity, affinity for cell membranes, low toxicity and high photooxidation activity. In this study, Hyp photophysical properties and photodynamic activity against melanoma B16-F10 cells were optimized using DPPC liposomes (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) as a drug delivery system. This nanoparticle is used as a cell membrane biomimetic model and solubilizes hydrophobic drugs. Hyp oxygen singlet lifetime (τ) in DPPC was approximately two-fold larger than that in P-123 micelles (Pluronic™ surfactants), reflecting a more hydrophobic environment provided by the DPPC liposome. On the other hand, singlet oxygen quantum yield values (ΦΔ1O2) in DPPC and P-123 were similar; Hyp molecules were preserved as monomers. The Hyp/DPPC liposome aqueous dispersion was stable during fluorescence emission and the liposome diameter remained stable for at least five days at 30 °C. However, the liposomes collapsed after the lyophilization/rehydration process, which was resolved by adding the lyoprotectant Trehalose to the liposome dispersion before lyophilization. Cell viability of the Hyp/DPPC formulation was assessed against healthy HaCat cells and high-metastatic melanoma B16-F10 cells. Hyp incorporated into the DPPC carrier presented a higher selectivity index than the Hyp sample previously solubilized in ethanol under the illumination effect. Moreover, the IC50 was lower for Hyp in DPPC than for Hyp pre-solubilized in ethanol. These results indicate the potential of the formulation of Hyp/DPPC for future biomedical applications in PDT treatment.

Hypericin photodynamic activity. Part III: in vitro evaluation in different nanocarriers against trypomastigotes of Trypanosoma cruzi.

Chagas is a parasitic endemic disease caused by the protozoan Trypanosoma cruzi. It represents a strong threat to public health due to its strong resistance against commonly available drugs. We studied the in vitro ability to inactivate the trypomastigote form of this parasite using photodynamic inactivation of microorganisms (or antimicrobial Photodynamic Therapy, aPDT). For this, we chose to use the photosensitizer hypericin (Hyp) formulated in ethanol/water (1% v/v) and Hyp loaded in the dispersion of different aqueous nanocarrier systems. These included polymeric micelles of F-127 and P-123 (both Pluronic™ surfactants), and liposomal vesicles of phospholipid 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). These systems with Hyp had their activity compared against trypomastigote forms under light and in the dark. Hyp revealed a high level of effectiveness to eradicate protozoa in vitro. Samples at concentrations higher than 0.8 µmol L-1 of Hyp in Pluronic micelles showed efficacy even in the dark, with the EC50 around (6-8) µmol L-1. Therefore, Hyp/Pluronics can be used also as a chemotherapeutic agent. The best result for EC50 is at approximately 0.31 µmol L-1 for illuminated systems of Hyp in F-127 micelles. For Hyp in P-123 micelles under light, the results also led to a low EC50 value of 0.36 µmol L-1. The highest value of EC50 was 2.22 µmol L-1, which was found for Hyp/DPPC liposomes under light. For the Hyp-free (ethanol/water, 1% v/v)/illuminated group, the EC50 value was 0.37 µmol L-1, which also is a value that shows effectiveness. However, in free-form, Hyp is not protected against blood components, unlike when Hyp is loaded into the nanocarriers.

Biomedical Platform Development of a Chlorophyll-Based Extract for Topic Photodynamic Therapy: Mechanical and Spectroscopic Properties.

