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A modular nanoplatform with an enhanced bio-imaging and chemodynamic efficiency was constructed using the core–shell synergistic effect of carbon dots.

The novel Mn-doped graphene quantum dot nanoplatform induces autophagy-associated tumour cell death by lysosome impairment.

This study aimed to characterize the aluminum phthalocyanine chloride (AlClPc) encapsulated in chitosan nanoparticles (CN) and apply it in antimicrobial photodynamic therapy (aPDT) on multispecies biofilm composed of Streptococcus mutans, Lactobacillus casei, and Candida albicans to analyze the antimicrobial activity and lactate production after treatment. Biofilms were formed in 24-well polystyrene plates at 37 °C for 48 h under microaerophilia. The following groups were evaluated (n = 9): as a positive control, 0.12% chlorhexidine gluconate (CHX); phosphate-buffered saline (PBS) as a negative control; 2.5% CN as release vehicle control; the dark toxicity control of the formulations used (AlClPc and AlClPc + CN) was verified in the absence of light; for aPDT, after 30 min incubation time, the photosensitizers at a final concentration of 5.8 × 10–3 mg/mL were photoirradiated for 1 min by visible light using a LED device (AlClPc + L and AlClPc + CN + L) with 660 nm at the energy density of 100 J/cm2. An in vitro kit was used to measure lactate. The biofilm composition and morphology were observed by scanning electron microscopy (SEM). The antimicrobial activity was analyzed by quantifying colony forming units per mL (CFU/mL) of each microorganism. Bacterial load between groups was analyzed by ANOVA and Tukey HSD tests (α = 0.05). A lower lactate dosage was observed in the aPDT AlClPc + CN + L and CHX groups compared to the CN and AlClPc groups. The aPDT mediated by the nanoconjugate AlClPc + CN + L showed a significant reduction in the viability of S. mutans (3.18 log10 CFU/mL), L. casei (4.91 log10 CFU/mL), and C. albicans (2.09 log10 CFU/mL) compared to the negative control PBS (p 0.05). The aPDT mediated by the nanoconjugate AlClPc + CN + L was efficient against the biofilm of S. mutans, L. casei, and C. albicans.

In natural systems like photosynthetic organisms and photo-active enzymes, the spatial organization of chromophores is critical for efficient light harvesting and bio-catalysis. Inspired by nature, a novel modular nanoplatform with both biological imaging and therapeutic functions is constructed by taking advantage of the intrinsic core-shell structure of Fe-decorated carbon dots. Light-harvesting chromophores with deep-red photoluminescence are densely packed into the carbon core. Simultaneously, the atomically dispersed Fe

A number of different metallic nanoparticles have been extensively investigated in recent years based on their diverse potential in biomedical and cancer applications, antibacterial activity, and chemical properties. Here, silver nitroprusside nanoparticles (AgNPs) were prepared from silver nitrate and sodium nitroprusside, and their anticancer activity was evaluated. AgNPs were prepared and characterized by Fourier transform infrared (FTIR), X-ray diffraction (DRX), scanning electron microscopy (SEM), X-ray dispersive energy spectroscopy (EDX), and transmission electron microscopy (TEM). In vitro tests were performed using two breast cancer cell lines, including a non-malignant breast epithelial cell line (MCF-10A) and a breast cancer cell line (MCF-7). The results obtained through cytotoxicity assays (MTT and resazurin) and bright-field microscopy revealed that AgNPs (3.0 mg/mL) exhibited specific selectivity for non-malignant breast epithelial cells. They were toxic to tumorigenic cells (MCF-7) with a lower selective toxicity to non-malignant breast epithelial cells (MCF-10A). In vivo imaging system (IVIS) imaging demonstrated the antitumor activity of AgNPs (3.0 mg/mL) compared to that of the control, and this was confirmed through histological analysis without any potential systemic toxicity. These results suggest that AgNPs possess potential anticancer applications in breast cancer treatment.

