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1.
Biomater Sci ; 9(18): 6183-6202, 2021 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-34346411

RESUMO

Among women, ovarian cancer is the fifth most frequent type of cancer, and despite benefiting from current standard treatment plans, 90% of patients relapse in the subsequent 18 months and, eventually, perish. As a result, via embracing nanotechnological advancements in the field of medical science, researchers working in the areas of cancer therapy and imaging are looking for the next breakthrough treatment strategy to ensure lower cancer recurrence rates and improved outcomes for patients. Herein, we design a novel phototheranostic agent with optical features in the biological window of the electromagnetic spectrum via encapsulating a newly synthesized phthalocyanine dye within biocompatible protein nanoparticles, allowing the targeted fluorescence imaging and synergistic dual therapy of ovarian cancer. The nanosized agent displays great biocompatibility and enhanced aqueous biostability and photothermal activity, as well as high reactive-oxygen-species generation efficiency. To achieve the active targeting of the desired malignant tissue and suppress the rapid clearance of the photosensitive agent from the peritoneal cavity, the nanoparticles are biofunctionalized with an anti-folate receptor antibody. A2780 ovarian cancer cells are employed to confirm the improved targeting capabilities and the in vitro cytotoxic efficiency of the theranostic nanoparticles after exposure to a 660 nm LED lamp; upon measurement via MTT and flow cytometry assays, a significant 95% decrease in the total number of viable cells is seen. Additionally, the therapeutic performance of our newly designed nanoparticles was evaluated in vivo, via real-time thermal monitoring and histopathological assays, upon the irradiation of tumour-bearing mice with a 660 nm LED lamp (0.05 W cm-2). Foremost, separately from steady-state fluorescence imaging, we found that, via utilizing FLIM investigations, the differences in fluorescence lifetimes of antibody biofunctionalized and non-functionalized nanoparticles can be correlated to different intracellular localization and internalization pathways of the fluorescent agent, which is relevant for the development of a cutting-edge method for the detection of cancer cells that overexpress folate receptors at their surfaces.


Assuntos
Nanopartículas , Neoplasias Ovarianas , Animais , Linhagem Celular Tumoral , Feminino , Humanos , Camundongos , Imagem Óptica , Neoplasias Ovarianas/diagnóstico por imagem , Neoplasias Ovarianas/tratamento farmacológico , Fototerapia , Medicina de Precisão , Nanomedicina Teranóstica
2.
Molecules ; 26(15)2021 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-34361832

RESUMO

In recent times, researchers have aimed for new strategies to combat cancer by the implementation of nanotechnologies in biomedical applications. This work focuses on developing protein-based nanoparticles loaded with a newly synthesized NIR emitting and absorbing phthalocyanine dye, with photodynamic and photothermal properties. More precisely, we synthesized highly reproducible bovine serum albumin-based nanoparticles (75% particle yield) through a two-step protocol and successfully encapsulated the NIR active photosensitizer agent, achieving a good loading efficiency of 91%. Making use of molecular docking simulations, we confirm that the NIR photosensitizer is well protected within the nanoparticles, docked in site I of the albumin molecule. Encouraging results were obtained for our nanoparticles towards biomedical use, thanks to their negatively charged surface (-13.6 ± 0.5 mV) and hydrodynamic diameter (25.06 ± 0.62 nm), favorable for benefitting from the enhanced permeability and retention effect; moreover, the MTT viability assay upholds the good biocompatibility of our NIR active nanoparticles. Finally, upon irradiation with an NIR 785 nm laser, the dual phototherapeutic effect of our NIR fluorescent nanoparticles was highlighted by their excellent light-to-heat conversion performance (photothermal conversion efficiency 20%) and good photothermal and size stability, supporting their further implementation as fluorescent therapeutic agents in biomedical applications.


Assuntos
Indóis/administração & dosagem , Nanopartículas/administração & dosagem , Neoplasias Ovarianas/tratamento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/administração & dosagem , Soroalbumina Bovina/química , Proliferação de Células , Feminino , Humanos , Indóis/química , Luz , Simulação de Acoplamento Molecular , Nanopartículas/química , Neoplasias Ovarianas/patologia , Fármacos Fotossensibilizantes/química , Espectroscopia de Luz Próxima ao Infravermelho , Células Tumorais Cultivadas
3.
Colloids Surf B Biointerfaces ; 194: 111213, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32622254