Photodynamic therapy (PDT) is a therapeutic modality that has shown effectiveness in the inactivation of cancer cell lines and microorganisms. Treatment consists of activating the photosensitizer (PS) upon light irradiation of adequate wavelength. After reaching the excited state, the PS can handle the intersystem conversion through energy transfer to the molecular oxygen, generating reactive oxygen species. This especially applies to singlet oxygen (1O2), which is responsible for the selective destruction of the sick tissue. Photosensitizing compounds (chlorophylls and derivatives) existing in the spinach extract have applicability for PDT. This study aimed to develop and characterize the thermoresponsive bioadhesive system composed of Pluronic F127 20.0%- and Carbopol 934P 0.2% (w/w) (FC)-containing chlorophyll-based extract 0.5% (w/w) (FC-Chl). Mechanical and rheological properties, in vitro release, sol-gel transition temperature, and ex vivo permeability of the spinach extract PS components (through pig ear skin) were investigated. Furthermore, photodynamic activity of the system was accessed through uric acid and time-solved measurements. The sol-gel transition temperature obtained for the FC-Chl system was 28.8 ± 0.3 °C. Rheological and texture properties of the platform were suitable for use as a dermatological system, exhibiting easy application and good characteristics of retention in the place of administration. In vitro release studies showed the presence of two distinct mechanisms that reasonably obey the zero-order and first-order kinetics models. PS components presented skin permeability and reached a permeation depth of 830 µm (between the epidermis and dermis). The photodynamic evaluation of the FC-Chl system was effective in the degradation of uric acid. The quantum yield (ΦΔ1O2) and life time (τ1O2) of singlet oxygen showed similar values for the spinach extract and the isolated chlorophyll a species in ethanol. These results allowed for the classification of the FC-Chl platform as potentially useful for the delivery of the chlorophyll-based extract in the topic PDT, suggesting that it is worthy for in vivo evaluation.

Antimicrobial Photodynamic Inactivation Mediated by Rose Bengal and Erythrosine Is Effective in the Control of Food-Related Bacteria in Planktonic and Biofilm States.

The thermal and chemical-based methods applied for microbial control in the food industry are not always environmentally friendly and may change the nutritional and organoleptic characteristics of the final products. Moreover, the efficacy of sanitizing agents may be reduced when microbial cells are enclosed in biofilms. The objective of this study was to investigate the effect of photodynamic inactivation, using two xanthene dyes (rose bengal and erythrosine) as photosensitizing agents and green LED as a light source, against Staphylococcus aureus, Listeria innocua, Enterococcus hirae and Escherichia coli in both planktonic and biofilm states. Both photosensitizing agents were able to control planktonic cells of all bacteria tested. The treatments altered the physicochemical properties of cells surface and also induced potassium leakage, indicating damage of cell membranes. Although higher concentrations of the photosensitizing agents (ranging from 0.01 to 50.0 µmol/L) were needed to be applied, the culturability of biofilm cells was reduced to undetectable levels. This finding was confirmed by the live/dead staining, where propidium iodide-labeled bacteria numbers reached up to 100%. The overall results demonstrated that photoinactivation by rose bengal and erythrosine may be a powerful candidate for the control of planktonic cells and biofilms in the food sector.

Functional Polymeric Systems as Delivery Vehicles for Methylene Blue in Photodynamic Therapy.

Antibiotic-resistant microorganisms have become a global concern, and the search for alternative therapies is very important. Photodynamic therapy (PDT) consists of the use of a nontoxic photosensitizer (PS), light, and oxygen. This combination produces reactive oxygen species and singlet oxygen, which can alter cellular structures. Methylene blue (MB) is a substance from the phenothiazine class often used as a PS. In this work, to facilitate the PS contact within the wounds, we have used Design of Experiments 2(3) plus central point to develop functional polymeric systems. The formulations were composed by poloxamer 407 [15.0, 17.5, or 20.0% (w/w)], Carbopol 934P [0.15, 0.20, or 0.25% (w/w)], and MB [0.25, 0.50, or 0.75% (w/w)]. The sol-gel transition temperature, flow rheometry, in vitro MB release, and ex vivo study of MB cutaneous permeation and retention were investigated. Moreover, the evaluation of photodynamic activity was also analyzed by in vitro degradation of tryptophan by singlet oxygen and using Artemia salina. The determination of the gelation temperature displayed values within the range of 25-37 °C, and the systems with better characteristics were subjected to rheological analysis and in vitro release profiling. The 20/0.15/0.25 formulation showed the best release profile (42.57% at 24 h). This system displayed no significant skin permeation (0.38% at 24 h), and the photooxidation of tryptophan test showed the production of reactive species of oxygen. The toxicity test using A. salina revealed that the MB associated with the light increased the mortality rate by 61.29%. Therefore, investigating the PDT efficacy of the functional polymeric system containing MB will be necessary in the future.

Photodynamic Inactivation Mediated by Erythrosine and its Derivatives on Foodborne Pathogens and Spoilage Bacteria.

