Vaterite microparticle-loaded methylene blue for photodynamic activity in macrophages infected with Leishmania braziliensis.

Calcium carbonate (CaCO3) exhibits a variety of crystalline phases, including the anhydrous crystalline polymorphs calcite, aragonite, and vaterite. Developing porous calcium carbonate microparticles in the vaterite phase for the encapsulation of methylene blue (MB) as a photosensitizer (PS) for use in photodynamic therapy (PDT) was the goal of this investigation. Using an adsorption approach, the PS was integrated into the CaCO3 microparticles. The vaterite microparticles were characterized by scanning electron microscopy (SEM) and steady-state techniques. The trypan blue exclusion method was used to measure the biological activity of macrophages infected with Leishmania braziliensis in vitro. The vaterite microparticles produced are highly porous, non-aggregated, and uniform in size. After encapsulation, the MB-loaded microparticles kept their photophysical characteristics. The carriers that were captured allowed for dye localization inside the cells. The results obtained in this study indicated that the MB-loaded vaterite microparticles show promising photodynamic activity in macrophages infected with Leishmania braziliensis.

Photodithazine photodynamic effect on viability of 9L/lacZ gliosarcoma cell line.

Even with the advances of conventional treatment techniques, the nervous system cancer prognosis is still not favorable to the patient which makes alternative therapies needed to be studied. Photodynamic therapy (PDT) is presented as a promising therapy, which employs a photosensitive (PS) agent, light wavelength suitable for the PS agent, and molecular oxygen, producing reactive oxygen species in order to induce cell death. The aim of this study is to observe the PDT action in gliosarcoma cell using a chlorin (Photodithazine, PDZ). The experiments were done with 9L/lacZ lineage cells, grown in a DMEM medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin solution and put in a culture chamber at 37 °C with an atmosphere of 5% CO2. The PS agent used was the PDZ to an LED light source device (Biopdi/IRRAD-LED 660) in the 660-nm region. The location of the PS agent was analyzed by fluorescence microscopy, and cell viability was analyzed by MTT assay (mitochondrial activity), exclusion by trypan blue (cell viability), and morphological examination through an optical microscope (Leica MD 2500). In the analysis of the experiments with PDZ, there was 100% cell death at different concentrations and clear morphological differences in groups with and without treatment. Furthermore, it was observed that the photodithazine has been focused on all nuclear and cytoplasmic extension; however, it cannot be said for sure whether the location is in the inside core region or on the plasma membrane. In general, the PDZ showed a promising photosensitive agent in PDT for the use of gliosarcoma.

Hydroxyapatite microspheres used as a drug delivery system for gliosarcoma strain 9l/Lacz treatment by photodynamic therapy protocols.

BACKGROUND: Hydroxyapatite (HAp) presents similarities with the human bone structure and presents properties such as biodegradability, biocompatibility, and osteoconductivity, which favors its use in prostheses implants and enables its use as a vehicle for the delivery of photosensitizers (PS) from systems of release (DDS) for photodynamic therapy applications Methods: In this work was to synthesized hydroxyapatite microspheres (meHAp), encapsulated with chloroaluminium phthalocyanine (ClAlPc), for DDS. meHAp was synthesized using vaterite as a template. The drug was encapsulated by mixing meHAp and a 50.0 mg.mL-1 ClAlPc solution. Photochemical, photophysical, and photobiological studies characterized the system. RESULTS: The images from the SEM analysis showed the spherical form of the particles. All spectroscopic results showed excellent photophysical parameters of the drug studied when served in the meHAp system. The incorporation efficiency was 57.8 %. The trypan blue exclusion test results showed a significant reduction (p < 0.05) in cell viability for the groups treated with PDT at all concentrations above 250 µg.mL-1. In 9 L/lacZ gliosarcoma cells, PDT mediated at concentrations from 250 to 62.5 µg.mL-1 reduced cell viability by more than 98 %. In the cell internalization study, it was possible to observe the internalization of phthalocyanines at 37 °C, with the accumulation of PS in the cytoplasm and inside the nucleus in the two tested concentrations. CONCLUSIONS: From all the results presented throughout the article, the meHAp system shows promise for use as a modified release system (DSD) in photodynamic therapy.

Antimicrobial Photodynamic Therapy Mediated by Fotenticine and Methylene Blue on Planktonic Growth, Biofilms, and Burn Infections of Acinetobacter baumannii.