Parkinson's disease (PD) is a progressive and chronic neurodegenerative disease of the central nervous system. Early treatment for PD is efficient; however, long-term systemic medication commonly leads to deleterious side-effects. Strategies that enable more selective drug delivery to the brain using smaller dosages, while crossing the complex brain-blood barrier (BBB), are highly desirable to ensure treatment efficacy and decrease/avoid unwanted outcomes. Our goal was to design and test the neurotherapeutic potential of a forefront nanoparticle-based technology composed of albumin/PLGA nanosystems loaded with dopamine (ALNP-DA) in 6-OHDA PD mice model. ALNP-DA effectively crossed the BBB, replenishing dopamine at the nigrostriatal pathway, resulting in significant motor symptom improvement when compared to Lesioned and L-DOPA groups. Notably, ALNP-DA (20 mg/animal dose) additionally up-regulated and restored motor coordination, balance, and sensorimotor performance to non-lesioned (Sham) animal level. Overall, ALNPs represent an innovative, non-invasive nano-therapeutical strategy for PD, considering its efficacy to circumvent the BBB and ultimately deliver the drug of interest to the brain.

Recently, photodynamic therapy (PDT) has been extensively applied in clinical and coadjuvant treatment of various kinds of tumors. However, the photosensitizer (PS) of PDT still lack of high production of singlet oxygen (1O2), low cytotoxicity and high biocompatibility. Herein, we propose a facile method for establishing a new core-shell structured Sn nanocluster@carbon dots (CDs) PS. Firstly, Sn4+@S-CDs complex is synthesized using the sulfur-doped CDs (S-CDs) and SnCl4 as raw materials, and subsequently the new PS (Sn nanocluster@CDs) is obtained after vaporization of Sn4+@S-CDs solution. Remarkably, the obtained Sn nanocluster@CDs show an enhanced fluorescence as well as a higher 1O2 quantum yield (QY) than S-CDs. The high 1O2 QY (58.3%) irradiated by the LED light (400–700 nm, 40 mW/cm2), induce the reduction of 4T1 cancer cells viability by 25%. More intriguingly, no visible damage happens to healthy cells, with little impact on liver tissue due to renal excretion, both in vitro and in vivo experiments demonstrate that Sn nanocluster@CDs may become a promising PS, owning a high potential for application in PDT.

The use of DNA as a functional biomaterial for therapeutic, diagnostic, and drug delivery applications has been prominent in recent years, but its use as a scaffold for tissue regeneration is still limited. This study aimed to evaluate the biocompatibility and interaction of DNA-based polymeric films (DNA-PFs) with primary human fibroblasts (PHF) for regenerative medicine and wound healing purposes. The morphological characterization of the films was performed by scanning electron microscopy, SEM-energy-dispersive X-ray spectroscopy, and atomic force microscopy analysis. Cell viability, cell cycle kinetics, oxidative stress, and migration studies were carried out at 48 and 72 hr of incubation and compared to control cells. Cell adhesion was impaired in the first 24 hr, DNA-PFs with higher concentrations of DNA (1.0 and 2.0 g/L) this effect was not seen in DNA-PFs (0.5 g/L), explained by the difference in topography and roughness of DNA-PFs, but it was overcome after 48 hr of incubation. PHF seeded on DNA films showed higher proliferation and migration rates than the control after 48 hr of incubation, with the maintenance of cell morphology and lower cytotoxicity and oxidative stress during the evaluation time. Therefore, these results indicate that DNA-PFs are highly biocompatible and provide a suitable microenvironment for dermal fibroblasts to maintain their activity, helping build new and more complex biomaterials suitable for future tissue repair applications.

Photodynamic therapy (PDT) is a minimally invasive clinical protocol that combines a nontoxic photosensitizer (PS), appropriate visible light, and molecular oxygen for cancer treatment. This triad generates reactive oxygen species (ROS)

Carbonized polymer dots (CPDs) are promising fluorescent probes for bioimaging, biomarkers, sensors, etc. Here a type of thermosensitive CPD (ts-CPD) was synthesized via a solvothermal method from ...

Melanoma spheroid-loaded 3D skin models allow for the study of crucial tumor characteristics and factors at a superior level because the neoplastic cells are integrated into essential human skin components, permitting tumor-skin model communication. Herein, we designed a melanoma-containing artificial dermis by inserting multicellular tumor spheroids from the metastatic phase of WM 1617 melanoma cells into an artificial dermis. We cultured multicellular melanoma spheroids by hanging drop method (250 cells per drop) with a size of 420 μm in diameter after incubation for 14 days. These spheroids were integrated into the dermal equivalents that had been previously preparedwith a type-I collagen matrix and healthy fibroblasts. The melanoma spheroid cells invaded and proliferated in the artificial dermis. Spheroids treated with a 1.0 μmol/L aluminum chloride phthalocyanine nanoemulsion in the absence of light showed high cell viability. In contrast, under irradiation with visible red light (660 nm) at 25 J/cm

