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Technique measures quantity, risks of engineered nanomaterials delivered to cells

Date:
March 28, 2014
Source:
Harvard School of Public Health
Summary:
Scientists have discovered a way to measure the effective density of engineered nanoparticles in physiological fluids, making it possible to determine the amount of nanomaterials that come into contact with cells and tissue in culture.

Thousands of consumer products containing engineered nanoparticles -- microscopic particles found in everyday items from cosmetics and clothing to building materials -- enter the market every year. Concerns about possible environmental health and safety issues of these nano-enabled products continue to grow with scientists struggling to come up with fast, cheap, and easy-to-use cellular screening systems to determine possible hazards of vast libraries of engineered nanomaterials. However, determining how much exposure to engineered nanoparticles could be unsafe for humans requires precise knowledge of the amount (dose) of nanomaterials interacting with cells and tissues such as lungs and skin.

With chemicals, this is easy to do but when it comes to nanoparticles suspended in physiological media, this is not trivial. Engineered nanoparticles in biological media interact with serum proteins and form larger agglomerates which alter both their so called effective density and active surface area and ultimately define their delivery to cell dose and bio-interactions. This behavior has tremendous implications not only in measuring the exact amount of nanomaterials interacting with cells and tissue but also in defining hazard rankings of various engineered nanomaterials (ENMs). As a result, thousands of published cellular screening assays are difficult to interpret and use for risk assessment purposes.

Scientists at the Center for Nanotechnology and Nanotoxicology at Harvard School of Public Health (HSPH) have discovered a fast, simple, and inexpensive method to measure the effective density of engineered nanoparticles in physiological fluids, thereby making it possible to accurately determine the amount of nanomaterials that come into contact with cells and tissue in culture.

The method, referred to as the Volumetric Centrifugation Method (VCM), will be published in the March 28, 2014 Nature Communications.

The new discovery will have a major impact on the hazard assessment of engineered nanoparticles, enabling risk assessors to perform accurate hazard rankings of nanomaterials using cellular systems. Furthermore, by measuring the composition of nanomaterial agglomerates in physiologic fluids, it will allow scientists to design more effective nano-based drug delivery systems for nanomedicine applications.

"The biggest challenge we have in assessing possible health effects associated with nano exposures is deciding when something is hazardous and when it is not, based on the dose level. At low levels, the risks are probably miniscule," said senior author Philip Demokritou, associate professor of aerosol physics in the Department of Environmental Health at HSPH. "The question is: At what dose level does nano-exposure become problematic? The same question applies to nano-based drugs when we test their efficiency using cellular systems. How much of the administered nano-drug will come in contact with cells and tissue? This will determine the effective dose needed for a given cellular response," Demokritou said.

Federal regulatory agencies do not require manufacturers to test engineered nanoparticles, if the original form of the bulk material has already been shown to be safe. However, there is evidence that some of these materials could be more harmful in the nanoscale -- a scale at which materials may penetrate cells and bypass biological barriers more easily and exhibit unique physical, chemical, and biological properties compared to larger size particles. Nanotoxicologists are struggling to develop fast and cheap toxicological screening cellular assays to cope with the influx of vast forms of engineered nanomaterials and avoid laborious and expensive animal testing. However, this effort has been held back due to the lack of a simple-to-use, fast, method to measure the dose-response relationships and possible toxicological implications. While biological responses are fairly easy to measure, scientists are struggling to develop a fast method to assess the exact amount or dose of nanomaterials coming in contact with cells in biological media.

"Dosimetric considerations are too complicated to consider in nano-bio assessments, but too important to ignore," Demokritou said. "Comparisons of biological responses to nano-exposures usually rely on guesstimates based on properties measured in the dry powder form (e.g., mass, surface area, and density), without taking into account particle-particle and particle-fluid interactions in biological media. When suspended in fluids, nanoparticles typically form agglomerates that include large amounts of the suspending fluid, and that therefore have effective densities much lower than that of dry material. This greatly influences the particle delivery to cells, and reduces the surface area available for interactions with cells," said Glen DeLoid, research associate in the Department of Environmental Health, one of the two lead authors of the study. "The VCM method will help nanobiologists and regulators to resolve conflicting in vitro cellular toxicity data that have been reported in the literature for various nanomaterials. These disparities likely result from lack of or inaccurate dosimetric considerations in nano-bio interactions in a cellular screening system," said Joel Cohen, doctoral student at HSPH and one of the two lead authors of the study.

Wolfgang Kreyling, a nanotoxicologist at the German Research Center for Environmental Health who was not involved in the study, says this method should help toxicologists to understand the nano-bio interactions and address possible nano hazards for the vast libraries of engineered nanoparticles (ENPs) currently in use.

"The paper by DeLoid et al in Nature Communications is a major achievement which offers a solution to solve the pending issues of the apparent ENM density and an easy way to determine the latter by the application of the volumetric centrifugation method. Hence, this paper provides a versatile concept easy to achieve which allows for a rather precise estimate of ENM dosimetry to in vitro cell cultures which hopefully will improve the power of toxicological studies using in vitro cell cultures when comparing to in vivo studies. In this case this would be a major contribution in aiming to reduce in vivo experimental animal work," Kreyling said.

Other authors of the study include Georgios Pyrgiotakis, research fellow at HSPH, Liying Rojanasakul, and Raymond Derk from the National Institute for Occupational Safety and Health, and Wendel Wohlleben from BASF, Germany.

This research project was supported by NIEHS grant (ES-000002), NSF grant (ID 1235806) and the Center for Nanotechnology and Nanotoxicology at HSPH. This work was performed in part at the Harvard Center for Nanoscale Systems (CNS), a member of the National Nanotechnology Infrastructure Network (NNIN), which is supported by the National Science Foundation under NSF award number ECS-0335765.


Story Source:

The above story is based on materials provided by Harvard School of Public Health. Note: Materials may be edited for content and length.


Journal Reference:

  1. Glen DeLoid, Joel M. Cohen, Tom Darrah, Raymond Derk, Liying Rojanasakul, Georgios Pyrgiotakis, Wendel Wohlleben, Philip Demokritou. Estimating the effective density of engineered nanomaterials for in vitro dosimetry. Nature Communications, 2014; 5 DOI: 10.1038/ncomms4514

Cite This Page:

Harvard School of Public Health. "Technique measures quantity, risks of engineered nanomaterials delivered to cells." ScienceDaily. ScienceDaily, 28 March 2014. <www.sciencedaily.com/releases/2014/03/140328175044.htm>.
Harvard School of Public Health. (2014, March 28). Technique measures quantity, risks of engineered nanomaterials delivered to cells. ScienceDaily. Retrieved October 20, 2014 from www.sciencedaily.com/releases/2014/03/140328175044.htm
Harvard School of Public Health. "Technique measures quantity, risks of engineered nanomaterials delivered to cells." ScienceDaily. www.sciencedaily.com/releases/2014/03/140328175044.htm (accessed October 20, 2014).

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