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1.
J Mater Chem B ; 11(17): 3919-3928, 2023 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-37060145

RESUMO

Intracellular polarity in lipid droplets as well as other organelles may provide useful knowledge about various processes taking place in live cells. Therefore, small fluorophores capable of visualising polarity are undergoing rapid development. In this paper, we report new red-emitting polarity sensitive BODIPY probes that can distinguish between liquid-ordered and liquid-disordered phases and can internalise into lipid droplets of live cells. Our reported probes sense lipid environment not through solvatochromic shift of the fluorescence spectra but through the change in the fluorescence lifetime of their monoexponential decays. This makes them convenient for fluorescence lifetime imaging microscopy. The probes were synthesised by modifying viscosity-sensitive meso-phenyl BODIPY with electron-donating 2-thienyl moieties at the α- and ß-positions, significantly red-shifting absorption and fluorescence spectra of the dyes and improving sensitivity to polarity, while suppressing viscosity dependence. Finally, a novel probe - BP OC16 TP2 was suitable for sensing polarity in lipid droplets of live MCF-7 human breast cancer cells. We demonstrated that different chemotherapeutics affected lipid droplet polarity differently: cisplatin had no effect on lipid droplet polarity, whereas paclitaxel, depending on its concentration, either decreased or increased lipid droplet polarity.


Assuntos
Gotículas Lipídicas , Tiofenos , Humanos , Microscopia de Fluorescência/métodos , Compostos de Boro
2.
Int J Mol Sci ; 23(10)2022 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-35628497

RESUMO

BODIPY-based molecular rotors are highly attractive imaging tools for imaging intracellular microviscosity in living cells. In our study, we investigated the ability to detect the microviscosity of biological objects by using BDP-NO2 and BDP-H molecular rotors. We describe in detail the optical properties of BDP-NO2 and BDP-H molecular rotors in aqueous media with and without proteins, together with their accumulation dynamics and localization in live and fixed human breast cancer cells. Furthermore, we investigate the applicability of these molecules to monitor microviscosity in the organelles of human breast cancer cells by fluorescence lifetime imaging microscopy (FLIM). We demonstrate that the BDP-NO2 molecular rotor aggregates in aqueous media and is incompatible with live cell imaging. The opposite effect is observed with BDP-H which preserves its stability in aqueous media, diffuses through the plasma membrane and accumulates in lipid droplets (LDs) and the cytosol of both live and fixed MCF-7 and MDA-MB-231 cancer cells. Finally, by utilizing BDP-H we demonstrate that LD microviscosity is significantly elevated in more malignant MDA-MB-231 human breast cancer cells, as compared to MCF-7 breast cancer cells. Our findings demonstrate that BDP-H is a water-compatible probe that can be successfully applied to measure microviscosity in the LDs of living cells.


Assuntos
Neoplasias da Mama , Compostos de Boro , Neoplasias da Mama/diagnóstico por imagem , Feminino , Corantes Fluorescentes , Humanos , Dióxido de Nitrogênio
3.
Cancers (Basel) ; 14(8)2022 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-35454798

RESUMO

Personalized cancer theranostics has a potential to increase efficiency of early cancer diagnostics and treatment, and to reduce negative side-effects. Protein-stabilized gold nanoclusters may serve as theranostic agents. To make gold nanoclusters personalized and highly biocompatible, the clusters were stabilized with human plasma proteins. Optical properties of synthesized nanoclusters were investigated spectroscopically, and possible biomedical application was evaluated using standard cell biology methods. The spectroscopic investigations of human plasma proteins stabilized gold nanoclusters revealed that a wide photoluminescence band in the optical tissue window is suitable for cancer diagnostics. High-capacity generation of singlet oxygen and other reactive oxygen species was also observed. Furthermore, the cluster accumulation in cancer cells and the photodynamic effect were evaluated. The results demonstrate that plasma proteins stabilized gold nanoclusters that accumulate in breast cancer cells and are non-toxic in the dark, while appear phototoxic under irradiation with visible light. The results positively confirm the utility of plasma protein stabilized gold nanoclusters for the use in cancer diagnostics and treatment.

