Timea Simon

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)polymer, 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.

Biosynthesized silver nanoparticles performing as biogenic SERS-nanotags for investigation of C26 colon carcinoma cells

In this work, two classes of silver nanoparticles (AgNPs) were biosynthesized with the goal to assess their reliability in vitro as surface-enhanced Raman scattering (SERS) nanotags. Mycosynthesized silver nanoparticles (MAgNPs) and phytosynthesized silver nanoparticles (PAgNPs) were produced through environmentally friendly procedures by reduction of silver nitrate with Fusarium oxysporum cell filtrate and Azadirachta indica extract, respectively. Two cell lines, namely C26 murine colon carcinoma cells as example of cancer cells and human immortalized keratinocyte cells (HaCaT) as representative of healthy cell line, were selected for in vitro investigation. The in vitro toxicity studies show that M(P)AgNPs present lower cytotoxic effect on both cell lines as compared with standard citrate coated AgNPs. The internalization of M(P)AgNPs by colon carcinoma cells and structural alterations induced in the morphology of treated cells were analyzed by dark-field (DF) and differential interference contrast (DIC) microscopy, respectively. The most informative data about the cellular uptake and tracking potential of M(P)AgNPs were provided by scanning Confocal Raman Microscopy (CRM) and multivariate K-means cluster analysis of collected Raman spectra. The analysis reveals the subcellular components and the localization of AgNPs inside the cell via the intrinsic SERS signature of biogenic coating material. The use of unique biological material to perform synthesis, stability, biocompatibility and SERS tagging is relevant both from the point of view of encoding nanoparticles with Raman reporters and further applications in cell investigation via Raman/SERS imaging.

Pluronic-Nanogold hybrids: Synthesis and tagging with photosensitizing molecules

The design of polymeric-metal hybrid nanocomposites with multiple functionalities, i.e. from enabling detection and imaging to assisting diagnosis and therapy, is becoming an important research topic in nanomedicine. In this work, Pluronic-Nanogold hybrid nanoparticles (Au-PF127) were successfully prepared in aqueous solution in a single step reaction using Pluronic F127 block copolymer as both reducing and stabilizing agent. The ability of polymer to control the nanoparticle formation and stabilization was systematically investigated by several characterization techniques: UV-Vis absorption, transmission electron microscopy (TEM), dynamic light scattering (DLS) and zeta potential measurements. It was found that polymer concentrations higher than critical micelle concentration (CMC) provide stable nanoparticles even in high molarity NaCl solution. In view of biomedical applications, as prepared Au-PF127 nanoparticles were modified with Methylene Blue (MB) phenothiazinium based photosensitizing drug. Combined surface enhanced Raman scattering (SERS) and fluorescence detection of MB embedded within the polymer shell has revealed the dual functionality of MB-encoded Pluronic-Nanogold hybrids (Au-PF127-MB) to operate under biological conditions as both effective drug carriers and sensitive optical probes.

Gold nanoparticles enhance the effect of tyrosine kinase inhibitors in acute myeloid leukemia therapy

Background and aims Every year, in Europe, acute myeloid leukemia (AML) is diagnosed in thousands of adults. For most subtypes of AML, the backbone of treatment was introduced nearly 40 years ago as a combination of cytosine arabinoside with an anthracycline. This therapy is still the worldwide standard of care. Two-thirds of patients achieve complete remission, although most of them ultimately relapse. Since the FLT3 mutation is the most frequent, it serves as a key molecular target for tyrosine kinase inhibitors (TKIs) that inhibit FLT3 kinase. In this study, we report the conjugation of TKIs onto spherical gold nanoparticles. Materials and methods The internalization of TKI-nanocarriers was proved by the strongly scattered light from gold nanoparticles and was correlated with the results obtained by transmission electron microscopy and dark-field microscopy. The therapeutic effect of the newly designed drugs was investigated by several methods including cell counting assay as well as the MTT assay. Results We report the newly described bioconjugates to be superior when compared with the drug alone, with data confirmed by state-of-the-art analyses of internalization, cell biology, gene analysis for FLT3-IDT gene, and Western blotting to assess degradation of the FLT3 protein. Conclusion The effective transmembrane delivery and increased efficacy validate its use as a potential therapeutic.

LED‐activated methylene blue‐loaded Pluronic‐nanogold hybrids for in vitro photodynamic therapy

In this work we introduce a new class of multifunctional photodynamic agents based on the coupling of photosensitizer molecules with noble metal nanoparticles, which can be efficiently activated under low light intensity. The favourable modification of the photophysical properties of methylene blue (MB) in MB-loaded Pluronic-nanogold hybrids (Au-PF127-MB) increases the probability of singlet oxygen generation, which in turn allows the use of a light emitting diode (LED) irradiation source instead of commonly used, more invasive lasers. In this regard, Au-PF127-MB treated human lung carcinoma cells (HTB 177) were irradiated at different light doses, using a 660 nm LED source, the results indicating a dose dependent therapeutic effect, decreasing the cell viability down to 13%. Owing to their effectiveness, biocompatibility and integrated imaging and therapeutic functionalities, Au-PF127-MB could represent an important development in the field of biophotonic applications.

Gelatin-coated Gold Nanoparticles as Carriers of FLT3 Inhibitors for Acute Myeloid Leukemia Treatment.

This study presents the design of a gold nanoparticle (AuNPs)—drug system with improved efficiency for the treatment of acute myeloid leukemia. The system is based on four different FLT3 inhibitors, namely midostaurin, sorafenib, lestaurtinib, and quizartinib, which were independently loaded onto gelatin‐coated gold nanoparticles. Detailed investigation of the physicochemical properties of the formed complexes lead to the selection of quizartinib—loaded AuNPs for the in vitro evaluation of the biological effects of the formed complex against OCI‐AML3 acute myeloid leukemia cells. Viability tests by MTT demonstrated that the proposed drug complex has improved efficacy when compared with the drug alone. The obtained results constitute a premise for further in vivo investigation of such drug vehicles based on AuNPs. To the best of our knowledge, this is the first study that investigates the delivery of the above‐mentioned FLT3 inhibitors via gelatin‐coated gold nanoparticles.