Pluronic® P123/F127 mixed micelles delivering sorafenib and its combination with verteporfin in cancer cells

Here, we developed Pluronic® P123/F127 (poloxamer) mixed micelles for the intravenous delivery of the anticancer drug sorafenib (SRB) or its combination with verteporfin (VP), a photosensitizer for photodynamic therapy that should complement well the cytotoxicity profile of the chemotherapeutic. SRB loading inside the core of micelles was governed by the drug:poloxamer weight ratio, while in the case of the SRB-VP combination, a mutual interference between the two drugs occurred and only specific ratios could ensure maximum loading efficiency. Coentrapment of SRB did not alter the photophysical properties of VP, confirming that SRB did not participate in any bimolecular process with the photosensitizer. Fluorescence resonance energy-transfer measurement of micelles in serum protein-containing cell-culture medium demonstrated the excellent stability of the system in physiologically relevant conditions. These results were in line with the results of the release study showing a release rate of both drugs in the presence of proteins slower than in phosphate buffer. SRB release was sustained, while VP remained substantially entrapped in the micelle core. Cytotoxicity studies in MDA-MB231 cells revealed that at 24 hours, SRB-loaded micelles were more active than free SRB only at very low SRB concentrations, while at 24+24 hours a prolonged cytotoxic effect of SRB-loaded micelles was observed, very likely mediated by the block in the S phase of the cell cycle. The combination of SRB with VP under light exposure was less cytotoxic than both the free combination and VP-loaded micelles + SRB-loaded micelles combination. This behavior was clearly explainable in terms of micelle uptake and intracellular localization. Besides the clear advantage of delivering SRB in poloxamer micelles, our results provide a clear example that each photochemotherapeutic combination needs detailed investigations on their particular interaction, and no generalization on enhanced cytotoxic effects should be derived a priori

Biotin-targeted Pluronic® P123/F127 mixed micelles delivering niclosamide: A repositioning strategy to treat drug-resistant lung cancer cells

With the aim to develop alternative therapeutic tools for the treatment of resistant cancers, here we propose targeted Pluronic1 P123/F127 mixed micelles (PMM) delivering niclosamide (NCL) as a repositioning strategy to treat multidrug resistant non-small lung cancer cell lines. To build multifunctional PMM for targeting and imaging, Pluronic1 F127 was conjugated with biotin, while Pluronic1 P123 was fluorescently tagged with rhodamine B, in both cases at one of the two hydroxyl end groups. This design intended to avoid any interference of rhodamine B on biotin exposition on PMM surface, which is a key fundamental for cell trafficking studies. Biotin-decorated PMM were internalized more efficiently than non-targeted PMM in A549 lung cancer cells, while very low internalization was found in NHI3T3 normal fibroblasts. Biotin-decorated PMM entrapped NCL with good efficiency, displayed sustained drug release in protein-rich media and improved cytotoxicity in A549 cells as compared to free NCL (P < 0.01). To go in depth into the actual therapeutic potential of NCL-loaded PMM, a cisplatin-resistant A549 lung cancer cell line (CPr-A549) was developed and its multidrug resistance tested against common chemotherapeutics. Free NCL was able to overcome chemoresistance showing cytotoxic effects in this cell line ascribable to nucleolar stress, which was associated to a significant increase of the ribosomal protein rpL3 and consequent up-regulation of p21. It is noteworthy that biotin- decorated PMM carrying NCL at low doses demonstrated a significantly higher cytotoxicity than free NCL in CPr-A549. These results point at NCL-based regimen with targeted PMM as a possible second-line chemotherapy for lung cancer showing cisplatin or multidrug resistance

Evaluation of the physical, chemical and mechanical properties of starch/PVA/bentonite clay films modified with glycidyl methacrylate