Photodynamic therapy (PDT) is a promising and emerging method for the treatment of cancer. Usually, Type II PDT is used in the clinic, and mainly involves three key elements: a photosensitizer, molecular oxygen and laser light. However, it is known that tumor tissue is deficient in oxygen molecules which is why Type I PDT is mostly preferred in the therapy of tumors in which the hypoxic tissue plays a major role. Fluorescent carbon dots (CDs) have shown great potential in cancer theranostics, acting as bioimaging agents and photosensitizers. Herein, we have synthesized novel kinds of fluorine and nitrogen co-doped carbon dots (F,NCDs) that emit bright green fluorescence under ultra-violet light. The F,NCDs have excellent water solubility and low cytotoxicity. They can generate hydroxyl radicals (˙OH) and superoxide anions (˙O

Made available in DSpace on 2020-12-12T01:25:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-06-07 Here we report a novel strategy to crosslink the surface of sulfonated-carbon dots (S-CDs) by complexing SnCl4with sulfonate groups (-SO3-) on the CDs in aqueous solution. The S-CDs show an average photoluminescence (PL) quantum yield of 21% and a mean diameter of 3.8 nm. After being complexed with Sn4+, the as-obtained Sn@S-CDs present a reduced size of 1.8 nm and a higher PL quantum yield of 32%. More interestingly, the Sn@S-CDs show an enhanced singlet oxygen (1O2) quantum yield as high as 37% compared to that of the S-CDs (27%). In the HepG2 cell line as a model, the Sn@S-CDs exhibit a remarkable cell imaging effect andin vitroPDT efficiency. Therefore, our study proposes a simple but effective cross-linking strategy to synthesize CDs incorporated with metal ions, for the purpose of achieving an enhanced fluorescence intensity and a higher1O2quantum yield. School of Chemistry and Chemical Engineering Anhui Key Laboratory of Modern Biomanufacturing Anhui University Department of Bioprocess and Biotechnology Faculty of Pharmaceutical Sciences of FCF/UNESP Department of Chemistry Center of Nanotechnology and Tissue Engineering-Photobiology and Photomedicine Research Group Faculty of Philosophy Sciences and Letters of Ribeirão Preto University of São Paulo Changchun Institute of Applied Chemistry Chinese Academy of Sciences, 5625Renmin Street Department of Bioprocess and Biotechnology Faculty of Pharmaceutical Sciences of FCF/UNESP

Objective To evaluate the effects of photodynamic therapy (PDT) on the dura mater using the photosensitizers aluminum chloride phthalocyanine and methylene blue in in vivo assays. Methods Fifty-six male Wistar rats were divided into two groups; one submitted to PDT and the other submitted to the photosensitizers without their photoactivation (control). The photosensitizers were applied to the dura mater after laminectomy at the T10 level. The methods used for assessment were the Basso, Beattie and Bresnahan (BBB) functional evaluation scale and study of the dura mater by light microscopy. Results No changes in motor activity were observed in the animals submitted to PDT compared to control. Histological and pathological evaluation did not show any differences between the group exposed to activated photosensitizers and the control group with regard to the inflammatory process and tissue necrosis. Conclusion The joint use of PDT with the photosensitizing pharmaceuticals aluminum chloride phthalocyanine and methylene blue did not induce any clinical neurotoxic effects or histological changes in the dura mater of the animals studied. Level de evidence V; Expert Opnion. RESUMO Objetivo Avaliar os efeitos da terapia fotodinâmica (PDT) na dura-máter usando os fotossensibilizadores cloreto de alumínio ftalocianina e azul de metileno em ensaios in vivo. Métodos Cinquenta e seis ratos Wistar machos divididos em dois grupos; um submetido à PDT e o outro submetido aos fotossensibilizadores sem a fotoativação (controle). Os fotossensibilizadores foram aplicados sobre a dura-máter depois de laminectomia no nível T10. Os métodos de avaliação usados foram a escala de avaliação funcional de Basso, Beattie e Bresnahan (BBB) e o estudo da dura-máter por microscopia óptica. Resultados Não foram observadas alterações da atividade motora dos animais submetidos à PDT com relação ao grupo controle. A avaliação histológica e histopatológica não mostrou diferenças entre o grupo exposto aos fotossensibilizadores ativados e o grupo controle, com relação ao processo inflamatório e à necrose tecidual. Conclusões O uso conjunto de PDT e os fármacos fotossensibilizantes cloreto de alumínio ftalocianina e azul de metileno não induziu efeitos neurotóxicos clínicos e/ou alterações histológicas sobre a dura-máter dos animais estudados. Nível de evidência V; Opinião de Especialista. RESUMEN Objetivo Evaluar los efectos de la terapia fotodinámica (PDT) en la duramadre utilizando los fotosensibilizadores de ftalocianina de aluminio clorada y azul de metileno en ensayos in vivo. Métodos Cincuenta y seis ratas Wistar machos se dividieron en 2 grupos; uno fue sometido a PDT y el otro sometido a fotosensibilizadores sin fotoactivación (control). Los fotosensibilizadores se aplicaron a la duramadre después de la laminectomía en el nivel T10. Los métodos de evaluación utilizados fueron la escala de evaluación funcional de Basso, Beattie y Bresnahan (BBB) y el estudio de la duramadre mediante microscopía óptica. Resultados No hubo cambios en la actividad motora de los animales sometidos a PDT en relación con el grupo de control. La evaluación histológica e histopatológica no mostró diferencias entre el grupo expuesto a fotosensibilizadores activados y el grupo de control con respecto al proceso inflamatorio y la necrosis tisular. Conclusiones El uso conjunto de PDT con las sustancias fotosensibilizadores ftalocianina de aluminio clorada y azul de metileno no indujo efectos neurotóxicos clínicos o cambios histológicos en la duramadre de los animales estudiados. Nivel de evidencia V; Opinión del Especialista.

