RESUMO
Photodynamic therapy (PDT) represents an emerging strategy to treat various malignancies, including colorectal cancer (CC), the third most common cancer type. This work presents an engineered M13 phage retargeted towards CC cells through pentavalent display of a disulfide-constrained peptide nonamer. The M13CC nanovector was conjugated with the photosensitizer Rose Bengal (RB), and the photodynamic anticancer effects of the resulting M13CC-RB bioconjugate were investigated on CC cells. We show that upon irradiation M13CC-RB is able to impair CC cell viability, and that this effect depends on i) photosensitizer concentration and ii) targeting efficiency towards CC cell lines, proving the specificity of the vector compared to unmodified M13 phage. We also demonstrate that M13CC-RB enhances generation and intracellular accumulation of reactive oxygen species (ROS) triggering CC cell death. To further investigate the anticancer potential of M13CC-RB, we performed PDT experiments on 3D CC spheroids, proving, for the first time, the ability of engineered M13 phage conjugates to deeply penetrate multicellular spheroids. Moreover, significant photodynamic effects, including spheroid disruption and cytotoxicity, were readily triggered at picomolar concentrations of the phage vector. Taken together, our results promote engineered M13 phages as promising nanovector platform for targeted photosensitization, paving the way to novel adjuvant approaches to fight CC malignancies.
Assuntos
Bacteriófagos , Neoplasias do Colo , Fotoquimioterapia , Humanos , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Fotoquimioterapia/métodos , Morte Celular , Rosa Bengala/farmacologia , Rosa Bengala/química , Neoplasias do Colo/terapiaRESUMO
X-ray-triggered scintillators (Sc) and photosensitizers (Ps) have been developed for X-ray-induced photodynamic therapy (X-PDT) to selectively destruct deep tissue tumors with a low X-ray dose. This study designed terbium (Tb)-rose bengal (RB) coordination nanocrystals (T-RBNs) by a solvothermal treatment, aiming to reduce photon energy dissipation between Tb3+ and RB and thus increase the reactive oxygen species (ROS) production efficiency. T-RBNs synthesized at a molar ratio of [RB]/[Tb] = 3 exhibited a size of 6.8 ± 1.2 nm with a crystalline property. Fourier transform infrared analyses of T-RBNs indicated successful coordination between RB and Tb3+. T-RBNs generated singlet oxygen (1O2) and hydroxyl radicals (â¢OH) under low-dose X-ray irradiation (0.5 Gy) via scintillating and radiosensitizing pathways. T-RBNs produced â¼8-fold higher ROS amounts than bare RB and â¼3.6-fold higher ROS amounts than inorganic nanoparticle-based controls. T-RBNs did not exhibit severe cytotoxicity up to 2 mg/mL concentration in cultured luciferase-expressing murine epithelial breast cancer (4T1-luc) cells. Furthermore, T-RBNs were efficiently internalized into cultured 4T1-luc cells and induced DNA double strand damage, as evidenced by an immunofluorescence staining assay with phosphorylated γ-H2AX. Ultimately, under 0.5 Gy X-ray irradiation, T-RBNs induced >70% 4T1-luc cell death via simultaneous apoptosis/necrosis pathways. Overall, T-RBNs provided a promising Sc/Ps platform under low-dose X-PDT for advanced cancer therapy.
Assuntos
Neoplasias da Mama , Nanopartículas , Fotoquimioterapia , Humanos , Animais , Camundongos , Feminino , Rosa Bengala/farmacologia , Rosa Bengala/química , Térbio/farmacologia , Térbio/química , Térbio/uso terapêutico , Espécies Reativas de Oxigênio/metabolismo , Raios X , Nanopartículas/uso terapêutico , Nanopartículas/químicaRESUMO
Photodynamic therapy (PDT) is a promising anticancer strategy based on the light energy stimulation of photosensitizers (PS) molecules within a malignant cell. Among a multitude of recently challenged PS, Rose bengal (RB) has been already reported as an inducer of cytotoxicity in different tumor cells. However, RB displays a low penetration capability across cell membranes. We have therefore developed a short-term amino acids starvation protocol that significantly increases RB uptake in human astrocytoma cells compared to normal rat astrocytes. Following induced starvation uptake, RB is released outside cells by the exocytosis of extracellular vesicles (EVs). Thus, we have introduced a specific pharmacological treatment, based on the GW4869 exosomes inhibitor, to interfere with RB extracellular release. These combined treatments allow significantly reduced nanomolar amounts of administered RB and a decrease in the time interval required for PDT stimulation. The overall conditions affected astrocytoma viability through the activation of apoptotic pathways. In conclusion, we have developed for the first time a combined scheme to simultaneously increase the RB uptake in human astrocytoma cells, reduce the extracellular release of the drug by EVs, and improve the effectiveness of PDT-based treatments. Importantly, this strategy might be a valuable approach to efficiently deliver other PS or chemotherapeutic drugs in tumor cells.
