RESUMO
BACKGROUND: Photodynamic therapy (PDT) is an anticancer treatment that utilizes the interaction of light and a photosensitiser (PS), promoting tumour cell death mediated by generation of reactive oxygen species. In this study, we evaluated the in vitro photoactivity of four meso-substituted porphyrins and a porphyrin coupled to a fullerene. METHODS: The cell line employed was the LM3 mammary adenocarcinoma, and the PS with the best photokilling activity was administered to mice bearing the LM3 subcutaneously implanted adenocarcinoma. The TEMCP4+ porphyrin and its analogue TEMCC4+ chlorine contain four identical carbazoyl substituents at the meso positions of the tetrapyrrolic macrocycle and have A4 symmetry. The TAPP derivative also has A4 symmetry, and it is substituted at the meso positions by aminopropoxy groups. The DAPP molecule has ABAB symmetry with aminopropoxy and the trifluoromethyl substituents in trans positions. The TCP-C604+ dyad is formed by a porphyrin unit covalently attached to the fullerene C60. RESULTS: The PSs are taken up by the cells with the following efficiency: TAPP> TEMCP4+ = TEMCC4+ > DAPP >TCP-C604+, and the amount of intracellular PS correlates fairly with the photodamage degree, but also the quantum yields of singlet oxygen influence the PDT outcome. TAPP, DAPP, TEMCC4+ and TEMCP4+ exhibit high photoactivity against LM3 mammary carcinoma cells, being TAPP the most active. After topical application of TAPP on the skin of mice bearing LM3 tumours, the molecule is localized mainly in the stratum corneum, and at a lower extent in hair follicles and sebaceous glands. Systemic administration of TAPP produces a tumour: normal skin ratio of 31.4, and high accumulation in intestine and lung. CONCLUSION: The results suggest a potential use of topical TAPP for the treatment of actinic keratosis and skin adnexal neoplasms. In addition, selectivity for tumour tissue after systemic administration highlights the selectivity of and potentiality of TAPP as a new PS.
Assuntos
Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Porfirinas/uso terapêutico , Neoplasias Cutâneas/tratamento farmacológico , Animais , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia de Fluorescência , Fármacos Fotossensibilizantes/farmacocinética , Distribuição TecidualRESUMO
PURPOSE: Melanoma is an invasive and very aggressive skin cancer due to its multi-drug resistance that results in poor patient survival. There is a need to test new treatment approaches to improve therapeutic efficacy and reduce side effects of conventional treatments. METHODS: PLA/PVA nanoparticles carrying both Dacarbazine and zinc phthalocyanine was produced by double emulsion technique. The characterization was performed by dynamic light scattering and atomic force microscopy. In vitro photodynamic therapy test assay using MV3 melanoma cells as a model has been performed. In vitro cell viability (MTT) was performed to measure cell toxicity of of nanoparticles with and without drugs using human endothelial cells as a model. The in vivo assay (biodistribution/tissue deposition) has been performed using radiolabeled PLA/PVA NPs. RESULTS: The nanoparticles produced showed a mean diameter of about 259 nm with a spherical shape. The in-vitro photodynamic therapy tests demonstrated that the combination is critical to enhance the therapeutic efficacy and it is dose dependent. The in vitro cell toxicity assay using endothelial cells demonstrated that the drug encapsulated into nanoparticles had no significant toxicity compared to control samples. In-vivo results demonstrated that the drug loading affects the biodistribution of the nanoparticle formulations (NPs). Low accumulation of the NPs into the stomach, heart, brain, and kidneys suggested that common side effects of Dacarbazine could be reduced. CONCLUSION: This work reports a robust nanoparticle formulation with the objective to leveraging the synergistic effects of chemo and photodynamic therapies to potentially suppressing the drug resistance and reducing side effects associated with Dacarbazine. The data corroborates that the dual encapsulated NPs showed better in-vitro efficacy when compared with the both compounds alone. The results support the need to have a dual modality NP formulation for melanoma therapy by combining chemotherapy and photodynamic therapy.