Introduction: Prodrugs have been used to improve the selectivity and efficacy of cancer therapy by targeting unique abnormal markers that are overexpressed by cancer cells and are absent in normal tissues. In this context, different strategies have been exploited and new ones are being developed each year. Areas covered: In this review, an integrated view of the potential use of prodrugs in targeted cancer therapy is provided. Passive and active strategies are discussed in light of the advantages of each one and some successful examples are provided, as well as the clinical status of several prodrugs. Among them, antibody-drug conjugates (ADCs) are the most commonly used. However, several drawbacks, including limited prodrug uptake, poor pharmacokinetics, immunogenicity problems, difficulties in selective targeting and gene expression, and optimized bystander effects limit their clinical applications. Expert opinion: Despite the efforts of different companies and research groups, several drawbacks, such as the lack of relevant in vivo models, complexity of the human metabolism, and economic limitations, have hampered the development of new prodrugs for targeted cancer therapy. As a result, we believe that the combination of prodrugs with cancer nanotechnology and other newly developed approaches, such as aptamer-conjugated nanomaterials, are efficient strategies.

Made available in DSpace on 2019-10-06T16:21:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-05-01 Universidade Estadual Paulista Curcumin, a natural compound has several antineoplastic activities and is a promising natural photosensitizer used in photodynamic therapy. However, its low solubility in physiological medium limit the clinical use of curcumin. This study aimed to analyze the action of curcumin-nanoemulsion, a new and well-designed Drug Delivery System (DDS+) molecule, used as a photosensitizing agent in photodynamic therapy in an in vitro breast cancer model, MCF-7 cells. The empty nanoemulsion fulfils all necessary requirements to be an excellent DDS. Furthermore, the use of curcumin-nanoemulsion in photodynamic therapy resulted in a high phototoxic effect after activation at 440 nm, decreasing to

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.

1,8-cineole is a monoterpene commonly used by the food, cosmetic, and pharmaceutical industries owing to its flavor and fragrances properties. In addition, this bioactive monoterpene has demonstrated bactericidal and fungicidal activities. However, such activities are limited due to its low aqueous solubility and stability. This study aimed to develop nanoemulsion containing cineole and assess its stability and antibacterial activity in this context. The spontaneous emulsification method was used to prepare nanoemulsion (NE) formulations (F1, F2, F3, F4, and F5). Following the development of NE formulations, we chose the F1 formulation that presented an average droplet size (in diameter) of about 100 nm with narrow size distribution (PdI0.2) and negative zeta potential (∼ - 35 mV). According to the analytical centrifugation method with photometric detection, F1 and F5 formulations were considered the most stable NE with lower droplet migration velocities. In addition, F1 formulation showed high incorporation efficiency (80 %) and TEM analyses demonstrated nanosized oil droplets with irregular spherical shapes and without any aggregation tendency. Antibacterial activity assessment showed that F1 NE was able to enhance the cineole action against Staphylococcus aureus, Enterococcus faecalis, and Streptococcus pyogenes. Therefore, using a simple and reproducible method of low energy emulsification we designed a stable nanoemulsion containing 1,8-cineole with improved antibacterial activity against Gram-positive strains.