Assuntos
Astrocitoma , Exossomos , Fotoquimioterapia , Aminoácidos , Animais , Astrocitoma/tratamento farmacológico , Humanos , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Ratos , Rosa Bengala/química , Rosa Bengala/farmacologiaRESUMO
Photodynamic therapy is one of the best alternatives to chemo-, radio- or surgical therapy, as it is noninvasive and causes no severe side effects. The mechanism of photodynamic therapy involves activation of the drug (photosensitizer) with light of appropriate wavelength, which combined with molecular oxygen, leads to production of reactive oxygen species. This starts a cascade of reactions leading to cell death. Thus, the efficiency of this therapy is based mainly on the properties of a photosensitizer, including singlet oxygen yield and accumulation in the tumor area. Current research is aimed at applying nanosystems for the improvement of availability and photodynamic properties of photosensitizers. In order to improve the activity and increase photodynamic potential of rose bengal, one of the most promising drugs in anticancer photodynamic therapy, several drug delivery systems were developed. Among them, polymersomes represent a group of innovative polymeric vesicles mimicking membranous cell structures. Polymersomes are nanosystems made of amphiphilic block copolymers, possessing a spherical, liposome-like architecture. Within this study we present biophysical and in vitro biological characterization of this novel pH-stable nanosystem, which due to the improvement of singlet oxygen and reactive oxygen species (ROS) production by rose bengal is a good candidate for nanocarrier in photodynamic therapy.
Assuntos
Fotoquimioterapia , Linhagem Celular Tumoral , Concentração de Íons de Hidrogênio , Fármacos Fotossensibilizantes/química , Polímeros/química , Espécies Reativas de Oxigênio/metabolismo , Rosa Bengala/química , Oxigênio SingleteRESUMO
Rose bengal has been used in the diagnosis of ophthalmic disorders and liver function, and has been studied for the treatment of solid tumor cancers. To date, the antibacterial activity of rose bengal has been sporadically reported; however, these data have been generated with a commercial grade of rose bengal, which contains major uncontrolled impurities generated by the manufacturing process (80-95% dye content). A high-purity form of rose bengal formulation (HP-RBf, >99.5% dye content) kills a battery of Gram-positive bacteria, including drug-resistant strains at low concentrations (0.01-3.13 µg/mL) under fluorescent, LED, and natural light in a few minutes. Significantly, HP-RBf effectively eradicates Gram-positive bacterial biofilms. The frequency that Gram-positive bacteria spontaneously developed resistance to HP-RB is extremely low (less than 1 × 10-13). Toxicity data obtained through our research programs indicate that HP-RB is feasible as an anti-infective drug for the treatment of skin and soft tissue infections (SSTIs) involving multidrug-resistant (MDR) microbial invasion of the skin, and for eradicating biofilms. This article summarizes the antibacterial activity of pharmaceutical-grade rose bengal, HP-RB, against Gram-positive bacteria, its cytotoxicity against skin cells under illumination conditions, and mechanistic insights into rose bengal's bactericidal activity under dark conditions.
Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Rosa Bengala/química , Rosa Bengala/farmacologia , Animais , Antibacterianos/síntese química , Antibacterianos/uso terapêutico , Bactérias/efeitos dos fármacos , Bactérias/genética , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Farmacorresistência Bacteriana , Humanos , Cinética , Testes de Sensibilidade Microbiana , Mycobacterium/efeitos dos fármacos , Rosa Bengala/síntese química , Rosa Bengala/uso terapêuticoRESUMO
Cell nucleus-based photodynamic therapy is a highly effective method for cancer therapy, but it is still challenging to design nucleus-targeting photosensitizers. Here, we propose the "one treatment, multiple irradiations" strategy to achieve nucleus-based photodynamic therapy using the photosensitizer rose bengal (RB)-loaded and mesoporous silica-coated upconversion nanoparticles with the surface modification of amine group (UCNP/RB@mSiO2-NH2 NPs). After implementation into cancer cells, the rationally designed UCNP/RB@mSiO2-NH2 NPs could be specifically accumulated in the acidic lysosomes due to their amino group-decorated surface. Upon a short-term (3 min) irradiation of 980 nm near-infrared light, the reactive oxygen species produced by RB through the Förster resonance energy transfer between the upconversion nanoparticles and RB molecules could effectively destroy lysosomes, followed by the release of the UCNP/RB@mSiO2-NH2 NPs from the lysosomes. Subsequently, these released UCNP/RB@mSiO2-NH2 NPs could be transferred into the cell nucleus, where a second 980 nm light irradiation was conducted to achieve the nucleus-based photodynamic therapy. The rationally designed UCNP/RB@mSiO2-NH2 NPs showed excellent anticancer performance in both two-dimensional and three-dimensional cell models using the "one treatment, multiple irradiations" strategy.
