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Research Papers

Size-Dependent Nanoparticle Uptake by Endothelial Cells in a Capillary Flow System

[+] Author and Article Information
Patrick Jurney

Department of Mechanical Engineering,
The University of Texas at Austin,
Austin, TX 78712
e-mail: Jurney4@gmail.com

Rachit Agarwal

Department of Mechanical Engineering,
Georgia Institute of Technology,
Atlanta, GA 30332-0535
e-mail: rachit.agarwal@me.gatech.edu

Krishnendu Roy

Department of Biomedical Engineering,
Georgia Institute of Technology and
Emory University,
Atlanta, GA 30332-0535
e-mail: krish.roy@gatech.edu

S. V. Sreenivasan

Department of Mechanical Engineering,
The University of Texas at Austin,
Austin, TX 78712-1591
e-mail: sv.sreeni@mail.utexas.edu

Li Shi

Department of Mechanical Engineering,
The University of Texas at Austin,
Austin, TX 78712
e-mail: lishi@mail.utexas.edu

1Corresponding author.

Manuscript received July 2, 2015; final manuscript received October 16, 2015; published online November 6, 2015. Assoc. Editor: Feng Xu.

J. Nanotechnol. Eng. Med 6(1), 011007 (Nov 06, 2015) (6 pages) Paper No: NANO-15-1049; doi: 10.1115/1.4031856 History: Received July 02, 2015; Revised October 16, 2015

An in vitro cell culture system is developed for studying the uptake characteristics of nanoparticles (NPs) by endothelial cells under shear stress. Results show that the smaller polystyrene nanospheres are uptaken more than larger nanospheres for sizes ranging from 100 nm to 500 nm for 12, 24, and 48 hrs delivery times. While the result is similar to that found in static cultures, the observed trend is different from NP delivery behaviors to a simple glass surface in a flow, where no clear size dependence was observed because of repulsive electrostatic force on marginating NPs. The trend is also opposite to the behavior found in another study of the adhesion of labeled particles onto endothelial cells in whole blood flow. The comparison shows that the reduced zeta potential of NPs in a serum-containing cell medium and particle removal by cells results in reduced repulsive electrostatic force on marginating NPs. Consequently, the uptake behaviors are dominated by Brownian diffusion and cell membrane deformation process, which favor the uptake of NPs with reduced sizes.

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Figures

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Fig. 1

(Top) Picture of flow setup with a peristaltic pump providing a shear stress of 12 dyn/cm2 in glass capillary tubes. (Bottom) Schematic of the inlet and outlet capillary tubes of the dynamic cell culture system. Media and NPs (when used) are drawn through the inlet and flow over HUVECs cultured on the capillary surface, as indicated by thick dashed lines, then returned to the reservoir in a closed-loop system. O2 and CO2 are allowed to diffuse through the top surface of the reservoir.

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Fig. 2

(a) Optical cross section of a capillary tube showing confluent HUVECs cultured on the vessel walls. Confluent cells can be seen at the edge of the capillary in the focal plane of the image. The several apparent black spots are most likely aberrations from the cells adhered in the foreground of the capillary tube, out of the focal plane of the image. (b) HUVECs cultured in a flat-walled microchannel with no flow. (c) HUVECs cultured in a flat-walled microchannel with a shear stress of 12 dyn/cm2 and the flow direction moving left to right. The scale bars are 30 μm.

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Fig. 3

Fluorescence intensity profiles of HUVEC cell populations after 12 hrs of flow: (a) without NPs and (b) with 100 nm NPs

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Fig. 4

Orientation angle of cells in Fig. 2(b), cultured in static conditions (a). Orientation angle of cells in Fig. 2(c), cultured with a shear stress of 12 dyn/cm2 (b).

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Fig. 5

NP delivery to HUVECs in dynamic culture system at different time points up to 48 hrs. Error bars represent the random uncertainty with a 95% confidence interval.

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Fig. 6

NP zeta potential in serum-containing media (black) and base media without serum (gray)

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Fig. 7

Depiction of (a) NPs adhered to a saturated surface retarding new NP adhesion and (b) NPs on an active surface being trafficked and creating new surface for additional particles to adhere

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