The present study examines the designer of DNA polymeric films (DNA-PFs) associated with aluminum chloride phthalocyanine (AlClPc) (DNA-PFs-AlClPc), as a promising drug delivery system (DDS), applicable for breast cancer treatment and early-stage diagnosis using photodynamic therapy (PDT). This study starts evaluating (MCF7) as a model for breast cancer cell behavior associated with DNA-PFs. Analyses of the morphological behaviors, biochemical reaction, and MCF7 cell adhesion profile on DNA-PFs were evaluated. SEM and AFM analysis allowed the morphological characterization of the DNA-PFs. Cell viability and cell cycle kinetics studies indicate highly biocompatible material capable of anchoring MCF7 cells, allowing the attachment and support of cell in the same structure where the insertion of AlClPc (DNA-PFs-AlClPc). The application of visible light photoactivation based on classical PDT protocol over the DNA-PFs-AlClPc showed a reduction in cell viability with increased cell death proportional to the fluency energy range from 600, 900, and 1800 mJ cm−2. The 3D organoid system mimics the tumor microenvironment which was precisely observed in human breast cancer in early-stage progression in the body. The results observed indicate that the viability was reduced by more than 80% in monolayer culture and around 50% in the 3D organoid cell culture at the highest energy fluency (1800 mJ cm−2). We could also point out that with low energy fluency (100 mJ cm−2,), the DNA-PFs-AlClPc did not show a cytotoxic effect on MCF7 cells, enabling this user dose for the photodiagnosis of early-stage human breast cancer detection in the initial stage of progression.

Among several optical non-contact thermometry methods, luminescence thermometry is the most versatile approach. Lanthanide-based luminescence nanothermometers may exploit not only downshifting, but also upconversion (UC) mechanisms. UC-based nanothermometers are interesting for biological applications: they efficiently convert near-infrared radiation to visible light, allowing local temperatures to be determined through spectroscopic investigation. Here, we have synthesized highly crystalline Er3+, Yb3+ co-doped upconverting KGd3F10 nanoparticles (NPs) by the EDTA-assisted hydrothermal method. We characterized the structure and morphology of the obtained NPs by transmission electron microscopy, X-ray diffraction, Raman spectroscopy, and dynamic light scattering. Nonlinear spectroscopic studies with the Er3+, Yb3+: KGd3F10 powder showed intense green and red emissions under excitation at 980 and 1,550 nm. Two- and three-photon processes were attributed to the UC mechanisms under excitation at 980 and 1,550 nm. Strong NIR emission centered at 1,530 nm occurred under low 980-nm power densities. Single NPs presented strong green and red emissions under continuous wave excitation at 975.5 nm, so we evaluated their use as primary nanothermometers by employing the Luminescence Intensity Ratio technique. We determined the temperature felt by the dried NPs by integrating the intensity ratio between the thermally coupled 2H11/2→4I15/2 and 4S3/2→4I15/2 levels of Er3+ ions in the colloidal phase and at the single NP level. The best thermal sensitivity of a single Er3+, Yb3+: KGd3F10 NP was 1.17% at the single NP level for the dry state at 300 K, indicating potential application of this material as accurate nanothermometer in the thermal range of biological interest. To the best of our knowledge, this is the first promising thermometry based on single KGd3F10 particles, with potential use as biomarkers in the NIR-II region.