Assuntos
Antineoplásicos Alquilantes/administração & dosagem , Portadores de Fármacos/química , Melanoma/tratamento farmacológico , Fármacos Fotossensibilizantes/administração & dosagem , Neoplasias Cutâneas/tratamento farmacológico , Animais , Antineoplásicos Alquilantes/efeitos adversos , Antineoplásicos Alquilantes/farmacocinética , Linhagem Celular Tumoral , Sobrevivência Celular , Dacarbazina/administração & dosagem , Dacarbazina/farmacocinética , Composição de Medicamentos/métodos , Células Endoteliais , Humanos , Isoindóis/administração & dosagem , Isoindóis/farmacocinética , Masculino , Melanoma/patologia , Camundongos , Nanopartículas/química , Compostos Organometálicos/administração & dosagem , Compostos Organometálicos/farmacocinética , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacocinética , Poliésteres/química , Álcool de Polivinil/química , Neoplasias Cutâneas/patologia , Distribuição Tecidual , Compostos de Zinco/administração & dosagem , Compostos de Zinco/farmacocinéticaRESUMO
Glioblastoma multiforme (GBM) is an extremely aggressive malignant brain tumor. Despite advances in treatment modalities, it remains largely incurable. This unfavorable prognosis for GBM is at least partly due to the lack of a successful drug delivery system across the blood-brain barrier (BBB). The delivery of drugs through nanomedicines combined with less invasive alternative therapies represents an important hope for the future of these incurable brain tumors. Whey protein nanocarriers represent promising strategy for targeted drug delivery to tumor cells by enhancing the drug's bioavailability and distribution, and reducing the body's response towards drug resistance. They have been extensively studied to find new alternatives for capacity to encapsulate different drugs and no need for cross-linkers. In this study, we report for the first time the incorporation and administration of Aluminum phthalocyanine chloride (AlClPc)-loaded whey protein drug delivery system (AlClPc-PDDS) for the treatment of glioblastoma brain cancer. This system was designed and optimized (with the use of the spray drying technique) to obtain the required particle size (in the range of 100 to 300 nm), zeta potential and drug loading. Our results suggest that we have developed a drug delivery system from a low-cost raw material and preparation method that is capable of incorporating hydrophobic drugs which, in combination with irradiation, cause photodamage to neoplasic cells, working as an effective adjuvant treatment for malignant glioma.
Assuntos
Neoplasias Encefálicas/tratamento farmacológico , Portadores de Fármacos , Glioblastoma/tratamento farmacológico , Nanopartículas , Fármacos Fotossensibilizantes , Proteínas do Soro do Leite , Barreira Hematoencefálica/metabolismo , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacologia , Glioblastoma/metabolismo , Glioblastoma/patologia , Humanos , Indóis/química , Indóis/farmacocinética , Indóis/farmacologia , Nanopartículas/química , Nanopartículas/uso terapêutico , Compostos Organometálicos/química , Compostos Organometálicos/farmacocinética , Compostos Organometálicos/farmacologia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacocinética , Fármacos Fotossensibilizantes/farmacologia , Proteínas do Soro do Leite/química , Proteínas do Soro do Leite/farmacocinética , Proteínas do Soro do Leite/farmacologiaRESUMO
Monomeric zinc phthalocyanine has been studied as a promising active photosensitizer in photodynamic therapy against cancer, in which its aggregate form is non-active. This paper aimed to describe the monomer/aggregates equilibrium of zinc phthalocyanine in binary water/DMSO mixtures. To reach this aim theoretical calculation, electronic absorption, static and time-resolved fluorescence, and resonance light scattering was used. Zinc phthalocyanine shows a complex water dependence behavior in the mixture. At least three distinct steps were observed: (i) until 30% water zinc phthalocyanine is essentially in the monomeric form, changing to (ii) small slipped cofacial-aggregates around 30% to 40% water and finally to (iii) a staircase arrangement of large aggregates at higher water percent. The staircase arrangement is driven by the intermolecular coordination between the pyrrolic nitrogen lone-pairs and the central metal zinc. The water-Zn coordination governs the fluorescence quenching by a static mechanism. These results have direct relevance in the better understanding on the behavior of zinc phthalocyanine in vivo and when incorporated in drug delivery systems for clinical applications in photodynamic therapy.