4.
J Mater Chem B ; 10(4): 625-636, 2022 01 26.
Artigo em Inglês | MEDLINE | ID: mdl-34989749

RESUMO

Upconverting nanoparticles (UCNPs) possess great potential for biomedical application. UCNPs absorb and convert near-infrared (NIR) radiation in the biological imaging window to visible (Vis) and even ultraviolet (UV) radiation. NIR excitation offers reduced scattering and diminished autofluorescence in biological samples, whereas the emitted UV-Vis and NIR photons can be used for cancer treatment and imaging, respectively. However, UCNPs are usually synthesized in organic solvents and are not readily suitable for biomedical application due to the hydrophobic nature of their surface. Herein, we have removed the hydrophobic ligands from the synthesized UCNPs and coated the bare UCNPs with two custom-made hydrophilic polyelectrolytes (synthesized via the reversible addition-fragmentation chain transfer (RAFT) polymerization method). Polymers containing different amounts of PEGylated and carboxylic groups were studied. Coating with both polymers increased the upconversion (UC) emission intensity and photoluminescence lifetime values of the UCNPs, which directly translates to more efficient cancer cell labeling nanoprobes. The polymer composition plays a crucial role in the modification of UCNPs, not only with respect to their colloidal stability, but also with respect to the cellular uptake. Colloidally unstable bare UCNPs aggregate in cell culture media and precipitate, rendering themselves unsuitable for any biomedical use. However, stabilization with polymers prevents UCNPs from aggregation, increases their uptake in cells, and improves the quality of cellular labeling. This investigation sheds light on the appropriate coating for UCNPs and provides relevant insights for the rational development of imaging and therapeutic tools.


Assuntos
Materiais Revestidos Biocompatíveis/química , Nanopartículas/química , Polímeros/química , Linhagem Celular Tumoral , Coloides/química , Humanos , Interações Hidrofóbicas e Hidrofílicas , Teste de Materiais , Tamanho da Partícula
5.
Chemistry ; 27(67): 16768-16775, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34553449

RESUMO

Viscosity imaging at a microscopic scale can provide important information about biosystems, including the development of serious illnesses. Microviscosity imaging is achievable with viscosity-sensitive fluorophores, the most popular of which are based on the BODIPY group. However, most of the BODIPY probes fluoresce green light, whereas the red luminescence is desired for the imaging of biological samples. Designing a new viscosity probe with suitable spectroscopic properties is a challenging task because it is difficult to preserve viscosity sensitivity after modifying the molecular structure. Here we describe how we developed a new red-emitting, viscosity-sensitive, BODIPY fluorophore BP-PH-2M-NO2 that is suitable for reliable intracellular viscosity imaging of lipid droplets in MCF-7 breast cancer cells. The design of BP-PH-2M-NO2 was aided by DFT calculations that allowed a successful prediction of the viscosity sensitivity of fluorophores before synthesis. In summary, we report a new red viscosity probe possessing monoexponential fluorescence decay that makes it attractive for lifetime-based viscosity imaging.


Assuntos
Compostos de Boro , Corantes Fluorescentes , Imagem Óptica , Viscosidade
6.
ACS Appl Mater Interfaces ; 13(37): 43937-43951, 2021 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-34499462

RESUMO

Nanotechnology has emerged as a promising solution to permanent elimination of cancer. However, nanoparticles themselves lack specificity to tumors. Due to enhanced migration to tumors, mesenchymal stem cells (MSCs) were suggested as cell-mediated delivery vehicles of nanoparticles. In this study, we have constructed a complex composed of photoluminescent quantum dots (QDs) and a photosensitizer chlorin e6 (Ce6) to obtain multifunctional nanoparticles, combining cancer diagnostic and therapeutic properties. QDs serve as energy donors-excited QDs transfer energy to the attached Ce6 via Förster resonance energy transfer, which in turn generates reactive oxygen species. Here, the physicochemical properties of the QD-Ce6 complex and singlet oxygen generation were measured, and the stability in protein-rich media was evaluated, showing that the complex remains the most stable in protein-free medium. In vitro studies on MSC and cancer cell response to the QD-Ce6 complex revealed the complex-loaded MSCs' potential to transport theranostic nanoparticles and induce cancer cell death. In vivo studies proved the therapeutic efficacy, as the survival of tumor-bearing mice was statistically significantly increased, while tumor progression and metastases were slowed down.