Made available in DSpace on 2021-06-25T10:37:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-01 Objective: Assess a single local application of curcumin-loaded nanoparticles as an adjunct to scaling and root planing (SRP) in nonsurgical periodontal treatment (NPT). Materials and methods: Twenty healthy subjects with periodontitis received SRP+PLGA/PLA nanoparticles loaded with 50 μg of curcumin (N-Curc) or SRP+empty nanoparticles. Probing pocket depth (PPD), clinical attachment level (CAL), and bleeding on probing (BOP) were monitored at baseline, 30, 90, and 180 days. IL-1α, IL-6, TNFα, and IL-10 in the gingival crevicular fluid (GCF) were assessed by ELISA, and counts of 40 bacterial species were determined by DNA hybridization at baseline, 3, 7, and 15 days post-therapy. Results: PPD, CAL, and BOP were similarly and significantly improved in both experimental groups. There was no difference in GCF cytokine levels between experimental groups, although IL-6 was decreased at 3 days only in the N-Curc group. NPT reduced counts of red complex bacterial species in both groups. Veillonella Parvula counts increased significantly only in N-Curc group at 7 days, whereas Aggregatibacter actinomycetemcomitans counts increased significantly only in the control group from day 3 to day 15. Conclusion: We conclude that a single local administration of nanoencapsulated curcumin in periodontally diseased sites had no additive benefits to NPT. Clinical relevance: Our results showed that a single local application of curcumin-loaded nanoparticles associated with nonsurgical periodontal therapy did not improve clinical outcomes. Hence, our findings do not support the use of curcumin as an adjunct to nonsurgical periodontal therapy. Department of Diagnosis and Surgery School of Dentistry at Araraquara Sao Paulo State University (UNESP) Department of Bioprocess and Biotechnology Faculty of Pharmaceutical Sciences of Araraquara Sao Paulo State University (UNESP) Department of Chemistry Center of Nanotechnology and Tissue Engineering- Photobiology and Photomedicine Research Group Faculty of Philosophy Sciences and Letters of Ribeirao Preto University of Sao Paulo Biostatistics University of Michigan School of Public Health Department of Periodontology and Oral Implantology Dental Research Division Universidade de Guarulhos - UNG Department of Diagnosis and Surgery School of Dentistry at Araraquara Sao Paulo State University (UNESP) Department of Bioprocess and Biotechnology Faculty of Pharmaceutical Sciences of Araraquara Sao Paulo State University (UNESP)

Made available in DSpace on 2021-06-25T15:04:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Project PRONON-SIPAR Breast cancer is the most common neoplasm among women but thanks to innovative therapies, patients' prognosis has considerably improved. In this aspect, nanotechnology has been applied for cancer therapy aiming to reduce its usual side effects. In this study we aimed to evaluate the effects of nanoemulsions containing photosensitizer and chemotherapeutic agents associated with photodynamic therapy in a breast cancer in vivo model. Our results showed that synergistic treatments in which chloroaluminum phthalocyanine (NE-Pc) administered together with Doxorubicin (Dox) in the presence of laser irradiation (NE-PcDoxo + PDT) led to a reduction of 4 T1 induced breast cancer in mice, decline of tumor VEGF expression, increase in Caspase-3 expression, tissue necrosis and massive decrease in proliferative cells, as shown by Ki67 immunostaining. Furthermore, this associated treatment induced overexpression of apoptotic genes ABL1, CD70, CRADD, FASL, and NME5 and a reduction in expression of anticancer drug target genes CDK2, ERBB2, FIGF, IGF2, PARP4 and PGR. These results validate this treatment as a promising alternative to improve the currently applied anticancer strategies. Sao Paulo State Univ, Lab Genom Studies, BR-15054000 Sao Jose Do Rio Preto, SP, Brazil Sao Paulo State Univ, Lab Microscopy & Microanal, BR-15054000 Sao Jose Do Rio Preto, SP, Brazil Univ Sao Paulo, Ctr Nanotechnol & Tissue Engn, Chem Dept, Photobiol & Photomed Res Grp, BR-14040901 Ribeirao Preto, SP, Brazil Sao Paulo State Univ, Lab Genom Studies, BR-15054000 Sao Jose Do Rio Preto, SP, Brazil Sao Paulo State Univ, Lab Microscopy & Microanal, BR-15054000 Sao Jose Do Rio Preto, SP, Brazil FAPESP: 2013/50181-1 FAPESP: 2020/04009-6 Project PRONON-SIPAR: 25000.077093/201586

This study investigated the ability of cholesterol-phosphatidylcholine liposomes loaded with chloride aluminum phthalocyanine (CL-AlClPc) to discriminate between healthy (MCF-10A) and neoplastic (MCF-7 and MDA-MB-231) breast cells for breast cancer diagnosis and treatment by photodynamic therapy (PDT) using a new drug delivery system consisting of CL-AlClPc. When PDT treatment was applied at an energy fluence of 700 mJ/cm², CL-AlClPc was more cytotoxic to neoplastic cells than to healthy breast cells because CL-AlClPc was better internalized by the tumor cells. An even higher fluorescence signal is expected for neoplastic cells during clinical treatment than for healthy cells, which will be useful for precise and targeted tumor cell detection. CL-AlClPc also facilitated better drug distribution and targeting of essential organelles inside the cells. This selectivity is critical for future in vivo diagnosis and treatment; it prevents side effects because it prioritizes tumor cells and tissues instead of healthy ones. The CL-AlClPc system designed herein had a small size (150 nm), low zeta potential (-6 mV), low polydispersity (0.16), high encapsulation rate efficiency (82.83%), and high shelf stability (12 months).