Assuntos
Complexos de Coordenação/química , Indóis/química , Modelos Moleculares , Fármacos Fotossensibilizantes/química , Zinco/química , Complexos de Coordenação/farmacocinética , Complexos de Coordenação/farmacologia , Sistemas de Liberação de Medicamentos , Humanos , Indóis/farmacocinética , Indóis/farmacologia , Isoindóis , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacocinética , Fármacos Fotossensibilizantes/farmacologia , Zinco/farmacocinética , Zinco/farmacologiaRESUMO
Oral therapy with 8-methoxypsoralen (8-MOP) may cause major side effects, whereas the topical treatment might not be much effective due to the low penetration induced by typical formulations. Therefore, the objectives of this work are the development and characterization of a nanoemulsion (NE) containing 8-MOP together with an ex vivo permeation study, monitored by a validated HPLC-Fluo method, to determine the amount of drug retained in viable skin (epidermis (E) and dermis (D)) and in stratum corneum (SC). The optimized conditions for NE formulation were achieved by full factorial designs (25 and 32): 60â¯s and 60% of ultrasound time and potency, respectively; 10â¯mL of final volume; 2% v/v of oil phase (clove essential oil); and 10% m/v of Poloxamer 407. The NE showed mean droplet diameter of 24.98⯱â¯0.49â¯nm, polydispersity index (PDI) of 0.091⯱â¯0.23, pH values of 6.54⯱â¯0.06, refractive index of 1.3525⯱â¯0.0001 and apparent viscosity of 51.15⯱â¯3.66â¯mPa at 20⯰C. Droplets with nanospherical diameters were also observed by transmission electron microscopy (TEM). Ex vivo permeation study showed that 8.5% of the applied 8-MOP dose permeated through the biological membranes, with flux (J) of 1.35⯵gâ¯cm-2â¯h-1. The drug retention in Eâ¯+â¯D and in SC was 10.15⯱â¯1.36 and 1.95⯱â¯0.71⯵gâ¯cm-2, respectively. Retention in viable skin induced by the NE was almost two-fold higher than a compounded cream (5.04⯱â¯0.30⯵gâ¯cm-2). These results suggested that the developed NE is a promising alternative for 8-MOP topical therapy when compared to commercial formulations.
Assuntos
Metoxaleno/administração & dosagem , Nanopartículas/administração & dosagem , Fármacos Fotossensibilizantes/administração & dosagem , Pele/metabolismo , Administração Cutânea , Animais , Óleo de Cravo/administração & dosagem , Óleo de Cravo/química , Óleo de Cravo/farmacocinética , Composição de Medicamentos , Sistemas de Liberação de Medicamentos , Estabilidade de Medicamentos , Emulsões , Metoxaleno/química , Metoxaleno/farmacocinética , Nanopartículas/química , Permeabilidade , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacocinética , Poloxâmero/administração & dosagem , Poloxâmero/química , Poloxâmero/farmacocinética , Absorção Cutânea , Solubilidade , SuínosRESUMO
The use of layer-by-layer (LbL) deposition technique allows materials, such as drugs, to be self-assembled in multilayers with other electrolytes by combining their properties in a nanostructured system. Triclosan (TCS) is commonly used as a drug because of its bactericidal action, while erythrosine (ERY) has been used as a photosensitizer in photodynamic therapies because of its high light absorptivity in the visible region of the electromagnetic spectrum. The major advantage of investigating systems immobilized in LbL films is the benefit of characterizing the interaction through available substances in solid state techniques. It was possible to immobilize in LbL films, ERY, and ERYâ¯+â¯TCS. The results show that the growth of the films was linear, indicating the deposition of the same amount of material from the first bilayer without substrate interference. The release analysis showed slow kinetics, which occurred more rapidly for ERY LbL films, probably due to apparent activation energy, which were higher for films with TCS. The combination of TCS, ERY, and laser light (532â¯nm) for photodynamic inactivation of the fungus Candida albicans was analyzed, and the results were promising for future studies in applications, such as coating surfaces of dental implants.
