Cancer Cell Targeting Via Selective Transferrin Receptor Labeling Using Protein-Derived Carbon Dots.

Carbon dot (CD) nanoparticles offer tremendous advantages as fluorescent probes in bioimaging and biosensing; however, they lack specific affinity for biomolecules, limiting their practical applications in selective targeting. Nanoparticles with intrinsic affinity for a target have applications in imaging, cytometry, therapeutics, etc. Toward that end, we report the transferrin receptor (CD71) targeting CDs, synthesized for the first time. The formation of these particles is truly groundbreaking, as direct tuning of nanoparticle affinity was achieved by simple and careful precursor selection of a protein, which has the targeting characteristic of interest. We hypothesized that the retention of the original protein's peptides on the nanoparticle surface provides the CDs with some of the function of the precursor protein, enabling selective binding to the protein's receptor. This was confirmed with FTIR (Fourier transform infrared) data and subsequent affinity-based cell assays. These transferrin (Tf)-derived CDs have been shown to possess an affinity for CD71, a cancer biomarker that is ubiquitously expressed in nearly every cancer cell line due to its central role mediating the uptake of cellular iron. The CDs were tested using the human leukemia cell line HL60 and demonstrated the selective targeting of CD71 and specific triggering of transferrin-mediated endocytosis via clathrin-coated pits. The particle characterization results reflect a carbon-based nanoparticle with bright violet fluorescence and 7.9% quantum yield in aqueous solution. These unpresented CDs proved to retain the functional properties of the precursor protein. Indicating that this process can be repeated for other disease biomarkers for applications ranging from biosensing and diagnostic bioimaging to targeted therapeutics.

Novel synthesis of fibronectin derived photoluminescent carbon dots for bioimaging applications.

Fibronectin (FN) derived from human plasma has been used for the first time as the carbon precursor in the top-down, microwave-assisted hydrothermal synthesis of nitrogen doped carbon dots (CDs). FN is a large glycoprotein primarily known for its roles in cell adhesion and cell growth. Due to these properties FN can be over expressed in the extracellular matrix (ECM) of some cancers allowing FN to be used as an indicator for the detection of cancerous cells over non-cancerous cells. These FN derived CDs display violet photoluminescence with UV excitation and appear to possess similar functional groups on their surface to their carbon precursor (-COOH and -NH2). This is believed to be due to the self-passivation of the CDs' nitrogen-containing surface functional groups during the heating process. These CDs were then used to stain MCF-7 and MDA-231 breast cancer cells and were observed to interact primarily with the cell membrane rather than intercalating into the cell like the many other types of CDs. This led to the hypothesis that the CDs are selectively binding to the FN overexpressed within the cancer cells' ECM via amide linkages. This is in agreement with the EDX and FTIR spectra of the FN CDs which indicate the presence of -COOH and nitrogen containing surface groups like -NH3. The inherent selectivity of the CDs combined with their ability to photoluminesce enables their use as a fluorophore for bioimaging applications.

Synthesis and Characterization of Dye-Doped Au@SiO2 Core-Shell Nanoparticles for Super-Resolution Fluorescence Microscopy.

Dye-doped nanoparticles have been investigated as bright, fluorescent probes for localization-based super-resolution microscopy. Nanoparticle size is important in super-resolution microscopy to get an accurate size of the object of interest from image analysis. Due to their self-blinking behavior and metal-enhanced fluorescence (MEF), Ag@SiO2 and Au@Ag@SiO2 nanoparticles have shown promise as probes for localization-based super-resolution microscopy. Here, several noble metal-based dye-doped core-shell nanoparticles have been investigated as self-blinking nanomaterial probes. It was observed that both the gold- and silver-plated nanoparticle cores exhibit weak luminescence under certain conditions due to the surface plasmon resonance bands produced by each metal, and the gold cores exhibit blinking behavior which enhances the blinking and fluorescence of the dye-doped nanoparticle. However, the silver-plated nanoparticle cores, while weakly luminescent, did not exhibit any blinking; the dye-doped nanoparticle exhibited the same behavior as the core fluorescent, but did not blink. Because of the blinking behavior, stochastic optical reconstruction microscopy (STORM) super-resolution analysis was able to be performed with performed on the gold core nanoparticles. A preliminary study on the use of these nanoparticles for localization-based super-resolution showed that these nanoparticles are suitable for use in STORM super resolution. Resolution enhancement was two times better than the diffraction limited images, with core sizes reduced to 15 nm using the hybrid Au-Ag cores.

A Review of Fluorescent Carbon Dots, Their Synthesis, Physical and Chemical Characteristics, and Applications.

Carbon dots (CDs) are a particularly useful type of fluorescent nanoparticle that demonstrate biocompatibility, resistance to photobleaching, as well as diversity in composition and characteristics amongst the different types available. There are two main morphologies of CDs: Disk-shaped with 1-3 stacked sheets of aromatic carbon rings and quasi-spherical with a core-shell arrangement having crystalline and amorphous properties. They can be synthesized from various potentially environmentally friendly methods including hydrothermal carbonization, microwaving, pyrolysis or combustion, and are then purified via one or more methods. CDs can have either excitation wavelength-dependent or -independent emission with each having their own benefits in microscopic fluorescent imaging. Some CDs have an affinity for a particular cell type, organelle or chemical. This property allows the CDs to be used as sensors in a biological environment and can even provide quantitative information if the quenching or intensity of their fluorescence is dependent on the concentration of the analyte. In addition to fluorescent imaging, CDs can also be used for other applications including drug delivery, quality control, photodynamic therapy, and photocatalysis.