Assuntos
Antineoplásicos/uso terapêutico , Carcinoma Pulmonar de Lewis/diagnóstico por imagem , Carcinoma Pulmonar de Lewis/tratamento farmacológico , Células-Tronco Mesenquimais/metabolismo , Nanopartículas Multifuncionais/uso terapêutico , Animais , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/efeitos da radiação , Compostos de Cádmio/química , Compostos de Cádmio/metabolismo , Compostos de Cádmio/efeitos da radiação , Compostos de Cádmio/uso terapêutico , Carcinoma Pulmonar de Lewis/metabolismo , Linhagem Celular Tumoral , Clorofilídeos/química , Clorofilídeos/metabolismo , Clorofilídeos/efeitos da radiação , Clorofilídeos/uso terapêutico , Feminino , Humanos , Luz , Camundongos Endogâmicos C57BL , Nanopartículas Multifuncionais/química , Nanopartículas Multifuncionais/metabolismo , Nanopartículas Multifuncionais/efeitos da radiação , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/metabolismo , Fármacos Fotossensibilizantes/efeitos da radiação , Fármacos Fotossensibilizantes/uso terapêutico , Medicina de Precisão/métodos , Pontos Quânticos/química , Pontos Quânticos/metabolismo , Pontos Quânticos/efeitos da radiação , Pontos Quânticos/uso terapêutico , Compostos de Selênio/química , Compostos de Selênio/metabolismo , Compostos de Selênio/efeitos da radiação , Compostos de Selênio/uso terapêutico , Oxigênio Singlete/metabolismo , Sulfetos/química , Sulfetos/metabolismo , Sulfetos/efeitos da radiação , Sulfetos/uso terapêutico , Compostos de Zinco/química , Compostos de Zinco/metabolismo , Compostos de Zinco/efeitos da radiação , Compostos de Zinco/uso terapêutico
7.
ACS Appl Mater Interfaces ; 13(33): 39076-39087, 2021 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-34378375

RESUMO

Fluorophores with multifunctional properties known as rare-earth-doped nanoparticles (RENPs) are promising candidates for bioimaging, therapy, and drug delivery. When applied in vivo, these nanoparticles (NPs) have to retain long blood-circulation time, bypass elimination by phagocytic cells, and successfully arrive at the target area. Usually, NPs in a biological medium are exposed to proteins, which form the so-called "protein corona" (PC) around the NPs and influence their targeted delivery and accumulation in cells and tissues. Different surface coatings change the PC size and composition, subsequently deciding the fate of the NPs. Thus, detailed studies on the PC are of utmost importance to determine the most suitable NP surface modification for biomedical use. When it comes to RENPs, these studies are particularly scarce. Here, we investigate the PC composition and its impact on the cellular uptake of citrate-, SiO2-, and phospholipid micelle-coated RENPs (LiYF4:Yb3+,Tm3+). We observed that the PC of citrate- and phospholipid-coated RENPs is relatively stable and similar in the adsorbed protein composition, while the PC of SiO2-coated RENPs is larger and highly dynamic. Moreover, biocompatibility, accumulation, and cytotoxicity of various RENPs in cancer cells have been evaluated. On the basis of the cellular imaging, supported by the inhibition studies, it was revealed that RENPs are internalized by endocytosis and that specific endocytic routes are PC composition dependent. Overall, these results are essential to fill the gaps in the fundamental understanding of the nano-biointeractions of RENPs, pertinent for their envisioned application in biomedicine.


Assuntos
Materiais Revestidos Biocompatíveis/química , Corantes Fluorescentes/química , Compostos de Lítio/química , Nanopartículas Metálicas/química , Coroa de Proteína/metabolismo , Dióxido de Silício/química , Ítrio/química , Adsorção , Neoplasias da Mama , Linhagem Celular Tumoral , Permeabilidade da Membrana Celular , Ácido Cítrico/química , Materiais Revestidos Biocompatíveis/metabolismo , Endocitose , Corantes Fluorescentes/metabolismo , Humanos , Tamanho da Partícula , Fosfolipídeos/química , Propriedades de Superfície
8.
J Photochem Photobiol B ; 204: 111802, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31981990