Assuntos
Candida albicans/efeitos dos fármacos , Eritrosina/uso terapêutico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Triclosan/uso terapêutico , Preparações de Ação Retardada , Relação Dose-Resposta a Droga , Eritrosina/administração & dosagem , Eritrosina/farmacocinética , Luz , Fármacos Fotossensibilizantes/administração & dosagem , Fármacos Fotossensibilizantes/farmacocinética , Triclosan/administração & dosagem , Triclosan/farmacocinéticaRESUMO
Bacterial infections have been a major challenge to health. Increasing resistance to antimicrobial agents, according to World Health Organization, could be the major cause of death until 2050. Photodynamic therapy emerges as an alternative in microbial inactivation, due to its selectivity and to decreasing or dismissing antibiotic use. This study aimed at evaluating, in vitro, the internalization of the Methylene Blue and its photodynamic activity against a clinical and ATCC strain of Pseudomonas aeruginosa and Staphyloccocus aureus. Thus, the strains were incubated with MB in concentrations of 100, 300 e 500⯵g/ml and then irradiated with a LED (±660â¯nm) at fluence of 10 and 25â¯J/cm2. The MB internalization was evaluated using a confocal microscope (Zeiss LSM 700), to capture the MB and the DAPI (for DNA staining). It was possible to observe that the MB was internalized by the bacterial cells, in all concentrations tested. The CFU/ml count demonstrated significant reduction (pâ¯≤â¯0,01) at the average 5.0 logs comparing with control group for the two species in all the tested concentrations. In conclusion, the strains tested were capable of internalizing the MB. PDT with MB was able to decrease the growth of the tested strains in vitro, being a promising alternative to the future treatment of infections caused by these species.
Assuntos
Azul de Metileno/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Relação Dose-Resposta a Droga , Azul de Metileno/farmacocinética , Viabilidade Microbiana , Microscopia Confocal , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacocinéticaRESUMO
Metal phthalocyanines are promising components in photodynamic therapy. Aluminum phthalocyanine chloride (AlClPc) has been used to treat oral cancer in mice, human carious tissue, lung cancer cells and other conditions. To overcome the high hydrophobicity of AlClPc, phthalocyanine is often encapsulated in nanoformulations. Despite increased usage, little is known about the pharmacokinetics and biodistribution of AlClPc. The aim of this study was the development and validation of a UHPLC-MS method for the determination of AlClPc in solution after extraction from nanoformulations and biological matrices such as plasma and tissue. The described method has been assayed as to selectivity, linearity, limits of detection and quantification, precision and recovery. The present study is the first to describe the behavior of AlClPc in biological matrices with mass spectrometry as well as the first to describe the chromatographic behavior of AlClPc contaminants. Molecular mass analysis identified dechlorination of AlClPc by both LC/MS and MALDI-MS and an adduct formation in LC/MS. The parameters observed indicated that the method has applicability and robustness for use in biodistribution studies.
