4 research outputs found
Doxorubicin-Incorporated Nanotherapeutic Delivery System Based on Gelatin-Coated Gold Nanoparticles: Formulation, Drug Release, and Multimodal Imaging of Cellular Internalization
In
this work, we developed a new pH- and temperature-responsive nanochemotherapeutic
system based on Doxorubicin (DOX) noncovalently bound to biosynthesized
gelatin-coated gold nanoparticles (DOX-AuNPs@gelatin). The real-time
release profile of DOX was evaluated at different pH values (7.4,
5.3, and 4.6) and temperatures (22–45 °C) in aqueous solutions,
and its therapeutic performance was examined <i>in vitro</i> against MCF-7 breast cancer cells. TEM, dark-field scattering, and
wide-field fluorescence microscopy indicated the effective uptake
of nanochemotherapeutics with the subsequent release and progressive
accumulation of DOX in cell nuclei. MTT assays clearly showed the
effectiveness of the treatment by inhibiting the growth of MCF-7 breast
cancer cells for a loaded drug concentration of 5 ÎĽg/mL. The
most informative data about the dynamic release and localization were
provided by scanning confocal microscopy using time-resolved fluorescence
and surface-enhanced Raman scattering (SERS) techniques. In particular,
fluorescence-lifetime imaging (FLIM) recorded under 485 nm pulsed
diode laser excitation revealed the bimodal distribution of DOX in
cells. The shorter fluorescence lifetime of DOX localized in nuclei
(1.52 ns) than in the cytoplasm (2.4 ns) is consistent with the cytotoxic
mechanism induced by DOX–DNA intercalation. Remarkably, the
few DOX molecules captured between nanoparticles (“electromagnetic
hotspots”) after most drug is released act as SERS reporters
for the localization of plasmonic nanocarriers in MCF-7 cells. The
high drug loading capacity and effective drug release under pH control
combined with the advantage of multimodal visualization inside cells
clearly indicate the high potential of our DOX–AuNPs@gelatin
delivery system for implementation in nanomedicine
Designing Theranostic Agents Based on Pluronic Stabilized Gold Nanoaggregates Loaded with Methylene Blue for Multimodal Cell Imaging and Enhanced Photodynamic Therapy
At present, multifunctional noble
metal-based nanocomposites are extensively investigated for their
potential in performing cellular imaging, diagnostics, and therapy
by integration of unique plasmonic properties with the spectroscopic
expression and therapeutic activity of appropriate drug. In this work,
we report the fabrication of 3-dimensional (3-D) close-packed nanoassemblies
of gold nanoparticles by controlling the aggregation of individual
nanoparticles in solution and subsequent stabilization of formed aggregates
by Pluronic block copolymer (F127) coating. Besides conferring high
stability, Pluronic mediates the loading of Methylene Blue (MB) molecules
which exhibit interesting spectroscopic and photochemical properties
to be employed as both optical label and photosensitizing drug. Indeed,
here we demonstrate the pertinence of the fabricated nanoassemblies
to provide optical imaging of murine colon carcinoma cells (C-26)
via both Raman and fluorescence signals collected from MB molecules,
specifically by using scanning confocal surface-enhanced resonant
raman spectroscopy (SERRS) and fluorescence lifetime imaging microscopy
(FLIM) techniques. The specific configuration of as fabricated nanoassemblies
allows a small population of MB molecules to be located in very small
areas between the aggregated nanoparticles (“hot spots”)
to provide SERRS signal while the other population remains captured
in Pluronic coating and preserves both its fluorescence signal and
singlet-oxygen generation capability. Remarkably, we demonstrate an
enhanced photodynamic therapeutic activity of MB-loaded gold nanoaggregates
against murine colon carcinoma cells (C-26), as compared to the free
photosensitizer. To our knowledge, this is the first report on plasmonic
nanoplatforms conveying photosensitizing drug into cells to operate
as optical label via both SERÂ(R)S and FLIM and to perform enhanced
photodynamic therapy
Folic Acid-Conjugated, SERS-Labeled Silver Nanotriangles for Multimodal Detection and Targeted Photothermal Treatment on Human Ovarian Cancer Cells
The
effectiveness of a therapeutic agent for cancer stands in its
ability to reduce and eliminate tumors without harming the healthy
tissue nearby. Nanoparticles peripherally conjugated with targeting
moieties offer major improvements in therapeutics through site specificity.