RESUMO

Suitable properties as well as eco-friendly synthesis of photoluminescent Au nanoclusters (NCs) make them promising compounds for biomedical diagnostics and visualization applications. However, the potential photochemical activity of such agents on cancerous cells is largely unknown. The nanoclusters (BSA-Au NCs) were synthetized in the presence of BSA (an average hydrodynamic diameter was about 9.4 nm, while the size of the metal cluster was <1.3 nm according to atomic force microscopy measurements) and possessed a broad photoluminescence band at 680 nm in buffered (pH 7.2) aqueous medium. The photochemical activity was studied by adding two fluorescent probes (dihydrorhodamine or Singlet Oxygen Sensor Green) for detection of reactive oxygen species in samples irradiated at 405 nm to minimize direct excitation of the probes. The photoluminescence measurements evidenced the capability of BSA-Au NCs to generate reactive oxygen species upon light exposure, while the observed sensitivity of the photoluminescence properties might be used to indicate photooxidative processes in the medium. The viability test performed on breast cancer cells after incubation with BSA-Au NCs and subsequent irradiation revealed notable difference in induced phototoxicity between two cell lines, which was not the case after the corresponding treatment using the photosensitizer chlorin e6.


Assuntos
Ouro/química , Nanopartículas Metálicas/química , Espécies Reativas de Oxigênio/metabolismo , Soroalbumina Bovina/química , Oxigênio Singlete/metabolismo , Animais , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Bovinos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Feminino , Corantes Fluorescentes/química , Humanos , Lasers Semicondutores , Nanopartículas Metálicas/uso terapêutico , Nanopartículas Metálicas/toxicidade , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Espécies Reativas de Oxigênio/química , Oxigênio Singlete/química , Espectrometria de Fluorescência
9.
J Nanobiotechnology ; 17(1): 39, 2019 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-30866960

RESUMO

BACKGROUND: Human mesenchymal stem cells (MSCs) have drawn much attention in the field of regenerative medicine for their immunomodulatory and anti-inflammatory effects. MSCs possess specific tumor-oriented migration and incorporation highlighting the potential for MSCs to be used as an ideal carrier for anticancer agents. Bone marrow is the main source of MSCs for clinical applications. MSCs tracking in vivo is a critical component of the safety and efficacy evaluation of therapeutic cell products; therefore, cells must be labeled with contrast agents to enable visualization of the MSCs migration in vivo. Due to their unique properties, quantum dots (QDs) are emerging as optimal tools in long-term MSC optical imaging applications. The aim of this study was to investigate the uptake dynamics, cytotoxity, subcellular and extracellular distribution of non-targeted carboxylated quantum dots in human bone marrow MSCs at different cell growing densities. RESULTS: QDs had no negative impact on MSC viability throughout the experiment and accumulated in all observed cells efficiently; however, in some MSCs QDs induced formation of lipid droplets. At low cell growing densities QDs distribute within MSCs cytoplasm already after 1 h of incubation reaching saturation after 6 h. After 24 h QDs localize mainly in the perinuclear region of the cells in endosomes. Interestingly, in more confluent culture QDs localize mostly outside MSCs. QDs abundantly mark MSC long filopodia-like structures attaching neighboring cells. At high cell density cultivation, we for the first time demonstrated that carboxylated QDs localize in human bone marrow MSC extracellular matrix. Moreover, we observed that average photoluminescence lifetime of QDs distributed in extracellular matrix are longer than lifetimes of QDs entrapped in endocytic vesicles; thus, for the first time showing the possibility to identify and distinguish localization of QDs in various extracellular and intracellular structures using fluorescence-lifetime imaging microscopy without additional staining assays. CONCLUSION: Carboxylated QDs can be used as nonspecific and effective dye for staining of human bone marrow MSCs and their specific extracellular structures. These results are promising in fundamental stem cell biology as well as in cellular therapy, anticancer drug delivery and tissue engineering.