Assuntos
Antineoplásicos/administração & dosagem , Metais Terras Raras/administração & dosagem , Nanopartículas/administração & dosagem , Fármacos Fotossensibilizantes/administração & dosagem , Rosa Bengala/administração & dosagem , Dióxido de Silício/administração & dosagem , Antineoplásicos/química , Antineoplásicos/efeitos da radiação , Núcleo Celular/química , Núcleo Celular/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Humanos , Luz , Lisossomos/química , Células MCF-7 , Metais Terras Raras/química , Metais Terras Raras/efeitos da radiação , Nanopartículas/química , Nanopartículas/efeitos da radiação , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/efeitos da radiação , Espécies Reativas de Oxigênio/química , Rosa Bengala/química , Rosa Bengala/efeitos da radiação , Dióxido de Silício/química , Dióxido de Silício/efeitos da radiação , Esferoides Celulares/efeitos dos fármacos , Células Tumorais CultivadasRESUMO
We designed a biodegradable hybrid nanostructure for near-infrared (NIR)-induced photodynamic therapy (PDT) using an ultrasmall upconversion (UC) phosphor (ß-NaYF4:Yb3+, Er3+ nanoparticle: NPs) and a hydrocarbonized rose bengal (C18RB) dye, a hydrophobized rose bengal (RB) derivative. The UC-NPs were encapsulated along with C18RB in the hydrophobic core of the micelle composed of poly(ethylene glycol) (PEG)-block-poly(ε-caprolactone) (PCL). The UC-NPs were well shielded from the aqueous environment, owing to the encapsulation in the hydrophobic PCL core, to efficiently emit green UC luminescence by avoiding the quenching by the hydroxyl groups. The hydrophobic part of C18 of C18RB worked well to be involved in the PCL core and located RB on the surface of the PCL core, making the efficient absorption of green light and the emission of singlet oxygen to surrounding water possible. Moreover, as the location is covered by PEG, the direct contact of RB to cells is prohibited to avoid their irradiation-free toxic effect on the cells. The hybrid nanostructure proved to be degradable by the hydrolysis of PEG-b-PCL. This degradation potentially results in renal excretion by the decomposition of the nanostructure into sub-10 nm size particles and makes them viable for clinical uses. These nanostructures can potentially be used for PDT of cancer in deep tissues.
Assuntos
Antineoplásicos/farmacologia , Materiais Biocompatíveis/farmacologia , Substâncias Luminescentes/farmacologia , Nanoestruturas/química , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Rosa Bengala/química , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Materiais Biocompatíveis/síntese química , Materiais Biocompatíveis/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Raios Infravermelhos , Substâncias Luminescentes/síntese química , Substâncias Luminescentes/química , Teste de Materiais , Tamanho da Partícula , Fármacos Fotossensibilizantes/síntese química , Fármacos Fotossensibilizantes/químicaRESUMO
The use of nanocarriers for intracellular transport of actives has been extensively studied in recent years and represents a central area of nanomedicine. The main novelty of this paper lies on the use of nanogels formed by a low-molecular-weight gelator (1). Here, non-polymeric, molecular nanogels are successfully used for intracellular transport of two photodynamic therapy (PDT) agents, Rose Bengal (RB) and hypericin (HYP). The two photosensitizers (PSs) exhibit different drawbacks for their use in clinical applications. HYP is poorly water-soluble, while the cellular uptake of RB is hindered due to its dianionic character at physiological pH values. Additionally, both PSs tend to aggregate precluding an effective PDT. Despite the different nature of these PSs, nanogels from gelator 1 provide, in both cases, an efficient intracellular transport into human colon adenocarcinoma cells (HT-29) and a notably improved PDT efficiency, as assessed by confocal laser scanning microscopy and flow cytometry. Furthermore, no significant dark toxicity of the nanogels is observed, supporting the biocompatibility of the delivery system. The developed nanogels are highly reproducible due to their non-polymeric nature, and their synthesis is easily scaled up. The results presented here thus confirm the potential of molecular nanogels as valuable nanocarriers, capable of entrapping both hydrophobic and hydrophilic actives, for PDT of cancer.