The purpose of the present study was to evaluate the efficacy of photodynamic inactivation (PDI) mediated by erythrosine (ERY) and its ester derivatives erythrosine methyl ester (ERYMET) and erythrosine butyl ester (ERYBUT) on foodborne pathogens and spoilage bacteria. We evaluated Staphylococcus aureus ATCC 25923, Aeromonas hydrophila ATCC 7966, Salmonella enterica serotype Typhimurium ATCC 14028, Escherichia coli ATCC 25922, and Pseudomonas aeruginosa ATCC 27853. The toxicity of all of the compounds was assessed in VERO cells. PDI mediated by ERY and its derivatives combined with a light-emitting diode was performed at different concentrations and exposure times. S. aureus was more photosensitive than Gram-negative bacteria to ERY, ERYMET, and ERYBUT. The ERY-mediated PDI of S. aureus induced a significant reduction of 4.0 log CFU/ml at a light dose of 40 J/cm(2). ERYMET and ERYBUT at lower light doses than ERY completely eradicated S. aureus. When photoirradiated with ERY at light doses of 156 and 234 J/cm(2), A. hydrophila was completely eradicated. ERYBUT was more efficient in the PDI of A. hydrophila than ERYMET, even at 1 x 10(-5) M and lower light doses. Salmonella Typhimurium, E. coli, and P. aeruginosa required higher concentrations of photosensitizers to reduce cell survival. ERYBUT and ERY may be promising photosensitizing agents against A. hydrophila and S. aureus. They were effective at reducing bacterial counts at nontoxic concentrations. The photoinactivation rate of the evaluated bacteria decreased in the following order: S. aureus > A. hydrophila > E. coli > S. Typhimurium > P. aeruginosa.

An efficient rose bengal based nanoplatform for photodynamic therapy.

Organically modified mesoporous silica nanoparticles (MSNs) containing rose bengal (RB), a xanthene dye, were successfully synthesized. RB-modified MSNs have shown a relevant photostability and a high efficiency in the photoproduction and delivery of singlet oxygen ((1)O2), which is particularly promising for photodynamic therapy (PDT) applications. In vitro tests have evidenced that RB-MSNs are able to reduce cell proliferation in one of the most aggressive skin cancer types (SK-MEL-28) after green-light irradiation.

Evaluation of the photodynamic activity of xanthene dyes on Artemia salina described by chemometric approaches.

The development of drugs for photodynamic therapy (PDT) is an important area of research due to their growing use in medical applications. Therefore, it is important to develop new bioassay methods for PDT photosensitizers that are inexpensive, easy to handle and highly sensitive to environmental conditions. Xanthene dyes (fluorescein, rose bengal B, erythrosine B and eosin Y) with LED light sources were investigated using Artemia salina as a bioindicator of photodynamic activity. In this study, three factors were investigated: (i) photosensitizers concentration, (ii) the LED irradiation time and (iii) the waiting time between the addition of the photosensitizers and the beginning of the irradiation. To analyze the photo-killing of A. salina, it was employed a 2³ full factorial design. The death of A. salina was related to dye structure and the interaction between the irradiation time and the photosensitizers concentration. About 60% of crustaceans death was obtained using rose bengal B, which presentes the highest quantum yield of singlet oxygen due to the number of iodide substituents in the xanthenes ring. The proposed bioassay using A. salina, xanthene dyes and LED irradiation was found suitable for quantitative PDT drug evaluation.

Proteomic Investigation over the Antimicrobial Photodynamic Therapy Mediated by Rose Bengal Against Staphylococcus aureus.

In order, understanding the antimicrobial action of photodynamic therapy and how this technique can contribute to its application in the control of pathogens. The objective of the study was to employ a proteomic approach to investigate the protein profile of Staphylococcus aureus after antimicrobial photodynamic therapy mediated by rose bengal (RB-aPDT). S. aureus was treated with RB (10 nmoL L-1 ) and illuminated with green LED (0.17 J cm-2 ) for cell viability evaluation. Afterward, proteomic analysis was employed for protein identification and bioinformatic tools to classify the differentially expressed proteins. The reduction in S. aureus after photoinactivation was ~2.5 log CFU mL-1 . A total of 12 proteins (four up-regulated and eight down-regulated) correspond exclusively to alteration by RB-aPDT. Functionally, these proteins are distributed in protein binding, structural constituent of ribosome, proton transmembrane transporter activity and ATPase activity. The effects of photodamage include alterations of levels of several proteins resulting in an activated stress response, altered membrane potential and effects on energy metabolism. These 12 proteins required the presence of both light and RB suggesting a unique response to photodynamic effects. The information about this technique contributes valuable insights into bacterial mechanisms and the mode of action of photodynamic therapy.