Assuntos
Ácidos Carboxílicos/química , Corantes Fluorescentes/metabolismo , Células-Tronco Mesenquimais/metabolismo , Pontos Quânticos/metabolismo , Transporte Biológico , Movimento Celular , Sobrevivência Celular , Matriz Extracelular/metabolismo , Humanos , Imagem Óptica , Coloração e Rotulagem , Fatores de Tempo
10.
Nanomaterials (Basel) ; 9(1)2018 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-30583495

RESUMO

Nanomaterials have permeated various fields of scientific research, including that of biomedicine, as alternatives for disease diagnosis and therapy. Among different structures, quantum dots (QDs) have distinctive physico-chemical properties sought after in cancer research and eradication. Within the context of cancer therapy, QDs serve the role of transporters and energy donors to photodynamic therapy (PDT) drugs, extending the applicability and efficiency of classic PDT. In contrast to conventional PDT agents, QDs' surface can be designed to promote cellular targeting and internalization, while their spectral properties enable better light harvesting and deep-tissue use. Here, we investigate the possibility of complex formation between different amphiphilic coating bearing QDs and photosensitizer chlorin e6 (Ce6). We show that complex formation dynamics are dependent on the type of coating-phospholipids or amphiphilic polymers-as well as on the surface charge of QDs. Förster's resonant energy transfer occurred in every complex studied, confirming the possibility of indirect Ce6 excitation. Nonetheless, in vitro PDT activity was restricted only to negative charge bearing QD-Ce6 complexes, correlating with better accumulation in cancer cells. Overall, these findings help to better design such and similar complexes, as gained insights can be straightforwardly translated to other types of nanostructures-expanding the palette of possible therapeutic agents for cancer therapy.

11.
Biochim Biophys Acta Gen Subj ; 1862(4): 914-923, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29288773

RESUMO

BACKGROUND: Monolayer cell cultures have been considered the most suitable technique for in vivo cellular experiments. However, a lot of cellular functions and responses that are present in natural tissues are lost in two-dimensional cell cultures. In this context, nanoparticle accumulation data presented in literature are often not accurate enough to predict behavior of nanoparticles in vivo. Cellular spheroids show a higher degree of morphological and functional similarity to the tissues. METHODS: Accumulation and distribution of carboxylated CdSe/ZnS quantum dots (QDs), chosen as model nanoparticles, was investigated in cellular spheroids composed of different phenotype mammalian cells. The findings were compared with the results obtained in in vivo experiments with human tumor xenografts in immunodeficient mice. The diffusive transport model was used for theoretical nanoparticles distribution estimation. RESULTS: QDs were accumulated only in cells, which were localized in the periphery of cellular spheroids. CdSe/ZnS QDs were shown to be stable and inert; they did not have any side-effects for cellular spheroids formation. Penetration of QDs in both cellular spheroids and in vivo tumor model was limited. The mathematical model confirmed the experimental results: nanoparticles penetrated only 25µm into cellular spheroids after 24h of incubation. CONCLUSIONS: Penetration of negatively charged nanoparticles is limited not only in tumor tissue, but also in cellular spheroids. GENERAL SIGNIFICANCE: The results presented in this paper show the superior applicability of cellular spheroids to cell monolayers in the studies of the antitumor effect and penetration of nanomedicines.


Assuntos
Ácidos Carboxílicos/química , Nanopartículas/química , Pontos Quânticos , Esferoides Celulares/química , Animais , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Compostos de Cádmio/química , Compostos de Cádmio/metabolismo , Ácidos Carboxílicos/metabolismo , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Humanos , Células MCF-7 , Camundongos , Microscopia Confocal , Células NIH 3T3 , Nanopartículas/metabolismo , Compostos de Selênio/química , Compostos de Selênio/metabolismo , Esferoides Celulares/metabolismo , Sulfetos/química , Sulfetos/metabolismo , Transplante Heterólogo , Compostos de Zinco/química , Compostos de Zinco/metabolismo
12.
Beilstein J Nanotechnol ; 8: 1734-1741, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28904834