Antimicrobial photodynamic therapy (aPDT) is considered a promising alternative strategy to control Acinetobacter baumannii infections. In this study, we evaluated the action of aPDT mediated by a new photosensitizer derivative from chlorin e-6 (Fotoenticine-FTC) on A. baumannii, comparing its effects with methylene blue (MB). For this, aPDT was applied on A. baumannii in planktonic growth, biofilms, and burn infections in Galleria mellonella. The absorption of FTC and MB by bacterial cells was also evaluated using microscopic and spectrophotometric analysis. The results of planktonic cultures showed that aPDT reduced the number of viable cells compared to the non-treated group for the reference and multidrug-resistant A. baumannii strains. These reductions varied from 1.4 to 2 log10 CFU for FTC and from 2 log10 CFU to total inhibition for MB. In biofilms, aPDT with MB reduced 3.9 log10 CFU of A. baumannii, whereas FTC had no effect on the cell counts. In G. mellonella, only MB-mediated aPDT had antimicrobial activity on burn injuries, increasing the larvae survival by 35%. Both photosensitizers were internalized by bacterial cells, but MB showed a higher absorption compared to FTC. In conclusion, MB had greater efficacy than FTC as a photosensitizer in aPDT against A. baumannii.

Synthesis, characterization, and evaluation of chloroaluminium phthalocyanine incorporated in poly(ε-caprolactone) nanoparticles for photodynamic therapy.

BACKGROUND: The use of nanotechnology has been widely used in biomedical science, including orthopedic implants, tissue engineering, cancer therapy and drug elution from nanoparticle systems, such as poly-caprolactone (PCL) nanoparticles, which stand out mainly for their biocompatibility, being considered as effective carriers for photosensitizing drugs (PS) in photodynamic therapy (PDT) protocols. METHODS: This manuscript describes the synthesis and characterization of PCL nanoparticles for controlled release of the drug chloro-aluminum phthalocyanine (ClAlPc) as a photosensitizer for application in PDT. The PCL-ClAlPc nanoparticles were developed by the nanoprecipitation process. The structure and morphology of the nanoparticles were studied with scanning electron microscopy (SEM) and with Fourier transform infrared (FTIR). The size of nanomaterials was studied using the Dynamic Light Scattering (DLS) method. Photophysical and photochemical characterizations were performed. Subsequently, photobiological studies were also used to characterize the system. RESULTS: The nanoparticles had an average diameter of 384.7 ± 138.6 nm and a polydispersity index of 0.153. SEM analysis revealed that the system formed a spherical shape typical of these delivery systems. Charging efficiency was 82.1% ± 1.2%. The phthalocyanine-loaded PCL nanoparticles maintained their photophysical behavior after encapsulation. Cell viability was determined after the dark toxicity test, and it was possible to observe that there was no evidence of toxicity in the dark, for all concentrations tested. The assay also revealed that adenocarcinoma cells treated with free ClAlPc and in the nanoformulation showed 100% cell death when subjected to PDT protocols. The intracellular location of the photosensitizer indicated a high potential for accumulation in the cytoplasm and nucleus. CONCLUSIONS: From the photophysical, photochemical and photobiological analyzes obtained, it was possible to observe that the development of PCL nanoparticles encapsulated with ClAlPc, by the nanoprecipitation method was adequate and that the in vivo release study is efficient to reduce the release rate and attenuate the burst of PS loaded on PCL nanoparticles. The results reinforce that the use of this system as drug delivery systems is useful in PDT protocols.

Synthesis and characterization of photosensitive gelatin-based hydrogels for photodynamic therapy in HeLa-CCL2 cell line.

BACKGROUND: Hydrogel systems are increasingly gaining visibility involving biomedicine, tissue engineering, environmental treatments, and drug delivery systems. These systems have a three-dimensional network composition and high-water absorption capacity, are biocompatible, allowing them to become an option as photosensitizer carriers (PS) for applications in Photodynamic Therapy (PDT) protocols. METHODS: A nanohydrogel system (NAHI), encapsulated with chloroaluminium phthalocyanine (ClAlPc) was synthesized for drug delivery.. NAHI was synthesized using gelatin as based polymer by the chemical cross-linking technique. The drug was encapsulated by immersing the hydrogel in a 1.0 mg.mL-1 ClAlPc solution. The external morphology of NAHI was examined by scanning electron microscopy (SEM). The degree of swelling of the synthesized system was evaluated to determine the water absorption potential. The produced nanohydrogel system was characterized by photochemical, photophysical and photobiologial studies. RESULTS: The images from the SEM analysis showed the presence of three-dimensional networks in the formulation. The swelling test demonstrated that the nanohydrogel freeze-drying process increases its water holding capacity. All spectroscopic results showed excellent photophysical parameters of the drug studied when served in the NAHI system. The incorporation efficiency was 70%. The results of trypan blue exclusion test have shown significant reduction (p < 0.05) in the cell viability for all groups treated with PDT, in all concentrations tested. In HeLa cells, PDT mediated by 0,5 mg.mL-1 ClAlPc encapsulated in NAHI showed a decrease in survival close to 95%. In the internalization cell study was possible to observe the internalization of phthalocyanine after one hour of incubation, at 37 °C, with the the accumulation of PS in the cytoplasm and inside the nucleus at both concentrations tested. CONCLUSIONS: Given the peculiar performance of the selected system, the resulting nanohydrogel is a versatile platform and display potential applications as controlled delivery systems of photosensitizer for photodynamic therapy application.