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.

Background and aim Oncological photodynamic therapy (PDT) relies on photosensitizers (PSs) to photo-oxidatively destroy tumor cells. Currently approved PSs yield satisfactory results in superficial and easy-to-access tumors but are less suited for solid cancers in internal organs such as the biliary system and the pancreas. For these malignancies, second-generation PSs such as metallated phthalocyanines are more appropriate. Presently it is not known which of the commonly employed metallated phtahlocyanines, namely aluminum phthalocyanine (AlPC) and zinc phthalocyanine (ZnPC) as well as their tetrasulfonated derivatives AlPCS4 and ZnPCS4, is most cytotoxic to tumor cells. This study therefore employed an attritional approach to ascertain the best metallated phthalocyanine for oncological PDT in a head-to-head comparative analysis and standardized experimental design. Methods ZnPC and AlPC were encapsulated in PEGylated liposomes. Analyses were performed in cultured A431 cells as a template for tumor cells with a dysfunctional P53 tumor suppressor gene and EGFR overexpression. First, dark toxicity was assessed as a function of PS concentration using the WST-1 and sulforhodamine B assay. Second, time-dependent uptake and intracellular distribution were determined by flow cytometry and confocal microscopy, respectively, using the intrinsic fluorescence of the PSs. Third, the LC50 values were established for each PS at 671 nm and a radiant exposure of 15 J/cm2 following 1-h PS exposure. Finally, the mode of cell death as a function of post-PDT time and cell cycle arrest at 24 h after PDT were analyzed. Results In the absence of illumination, AlPC and ZnPC were not toxic to cells up to a 1.5-μM PS concentration and exposure for up to 72 h. Dark toxicity was noted for AlPCS4 at 5 μM and ZnPCS4 at 2.5 μM. Uptake of all PSs was observed as early as 1 min after PS addition to cells and increased in amplitude during a 2-h incubation period. After 60 min, the entire non-nuclear space of the cell was photosensitized, with PS accumulation in multiple subcellular structures, especially in case of AlPC and AlPCS4. PDT of cells photosensitized with ZnPC, AlPC, and AlPCS4 yielded LC50 values of 0.13 μM, 0.04 μM, and 0.81 μM, respectively, 24 h post-PDT (based on sulforhodamine B assay). ZnPCS4 did not induce notable phototoxicity, which was echoed in the mode of cell death and cell cycle arrest data. At 4 h post-PDT, the mode of cell death comprised mainly apoptosis for ZnPC and AlPC, the extent of which was gradually exacerbated in AlPC-photosensitized cells during 8 h. ZnPC-treated cells seemed to recover at 8 h post-PDT compared to 4 h post-PDT, which had been observed before in another cell line. AlPCS4 induced considerable necrosis in addition to apoptosis, whereby most of the cell death had already manifested at 2 h after PDT. During the course of 8 h, necrotic cell death transitioned into mainly late apoptotic cell death. Cell death signaling coincided with a reduction in cells in the G0/G1 phase (ZnPC, AlPC, AlPCS4) and cell cycle arrest in the S-phase (ZnPC, AlPC, AlPCS4) and G2 phase (ZnPC and AlPC). Cell cycle arrest was most profound in cells that had been photosensitized with AlPC and subjected to PDT. Conclusions Liposomal AlPC is the most potent PS for oncological PDT, whereas ZnPCS4 was photodynamically inert in A431 cells. AlPC did not induce dark toxicity at PS concentrations of up to 1.5 μM, i.e., > 37 times the LC50 value, which is favorable in terms of clinical phototoxicity issues. AlPC photosensitized multiple intracellular loci, which was associated with extensive, irreversible cell death signaling that is expected to benefit treatment efficacy and possibly immunological long-term tumor control, granted that sufficient AlPC will reach the tumor in vivo. Given the differential pharmacokinetics, intracellular distribution, and cell death dynamics, liposomal AlPC may be combined with AlPCS4 in a PS cocktail to further improve PDT efficacy.