Photo-responsive polymeric micelles for the light-triggered release of curcumin targeting antimicrobial activity

Nanocarriers have been successfully used to solubilize, deliver, and increase the bioavailability of curcumin (CUR), but slow CUR release rates hinder its use as a topical photosensitizer in antimicrobial photodynamic therapy. A photo-responsive polymer (PRP) was designed for the light-triggered release of CUR with an effective light activation-dependent antimicrobial response. The characterization of the PRP was compared with non-responsive micelles comprising Pluronics™ P123 and F127. According to the findings, the PRP formed photo-responsive micelles in the nanometric scale (< 100 nm) with a lower critical micelle concentration (3.74 × 10−4 M−1, 5.8 × 10−4 M−1, and 7.2 × 10−6 M−1 for PRP, F127, P123, respectively, at 25°C) and higher entrapment efficiency of CUR (88.7, 77.2, and 72.3% for PRP, F127, and P123 micelles, respectively) than the pluronics evaluated. The PRP provided enhanced protection of CUR compared to P123 micelles, as demonstrated in fluorescence quenching studies. The light-triggered release of CUR from PRP occurred with UV light irradiation (at 355 nm and 25 mW cm−2) and a cumulative release of 88.34% of CUR within 1 h compared to 80% from pluronics after 36 h. In vitro studies showed that CUR-loaded PRP was non-toxic to mammal cell, showed inactivation of the pathogenic microorganisms Candida albicans, Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus aureus, and decreased biofilm biomass when associated with blue light (455  nm, 33.84 J/cm2). The findings show that the CUR-loaded PRP micelle is a viable option for antimicrobial activity

Preparation, characterization and properties of liposomes containing the a-cyano-4-hydroxycinnamic acid and the photosensitizer agent AlClPc: A new specific carrier system with synergistic action applied to photodynamic therapy

Ftalocianina de alumínio-cloro (AlClPc) é um fotossensibilizador de segunda geração em terapia fotodinâmica (TFD) caracterizado por seu caráter anfifílico e tendência de auto-agregação em meio aquoso, o que prejudica seu potencial de aplicação. O aCHC é um substrato de transportadores de monocarboxilato (MCT) superexpresso em células de MCF-7. Objetivando a solubilização da AlClPc e aumento de internalização em tecidos neoplásicos nos propomos aqui o uso de DSPC e DOPC em diferentes proporções para formar vesículas lipidicas mistas (LV) na presença de aCHC como sistemas veiculadores de fármaco. Lv foi preparado pelo método de injeção etanólica e formou vesículas de dimensões nanométricas (aproximadamente 100 nm) com bom índice de polidispersão, valores negativos de potencial zeta e estáveis em meio aquoso por mais de 50 dias. AlClPc se complexou com o fosfato das LV o que conferiu uma localização interfacial às moléculas de AlClPc como demonstrado pelos resultados de supressão de fluorescência. Medidas de anisotropia, fluorescência estática e resolvida no tempo corroboram com estes resultados e demonstram que a auto-agregação da AlClPc ocorre mesmo em lipossomas. Entretanto, a veiculação da AlClPc por LV em carcinoma de células escamosas oral (OSCC) levou a um processo de desagregação demonstrado por (FLIM). Este incrível comportamento é novo e aumenta o conhecimento científico sobre o mecanismo intracelular de ação de fotossensibilizadores em TFD. Em TFD, ambos os sistemas LVIII+AlClPc e LVIII+AlClPc+aCHC não apresentaram toxicidade no escuro no período de incubação de 3 h com as concentrações de lipídios, AlClPc e aCHC iguais a 0,15 mmol/L, 0,5 umol/L e 10,0 umol/L, respectivamente. De maneira inesperada, o sistema LVIII+AlClPc foi mais eficiente em TFD que o sistema LVIII+AlClPc+aCHC, devido ao caráter antioxidante do aCHC. Estes resultados abrem uma nova perspectiva do potencial uso de LV-AlClPc para o tratamento fotodinâmico.Aluminum phthalocyanine chloride (AlClPc) is a second-generation photodynamic therapy (PDT) photosensitizer characterized for its high hydrophobicity and self-aggregation tendency in aqueous media, which hampers its potential application. The aCHC is a monocarboxylate transporter substrate (MCT) overexpressed in MCF-7 line cell. Aiming at AlClPc solubilization and uptake increase we proposed here the use of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) at different proportions to form mixed lipid vesicles (LV) in presence of aCHC as a drug delivery system. LV were prepared by ethanol injection method and formed nano-sized vesicles (about 100 nm) with suitable polydispersity index, negative zeta potential, and stable in aqueous medium for at least 50 days. AlClPc strongly interacts with LV (high binding constant values), especially due to aluminum-phosphate specific interactions, which gives a surface localization to AlClPc molecules as demonstrated by fluorescence quenching data. Anisotropy, static and time-resolved fluorescence measurements corroborated with these results and demonstrated that AlClPc self-aggregation occurred even in the liposomes. However, formulation uptake by oral squamous cell carcinoma (OSCC) the AlClPc suffered a disaggregation process demonstrated by fluorescence life-time imaging microscopy. This amazing behavior is new and increases the scientific knowledge about the intracellular mechanism of action of PDT photosensitizers. In PDT, both systems LVIII+AlClPc e LVIII+AlClPc+aCHC did not show toxicity in the dark at 3 h incubation period with lipid, AlClPc, and aCHC concentrations equal at 0,15 mmol/L, 0,5 umol/L, and 10,0 umol/L, respectively. Unexpectedly, the system LVIII+AlClPc was more effective in PDT that the LVIII+AlClPc+aCHC, devido ao caráter antioxidante do aCHC due to the antioxidant character of aCHC. These results open a new perspective to the potential use of ALClPc-LV formulations for photodynamic treatment