Assuntos
Antracenos/química , Nanogéis/química , Perileno/análogos & derivados , Fármacos Fotossensibilizantes/química , Rosa Bengala/química , Antracenos/metabolismo , Antracenos/farmacologia , Materiais Biocompatíveis/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , Humanos , Luz , Microscopia Confocal , Perileno/química , Perileno/metabolismo , Perileno/farmacologia , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/metabolismo , Fármacos Fotossensibilizantes/farmacologia , Rosa Bengala/metabolismo , Rosa Bengala/farmacologia , Oxigênio Singlete/metabolismoRESUMO
In photodynamic therapy (PDT), photosensitizer (PS) molecules are irradiated by light to generate reactive oxygen species (ROS), the presence of which subsequently leads to cell death. At present, the modality is limited to the treatment of skin diseases because of the low tissue penetration of visible or ultraviolet light required for producing ROS. To increase tissue penetration and extend the therapeutic possibilities of PDT to the treatment of deep-seated cancer, rare-earth doped nanoparticles capable of up-converting infrared to visible light are investigated. These up-converting nanoparticles (UCNPs) are conjugated with PS molecules to efficiently generate ROS. In this work, we employ hexagonal ß-NaYF4:Yb3 + ,Er3 + as UCNPs and Rose Bengal (RB) as PS molecules and demonstrate efficient in vitro PDT using this nanoformulation. Covalent bonding of the RB molecules is accomplished without their functionalization-an approach which is expected to increase the efficiency of ROS generation by 30%. Spectroscopic studies reveal that our approach results in UCNP surface fully covered with RB molecules. The energy transfer from UCNPs to RB is predominantly non-radiative as evidenced by luminescence lifetime measurements. As a result, ROS are generated as efficiently as under visible light illumination. The in vitro PDT is tested on murine breast 4T1 cancer cells incubated with 250 µg ml-1 of the nanoparticles and irradiated with NIR light under power density of 2 W cm-2 for 10 minutes. After 24 hours, the cell viability decreased to 33% demonstrating a very good treatment efficiency. These results are expected to simplify the protocols for preparation of the PDT agents and lead to improved therapeutic effects.
Assuntos
Érbio/farmacologia , Fluoretos/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Rosa Bengala/farmacologia , Itérbio/farmacologia , Ítrio/farmacologia , Animais , Linhagem Celular Tumoral , Érbio/química , Feminino , Fluoretos/química , Neoplasias Mamárias Animais/tratamento farmacológico , Neoplasias Mamárias Experimentais/tratamento farmacológico , Camundongos , Nanopartículas/química , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Rosa Bengala/química , Itérbio/química , Ítrio/químicaRESUMO
A desirable cancer therapeutic strategy is supposed to have effective ability to not only exert maximum anticancer ability but also inspire antitumor immunity for preventing tumor relapse and metastasis. During this research, multifunctional upconversion nanoparticles (UCNPs) coated by ROS-responsive micelles are prepared for tumor targeting and near-infrared (NIR)-triggered photodynamic therapy (PDT)-combined synergistic effect of chemotherapy. Moreover, both PDT and chemotherapy agents could activate antitumor immunity via inducing immunogenic cell death with CD8+ and CD4+ T cells infiltrating in tumors. Through the experiments, intravenous administration of multifunctional nanocarriers with noninvasive NIR irradiation destroys the orthotopic tumors and efficiently suppresses lung metastasis in a metastatic triple-negative breast cancer model by cascade-amplifying chemo-PDT and systemic antitumor immunity. In conclusion, this study provides prospective chemo-PDT with inspired antitumor immunity for metastatic cancer treatment.
Assuntos
Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Nanopartículas/química , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Rosa Bengala/farmacologia , Animais , Antineoplásicos/química , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Morte Celular/efeitos dos fármacos , Morte Celular/imunologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Doxorrubicina/química , Portadores de Fármacos/química , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Raios Infravermelhos , Camundongos , Camundongos Endogâmicos BALB C , Tamanho da Partícula , Fármacos Fotossensibilizantes/química , Rosa Bengala/química , Propriedades de SuperfícieRESUMO
Photodynamic therapy (PDT) is a skin cancer treatment alternative to chemotherapy and radiotherapy. This method exploits three elements: a phototoxic compound (photosensitizer), light source and oxygen. Upon irradiation by light of a specific wavelength, the photosensitizer generates reactive oxygen species triggering the cascade of reactions leading to cell death. The positive therapeutic effect of PDT may be limited due to low solubility, low tumor specificity and inefficient cellular uptake of photosensitizers. A promising approach to overcome these obstacles involves the use of nanocarrier systems. The aim of this initial study was to determine the potential of the application of phosphorus dendrimers as carriers of a photosensitizer-rose bengal (RB). The primary goal involved the synthesis and in vitro studies of covalent drug-dendrimer conjugates. Our approach allowed us to obtain RB-dendrimer conjugates with the use of tyramine as an aromatic linker between the carrier and the drug. The compounds were characterized by FT-IR, 1H NMR, 13C NMR, 31P NMR, size and zeta potential measurements and spectrofluorimetric analysis. The dialysis to check the drug release from the conjugate, flow cytometry to specify intracellular uptake, and singlet oxygen generation assay were also applied. Finally, we used MTT assay to determine the biological activity of the tested compounds. The results of our experiments indicate that the conjugation of RB to phosphorus dendrimers via the tyramine linker decreases photodynamic activity of RB.
