Featured Research

from universities, journals, and other organizations

Inverting a standard experiment sometimes produces different results

Date:
May 1, 2011
Source:
Washington University in St. Louis
Summary:
The standard experimental setup for measuring the cellular uptake of nanoparticles is to place cells in a well on a culture plate and cover them with culture medium containing nanoparticles. The assumption underlying these experiments is that the particles remain well-dispersed. But when a scientist turned cell cultures upside down, he discovered that this assumption doesn't always hold. Some experiments preparing for the clinical use of nanoparticles may therefore need to be redone.

The experiments in Xia’s lab compared the usual experimental setup (top) to an upside-down setup (bottom). Nanoparticle uptake in the two setups differs only if the ratio of the forces driving sedimentation (S) to those driving diffusion (D) are different. In the situation shown here the upright cells have taken up more nanoparticles than the upside-down ones because there is sedimentation.
Credit: Younan Xia/WUSTL

Nanoparticles will soon be used as tiny shuttles to deliver genes to cells and drugs to tumors in a more targeted way than was possible in the past. But as the scientists prepare to use the nanoparticles in medicine, concerns have arisen about their potential toxicity. Studies of both the applications of nanoparticles and their toxicity rely on the ability of scientists to quantify the interaction between the nanoparticles and cells, particularly the uptake (ingestion) of nanoparticles by cells.

In the standard laboratory tests of the biological activity of nanoparticles, cells are plated on the bottom of a dish and culture medium containing nanoparticles is poured on top of them.

It seems straightforward enough. But recently Washington University in St. Louis scientist Younan Xia started to worry about the in vitro experiments his lab was doing with gold nanoparticles.

What if the cells were upside down, he wondered? Would that make a difference? Would it change their uptake rate?

"People assumed that if they prepared a suspension, the suspension was going to have the same concentration everywhere, including at the surface of the cells," says Xia, PhD, the James M. McKelvey Professor in the Department of Biomedical Engineering.

A battery of experiments in Xia's lab with both the standard and upside-down setups showed that nanoparticles above certain sizes and weights will settle out. So concentrations of the nanoparticles near the cell surfaces are different from those in the bulk solution and cellular uptake rates are higher.

As the scientists conclude in the Nature Nanotechnology article describing the experiments, "Studies on the cellular uptake of nanoparticles that have been conducted with cells in the upright configuration may have given rise to erroneous and misleading data."

Topsies and Turveys

Scientists have felt they could safely assume that the concentration of nanoparticles in the fluid next to the cells, which drives cellular uptake, was the same as the initial concentration of nanoparticles in the medium because the particles are small enough to be easily lofted by Brownian motion, the random motion of the molecules in the liquid.

Gravity, by this accounting, did not override this force for diffusion and the nanoparticles stayed in solution instead of settling out.

"We started to wonder, however, because our nanoparticles are made of gold," Xia says. "Gold is nontoxic but it is also very heavy, so it was conceivable relatively large nanoparticles might settle."

Since it is impossible to measure the exact concentration of gold nanoparticles at the surface of a cell, Xia and coworkers designed a simple experiment to test whether settling changed the concentration there and the cellular uptake.

Xia's lab tested gold nanospheres of three sizes, nanocages of two edge lengths, and nanorods, some with surface coatings that picked up serum proteins in solution and others coated with a chemical that acts as an antifouling agent.

After the cells were incubated in the nanoparticle-bearing medium, the concentration of the nanoparticles in the medium was measured spectroscopically and the number of particles each cell had taken up was calculated from the difference in the concentrations.

In the literature, Xia says, there are reports that the cellular uptake of nanoparticles depends on the nanoparticles' size, shape and surface coating.

His lab's experiments showed that these characteristics are secondary, relevant only insofar as they affect the sedimentation and diffusion velocities of the nanoparticles.

For small, light particles, there was no disparity between the cells in the upright and the upside-down configurations. In the case of larger, heavier particles, however, sedimentation dominated, and the upright cells took in more nanoparticles than the upside-down cells.

