Decagram-Scale Synthesis of Multicolor Carbon Nanodots: Self-Tracking Nanoheaters with Inherent and Selective Anticancer Properties.

Carbon nanodots (CDs) are a new class of carbon-based nanoparticles endowed with photoluminescence, high specific surface area, and good photothermal conversion, which have spearheaded many breakthroughs in medicine, especially in drug delivery and cancer theranostics. However, the tight control of their structural, optical, and biological properties and the synthesis scale-up have been very difficult so far. Here, we report for the first time an efficient protocol for the one-step synthesis of decagram-scale quantities of N,S-doped CDs with a narrow size distribution, along with a single nanostructure multicolor emission, high near-infrared (NIR) photothermal conversion efficiency, and selective reactive oxygen species (ROS) production in cancer cells. This allows achieving targeted and multimodal cytotoxic effects (i.e., photothermal and oxidative stresses) in cancer cells by applying biocompatible NIR laser sources that can be remotely controlled under the guidance of fluorescence imaging. Hence, our findings open up a range of possibilities for real-world biomedical applications, among which is cancer theranostics. In this work, indocyanine green is used as a bidentate SOx donor which has the ability to tune surface groups and emission bands of CDs obtained by solvothermal decomposition of citric acid and urea in N,N-dimethylformamide. The co-doping implies various surface states providing transitions in the visible region, thus eliciting a tunable multicolor emission from blue to red and excellent photothermal efficiency in the NIR region useful in bioimaging applications and image-guided anticancer phototherapy. The fluorescence self-tracking capability of SOx-CDs reveals that they can enter cancer cells more quickly than healthy cell lines and undergo a different intracellular fate after cell internalization. This could explain why sulfur doping entails pro-oxidative activities by triggering more ROS generation in cancer cells when compared to healthy cell lines. We also find that oxidative stress can be locally enhanced under the effects of a NIR laser at moderate power density (2.5 W cm-2). Overall, these findings suggest that SOx-CDs are endowed with inherent drug-independent cytotoxic effects toward cancer cells, which would be selectively enhanced by external NIR light irradiation and helpful in precision anticancer approaches. Also, this work opens a debate on the role of CD surface engineering in determining nanotoxicity as a function of cell metabolism, thus allowing a rational design of next-generation nanomaterials with targeted anticancer properties.

Pressure-Dependent Tuning of Photoluminescence and Size Distribution of Carbon Nanodots for Theranostic Anticancer Applications.

Carbon nanodots (CDs) have recently attracted attention in the field of nanomedicine because of the biocompatibility, cost-effective nature, high specific surface, good near infrared (NIR) photothermal conversion into heat and tunable fluorescence properties, which have paved the way toward incorporating use of CDs into innovative anticancer theranostic platforms. However, a reliable synthesis of CDs with established and controlled physiochemical proprieties is precluded owing to the lack of full manipulation of thermodynamic parameters during the synthesis, thus limiting their use in real world medical applications. Herein, we developed a robust solvothermal protocol which allow fine controlling of temperature and pressure in order to obtain CDs with tunable properties. We obtained different CDs by modulating the operating pressure (from 8 to 18.5 bar) during the solvothermal decomposition of urea and citric acid in N,N-dimethylformamide at fixed composition. Atomic force microscopy (AFM), Fourier transform infrared (FTIR), ultraviolet-visible (UV-vis) and fluorescence spectroscopy were used to assess the role of pressure in influencing size, optical and surface properties of the obtained CDs. While preliminary biological and anticancer performance of CDs was established on the MDA-MB-231 cell line, used as triple negative breast cancer model. Our results indicate that pressure impinge on the formation of carbon nanoparticles under solvothermal conditions and impart desired optical, size distribution, surface functionalization and anticancer properties in a facile way. However, we have highlighted that a strategic surface engineering of these CDs is needed to limit the adsorption of corona proteins and also to increase the average surface diameter, avoiding a rapid renal clearance and improving their therapeutic efficacy in vivo.

Highly Homogeneous Biotinylated Carbon Nanodots: Red-Emitting Nanoheaters as Theranostic Agents toward Precision Cancer Medicine.

Very recent red-emissive carbon nanodots (CDs) have shown potential as near-infrared converting tools to produce local heat useful in cancer theranostics. Besides, CDs seem very appealing for clinical applications combining hyperthermia, imaging, and drug delivery in a single platform capable of selectively targeting cancer cells. However, CDs still suffer from dramatic dot-to-dot variability issues such that a rational design of their structural, optical, and chemical characteristics for medical applications has been impossible so far. Herein, we report for the first time a simple and highly controllable layer-by-layer synthesis of biotin-decorated CDs with monodisperse size distribution, well established polymeric shell thickness, and degree of surface functionalization, endowed with strong red luminescence and the ability to convert NIR light into heat. These CDs, henceforth named CDs-PEG-BT, consist of a carbonaceous core passivated with biotin-terminated PEG2000 chains, which we demonstrate as active targeting groups to recognize cancer cells. The CDs-PEG-BT are designed to efficiently incorporate a high amount of anticancer drugs such as irinotecan (16-28%) and to act as NIR-activated nanoheaters capable of triggering local hyperthermia and massive drug release inside tumors, thus provoking sudden and efficient tumor death. The potential of the irinotecan-loaded CDs-PEG-BT (CDs-PEG-BT@IT) in fluorescence imaging was studied on 2D cultures and on complex 3D spheroids mimicking in vivo tumor architectures, showing their capability of selectively entering cancer cells through biotin receptors overexpressed in cell membranes. The efficient anticancer effect of these CDs was thoroughly assessed on multicellular 3D spheroids and patient organoids (tumor-on-a-dish preclinical models) to predict the drug response in humans in view of personalized medicine applications. CDs-PEG-BT@IT have a smart combination of properties, which pave the way to their real-world use as anticancer theranostic agents for image-guided photothermal applications.