Assuntos
Carcinoma Basocelular/tratamento farmacológico , Dendrímeros/química , Fósforo/química , Fármacos Fotossensibilizantes/farmacologia , Rosa Bengala/química , Neoplasias Cutâneas/tratamento farmacológico , Tiramina/química , Animais , Carcinoma Basocelular/patologia , Morte Celular , Portadores de Fármacos/química , Corantes Fluorescentes/química , Camundongos , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Espécies Reativas de Oxigênio/metabolismo , Oxigênio Singlete , Neoplasias Cutâneas/patologia , Células Tumorais CultivadasRESUMO
Combination therapy based on nanomedicine has gained momentum in oncology in recent years, offering superior safety and efficacy over monotherapies. It is critical to design theranostics that are composed of imaging and therapeutic agents already approved. Herein, gadolinium (Gd)-rose bengal coordination polymer nanodots (GRDs) are reported. The GRDs exhibit a unique absorption property and 7.7-fold luminescence enhancement, as well as a 1.9-fold increase in singlet oxygen generation efficiency over free rose bengal. Meanwhile, GRDs exhibit a twofold increase in r1 relaxivity over gadopentetic acid (Gd-DTPA) and have better X-ray absorption ability than rose bengal alone. These excellent properties of the GRDs are verified both in vitro and in vivo. The combination of photodynamic therapy (PDT) and radiation therapy (RT) more significantly inhibits tumor growth than monotherapies (i.e., PDT or RT). This work offers a new route to designing and synthesizing Gd-based nanotheranostics for image-guided cancer therapy.
Assuntos
Complexos de Coordenação/química , Gadolínio/química , Fotoquimioterapia/métodos , Polímeros/química , Polímeros/uso terapêutico , Radioterapia Guiada por Imagem/métodos , Rosa Bengala/química , Animais , Linhagem Celular Tumoral , Imageamento por Ressonância Magnética , Camundongos , Nanomedicina , Nanopartículas/química , Imagem ÓpticaRESUMO
Though emerging as a promising therapeutic approach for cancers, the crucial challenge for photodynamic therapy (PDT) is activatable phototoxicity for selective cancer cell destruction with low "off-target" damage and simultaneous therapeutic effect prediction. Here, we design an upconversion nanoprobe for intracellular cathepsin B (CaB)-responsive PDT with in situ self-corrected therapeutic effect prediction. The upconversion nanoprobe is composed of multishelled upconversion nanoparticles (UCNPs) NaYF4:Gd@NaYF4:Er,Yb@NaYF4:Nd,Yb, which covalently modified with an antenna molecule 800CW for UCNPs luminance enhancement under NIR irradiation, photosensitizer Rose Bengal (RB) for PDT, Cy3 for therapeutic effect prediction, and CaB substrate peptide labeled with a QSY7 quencher. The energy of UCNPs emission at 540 nm is transferred to Cy3/RB and eventually quenched by QSY7 via two continuous luminance resonance energy transfer processes from interior UCNPs to its surface-extended QSY7. The intracellular CaB specifically cleaves peptide to release QSY7, which correspondingly activates RB with reactive oxygen species (ROS) generation for PDT and recovers Cy3 luminance for CaB imaging. UCNPs emission at 540 nm remains unchanged during the peptide cleavage process, which is served as an internal standard for Cy3 luminance correction, and the fluorescence intensity ratio of Cy3 over UCNPs (FI583/FI540) is measured for self-corrected therapeutic effect prediction. The proposed self-corrected upconversion nanoprobe implies significant potential in precise tumor therapy.
