Micro Versus Macro – The Effect of Environmental Confinement on Cellular Nanoparticle Uptake

Abstract

While the microenvironment is known to alter the cellular behavior in terms of metabolism, growth and the degree of endoplasmic reticulum stress, its influence on the nanoparticle uptake is not yet investigated. Specifically, it is not clear if the cells cultured in a microenvironment ingest different amounts of nanoparticles than cells cultured in a macroenvironment (for example a petri dish). To answer this question, here we used J774 murine macrophages and fluorescent nanodiamonds (FND) as a model system to systematically compare the uptake efficiency of cells cultured in a petri dish and in a microfluidic channel. Specifically, equal numbers of cells were cultured in two devices followed by the FND incubation. Then cells were fixed, stained and imaged to quantify the FND uptake. We show that the FND uptake in the cells cultured in petri dishes is significantly higher than the uptake in a microfluidic chip where the alteration in CO 2 environment, the cell culture medium pH and the surface area to volume ratio seem to be the underlying causes leading to this observed difference.

pH Sensitive Dextran Coated Fluorescent Nanodiamonds as a Biomarker for HeLa Cells Endocytic Pathway and Increased Cellular Uptake

“…Nanodiamonds have excellent biocompatibility in many different cell lines (in vitro) like HT-29 [ 5 ], macrophages, yeast cells [ 6 ], BHK-21 [ 7 ], MCF-7 [ 8 ]. Prabhakar et al showed no toxicity after incubating MDA-MB-231 (human breast adenocarcinoma) cells with FNDs for 120 h. While other carbon nanomaterials have been reported to cause proinflammatory responses [ 9 ] this is not the case for nanodiamonds [ 10 ].…”

Diamond Color Centers in Diamonds for Chemical and Biochemical Analysis and Visualization

“…(b) Overview of FNDs destination throughout cellular processes, such as cell−cell communication (14), proliferation (15), migration (16), and trafficking inside cells (17). Representative literature can be found for each image: (1), 11,142,143 (2) and (3), 144 (4), 90 (5), 142,147,148 (6) and ( 7), 150 (8), 47,157 (9), 155,157 (10), 155 (11), 158 (12), 168,173 (13), 161 (14), 168 (15), 179 (16), 141 Better understanding of FNDs trafficking via endosomes requires co-localization with other biomarkers which still seems to be a challenge on the labeling and image processing levels. Another challenge is to understand what the recorded trajectories mean.…”

“…(b) Overview of FNDs destination throughout cellular processes, such as cell−cell communication (14), proliferation (15), migration (16), and trafficking inside cells (17). Representative literature can be found for each image: (1), 11,142,143 (2) and (3), 144 (4), 90 (5), 142,147,148 (6) and ( 7), 150 (8), 47,157 (9), 155,157 (10), 155 (11), 158 (12), 168,173 (13), 161 (14), 168 (15), 179 nanoparticles that contain high concentrations of SiV centers. While this might be beneficial for some of the quantum sensing applications presented in NV Center-Based Sensing and Magnetometry, this is not desirable in labeling applications where high brightness is needed.…”

“…Liposomes cationic polymers or dendrimers also support uptake of FNDs by mammalian cells. It has been shown that shear stress and other types of mechanical stimulation may increase the FNDs uptake by cells . Nevertheless, these techniques are associated with transport of particles within endosomes.…”

“…(b) Overview of FNDs destination throughout cellular processes, such as cell−cell communication(14), proliferation(15), migration(16), and trafficking inside cells(17). Representative literature can be found for each image: (1),11,142,143 (2) and (3),144 (4),90 (5),142,147,148 (6) and (7), 150 (8),47,157 (9),155,157 (10),155 (11),158 (12),168,173 (13),161 (14),168 (15),179 (16),141 and (17) 156.…”