RESUMO

A great amount of effort is directed towards the progress of cancer treatment approaches aspiring to develop non-invasive, targeted and highly efficient therapies. In this context, Photothermal (PTT) and Photodynamic (PDT) Therapies were proven as promising. This work aims to integrate the therapeutic activities of two near-infrared (NIR) photoactive biomaterials - gold nano-bipyramids (AuBPs) and Indocyanine Green (ICG) - into one single targeted hybrid nanosystem able to operate as dual PTT-PDT agent with higher efficiency compared with each one alone. Firstly, different aspect ratio' AuBPs were systematically investigated in water solution for their intrinsic ability to efficiently generate toxic reactive oxygen species, namely oxygen singlet (1O2), under NIR laser irradiation, as this effect is less investigated in literature. Interestingly, the photodynamic activity of AuBPs measured by monitoring the photooxidation of 9,10-Anthracenediyl-bis(methylene)dimalonic acid (ABDA) - a well-known 1O2 sensor, is important, counting for 30 % decrease in ABDA optical absorbance for the most active AuBPs, well-correlating with the previously determined photothermal conversion efficiency. Furthermore, ICG was successfully grafted onto the Poly-lactic acid (PLA) coating of plasmonic nanoparticles and, consequently, the as-designed fully integrated hybrid nanosystem shows improved PTT-PDT performance in solution. Specifically, by triggering simultaneous PTT-PDT activities, the 1O2 amount is doubled, while the heating monitoring shows higher and faster increase in temperature compared to AuBPs alone. Finally, the efficiency of the combined PTT-PDT therapeutic activity was validated in vitro against B16-F10 cell line by covalent conjugation of the nanosystem with Folic Acid, which ensures the cellular recognition by overexpression of folate receptor.


Assuntos
Melanoma , Fotoquimioterapia , Ouro , Humanos , Verde de Indocianina , Triazenos
4.
Molecules ; 25(14)2020 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-32668589

RESUMO

Nowadays, thanks to nanotechnological progress, which itself guides us more and more closely toward not only the efficient design of innovative nanomaterials or nanostructures, but to the improvement of their functionality, we benefit from an important asset in the battle against pathogenic illnesses. Herein, we report a versatile biocompatible plasmonic nanoplatform based on a Whatman paper incorporating positively-charged gold nanospherical particles via the immersion approach. The morphological characterization of the as-engineered-plasmonic paper was examined by SEM (scanning electron microscopy) and HRTEM (high-resolution transmission electron microscopy) investigations, while its surface chemical modification with a synthetic polypeptide, specifically RRWHRWWRR-NH2 (P2), was proved by monitoring the plasmonic response of loaded gold nanospheres and the emission signal of P2 via fluorescence spectroscopy. The as-functionalized plasmonic paper is non-cytotoxic towards BJ fibroblast human cells at bactericidal concentrations. Finally, the antimicrobial activity of the P2-functionalized plasmonic paper on both planktonic bacteria and biofilms was tested against two reference strains: Gram-positive Bacteria, i.e., Staphylococcus aureus and the Gram-negative Bacteria, i.e., Escherichia coli, determining microbial inhibition of up to 100% for planktonic bacteria. In line with the above presented nanoplatform's proper design, followed by their functionalization with active antimicrobial peptides, new roads can be open for determining antibiotic-free treatments against different relevant pathogens.


Assuntos
Antibacterianos , Materiais Biocompatíveis , Escherichia coli/efeitos dos fármacos , Ouro/farmacologia , Nanopartículas Metálicas/química , Peptídeos/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/farmacologia , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Biofilmes/efeitos dos fármacos , Linhagem Celular , Humanos , Papel
5.
Nanomaterials (Basel) ; 10(6)2020 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-32471140

RESUMO

Designing innovative (nano)detection platforms, respecting their low-cost and fabrication simplicity, capable to chemically detect multiple target analytes by employing the same engineered device, is still a great challenge in the multiplexed biosensor development. In this scientific context, in the current manuscript, we exploit the low-cost plasmonic calligraphy as a versatile approach to directly draw continuous plasmonic lines on Whatman paper using a regular ballpoint pen successively filled with two different anisotropic nanoparticles shapes (gold bipyramids-AuBPs and gold nanorods-AuNRs) as colloidal inks. After the efficient immobilization of the positively-charged AuBPs and AuNRs onto the paper fibres, proved by Scanning Electron Microscopy (SEM) investigations, the specificity of our as-calligraphed-paper platform is ensured by coating the selected lines with a thin layer of anionic poly(styrene sulfonate) polyelectrolyte, creating, consequently, a well-defined plasmonic array of charge-selective regions. Finally, the functionality of the well-isolated and as-miniaturized active plasmonic array is, subsequently, tested using the anionic Rose-Bengal and cationic Rhodamine 6G target analytes and proved by complementary dual optical "ON/OFF Switch" sensing (i.e. Surface-enhanced Raman Scattering sensing/metal-enhanced fluorescence sensing) onto the same plasmonic line, developing thus a simple multiplexed plasmonic array platform, which could further facilitate the well-desired biomarker detection in complex mixtures.