Assuntos
Cromatografia Líquida de Alta Pressão/normas , Indóis/sangue , Nanoestruturas/química , Compostos Organometálicos/sangue , Fármacos Fotossensibilizantes/sangue , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/normas , Animais , Disponibilidade Biológica , Biotransformação , Óleo de Rícino/química , Sistemas de Liberação de Medicamentos , Emulsões , Humanos , Interações Hidrofóbicas e Hidrofílicas , Indóis/farmacocinética , Indóis/farmacologia , Rim/efeitos dos fármacos , Rim/metabolismo , Limite de Detecção , Fígado/efeitos dos fármacos , Fígado/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Nanoestruturas/administração & dosagem , Compostos Organometálicos/farmacocinética , Compostos Organometálicos/farmacologia , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacocinética , Fármacos Fotossensibilizantes/farmacologia , Polietilenoglicóis/química , Baço/efeitos dos fármacos , Baço/metabolismo , Distribuição TecidualRESUMO
BACKGROUND: Photodynamic therapy (PDT) may have topical indications. In those cases it is important for a topical photosensitizer to penetrate into the tissue to which it has been applied. This study aimed to compare the penetration of two different concentrations of erythrosine into intact and in vitro decayed dentin samples. METHODS: This in vitro study evaluated erythrosine (0.3 and 5%) penetration into sound (intact) and decayed dentin. A total of 11 dentin discs were prepared and divided into two equal halves, in order to keep one half sound while the other half was submitted to sterilization and an in vitro demineralization model for 5 days. Before erythrosine application, the organic and inorganic composition of all samples was evaluated by Fourier Transform Raman spectroscopy, and after erythrosine application for 30â¯min, the penetration depth was determined by Photoacoustic spectroscopy technique. RESULTS: The results indicated that 0.3% erythrosine showed a higher penetration depth into sound dentin (pâ¯=â¯0.002); and 5% erythrosine higher penetration into decayed dentin (pâ¯<â¯0.001). However considering clinical parameters, no statistically significant difference was found between any of the conditions tested. CONCLUSIONS: Erythrosine demonstrated ability to penetrate into dentin, irrespective of sound or decayed condition. Photoacoustic spectroscopy can be considered a method for estimating the penetration into hard tissues, and in conjunction with Raman spectroscopy, these are effective methods for evaluating the spectral response of dentin. Considering that erythrosine is capable of penetrating into decayed dentin, clinical trials are needed to test the effectiveness of this photosensitizer in Photodynamic therapy and Antimicrobial Photodynamic therapy.
Assuntos
Dentina/metabolismo , Eritrosina/farmacocinética , Técnicas Fotoacústicas/métodos , Fármacos Fotossensibilizantes/farmacocinética , Análise Espectral/métodos , Biofilmes/efeitos dos fármacos , Cárie Dentária/patologia , Relação Dose-Resposta a Droga , Humanos , Dente Serotino , Análise Espectral Raman/métodosRESUMO
In vivo spectrofluorometric analysis during photodynamic therapy (PDT) is a fundamental tool to obtain information about drug bleaching kinetics. Using a portable spectrofluorometer with an excitation source emitting at 400nm wavelength and a spectral analyzer ranging from 500nm to 800nm, the evolution of the meta-tetra(hydroxyphenyl) chlorin (m-THPC) photosensitizer fluorescence spectrum at the tumoral tissue of BALB/c murines with fibrosarcoma located at their flank was followed up. Ex vivo fluorescence measurements of the tumor and skin were also performed with the aim of better characterizing the in vivo signal at different parts of the tumor. PDT was performed employing a LED 637nm light source. Fluorescence at different parts of the tumor and at the tail and armpit of mice was measured immediately after injection and followed daily. The average fluorescence intensity in the tumor reached a maximum after 24-72h. Subsequently, illuminations 24, 48, 72 and 96h post-injection were performed, and the fluorescence was measured immediately before and after each illumination. Eventually, 24h post-illumination, the fluorescence at certain parts of the tumor increased in comparison with that measured immediately after illumination. This effect, named "rebound effect", was due to the new local accumulation of the drug, and was used to perform a second illumination on some mice to increase the amount of photodynamic reaction and significantly improve the PDT outcome. These results are encouraging to optimize PDT in the proposed animal model, thinking about the possible translation to humans.