B6 vitamers as generators and scavengers of reactive oxygen species

B6 vitamers are important natural compounds for human life, principally for amino acid metabolism, and have been reported as potent scavenger molecules of both endogenous and exogenous reactive oxygen species (ROS). However, under UV-light excitation from UVC (200–280 nm) to UVA (315–400 nm), B6 vitamers act as endogenous photosensitizers that promote both cellular photodamage and enzyme inactivation. This occurs through classical photochemical mechanisms mediated by ROS, toxic photoproducts, and the formation of adducts with active protein sites. This minireview aimed to present and discuss the dual roles of B6 vitamers as generators and scavengers of singlet oxygen (an important reactive species in photodynamic processes) and various other ROS. We also examine the basic photophysical, photochemical, and protolytic equilibria principles of B6 vitamers and their role as photodamaging and photoprotecting compounds in UV-light mediated photobiological processes

Characterization of glioblastoma spheroid models for drug screening and phototherapy assays

Glioblastoma (GBM) spheroids present as a promising alternative testing platform for in vivo models of brain cancer, aiming at the 3Rs principle of animal use in pre-clinical trials. However, the poor characterization and irreproducibility of various GBM-based spheroids have hampered the achievement of statistically relevant data for drug screening and photodynamic therapy (PDT) assays. In a previous study (Calori et al., 2022), we defined the conditions for inducing tight compaction of U87MG, T98G, UW473, A172, and U251 GBM cell lines into reproducible 3D spheroids using type I collagen as an extracellular matrix. Herein, GBM spheroids were characterized in their growth profile, proliferation rate, cell viability, necrotic nucleus, and cell death mechanisms and applied in PDT assays. Driven by proliferation, U87MG and A172 spheroids grew with time, at least up to day 12, whereas T98G and U251 demonstrated little change. The order of cell viability of spheroids over time was U87MG > A172 > U251 > T98G. A necrotic nucleus was formed over time for all cell lines, as shown by histological assays and the inverse size-cell viability relationship. When subjected to PDT, all spheroids showed a dose-dependent response, with an excellent dose-light-cell viability correlation. The higher light dose required for PDT of GBM 3D spheroids compared with the simplest 2D monolayers and its proximity to in vivo dose response in the literature is proof of its applicability in pre-clinical tests to reduce the use of animals in research

Multifunctional theranostic Pluronic mixed micelles improve targeted photoactivity of Verteporfin in cancer cells

Nanotechnology development provides new strategies to treat cancer by integration of different treatment modalities in a single multifunctional nanoparticle. In this scenario, we applied the multifunctional Pluronic P123/F127 mixed micelles for Verteporfin-mediated photodynamic therapy in PC3 and MCF-7 cancer cells. Micelles functionalization aimed the targeted delivery by the insertion of biotin moiety on micelle surface and fluorescence image-based through rhodamine-B dye conjugation in the polymer chains. Multifunctional Pluronics formed spherical nanoparticulated micelles that efficiently encapsulated the photosensitizer Verteporfin maintaining its favorable photophysical properties. Lyophilized formulations were stable at least for 6months and readily reconstituted in aqueous media. The multifunctional micelles were stable in protein-rich media due to the dual Pluronic mixed micelles characteristic: high drug loading capacity provided by its micellar core and high kinetic stability due its biocompatible shell. Biotin surface functionalized micelles showed higher internalization rates due biotin-mediated endocytosis, as demonstrated by competitive cellular uptake studies. Rhodamine B-tagged micelles allowed monitoring cellular uptake and intracellular distribution of the formulations. Confocal microscopy studies demonstrated a larger intracellular distribution of the formulation and photosensitizer, which could drive Verteporfin to act on multiple cell sites. Formulations were not toxic in the dark condition, but showed high Verteporfin-induced phototoxicity against both cancer cell lines at low drug and light doses. These results point Verteporfin-loaded multifunctional micelles as a promising tool to further developments in photodynamic therapy of cancer

Data_Sheet_1_Photo-responsive polymeric micelles for the light-triggered release of curcumin targeting antimicrobial activity.docx

Nanocarriers have been successfully used to solubilize, deliver, and increase the bioavailability of curcumin (CUR), but slow CUR release rates hinder its use as a topical photosensitizer in antimicrobial photodynamic therapy. A photo-responsive polymer (PRP) was designed for the light-triggered release of CUR with an effective light activation-dependent antimicrobial response. The characterization of the PRP was compared with non-responsive micelles comprising Pluronics™ P123 and F127. According to the findings, the PRP formed photo-responsive micelles in the nanometric scale (−4 M−1, 5.8 × 10−4 M−1, and 7.2 × 10−6 M−1 for PRP, F127, P123, respectively, at 25°C) and higher entrapment efficiency of CUR (88.7, 77.2, and 72.3% for PRP, F127, and P123 micelles, respectively) than the pluronics evaluated. The PRP provided enhanced protection of CUR compared to P123 micelles, as demonstrated in fluorescence quenching studies. The light-triggered release of CUR from PRP occurred with UV light irradiation (at 355 nm and 25 mW cm−2) and a cumulative release of 88.34% of CUR within 1 h compared to 80% from pluronics after 36 h. In vitro studies showed that CUR-loaded PRP was non-toxic to mammal cell, showed inactivation of the pathogenic microorganisms Candida albicans, Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus aureus, and decreased biofilm biomass when associated with blue light (455  nm, 33.84 J/cm2). The findings show that the CUR-loaded PRP micelle is a viable option for antimicrobial activity.</p