RESUMO

Biocompatible superparamagnetic iron oxide nanoparticles (NPs) through smart chemical functionalization of their surface with fluorescent species, therapeutic proteins, antibiotics, and aptamers offer remarkable potential for diagnosis and therapy of disease sites at their initial stage of growth. Such NPs can be obtained by the creation of proper linkers between magnetic NP and fluorescent or drug probes. One of these linkers is gold, because it is chemically stable, nontoxic and capable to link various biomolecules. In this study, we present a way for a simple and reliable decoration the surface of magnetic NPs with gold quantum dots (QDs) containing more than 13.5% of Au+. Emphasis is put on the synthesis of magnetic NPs by co-precipitation using the amino acid methionine as NP growth-stabilizing agent capable to later reduce and attach gold species. The surface of these NPs can be further conjugated with targeting and chemotherapy agents, such as cancer stem cell-related antibodies and the anticancer drug doxorubicin, for early detection and improved treatment. In order to verify our findings, high-resolution transmission electron microscopy (HRTEM), atomic force microscopy (AFM), FTIR spectroscopy, inductively coupled plasma mass spectroscopy (ICP-MS), and X-ray photoelectron spectroscopy (XPS) of as-formed CoFe2O4 NPs before and after decoration with gold QDs were applied.

13.
Beilstein J Nanotechnol ; 8: 1815-1824, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28904843

RESUMO

Upconverting nanoparticles (UCNPs) are promising, new imaging probes capable of serving as multimodal contrast agents. In this study, monodisperse and ultrasmall core and core-shell UCNPs were synthesized via a thermal decomposition method. Furthermore, it was shown that the epitaxial growth of a NaGdF4 optical inert layer covering the NaGdF4:Yb,Er core effectively minimizes surface quenching due to the spatial isolation of the core from the surroundings. The mean diameter of the synthesized core and core-shell nanoparticles was ≈8 and ≈16 nm, respectively. Hydrophobic UCNPs were converted into hydrophilic ones using a nonionic surfactant Tween 80. The successful coating of the UCNPs by Tween 80 has been confirmed by Fourier transform infrared (FTIR) spectroscopy. Scanning electron microscopy (SEM), powder X-ray diffraction (XRD), photoluminescence (PL) spectra and magnetic resonance (MR) T1 relaxation measurements were used to characterize the size, crystal structure, optical and magnetic properties of the core and core-shell nanoparticles. Moreover, Tween 80-coated core-shell nanoparticles presented enhanced optical and MR signal intensity, good colloidal stability, low cytotoxicity and nonspecific internalization into two different breast cancer cell lines, which indicates that these nanoparticles could be applied as an efficient, dual-modal contrast probe for in vivo bioimaging.

14.
Int J Mol Sci ; 17(8)2016 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-27548152

RESUMO

The uptake and distribution of negatively charged superparamagnetic iron oxide (Fe3O4) nanoparticles (SPIONs) in mouse embryonic fibroblasts NIH3T3, and magnetic resonance imaging (MRI) signal influenced by SPIONs injected into experimental animals, were visualized and investigated. Cellular uptake and distribution of the SPIONs in NIH3T3 after staining with Prussian Blue were investigated by a bright-field microscope equipped with digital color camera. SPIONs were localized in vesicles, mostly placed near the nucleus. Toxicity of SPION nanoparticles tested with cell viability assay (XTT) was estimated. The viability of NIH3T3 cells remains approximately 95% within 3-24 h of incubation, and only a slight decrease of viability was observed after 48 h of incubation. MRI studies on Wistar rats using a clinical 1.5 T MRI scanner were showing that SPIONs give a negative contrast in the MRI. The dynamic MRI measurements of the SPION clearance from the injection site shows that SPIONs slowly disappear from injection sites and only a low concentration of nanoparticles was completely eliminated within three weeks. No functionalized SPIONs accumulate in cells by endocytic mechanism, none accumulate in the nucleus, and none are toxic at a desirable concentration. Therefore, they could be used as a dual imaging agent: as contrast agents for MRI and for traditional optical biopsy by using Prussian Blue staining.