6.
Nanotechnology ; 31(33): 335502, 2020 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-32348974

RESUMO

In this work, we propose a novel approach to design robust microfluidic devices with integrated plasmonic transducers allowing portability, reduced analysis time through dynamic measurements and high sensitivity. Specifically, the strategy we apply involves two steps: (i) the controlled deposition of gold bipyramidal nanoparticles (AuBPs) onto a functionalized solid glass substrate and (ii) the integration of the as-fabricated plasmonic substrate into a polydimethylsiloxane (PDMS) microfluidic circuit. The localized surface plasmon resonance (LSPR) sensitivity of the plasmonic-microfluidic device was evaluated by monitoring the optical responses at refractive index changes, proving a bulk sensitivity of 243 nm RIU-1 for the longitudinal LSPR band of isolated AuBPs and 150 nm RIU-1 for the band assigned to end-to-end linked nanoparticles. A strong electric field generated in the gaps between AuBPs-due to the generation of the so-called extrinsic 'hot-spots'-was subsequently proved by the volumetric surface enhanced Raman scattering (SERS) detection of molecules in continuous flow conditions by loading the analyte into the microfluidic channel via a syringe pump. In conclusion, our miniaturized portable microfluidic system aims to detect and identify, in real-time with high accuracy, analyte molecules in laminal flow, thus providing a groundwork for further complex biosensing applications.

7.
Nanotechnology ; 30(40): 405701, 2019 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-31247611

RESUMO

In this work, we present a thorough study on the evaluation of the photothermal conversion efficiencies of gold nanobipyramids (AuBPs) under irradiation by two phototherapeutic laser lines at 785 and 808 nm. Due to fine tunability of the longitudinal localized surface plasmon resonance (LSPR) of AuBPs along the entire biological window, AuBPs have great potential to be applied as efficient photothermal agents in specific hyperthermia applications. Aiming to identify the most suitable AuBPs for each laser line, here we synthetized AuBPs of six different aspect ratios with longitudinal LSPR ranging from 662 to 929 nm and compared their intrinsic photothermal properties in colloidal solutions under laser irradiation at various experimental parameters such as sample volume, optical density and laser power. In addition, the experimental plasmonic resonances of the as-prepared AuBPs were perfectly simulated and their theoretical extinction and absorption cross-sections provided by finite-difference time-domain technique. Finally, we found photothermal conversion efficiencies ranging from 40% to 97% for all AuBPs systems under both NIR irradiation laser lines concluding that for the 785 nm excitation wavelength the AuBPs with longitudinal LSPR at 802 nm are most efficient, whereas in the case of the 808 nm laser line the AuBPs with optical response at 812 nm exhibit the best thermal performance. These studies are crucial for designing AuBPs as effective phototherapy agents acting alone or in combination with other plasmon-based or plasmon-assisted therapies.

8.
Front Chem ; 7: 55, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30800650

RESUMO

In this work, we design new plasmonic paper-based nanoplatforms with interesting capabilities in terms of sensitivity, efficiency, and reproducibility for promoting multimodal biodetection via Localized Surface Plasmon Resonance (LSPR), Surface Enhanced Raman Spectroscopy (SERS), and Metal Enhanced Fluorescence (MEF). To succeed, we exploit the unique optical properties of gold nanobipyramids (AuBPs) deposited onto the cellulose fibers via plasmonic calligraphy using a commercial pen. The first step of the biosensing protocol was to precisely graft the previously chemically-formed p-aminothiophenol@Biotin system, as active recognition element for target streptavidin detection, onto the plasmonic nanoplatform. The specific capture of the target protein was successfully demonstrated using three complementary sensing techniques. As a result, while the LSPR based sensing capabilities of the nanoplatform were proved by successive 13-18 nm red shifts of the longitudinal LSPR associated with the change of the surface RI after each step. By employing the ultrasensitive SERS technique, we were able to indirectly confirm the molecular identification of the biotin-streptavidin interaction due to the protein fingerprint bands assigned to amide I, amide III, and Trp vibrations. Additionally, the formed biotin-streptavidin complex acted as a spacer to ensure an optimal distance between the AuBP surface and the Alexa 680 fluorophore for achieving a 2-fold fluorescence emission enhancement of streptavidin@Alexa 680 on the biotinylated nanoplatform compared to the same complex on bare paper (near the plasmonic lines), implementing thus a novel MEF sensing nanoplatform. Finally, by integrating multiple LSPR, SERS, and MEF nanosensors with multiplex capability into a single flexible and portable plasmonic nanoplatform, we could overcome important limits in the field of portable point-of-care diagnostics.