"All earlier work may need to be re-evaluated to account for the effects of sedimentation on nanoparticle dosimetry," the authors conclude.

"It's no different from medicines that have to be shaken to suspend a powder in a water. If you don't shake the bottle," Xia says, "you end up under- or overdosing yourself."


Story Source:

The above story is based on materials provided by Washington University in St. Louis. Note: Materials may be edited for content and length.


Journal Reference:

  1. Eun Chul Cho, Qiang Zhang, Younan Xia. The effect of sedimentation and diffusion on cellular uptake of gold nanoparticles. Nature Nanotechnology, 2011; DOI: 10.1038/nnano.2011.58

Cite This Page:

Washington University in St. Louis. "Inverting a standard experiment sometimes produces different results." ScienceDaily. ScienceDaily, 1 May 2011. <www.sciencedaily.com/releases/2011/04/110428132231.htm>.
Washington University in St. Louis. (2011, May 1). Inverting a standard experiment sometimes produces different results. ScienceDaily. Retrieved September 15, 2014 from www.sciencedaily.com/releases/2011/04/110428132231.htm
Washington University in St. Louis. "Inverting a standard experiment sometimes produces different results." ScienceDaily. www.sciencedaily.com/releases/2011/04/110428132231.htm (accessed September 15, 2014).

Share This



More Matter & Energy News

Monday, September 15, 2014

Featured Research

from universities, journals, and other organizations


Featured Videos

from AP, Reuters, AFP, and other news services

Manufacturer Prints 3-D Car In Record Time

Manufacturer Prints 3-D Car In Record Time

Newsy (Sep. 15, 2014) Automobile manufacturer Local Motors created a drivable electric car using a 3-D printer. Printing the body only took 44 hours. Video provided by Newsy
Powered by NewsLook.com
Refurbished New York Subway Tunnel Unveiled After Sandy Damage

Refurbished New York Subway Tunnel Unveiled After Sandy Damage

Reuters - US Online Video (Sep. 15, 2014) New York officials unveil subway tunnels that were refurbished after Superstorm Sandy. Nathan Frandino reports. Video provided by Reuters
Powered by NewsLook.com
Frustration As Drone Industry Outpaces Regulation In U.S.

Frustration As Drone Industry Outpaces Regulation In U.S.

Newsy (Sep. 14, 2014) U.S. firms worry they’re falling behind in the marketplace as the FAA considers how to regulate commercial drones. Video provided by Newsy
Powered by NewsLook.com
Smart Gun Innovators Fear Backlash From Gun Rights Advocates

Smart Gun Innovators Fear Backlash From Gun Rights Advocates

Newsy (Sep. 14, 2014) Winners of a contest for smart gun design are asking not to be named after others in the industry received threats for marketing similar products. Video provided by Newsy
Powered by NewsLook.com

Search ScienceDaily

Number of stories in archives: 140,361

Find with keyword(s):
Enter a keyword or phrase to search ScienceDaily for related topics and research stories.

Save/Print:
Share:

Breaking News:
from the past week

In Other News

... from NewsDaily.com

Science News

Health News

Environment News

Technology News



Save/Print:
Share:

Free Subscriptions


Get the latest science news with ScienceDaily's free email newsletters, updated daily and weekly. Or view hourly updated newsfeeds in your RSS reader:

Get Social & Mobile


Keep up to date with the latest news from ScienceDaily via social networks and mobile apps:

Have Feedback?


Tell us what you think of ScienceDaily -- we welcome both positive and negative comments. Have any problems using the site? Questions?
Mobile: iPhone Android Web
Follow: Facebook Twitter Google+
Subscribe: RSS Feeds Email Newsletters
Latest Headlines Health & Medicine Mind & Brain Space & Time Matter & Energy Computers & Math Plants & Animals Earth & Climate Fossils & Ruins