Assuntos
Compostos Férricos/metabolismo , Compostos Férricos/toxicidade , Nanopartículas Metálicas/toxicidade , Animais , Meios de Contraste/metabolismo , Meios de Contraste/toxicidade , Injeções Intramusculares , Imageamento por Ressonância Magnética , Espectroscopia de Ressonância Magnética , Camundongos , Microscopia de Força Atômica , Células NIH 3T3 , Ratos , Ratos Wistar
15.
J Biomed Nanotechnol ; 11(5): 841-53, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-26349396

RESUMO

Quantum dots (QDs) play an active role in triggering biological effects and should not be viewed as ordinary carriers for biomedical applications; therefore, the aim of this study was to investigate the molecular mechanisms involved in the saturating accumulation of non-targeted, carboxylated QDs, the related specific internalization pathways and the induced changes in the endocytotic cycle in NIH3T3 cells. We determined that the saturating accumulation of QDs suppressed the internalization of subsequently introduced QDs that had an identical chemical composition. However, the reinitiation of uptake was detected in the NIH3T3 cells after 8 h of incubation in medium without QDs. A very small suppressive effect of accumulated QDs was observed on uptake via the clathrin-mediated endocytosis pathway and macropinocytosis. In contrast, uptake via the caveolin-mediated pathway was almost completely prevented. Deeper insight into the suppression mechanism was obtained by transiently transfecting NIH3T3 cells with the plasmid pEGFP-C1-Caveolin-1. In these transfected cells, the usual intracellular presence of Caveolin-1 near the plasma membrane was not observed after long-term incubation with QDs. The putative application of QDs for diagnostic visualization in combination with certain anticancer substances was also evaluated. The QDs did not affect the intracellular photosensitization pattern of the amphiphilic molecule chlorin e6 in the case of photodynamic therapy. However, the saturating accumulation of QDs increased the resistance of NIH3T3 cells to the widely used anticancer drug cisplatin.


Assuntos
Endocitose/efeitos dos fármacos , Pontos Quânticos/toxicidade , Animais , Caveolina 1/genética , Caveolina 1/metabolismo , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Redes e Vias Metabólicas/efeitos dos fármacos , Redes e Vias Metabólicas/genética , Camundongos , Células NIH 3T3 , Pontos Quânticos/metabolismo , Transfecção
16.
Medicina (Kaunas) ; 50(4): 237-44, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25458961

RESUMO

BACKGROUND AND OBJECTIVE: Superparamagnetic iron oxide nanoparticles (SPIONs) emerge as a promising tool for early cancer diagnostics and targeted therapy. However, both toxicity and biological activity of SPIONs should be evaluated in detail. The aim of this study was to synthesize superparamagnetic cobalt ferrite nanoparticles (Co-SPIONs), and to investigate their uptake, toxicity and effects on cancer stem-like properties in human pancreatic cancer cell line MiaPaCa2 and human ovarian cancer cell line A2780. MATERIALS AND METHODS: Co-SPIONs were produced by Massart's co-precipitation method. The cells were treated with Co-SPIONs at three different concentrations (0.095, 0.48, and 0.95µg/mL) for 24 and 48h. Cell viability and proliferation were analyzed after treatment. The stem-like properties of cells were assessed by investigating the cell clonogenicity and expression of cancer stem cell-associated markers, including CD24/ESA in A2780 cell line and CD44/ALDH1 in MiaPaCa2 cell line. Magnetically activated cell sorting was used for the separation of magnetically labeled and unlabeled cells. RESULTS: Both cancer cell lines accumulated Co-SPIONs, however differences in response to nanoparticles were observed between MiaPaCa2 and A2780 cell. In particular, A2780 cells were more sensitive to exposition to Co-SPIONs than MiaPaCa2 cells, indicating that a safe concentration of nanoparticles must be estimated individually for a particular cell type. Higher doses of Co-SPIONs decreased both the clonogenicity and ESA marker expression in A2780 cells. CONCLUSIONS: Co-SPIONs are not cytotoxic to cancer cells, at least when used at a concentration of up to 0.95µg/mL. Co-SPIONs have a dose-dependent effect on the clonogenic potential and ESA marker expression in A2780 cells. Magnetic detection of low concentrations of Co-SPIONS in cancer cells is a promising tool for further applications of these nanoparticles in cancer diagnosis and treatment; however, extensive research in this field is needed.