Assuntos
Antineoplásicos/uso terapêutico , Nanopartículas Metálicas/uso terapêutico , Neoplasias/tratamento farmacológico , Fármacos Fotossensibilizantes/uso terapêutico , Animais , Antineoplásicos/química , Antineoplásicos/efeitos da radiação , Carbocianinas/química , Catepsina B/química , Corantes Fluorescentes/química , Fluoretos/química , Fluoretos/efeitos da radiação , Fluoretos/uso terapêutico , Células HeLa , Humanos , Elementos da Série dos Lantanídeos/química , Elementos da Série dos Lantanídeos/efeitos da radiação , Elementos da Série dos Lantanídeos/uso terapêutico , Luz , Nanopartículas Metálicas/química , Nanopartículas Metálicas/efeitos da radiação , Camundongos , Células NIH 3T3 , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismo , Rosa Bengala/química , Ensaios Antitumorais Modelo de Xenoenxerto , Ítrio/química , Ítrio/efeitos da radiação , Ítrio/uso terapêuticoRESUMO
Upconversion nanoparticles (UCNPs) are prospective platforms for bioimaging and phototherapy, but a critical bottleneck is the limited brightness due to the faint absorptivity of lanthanide ions and the low quantum yield. To circumvent this problem, we herein propose our strategy to reconstruct the energy cascade of UCNPs using semiconductor quantum dots (QDs) as light sensitizer of Nd3+/Yb3+ codoped UCNPs. Ag2Se QDs with strong absorption at 808 nm acted as efficient antenna and transferred their energy to Yb3+ via a resonance energy transfer process, significantly enhancing the luminescence of UCNPs. This nanocomposite was then combined with Rose Bengal and applied for photodynamic therapy. Both in vitro and in vivo studies revealed the introduction of QDs improved the therapeutic performance remarkably. Our study suggests Ag2Se QDs with excellent photophysical properties can be promising agents to overcome the shortcomings of UCNPs and further strengthen their applications.
Assuntos
Nanopartículas de Magnetita/química , Neoplasias/tratamento farmacológico , Fármacos Fotossensibilizantes/uso terapêutico , Pontos Quânticos/química , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Transferência de Energia , Humanos , Raios Infravermelhos , Nanopartículas de Magnetita/toxicidade , Camundongos , Camundongos Nus , Microscopia Confocal , Neodímio/química , Neoplasias/patologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacologia , Pontos Quânticos/toxicidade , Rosa Bengala/química , Rosa Bengala/farmacologia , Rosa Bengala/uso terapêutico , Semicondutores , Itérbio/químicaRESUMO
Radio- and photodynamic therapies are the first line of cancer treatments but suffer from poor light penetration and less radiation accumulation in soft tissues with high radiation toxicity. Therefore, a multifunctional nanoplatform with diagnosis-assisted synergistic radio- and photodynamic therapy and tools facilitating early prognosis are urgently needed to fight the war against cancer. Further, integrating cancer therapy with untargeted metabolomic analysis would collectively offer clinical pertinence through facilitating early diagnosis and prognosis. Here, we enriched scintillation of CeF3 nanoparticles (NPs) through codoping Tb3+ and Gd3+ (CeF3:Gd3+,Tb3+) for viable clinical approach in the treatment of deep-seated tumors. The codoped CeF3:Gd3+,Tb3+ scintillating theranostic NPs were then coated with mesoporous silica, followed by loading with rose bengal (CGTS-RB) for later computed tomography (CT)- and magnetic resonance image (MRI)-guided X-ray stimulated synergistic radio- and photodynamic therapy (RT+XPDT) using low-dose, one-time X-ray irradiation. The results corroborated an efficient tumor regression with synergistic RT+XPDT relative to single RT. Global untargeted metabolome shifts highlighted the mechanism behind this efficient tumor regression using RT, and synergistic RT+XPDT treatment is due to the starvation of nonessential amino acids involved in protein and DNA synthesis and energy regulation pathways necessary for growth and progression. Our study also concluded that tumor and serum metabolites shift during disease progression and regression and serve as robust biomarkers for early assessment of disease state and prognosis. From our results, we propose that codoping is an effective and extendable technique to other materials for gaining high optical yield and multifunctionality and for use in diagnostic and therapeutic applications. Critically, the integration of multifunctional theranostic nanomedicines with metabolomics has excellent potential for the discovery of early metabolic biomarkers to aid in better clinical disease diagnosis and prognosis.
