Nanodiamond uptake in colon cancer cells: the influence of direction and trypsin-EDTA treatment

Abstract

Nanoparticles are routinely used in cell biology. They deliver drugs or function as labels or sensors. For many of these applications it is essential that the nanoparticles enter the cells. While some cell types readily ingest all kinds of particles, others just don't. We report that uptake can be enhanced for some cells if the particles are administered from the basolateral side of the cells (in this case from below).Compared to apical uptake (from above), we report an 8-fold increase in the number of fluorescent nanodiamonds internalized by the colon cancer cell line HT29. Up to 96% of the cells treated by a modified protocol contain at least one nanodiamond, whereas in the control group we could observe nanodiamonds in less than half of the cells. We were also able to show that simple treatment of cell clusters with trypsin-EDTA leads to the same enhancement of the nanodiamond uptake as seeding the cells on top of the nanoparticles. Although our study is focused on nanodiamonds in HT29 cells, we believe that this method could also be applicable for other nanoparticles and cells with a specific directionality. † Electronic supplementary information (ESI) available. See

Not all cells are created equal – endosomal escape in fluorescent nanodiamonds in different cells

“…Similarly as reported before [ 34 ], the FND objects were identified by the FIJI plugin "3D Object Counter" [ 40 ]. It's worth noting that these measurements only reveal an estimate rather than the precise amount of objects due to limitations of the conventional confocal resolution and FND brightness.…”

“…The quantification procedure was done in the same way as reported previously [ 34 ]. In this study we chose to investigate HeLa cells as well as HUVECs because of their relevance in nanoparticle uptake studies.…”

“…These processes can differ per cell type[ 43,44 ]. Some immune cells like macrophages are more likely to ingest the particles while HT-29 cells are not [ 34 ]. Besides, the shape and size of the cells vary even within the same cell line which is less explored [ 45 ].…”

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“…By both strategies, about 95% of colon carcinoma cells contained FNDs for a concentration of 5 µg.mL -1 , whereas it was 50% when directed to the apical side without enzymatic treatment. 40 This research is notable for not entailing any FNDs surface functionalization and its results could be taken into account in future theranostic applications.…”

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