9.
Sensors (Basel) ; 18(9)2018 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-30208609

RESUMO

Paper-based platforms can be a promising choice as portable sensors due to their low-cost and facile fabrication, ease of use, high sensitivity, specificity and flexibility. By combining the qualities of these 3D platforms with the optical properties of gold nanoparticles, it is possible to create efficient nanodevices with desired biosensing functionalities. In this work, we propose a new plasmonic paper-based dual localized surface plasmon resonance⁻surface-enhanced Raman scattering (LSPR-SERS) nanoplatform with improved detection abilities in terms of high sensitivity, uniformity and reproducibility. Specifically, colloidal gold nanorods (GNRs) with a well-controlled plasmonic response were firstly synthesized and validated as efficient dual LSPR-SERS nanosensors in solution using the p-aminothiophenol (p-ATP) analyte. GNRs were then efficiently immobilized onto the paper via the immersion approach, thus obtaining plasmonic nanoplatforms with a modulated LSPR response. The successful deposition of the nanoparticles onto the cellulose fibers was confirmed by LSPR measurements, which demonstrate the preserved plasmonic response after immobilization, as well as by dark-field microscopy and scanning electron microscopy investigations, which confirm their uniform distribution. Finally, a limit of detection for p-ATP as low as 10-12 M has been achieved by our developed SERS-based paper nanoplatform, proving that our optimized plasmonic paper-based biosensing design could be further considered as an excellent candidate for miniaturized biomedical applications.

10.
Anal Chem ; 90(14): 8567-8575, 2018 07 17.
Artigo em Inglês | MEDLINE | ID: mdl-29902917

RESUMO

In this work, we demonstrate the feasibility of gold bipyramidal-shaped nanoparticles (AuBPs) to be used as active plasmonic nanoplatforms for the detection of the biotin-streptavidin interaction in aqueous solution via both Localized Surface Plasmon Resonance and Surface Enhanced Raman Scattering (LSPR/SERS). Our proof of concept exploits the precise attachment of the recognition element at the tips of AuBPs, where the electromagnetic field is stronger, which is beneficial to the surface sensitivity of longitudinal LSPR on the local refractive index and to the electromagnetic enhancement of SERS activity, too. Indeed, successive red shifts of the longitudinal LSPR associated with increased local refractive index reveal the attachment of para-aminothiophenol (p-ATP) chemically labeled Biotin to the Au surface and the specific capture of the target protein by biotin-functionalized AuBPs. Finite-Difference Time-Domain simulations based on the reconstructed index of refraction confirm LSPR measurements. However, the molecular identification of the biotin-streptavidin interaction remains elusive by LSPR investigation alone. Remarkably, we succeeded to complement the LSPR detection with reliable SERS measurements which permitted to (a) certify the molecular identification of biotin-streptavidin interaction and (b) extend the limit of detection of streptavidin in solution toward 10-12 M. Finally, to further probe the possibility to implement the AuBPs as dual LSPR-SERS based immunoassays in solution for real clinical diagnostics, we additionally investigated the AuBP's performance to transduce the specific antihuman IgG- human IgG binding event, providing thus a reference design for building unique plasmonic immunoassays for dual-optical detection of target proteins in aqueous solution.


Assuntos
Técnicas Biossensoriais/instrumentação , Ouro/química , Imunoensaio/instrumentação , Imunoglobulina G/análise , Análise Espectral Raman/instrumentação , Ressonância de Plasmônio de Superfície/instrumentação , Biotina/química , Humanos , Estreptavidina/química
11.
Biosens Bioelectron ; 86: 728-735, 2016 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-27476053

RESUMO

In this manuscript we propose a simple and efficient strategy to improve the sensitivity of localized surface plasmon resonance (LSPR) shift-based biosensors using biotin-streptavidin recognition interaction as a proof-of-concept. Specifically, biotin molecules are immobilized on a low-cost plasmonic LSPR biosensor based on annealed self-assembled spherical gold nanoparticles (AuNSs) and successively incubated with increasing concentrations of streptavidin, achieving a limit of detection (LOD) of 5nM. Interestingly, when the detection is performed by the same biotin-functionalized plasmonic AuNSs substrate but against streptavidin previously conjugated to gold nanorods, the LSPR shift is 26-fold enhanced. Moreover, we confirm these results through numerical simulations and demonstrate that the proposed sensing architecture can operate as transducer not only to confirm the adsorption of bioanalyte but also to provide the chemical identity of the capture and targeted molecules from their vibrational Raman fingerprints. Therefore, we are confident that the development of such plasmonic biosensors that use metallic labels for improving the sensitivity of detection could become highly promising for future point-of-care diagnostic assays, pushing sensitivity towards single-molecule detection limit.


Assuntos
Reações Antígeno-Anticorpo/imunologia , Biotina/imunologia , Imunoensaio/instrumentação , Nanotubos/química , Estreptavidina/imunologia , Ressonância de Plasmônio de Superfície/instrumentação , Técnicas Biossensoriais/instrumentação , Biotina/análise , Desenho de Equipamento , Análise de Falha de Equipamento , Ouro , Nanotecnologia/instrumentação , Nanotubos/ultraestrutura , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Estreptavidina/análise
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