Assuntos
Cobalto/metabolismo , Compostos Férricos/metabolismo , Nanopartículas/metabolismo , Neoplasias Ovarianas/metabolismo , Neoplasias Pancreáticas/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Separação Celular , Sobrevivência Celular/efeitos dos fármacos , Cobalto/farmacologia , Feminino , Compostos Férricos/farmacologia , Citometria de Fluxo , Humanos , Tamanho da Partícula
17.
Int J Nanomedicine ; 8: 555-68, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23429995

RESUMO

BACKGROUND: The purpose of this study was to elucidate the mechanism of natural uptake of nonfunctionalized quantum dots in comparison with microinjected quantum dots by focusing on their time-dependent accumulation and intracellular localization in different cell lines. METHODS: The accumulation dynamics of nontargeted CdSe/ZnS carboxyl-coated quantum dots (emission peak 625 nm) was analyzed in NIH3T3, MCF-7, and HepG2 cells by applying the methods of confocal and steady-state fluorescence spectroscopy. Intracellular colocalization of the quantum dots was investigated by staining with Lysotracker(®). RESULTS: The uptake of quantum dots into cells was dramatically reduced at a low temperature (4 °C), indicating that the process is energy-dependent. The uptake kinetics and imaging of intracellular localization of quantum dots revealed three accumulation stages of carboxyl-coated quantum dots at 37 °C, ie, a plateau stage, growth stage, and a saturation stage, which comprised four morphological phases: adherence to the cell membrane; formation of granulated clusters spread throughout the cytoplasm; localization of granulated clusters in the perinuclear region; and formation of multivesicular body-like structures and their redistribution in the cytoplasm. Diverse quantum dots containing intracellular vesicles in the range of approximately 0.5-8 µm in diameter were observed in the cytoplasm, but none were found in the nucleus. Vesicles containing quantum dots formed multivesicular body-like structures in NIH3T3 cells after 24 hours of incubation, which were Lysotracker-negative in serum-free medium and Lysotracker-positive in complete medium. The microinjected quantum dots remained uniformly distributed in the cytosol for at least 24 hours. CONCLUSION: Natural uptake of quantum dots in cells occurs through three accumulation stages via a mechanism requiring energy. The sharp contrast of the intracellular distribution after microinjection of quantum dots in comparison with incubation as well as the limited transfer of quantum dots from vesicles into the cytosol and vice versa support the endocytotic origin of the natural uptake of quantum dots. Quantum dots with proteins adsorbed from the culture medium had a different fate in the final stage of accumulation from that of the protein-free quantum dots, implying different internalization pathways.


Assuntos
Espaço Intracelular/metabolismo , Microinjeções/métodos , Pontos Quânticos , Animais , Linhagem Celular Tumoral , Endocitose , Proteínas de Fluorescência Verde/metabolismo , Células Hep G2 , Humanos , Camundongos , Microscopia Confocal , Células NIH 3T3
18.
Medicina (Kaunas) ; 42(7): 542-58, 2006.
Artigo em Lituano | MEDLINE | ID: mdl-16861836

RESUMO

Nanotechnology is an area of scientific research and technology development dealing with the structures and devices with length scales in the 1- to 100-nanometer range. Multifunctional nanoparticles that can target, diagnose, and treat diseases such as cancer are designed and developed. Numerous studies have shown that anticancer drugs encapsulated into nanoparticles can be delivered to target tumor cells and tissue. This would increase antitumor efficacy and reduce systemic side effects. Quantum dots are kind of nanoparticles with unique photochemical and photophysical properties. They are several orders of magnitude brighter than conventional fluorophores, and their emission spectra are very narrow and can be tuned by adjusting the size of the dots. Quantum dots are a new class of fluorescent labels with improved brightness and resistance against photobleaching. These properties could improve the sensitivity of biological detection and imaging by at least 10- to 100-fold. In this review, data of tumor targeting with conventional and long-circulating nanoparticles as well as applications of semiconductor quantum dots for in vivo imaging are discussed.


Assuntos
Antineoplásicos/administração & dosagem , Nanomedicina , Nanoestruturas , Nanotecnologia , Neoplasias/diagnóstico , Neoplasias/tratamento farmacológico , Animais , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/uso terapêutico , Carcinoma Hepatocelular/tratamento farmacológico , Doxorrubicina/administração & dosagem , Doxorrubicina/uso terapêutico , Sistemas de Liberação de Medicamentos , Resistencia a Medicamentos Antineoplásicos , Feminino , Fluorescência , Previsões , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Masculino , Camundongos , Microscopia de Fluorescência , Fotoquímica , Pontos Quânticos , Ratos , Semicondutores
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