In this study we demonstrate this approach by targeting the folate
receptor of NIH:OVCAR-3 human ovary cancer cell line. Herein we used
silver nanotriangles which were biocompatibilized with chitosan (bio)Âpolymer,
labeled with para-aminothiophenol (pATP) Raman reporter molecule,
and conjugated with folic acid. The nanoparticles conjugation and
efficient labeling was investigated by localized surface plasmon resonance
(LSPR), zeta potential, and surface-enhanced Raman scattering (SERS)
measurements. Conjugated particles were proven to be highly stable
in aqueous and cellular medium. The targeted uptake of conjugated
nanoparticles by human ovary cancer cells was confirmed by dark field
microscopy and scattering spectra of the particles inside cells. Comparative
studies revealed specific internalization of the conjugated nanoparticles
in comparison with similar bare nanoparticles. Moreover, the SERS
identity of the particles was proven to be highly conserved inside
cells. Targeted cancer cell treatment conducted by irradiating the
nanoparticle-treated cells with a continuous wave-nearinfrared (cw-NIR)
laser in resonance with their plasmonic band proved an efficient therapeutic
response. By integrating the advantages of multimodal optical imaging
and SERS detection with hyperthermia capabilities through site specificity,
these nanoparticles can represent a real candidate for personalized
medicine
Carboplatin-Loaded, Raman-Encoded, Chitosan-Coated Silver Nanotriangles as Multimodal Traceable Nanotherapeutic Delivery Systems and pH Reporters inside Human Ovarian Cancer Cells
Ovarian
cancer is a common cause of cancer death in women and is associated
with the highest mortality rates of all gynecological malignancies.
Carboplatin (CBP) is the most used cytotoxic agent in the treatment
of ovarian cancer. Herein, we design and assess a CBP nanotherapeutic
delivery system which allows combinatorial functionalities of chemotherapy,
pH sensing, and multimodal traceable properties inside live NIH:OVCAR-3
ovarian cancer cells. In our design, a pH-sensitive Raman reporter,
4-mercaptobenzoic acid (4MBA) is anchored onto the surface of chitosan-coated
silver nanotriangles (chit-AgNTs) to generate a robust surface-enhanced
Raman scattering (SERS) traceable system. To endow this nanoplatform
with chemotherapeutic abilities, CBP is then loaded to 4MBA-labeled
chit-AgNTs (4MBA-chit-AgNTs) core under alkaline conditions. The uptake
and tracking potential of CBP-4MBA-chit-AgNTs at different <i>Z</i>-depths inside live ovarian cancer cells is evaluated by
dark-field and differential interference contrast (DIC) microscopy.
The ability of CBP-4MBA-chit-AgNTs to operate as near-infrared (NIR)-responsive
contrast agents is validated using two noninvasive techniques: two-photon
(TP)-excited fluorescence lifetime imaging microscopy (FLIM) and confocal
Raman microscopy (CRM). The most informative data about the precise
localization of nanocarriers inside cells correlated with intracellular
pH sensing is provided by multivariate analysis of Raman spectra collected
by scanning CRM. The <i>in vitro</i> cell proliferation
assay clearly shows the effectiveness of the prepared nanocarriers
in inhibiting the growth of NIH:OVCAR-3 cancer cells. We anticipate
that this class of nanocarriers holds great promise for application
in image-guided ovarian cancer chemotherapy