Conspectus Nanoparticles are widely used in various biomedical applications as drug delivery carriers, imaging probes, single-molecule tracking/detection probes, artificial chaperones for inhibiting protein aggregation, and photodynamic therapy materials. One key parameter of these applications is the ability of the nanoparticles to enter into the cell cytoplasm, target different subcellular compartments, and control intracellular processes. This is particularly the case because nanoparticles are designed to interact with subcellular components for the required biomedical performance. However, cells are protected from their surroundings by the cell membrane, which exerts strict control over entry of foreign materials. Thus, nanoparticles need to be designed appropriately so that they can readily cross the cell membrane, target subcellular compartments, and control intracellular processes. In the past few decades there have been great advancements in understanding the principles of cellular uptake of foreign materials. In particular, it has been shown that internalization of foreign materials (small molecules, macromolecules, nanoparticles) is size-dependent: endocytotic uptake of materials requires sizes greater than 10 nm, and materials with sizes of 10–100 nm usually enter into cells by energy-dependent endocytosis via biomembrane-coated vesicles. Direct access to the cytosol is limited to very specific conditions, and endosomal escape of material appears to be the most practical approach for intracellular processing. In this Account, we describe how cellular uptake and intracellular processing of nanoscale materials can be controlled by appropriate design of size and surface chemistry. We first describe the cell membrane structure and principles of cellular uptake of foreign materials followed by their subcellular trafficking. Next, we discuss the designed surface chemistry of a 5–50 nm particle that offers preferential lipid-raft/caveolae-mediated endocytosis over clathrin-mediated endocytosis with minimum endosomal/lysosomal trafficking or energy-independent direct cell membrane translocation (without endocytosis) followed by cytosolic delivery without endosomal/lysosomal trafficking. In particular, we emphasize that the zwitterionic–lipophilic surface property of a nanoparticle offers preferential interaction with the lipid raft region of the cell membrane followed by lipid raft uptake, whereas a lower number of affinity biomolecules (<25) on the nanoparticle surface offers caveolae/lipid-raft uptake, while an arginine/guanidinium-terminated surface along with a size of <10 nm offers direct cell membrane translocation. Finally, we discuss how nanoprobes can be designed by adapting these surface chemistry and size preference principles so that they can readily enter into the cell, label different subcellular compartments, and control intracellular processes such as trafficking kinetics, exocytosis, autophagy, amyloid aggregation, and clearance of toxic amyloid aggregates. The Account ends with a Conclusions ...

Although the piezoelectric property of a BaTiO3 nanoparticle is routinely used in energy harvesting application, it can also be exploited for wireless cell stimulation and cell therapy. However, such biomedical application is rare due to limited availability of colloidal BaTiO3 nanoparticles of <100 nm hydrodynamic size with good piezocatalytic property and efficient biolabeling performance. Here, we report a colloidal form of a piezocatalytic BaTiO3-based nanorod of <100 nm hydrodynamic size that can offer wireless cell stimulation. The nanorod is prepared using a TiO2 nanorod as the template, and the resultant TiO2–BaTiO3-based composite nanorod is coated with a hydrophilic polymer shell. These nanorods can label cells and, under the ultrasound exposure, produce reactive oxygen species inside cells via piezocatalysis, leading to cell death. These nanorods can be used for wireless modulation of intracellular processes.

Although the cytotoxic effect of iron oxide nanoparticle via peroxidase-like activity and associated Fenton reaction is well-known, its effective utilization in cancer therapy is limited because of poor availability of endogenous hydrogen peroxide. Similarly, the cytotoxic effect of vitamin C via hydrogen peroxide generation is well-known but its practical use in cancer therapy is limited due to requirement of high dose (typically above one milimolar) which is not achievable under physiological condition. Here, we show that pharmacologically achievable dose of vitamin C in the range of 0.1−1.0 mM can induce cell death via iron oxide nanoparticle-based intracellular Fenton reaction. Cells are exposed with exogenous vitamin C after labeling with Fe 3 O 4 nanoparticle or directly exposing with vitamin C-conjugated Fe 3 O 4 nanoparticle. Results show that intracellular Fe 3 O 4 nanoparticle can induce cell death by as low as 0.1 mM exogenous vitamin C via Fenton reaction-based intracellular reactive oxygen species (ROS) generation. Following this approach, selective killing of cancer cell has been achieved via functional Fe 3 O 4 nanoparticle-based cell targeting/labeling followed by exogenous vitamin C-based cell therapy. This approach can be extended for other nanoparticles with oxidase/peroxidase-like activity and in designing more effective nanoparticle for vitamin C-based cell therapy.

Two-photon imaging of endogenous bisulphite ions of the hippocampus region of mouse brain.

Fluoroquinolones are third-generation broad spectrum bactericidal antibiotics and work against both Gram-positive and Gram-negative bacteria. Levofloxacin (L), a fluoroquinolone, is widely used in anti-infective chemotherapy and treatment of urinary tract infection and pneumonia. The main pathogen for urinary tract infections is Escherichia coli, and Streptococcus pneumoniae is responsible for pneumonia, predominantly a lower respiratory tract infection. Poor permeability of L leads to the use of higher dose of this drug and excess drug in the outer cellular fluid leads to central nervous system (CNS) abnormality. One way to counter this is to improve the lipophilicity of the drug molecule, and accordingly, we have synthesized two new Levofloxacin derivatives, which participated in the spatiotemporal release of drug via disulfide bond cleavage induced by glutathione (GSH). Recent studies with Streptococcus mutants suggest that it is localized in epithelial lining fluid (ELF) of the normal lower respiratory tract and the effective [GSH] in ELF is ∼430 μM. E. coli typically cause urinary tract infections and the concentration of GSH in porcine bladder epithelium is reported as 0.6 mM for a healthy human. Thus, for the present study we have chosen two important bacteria (Gram + ve and Gram - ve), which are operational in regions having high extracellular GSH concentration. Interestingly, this supports our design of new lipophilic Levofloxacin based prodrugs, which released effective drug on reaction with GSH. Higher lipophilicity favored improved uptake of the prodrugs. Site specific release of the drug (L) could be achieved following a glutathione mediated biochemical transformation process through cleavage of a disulfide bond of these purpose-built prodrugs. Further, appropriate design helped us to demonstrate that it is possible also to control the kinetics of the drug release from respective prodrugs. Associated luminescence enhancement helps in probing the release of the drug from the prodrug in bacteria and helps in elucidating the mechanistic pathway of the transformation. Such an example is scarce in the contemporary literature.