Assuntos
Luminescência , Metabolômica , Nanopartículas/química , Neoplasias/metabolismo , Neoplasias/terapia , Animais , Materiais Biocompatíveis/química , Circulação Sanguínea , Meios de Contraste/química , Imageamento por Ressonância Magnética , Camundongos Endogâmicos BALB C , Nanopartículas/toxicidade , Nanopartículas/ultraestrutura , Neoplasias/diagnóstico por imagem , Fotoquimioterapia , Prognóstico , Rosa Bengala/química , Oxigênio Singlete/química , Tomografia Computadorizada por Raios X , Testes de Toxicidade , Raios XRESUMO
In this work, we report new formulations for the combined photo-chemotherapy of colon cancer. Fibers were fabricated via coaxial-electrospinning with the intent of targeting delivery of the anti-cancer drug carmofur (CAR) and the photosensitizer rose bengal (RB) selectively to the colon site. The fibers comprised a hydroxypropyl methylcellulose (HPMC) core loaded with the active ingredients, and a pH-sensitive Eudragit L100-55 shell. The fibers were found to be homogeneous and cylindrical and have visible core-shell structures. X-ray diffraction and differential scanning calorimetry demonstrated that both CAR and RB were present in the fibers in the amorphous physical form. In vitro drug release studies showed that the fibers have the potential to selectively deliver drugs to the colon, with only 10-15 % release noted in the acidic conditions of the stomach but sustained release at pH 7.4. Cytotoxicity studies were undertaken on human dermal fibroblast (HDF) and colon cancer (Caco-2) cells, and the influence of light on cell death was also explored. The fibers loaded with CAR alone showed obvious toxicity to both cell lines, with and without the application of light. The RB-loaded fibers led to high viability (ca. 80% for both cell types) in the absence of light, but much greater toxicity was noted (30-50%) with light. The same trends were observed with the formulation containing both CAR and RB, but with lower viabilities. The RB and RB/CAR loaded systems show clear selectivity for cancerous over non-cancerous cells. Finally, mucoadhesion studies revealed there were strong adhesive forces between the rat colonic mucosa and the fibers after they had passed through an acidic environment. Such electrospun fibers thus could have potential in the development of oral therapies for colon cancer.
Assuntos
Antineoplásicos/farmacologia , Portadores de Fármacos , Fluoruracila/análogos & derivados , Nanofibras/química , Fármacos Fotossensibilizantes/farmacologia , Rosa Bengala/farmacologia , Resinas Acrílicas/química , Administração Oral , Animais , Antineoplásicos/química , Células CACO-2 , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Combinação de Medicamentos , Técnicas Eletroquímicas , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fluoruracila/química , Fluoruracila/farmacologia , Humanos , Derivados da Hipromelose/química , Intestino Grosso/efeitos dos fármacos , Intestino Grosso/metabolismo , Luz , Nanofibras/administração & dosagem , Nanofibras/ultraestrutura , Especificidade de Órgãos , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/efeitos da radiação , Fototerapia/métodos , Ratos Sprague-Dawley , Rosa Bengala/química , Rosa Bengala/efeitos da radiação , Técnicas de Cultura de TecidosRESUMO
The effective strategy to treat superficial malignant tumors is challenging due to the complexed mechanisms for tumor growth inhibition. The proposed immune-enhancement and immune response activation pathways rely on the direct and massive implementation of therapeutic agents. In this manuscript, an alternative route has been developed to apply the multifunctional peptide amphiphile-Rose Bengal (RB) nanocapsules for noninvasive sonodynamic and photodynamic therapies in association with the targeted immune enhancement to tumor proliferation. The nanocapsules proved better RB loading capacity, comparable reactive oxygen species generation and improved B16 and HeLa cell killing capability. The proof-of-principle in-vivo nude mice therapy studies with routine blood examinations and cytokine analysis validated their effectiveness for tumor cell elimination, and a direct tumor vasculature damage for efficacious lesion necrosis, positive prognosis with minimized side-effects. This state-of-the-art strategy provides a promising route to merge tumor enhancement and tumor response activation methodologies, and is expected to be universalized for a broad spectra of cancer diseases.
Assuntos
Imunoterapia/métodos , Nanocápsulas/química , Peptídeos/química , Fotoquimioterapia/métodos , Rosa Bengala/química , Animais , Linhagem Celular Tumoral , Células HeLa , Humanos , Masculino , Camundongos , Camundongos Nus , Espécies Reativas de Oxigênio/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
X-ray-induced photodynamic therapy (X-PDT) combines both the advantages of radiotherapy (RT) and PDT, and has considerable potential applications in clinical deep-penetrating cancer therapy. However, it is still a major challenge to prepare monodisperse nanoscintillators with uniform size and high light yield. In this study, a general and rapid synthesis method is presented that can achieve large-scale preparation of monodisperse and uniform silicate nanoscintillators. By simply adjusting the metal dopants, silicate nanoscintillators with controllable size and X-ray-excited optical luminescence (450-900 nm) are synthesized by employing a general ion-incorporated silica-templating method. To make full use of external radiation, the silicate nanoscintillators are conjugated with photosensitizer rose bengal and arginylglycylaspartic acid (RGD) peptide, making them intrinsically dual-modal targeted imaging probes. Both in vitro and in vivo experiments demonstrate that the silicate nanosensitizers can accumulate effectively in tumors and achieve significant inhibitory effect on tumor progression under low-dose X-ray irradiation, while minimally affecting normal tissues. The insights gained in this study may provide an attractive route to synthesize nanosensitizers to overcome some of the limitations of RT and PDT in cancer treatment.