Filmes constituídos por amido de mandioca, álcool polivinílico (PVA) e argila Bentonita (BNT) e glicerol como plastificante foram obtidos utilizando o método casting. A fim de aumentar a compatibilidade dos polímeros ao mineral e sua hidrofobicidade, estes foram posteriormente modificados quimicamente utilizando metacrilato de glicidila (GMA) para a substituição dos grupamentos hidroxilas por vinílicos. Este procedimento gerou filmes com excelente estabilidade a sorção de água e permeabilidade ao vapor de água 15% menor quando comparados coms os filmes sem modificação química. Adicionalmente, esta estratégia também resultou na melhora de 25% na resistência a tração, mantendo o mesmo módulo de Young mas, com uma leve diminuição na estabilidade térmica (início de degradação 14ºC menor). A adição da argila BNT gera filmes compósitos com propriedades, mecânicas, térmicas e físicas diferenciadas, contudo há uma dependência entre a quantidade de reforço e a distribuição estrutural das lamelas de argila na matriz. As modificações químicas foram comprovadas por RMN H1, FTIR, DRX e os filmes foram caracterizados por DRX, sorção e permeabilidade a água, ensaios mecânicos (tração, alongamento e módulo de Young) e TGA. A utilização de 1% m/m deste reforço gera compósitos com BNT estruturalmente esfoliado, independentemente da prévia modificação com metacrilato de glicidila. Já a formulação com 5% m/m gerou filmes compósitos intercalados. Qualquer outro teor de reforço exibe estruturas lamelares aglomeradas (empacotadas). Destaca-se que nas formulações reforçadas com 1% de argila, houve melhora em cerca 30% na resistência a tração com permeabilidade a água 60% menor, para cargas maiores de argila. Estes fatos apontam que as modificações propostas neste trabalho geraram propriedades promissoras para a produção de materiais de embalagem biodegradáveis.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP: 2019/01604-

Avaliação das propriedades físicas, químicas e mecânicas de filmes de amido/PVA/argila bentonita modificados com metacrilato de glicidila

Filmes constituídos por amido de mandioca, álcool polivinílico (PVA) e argila Bentonita (BNT) e glicerol como plastificante foram obtidos utilizando o método casting. A fim de aumentar a compatibilidade dos polímeros ao mineral e sua hidrofobicidade, estes foram posteriormente modificados quimicamente utilizando metacrilato de glicidila (GMA) para a substituição dos grupamentos hidroxilas por vinílicos. Este procedimento gerou filmes com excelente estabilidade a sorção de água e permeabilidade ao vapor de água 15% menor quando comparados coms os filmes sem modificação química. Adicionalmente, esta estratégia também resultou na melhora de 25% na resistência a tração, mantendo o mesmo módulo de Young mas, com uma leve diminuição na estabilidade térmica (início de degradação 14ºC menor). A adição da argila BNT gera filmes compósitos com propriedades, mecânicas, térmicas e físicas diferenciadas, contudo há uma dependência entre a quantidade de reforço e a distribuição estrutural das lamelas de argila na matriz. As modificações químicas foram comprovadas por RMN H¹, FTIR, DRX e os filmes foram caracterizados por DRX, sorção e permeabilidade a água, ensaios mecânicos (tração, alongamento e módulo de Young) e TGA. A utilização de 1% m/m deste reforço gera compósitos com BNT estruturalmente esfoliado, independentemente da prévia modificação com metacrilato de glicidila. Já a formulação com 5% m/m gerou filmes compósitos intercalados. Qualquer outro teor de reforço exibe estruturas lamelares aglomeradas (empacotadas). Destaca-se que nas formulações reforçadas com 1% de argila, houve melhora em cerca 30% na resistência a tração com permeabilidade a água 60% menor, para cargas maiores de argila. Estes fatos apontam que as modificações propostas neste trabalho geraram propriedades promissoras para a produção de materiais de embalagem biodegradáveis.Palavras-chave: Bentonita, Amido, PVA, GMA, Compósitos

Síntese, caracterização e avaliação das propriedades fotocatalíticas do WO3 obtido através do método poliol

Esse trabalho mostra uma rota sintética mediada por tetraetilenoglicol (P. E. 325°C) em que utiliza o ácido túngstico, como material de partida.  A formação de uma suspensão estável de cor azul foi observada durante o curso da reação. O tamanho das partículas depende do tempo de síntese, sendo que partículas de tamanho em torno de 400 nm foram obtidas após 15 minutos de reação em tetraetilenoglicol. Os materiais particulados foram então recozidos em diferentes temperaturas: 150°, 350°C, 500°C e 800°C,  e caracterizados por  TGA / DSC, DRX, FT-RAMAN, MEV, Espectroscopia óptica UV-VIS e adsorção de N2 a 77K. Observou-se que a síntese proporcionou WO3 sob fase ortorrômbica que evoluiu para monoclínica após tratamento térmico na temperatura igual ou superior a 350°C. A caracterização morfológica indicou a formação de partículas ovoides quando tratadas em temperaturas em torno de 500°C, temperatura a partir da qual a área superficial diminui drasticamente.  A atividade fotocatalítica para a degradação de rodamina B (rhB, 10 mg.L-1) foi avaliada sob irradiação ultravioleta (6 Watts). Maior atividade foi encontrada para o material tratado a 500°C, sendo que descoramento foi total após 180 minutos na presença de 10 mg desse catalisador.Palavras-chave: Trióxido de tungstênio, Rodamina B, Fotocatálise, Método Poliol.