Elucidation the binding interaction of hypericin-loaded P84 copolymeric micelles by using 1D and 2D NMR techniques.

Hypericin (HYP) is an active compound of Hypericum perforatum. Associated with photodynamic therapy (PDT), HYP has shown a broad therapeutic potential against microorganisms and cancer cells. Due to the low water solubility of HYP, its application in the biological medium becomes limited. To solve this limitation, our research group has been used copolymeric micelles to carrier HYP with high efficiency. However, there is no elucidated mechanism for HYP delivery mediated by copolymeric micelles. In this sense, we believed that the study of binding-sites of copolymeric micelles and HYP is the first step to its understanding. For this purpose, in this work, we employed 1D and 2D NMR techniques to investigate the behaviour of HYP-loaded P84 micelles in different concentrations . 1D and 2D NMR analysis revealed that HYP molecules were arrangement in a π-stacked aggregation form with a specific location on the core of P84 micelles.

Interaction of triblock copolymers (Pluronic®) with DMPC vesicles: a photophysical and computational study.

Pluronic/lipid mix promises stealth liposomes with long circulation time and long-term stability for pharmaceutical applications. However, the influence of Pluronics on several aspects of lipid membranes has not been fully elucidated. Herein it was described the effect of Pluronics on the structured water, alkyl chain conformation, and kinetic stability of dimyristoylphosphatidylcholine (DMPC) liposomes using interfacial and deeper fluorescent probes along with computational molecular modeling data. Interfacial water changed as a function of Pluronics' hydrophobicity with polypropylene oxide (PPO) anchoring the copolymers in the lipid bilayer. Pluronics with more than 30-40 PO units had facilitated penetration at the bilayer while shorter PPO favored a more interfacial interaction. Low Pluronic concentrations provided long-term stability of vesicles by steric effects of polyethylene oxide (PEO), but high amounts destabilized the vesicles as a sum of water-bridge cleavage at the polar head group and the reduced alkyl-alkyl interactions among the lipids. The high kinetic stability of Pluronic/DMPC vesicles is a proof-of-concept of its advantages and applicability in nanotechnology over conventional liposome-based pharmaceutical products for future biomedical applications.

Spectrophotometric studies of charge-transfer complexes formed with ions N,N'-alkyldiyl-bis(pyridinium) derivatives and iodide.

The charge-transfer complexes (CTC) between electron donor iodide and pyridinium dimers and monomers as acceptors have been studied spectrophotometrically in acetonitrile, DMSO and water. The dimers were: N,N'-alkyldiyl-bis(4-cyanopyridinium) and N,N'-alkyldiyl-bis(2-bromopyridinium), and the monomers were: N-alkyl-4-cyanopyridinium and N-alkyl-2-bromopyridinium, bridged by n methylene spacers. The formation constant (KCTC), molar absorptivity (εCTC), fluorescence-quenching constant (KSV) were determined. The results indicate that the stoichiometry of the CTC for dimers and monomers is 1:1 (equimolar ratio), and its formation is favored for dimers more than for monomers, especially dimers with short methylene spacers forming a "sandwiched type-complex" in which the iodide is close to the two pyridinium rings. Solvents with low polarity favored the complex, which was destroyed by the presence of water. The experimental studies were complemented with theoretical studies with quantum mechanics (QM) calculations using density functional theory (DFT) and molecular mechanics (MM) simulations with Molecular Dynamics for identify the most stable conformers in acetonitrile solution. The electronic excitations were calculated with sequential QM/MM hybrid method, showing a charge-transfer wavelength in agreement with the UV-Vis absorption spectra obtained experimentally. It confirms the "sandwiched type-complex" conformers favoring the interaction of the iodide with one pyridinium rings and simultaneously forming one, or two, hydrogen bonds with the alkyl chain and additionally, for the case of dimers, with the other pyridinium ring.