Photodynamic activity of Photogem® in Leishmania promastigotes and infected macrophages.

Objectives: This study aimed to evaluate the effect of photodynamic therapy (PDT) with Photogem® in promastigotes of Leishmania braziliensis and Leishmania major, and in infected macrophages. Materials & methods: The following parameters were analyzed: Photogem® internalization, mitochondrial activity, viability, tubulin marking and morphological alterations in promastigotes and viability in infected macrophages. Results: Photogem® accumulated in the cytosol and adhered to the flagellum. Changes were observed in the mitochondrial activity in groups maintained in the dark, with no viability alteration. After PDT, viability decreased up to 80%, and morphology was affected. Conclusion: The results point out that PDT with Photogem® can reduce parasite and macrophage viability.

Vaterite submicron particles designed for photodynamic therapy in cells.

BACKGROUND: Calcium carbonate (CaCO3) is one of the most abundant materials in the world. It has several different crystalline phases as present in the minerals: calcite, aragonite and vaterite, which are anhydrous crystalline polymorphs. Regarding the preparation of these microparticles, the most important aspect is the control of the polymorphism, particle size and material morphology. This study aimed to develop porous microparticles of calcium carbonate in the vaterite phase for the encapsulation of chloro-aluminum phthalocyanine (ClAlPc) as a photosensitizer (PS) for application in Photodynamic Therapy (TFD). METHODS: In this study, spherical vaterite composed of microparticles are synthesized by precipitation route assisted by polycarboxylate superplasticizer (PSS). The calcium carbonate was prepared by reacting a mixed solution of Na2CO3 with a CaCl2 solution at an ambient temperature, 25 °C, in the presence of polycarboxylate superplasticizer as a stabilizer. The photosensitizer was incorporated by adsorption technique in the CaCO3 microparticles. The CaCO3 microparticles were studied by scanning electron microscopy, steady-state, and their biological activity was evaluated using in vitro cancer cell lines by trypan blue exclusion method. The intracellular localization of ClAlPc was examined by confocal microscopy. RESULTS: The CaCO3 microparticles obtained are uniform and homogeneously sized, non-aggregated, and highly porous microparticles. The calcium carbonate microparticles show an average size of 3 µm average pore size of about 30-40 nm. The phthalocyanine derivative loaded-microparticles maintained their photophysical behavior after encapsulation. The captured carriers have provided dye localization inside cells. The in vitro experiments with ClAlPc-loaded CaCO3 microparticles showed that the system is not cytotoxic in darkness, but exhibits a substantial phototoxicity at 3 µmol.L-1 of photosensitizer concentration and 10 J.cm-2 of light. These conditions are sufficient to kill about 80 % of the cells. CONCLUSIONS: All the performed physical-chemical, photophysical, and photobiological measurements indicated that the phthalocyanine-loaded CaCO3 microparticles are a promising drug delivery system for photodynamic therapy and photoprocesses.

Bacterial viability after antimicrobial photodynamic therapy with curcumin on multiresistant Staphylococcus aureus.

Staphylococcus aureus are multiresistant pathogens that causes superficial and systemic infections. Antimicrobial photodynamic therapy (APDT) is an alternative in the treatment of diseases caused by these bacteria. Aim: In this study the APDT response on growth, viability, formation of reactive oxygen species and adhesion of methicillin-sensitive strains of Staphylococcus aureus, strains of methicillin-resistant S. aureus and American-type culture collection (ATCC) of S. aureus were evaluated in vitro, after incubation with curcumin for 20 min, and irradiated with LED. Materials & methods: Bacterial growth was assessed by the number of colony-forming units, viability and adhesion were evaluated by confocal microscopy and ROS quantification was performed by fluorimetry. Results: Was observed increase in the production of ROS in APDT groups, besides a decrease in the 4 log growth and loss of the bacterial adhesion. Conclusion: APDT with Curcumin may be an interesting therapeutic alternative, due to its in vitro response, in the control multiresistant clinical S. aureus strains.