Biotransformation of Metal-Rich Effluents and Potential Recycle Applications

In this chapter, it was introduced about the metallurgic effluents, and their potential to be converted into some feasible coproducts for industries. Some possibilities to introduce circular economy in the context of metallurgic effluents, and in the same way, some techniques to promote bioremediation using microorganisms and products from them were also described. Reported studies, as well as some perspectives to use metal-rich effluents in agriculture and soil quality improvement, were also shown. Copper effluents were kept as the main candidate for sustainable use, as a potentially interesting material for circular economy approaches

Polymeric Nanoparticles for Cancer Photodynamic Therapy

In chemotherapy a fine balance between therapeutic and toxic effects needs to be found for each patient, adapting standard combination protocols each time. Nanotherapeutics has been introduced into clinical practice for treating tumors with the aim of improving the therapeutic outcome of conventional therapies and of alleviating their toxicity and overcoming multidrug resistance.Photodynamic therapy (PDT) is a clinically approved, minimally invasive procedure emerging in cancer treatment. It involves the administration of a photosensitizer (PS) which, under light irradiation and in the presence of molecular oxygen, produces cytotoxic species. Unfortunately, most PSs lack specificity for tumor cells and are poorly soluble in aqueous media, where they can form aggregates with low photoactivity. Nanotechnological approaches in PDT (nanoPDT) can offer a valid option to deliver PSs in the body and to solve at least some of these issues. Currently, polymeric nanoparticles (NPs) are emerging as nanoPDT system because their features (size, surface properties, and release rate) can be readily manipulated by selecting appropriate materials in a vast range of possible candidates commercially available and by synthesizing novel tailor-made materials. Delivery of PSs through NPs offers a great opportunity to overcome PDT drawbacks based on the concept that a nanocarrier can drive therapeutic concentrations of PS to the tumor cells without generating any harmful effect in non-target tissues. Furthermore, carriers for nanoPDT can surmount solubility issues and the tendency of PS to aggregate, which can severely affect photophysical, chemical, and biological properties. Finally, multimodal NPs carrying different drugs/bioactive species with complementary mechanisms of cancer cell killing and incorporating an imaging agent can be developed.In the following, we describe the principles of PDT use in cancer and the pillars of rational design of nanoPDT carriers dictated by tumor and PS features. Then we illustrate the main nanoPDT systems demonstrating potential in preclinical models together with emerging concepts for their advanced design

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

Pluronic^® mixed micelles as efficient nanocarriers for benzoporphyrin derivatives applied to photodynamic therapy in cancer cells

In this study we attempted to develop Pluronic micelles delivering the photodynamic therapy photosensitizers benzoporphyrin derivatives (BPD). The BPD A-ring (BPDMA or Verteporfin®, the active drug of FDA/USA approved Visudyne®), its regioisomer ring-B (BPDMB, not used in Visudyne® formulation due its poor solubility) and a BPDMA/BPDMB mixture (BPD-Mixt) were formulated in Pluronic P123 or F127 as well as P123/F127 mixed micelles at two different mass ratios. P123/F127 presented the lowest critical micelle concentration showing high stability due synergistic aggregation of P123 and F127. Mixed micelles allowed the encapsulation of BPD as monomers enhancing their photophysical properties and stability during time even under diluted conditions. High loading was attributed to the strong hydrophobic affinity of BPD for micelle core especially in the binary system due synergistic aggregation of P123 and F127 demonstrating the high potential of these micelles to encapsulate hydrophobic drugs. The in vitro assays showed a photo-activity of BPD-Mixt comparable to that of BPDMA against HeLa and A549 cancer cells under red light. The use of BPD-mixed formulations avoids the complex separation steps of these regioisomers and implies in cost reduction. The proposed system allies costs reduction and photodynamic efficiency, which stimulates further development on this nanosystem and may be of clinical interest for cancer PDT