Melanoma-targeted photodynamic therapy based on hypericin-loaded multifunctional P123-spermine/folate micelles.

Multifunctional P123 micelle linked covalently with spermine (SM) and folic acid (FA) was developed as a drug delivery system of hypericin (HYP). The chemical structures of the modified copolymers were confirmed by spectroscopy and spectrophotometric techniques (UV-vis, FTIR, and 1H NMR). The copolymeric micelles loading HYP were prepared by solid dispersion and characterized by UV-vis, fluorescence, dynamic light scattering (DLS), ζ potential, and transmission electron microscopy (TEM). The results provided a good level of stability for HYP-loaded P123-SM, P123-FA, and P123-SM/P123-FA in the aqueous medium. The morphology analysis showed that all copolymeric micelles are spherical. Well-defined regions of different contrast allow us to infer that SM and FA were localized on the surface of micelles, and the HYP molecules are located in the core region of micelles. The uptake potential of multifunctional P123 micelle was accessed by exposing the micellar systems loading HYP to two cell lines, B16-F10 and HaCaT. HYP-loaded P123 micelles reveal a low selectivity for melanoma cells, showing significant photodamage for HaCat cells. However, the exposition of B16-F10 cells to Hyp-loaded SM- and FA-functionalized P123 micelles under light irradiation revealed the lowest CC50 values. The interpretation of these results suggested that the combination of SM and FA on P123 micelles is the main factor in enhancing the HYP uptake by melanoma cells, consequently leading to its photoinactivation.

Photodynamic therapy for American cutaneous leishmaniasis: the efficacy of methylene blue in hamsters experimentally infected with Leishmania (Leishmania) amazonensis.

The aim of this study was to investigate the effectiveness of Photodynamic Therapy (PDT) using Methylene Blue (MB) as the photosensitizing compound and a Light-Emitting Diode (LED) in American cutaneous leishmaniasis (ACL). Hamsters were experimentally infected with Leishmania (Leishmania) amazonensis. After the development of the lesions in the footpad, the animals were treated with MB three times a week for 3 months. Ten minutes after each application of MB, the lesions were irradiated with LED for 1 h. The lesions were evaluated weekly by the measurement of the hamster footpad thickness. At the end of the treatment the parasitic load was quantified in the regional lymph node of the hamsters. The treatment promoted a decrease in the thickness of infected footpad (P=0.0001) and reduction in the parasitic load in the regional lymph node (P=0.0007) of the animals from group treated with MB+LED. PDT using MB+LED in ACL caused by L. amazonensis shows a strong photodynamic effect. This therapy is very promising, once it is an inexpensive system and the own patient can apply it in their wound and in their house without the need of technical assistance.

Mesoporous Silica Nanoparticles Functionalized with Amino Groups for Biomedical Applications.

The synthesis and characterization of amino-functionalized mesoporous silica nanoparticles are presented following two different synthetic methods: co-condensation and post-synthesis grafting of 3-aminopropyltriethoxysilane. The amino groups' distribution on the mesoporous silica nanoparticles was evaluated considering the aggregation state of a grafted photosensitizer (Verteporfin) by using spectroscopic techniques. The homogeneous distribution of amino groups within the silica network is a key factor to avoid aggregation during further organic functionalization and to optimize the performance of functionalized silica nanoparticles in biomedical applications. In addition, the formation of a protein corona on the external surface of both bare and amino-functionalized mesoporous silica was also investigated by adsorbing Bovine Serum Albumin (BSA) as a model protein. The adsorption of BSA was found to be favorable, reducing the aggregation phenomena for both bare and amino-modified nanoparticles. Nevertheless, the dispersant effect of BSA was much more evident in the case of amino-modified nanoparticles, which reached monodispersion after adsorption of the protein, thus suggesting that amino-modified nanoparticles can benefit from protein corona formation for preventing severe aggregation in biological media.

Photophysical characterization of Hypericin-loaded in micellar, liposomal and copolymer-lipid nanostructures based F127 and DPPC liposomes.

