Temporal and mechanistic tracking of cellular uptake dynamics with novel surface fluorophore-bound nanodiamonds.
Nanoparticles (NPs) offer promise for a multitude of biological applications including cellular probes at the bio-interface for targeted delivery of anticancer substances, Raman and fluorescent-based imaging and directed cell growth. Nanodiamonds (NDs), in particular, have several advantages compared to other carbon-based nanomaterials - including a rich surface chemistry useful for chemical conjugation, high biocompatibility with little reactive oxygen species (ROS) generation, physical and chemical stability that affords sterilization, high surface area to volume ratio, transparency and a high index of refraction. The visualization of ND internalization into cells is possible via photoluminescence, which is produced by direct dye conjugation or high energy irradiation that creates nitrogen vacancy centers. Here, we explore the kinetics and mechanisms involved in the intracellular uptake and localization of novel, highly-stable, fluorophore-conjugated NDs. Examination in a neuronal cell line (N2A) shows ND localization to early endosomes and lysosomes with eventual release into the cytoplasm. The addition of endocytosis and exocytosis inhibitors allows for diminished uptake and increased accumulation, respectively, which further corroborates cellular behavior in response to NDs. Ultimately, the ability of the NDs to travel throughout cellular compartments of varying pH without degradation of the surface-conjugated fluorophore or alteration of cell viability over extended periods of time is promising for their use in biomedical applications as stable, biocompatible, fluorescent probes.
Schrand
Amanda M
AM
Applied Biotechnology Branch, Human Effectiveness Directorate, AFRL/RHPB, 2729 R Street, Bldg 837, Wright-Patterson AFB, OH 45433-5707, USA. amanda.schrand@wpafb.af.mil
Lin
Jonathan B
JB
Hens
Suzanne Ciftan
SC
Hussain
Saber M
SM
D020126:Brefeldin A / Q000494:pharmacology; D045744:Cell Line, Tumor; D004705:Endocytosis / Q000187:drug effects; D005089:Exocytosis / Q000187:drug effects; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006835:Hydrazones / Q000494:pharmacology; D018613:Microscopy, Confocal; D058612:Nanodiamonds / Q000737:chemistry*; D009447:Neuroblastoma / Q000378:metabolism; D056951:Photoelectron Spectroscopy; D012235:Rhodamines / Q000737:chemistry
D005456:Fluorescent Dyes; D006835:Hydrazones; C511794:N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide; D058612:Nanodiamonds; D012235:Rhodamines; D020126:Brefeldin A; C005358:tetramethylrhodamine