Assuntos
Nanopartículas/química , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/síntese química , Silicatos/química , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Corantes Fluorescentes/química , Xenoenxertos , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Oligopeptídeos/química , Imagem Óptica , Permeabilidade , Fármacos Fotossensibilizantes/administração & dosagem , Rosa Bengala/química , Raios XRESUMO
Photodynamic therapy (PDT) is a well-established treatment of cancer in which cell toxic reactive oxygen species, including singlet oxygen (1O2), are produced by a photosensitiser drug following irradiation of a specific wavelength. Visible light is commonly used as the excitation source in PDT, although these wavelengths do have limited tissue penetration. In this research, upconverting nanoparticles (UCNPs) functionalised with the photosensitiser Rose Bengal (RB) have been designed and synthesised for PDT of breast cancer cells. The use of UCNPs shifts the required excitation wavelength for the production of 1O2 to near infrared light (NIR) thus allowing deeper tissue penetration. The system was designed to maximise the production of 1O2via efficient Förster resonance energy transfer (FRET) from the UCNPs to the photosensitiser. Highly luminescent NaYF4:Yb,Er,Gd@NaYF4 core-shell UCNPs were synthesised that exhibited two main anti-Stokes emission bands at 541 and 652 nm following 980 nm irradiation. RB was chosen as the photosensitiser since its absorption band overlaps with the green emission of the UCNPs. To achieve efficient energy transfer from the nanoparticles to the photosensitiser, the functionalised UCNPs included a short l-lysine linker to attach the RB to the nanocore yielding RB-lysine functionalised UCNPs. The efficient FRET from the UCNPs to the RB was confirmed by luminescence lifetime measurements. The light emitted by the UCNPs at 541 nm, following excitation at 980 nm, generates the 1O2via the RB. Multi-photon and confocal laser scanning microscopies confirmed the internalisation of the RB-lysine-UCNPs by SK-BR-3 breast cancer cells. Cell viability studies revealed that the RB-lysine-UCNPs induced low dark toxicity in cells prior to PDT treatment. Importantly, following irradiation at 980 nm, high levels of cell death were observed in cells loaded with the RB-lysine-UCNPs. Cell death following PDT treatment was also confirmed using propidium iodide and confocal microscopy. The high drug loading capacity (160 RB/nanoparticle) of the UCNPs, the efficient FRET from the UCNPs to the photosensitiser, the high level of accumulation inside the cells and their PDT cell kill suggest that the RB-lysine-UCNPs are promising for NIR PDT and hence suitable for the treatment of deep-lying cancer tumours.
Assuntos
Neoplasias da Mama/tratamento farmacológico , Corantes Fluorescentes/farmacologia , Nanopartículas , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Endocitose , Feminino , Corantes Fluorescentes/química , Corantes Fluorescentes/uso terapêutico , Humanos , Lisina/química , Lisina/farmacologia , Lisina/uso terapêutico , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacologia , Rosa Bengala/química , Rosa Bengala/farmacologia , Rosa Bengala/uso terapêutico , Oxigênio SingleteRESUMO
The limitation of light penetration depth invalidates the application of photodynamic therapy in deep-seated tumors. X-ray excited photodynamic therapy (X-PDT), which is based on X-rays excited luminescent nanoparticles (XLNP), provides a new strategy for PDT in deep tissues. However, the high X-ray dosage used and non-specific cytotoxicity of the nanoparticle-photosensitizer nanocomposite (NPs-PS) hamper in-vivo X-PDT applications. To address these problems, a simple and efficient NPs-PS nanocomposite using ß-NaGdF4: Tb3+ nanoparticles and widely used PS called Rose Bengal (RB) was designed. With perfectly matched spectrum of NPs emission and RB absorption upon X-ray excitation and covalent conjugation of a large amount of RB on NP surfaces to minimize the energy transfer distance, the system demonstrated ultra-high FRET efficiency up to 99.739%, which leads to maximum production of singlet oxygen for PDT with significantly increased anti-tumor efficacy. By 2-aminoethylphosphonic acid surface modification of NPs, excellent biocompatibility was achieved even at a high concentration of 1â¯mg/mL. The in-vivo X-PDT efficacy was found around 90% of HepG2 tumor growth inhibition with X-ray dose of only 1.5â¯Gy, which shows the best anti-tumor efficacy at same X-ray dose level reported so far. The present work provides a promising platform for in-vivo X-PDT in deep tumors.