Hypericin (Hy) compound presents a high photoactivity in photodynamic therapy (PDT), photodiagnosis and theranostics applications. The maintenance of this compound in monomeric form could undermine the potential benefits of its photophysical and photodynamic activity. In this study, we demonstrated that the Hy formulated in a system based on the use of the F127 copolymer and the 1,2-dipalmitoyl-sn-3-glycerol-phosphatidylcholine (DPPC) as micelles, liposomal vesicles and Copolymer-Lipid coated systems, have improved its photophysical properties for many clinical modalities. Based on the results of the triplet state lifetime values (τt), the singlet oxygen quantum yield (ΦΔ1O2), the fluorescence lifetime (τF) and the fluorescence quantum yield (ΦF), all Hy formulations had its photophysical properties described in different models of drug delivery systems (DDS). In addition, the transient spectra profile of those formulations was unaffected by the Hy incorporation process, except for the liposomal system, which demonstrated to be the less stable one by flash photolysis technique. The cytotoxic effects of those formulations were also investigated for CaCo-2 and HaCat cells line. The cytotoxic concentrations for 50% (CC50) were 0.56, 1.05, 1.33 and 4.80 µmol L-1 for Copolymer-Lipid/Hy, DPPC/Hy, F127/Hy and ethanol/Hy for CaCo-2 cells, respectively, and 0.69, 2.02, 1.45 and 1.16 µmol L-1 for Copolymer-Lipid/Hy, DPPC/Hy, F127/Hy and ethanol/Hy for HaCat cells, respectively. The F127 copolymer had a significant role in many photophysical parameters determined for Copolymer-Lipid/Hy coated system. Although all those formulations had shown satisfactory results, Copolymer-Lipid/Hy proved to be superior in many aspects, being the most promising formulation for PDT, photodiagnosis and theranostics applications.

Antiproliferative activity and energy calculations of a new triterpene isolated from the palm tree Acrocomia totai.

Acrocomia totai Mart (Arecaceae) is a palm tree native to South America, widely studied for biodiesel production. The aim of this work was to perform the first phytochemical study of A. totai leaves, as well as to do biological assays against human cancer cell lines. A new triterpene of the hopane class named totaiol (1), three known triterpenes (2-4), and two phytosteroids (5-6) were identified. The new natural product was characterized using 1 D and 2 D NMR, single crystal X-ray diffraction analises, and high resolution mass spectrometry. The intercontacts in the crystal packing were also analised. Complete stereochemical characterization of compound 1 revealed an unusual positioning pattern for methyl and isopropenyl groups in the polycyclic skeleton. Compounds 1-5 were evaluated for the first time in antiproliferative assays against Ca Ski, MCF-7 and MCF-10 cells. The new natural product was active against Ca Ski cells with IC50 ≤ 6.25 µg mL-1.

Hypericin-P123-photodynamic therapy in an ex vivo model as an alternative treatment approach for onychomycosis caused by Fusarium spp.

BackgroundFusarium has been considered an opportunistic pathogen, causing several infections in humans, including onychomycosis. In addition, a high resistance to conventional antifungals has been linked to this genus. Photodynamic Therapy (PDT), known as a non-invasive therapy, can be an alternative treatment for fungal infections, based on the excitation of a photosensitizing compound (PS) by a specific length of light, causing damage to the target. The aim of this study was to evaluate the effects of a formulation of Hypericin (Hyp) encapsulated in Pluronic™ (P123), via photodynamic therapy (PDT), on planktonic cells and biofilms in Fusarium spp. using in vitro and ex vivo assays. Materials & Methods epidemiology studies about Fusarium spp. in onychomycosis was perfomed, carried out molecular identification, compared the antifungal activity of the conventional antifungals with PDT with encapsulated Hypericin (Hyp-P123), carried out detection of reactive oxygen species, and measured the antibiofilm effect of the Hyp-P123-PDT in vitro and in an ex vivo model of onychomycosis. Results Hyp-P123-PDT exhibited a fungicidal effect in vitro with reductions ≥ 3 log10. ROS generation increased post-Hyp-P123-PDT in Fusarium spp. Hyp-P123-PDT showed a potent inhibitory effect on adhesion-phase and mature biofilms in vitro tests and an ex vivo model of onychomycosis (p<0.0001). Conclusion Hyp-P123-PDT had a potent effect against Fusarium spp., suggesting that photodynamic therapy with Hyp-P123 is a safe and promising treatment for onychomycosis in clinical practice.