Symposium Announcement
Toxicity Associated with
Nanomaterials
Monday, December 16, 2013
10:00AM - 3:00PM
Lister Hill Center Auditorium
8600 Rockville Pike, NIH Building 38A
National Institutes of Health, Bethesda, MD
(NIH Visitor information http://www.nih.gov/about/visitor/index.htm)
Speakers: Dr. Christopher Weis, Ph.D., D.A.B.T.
Dr. Scott McNeil, Ph.D.
Dr. Jennifer Sass, Ph.D.
Student/PostDocs: Georgina Harris, Peter Petrochenko,
Javiera Bahamonde, and CE McLoughlin.
For more information please contact Mark Miller at 919-541.7758 or mark.miller2@nih.gov
Security Information: Any individuals seeking access to the NIH campus to attend a conference/seminar will need to be prepared
to show two forms of identification such as a driver’s license plus a company ID, a government ID, or a university ID. You will also
be asked to name the title or sponsor of the event you wish to attend. Thank you for your cooperation.
Individuals with disabilities who need accommodation to participate in this event should contact Mark Miller at 919-541.7758 or
mark.miller2@nih.gov. Requests should be made at least 5 business days in advance of the event.
Agenda:
09:45 - 10:00
10:00 - 10:15
10:15 - 11:00
11:00 - 11:20
11:20 - 11:40
11:40 - 12:30
12:30 - 13:15
13:15 - 13:35
13:35 - 13:55
14:00 - 14:45
14:45 - 15:00
Poster set-up
Welcome/Introduction (Fowler/Miller)
Plenary speaker 1 (Weis)
Student Presentation 1
Student Presentation 2
Lunch/ Poster Session
Plenary speaker 2 (McNeil)
Student Presentation 3
Student Presentation 4
Plenary speaker 3 (Sass)
Discussion/Closing Remarks (Fowler)
Plenary Speakers
Christopher Weis, Ph.D., D.A.B.T.
Chris Weis, Ph.D., joined NIEHS in August 2010. He brings decades of
practical experience working in the fields of rapid exposure and risk
evaluation for environmental emergencies and environmental forensics.
Weis’ office is located on the NIH campus in Bethesda, Md.
As the NIEHS toxicology liaison, Weis serves as a senior advisor to NIEHS/NTP
Director Linda Birnbaum, Ph.D., and also represents the Institute and the NTP on
national and international committees, subcommittees, task forces, and ad hoc
working groups. This allows Weis the opportunity to not only communicate the goals
and priorities of the director to key officials at other Federal research, regulatory, and
health agencies, but also to hear from them and others to help guide the development
of the Institute’s programs and functions. Weis also serves as a liaison to external constituencies, stakeholders,
and advocacy groups, as well as members of the NTP community.
As part of his responsibilities, Weis was recently named to represent NIEHS as a co-chair on the Toxics and
Risks Subcommittee of the White House National Science and Technology Council Committee on Environment,
Natural Resources, and Sustainability.
Prior to joining NIEHS in August 2010, Weis served as a senior toxicologist at the U.S. Environmental Protection
Agency National Enforcement Investigations Center (NEIC) in Denver. While at NEIC, he worked in criminal
forensics and established a laboratory for trace analysis of biomolecular evidence. Weis is well versed in a wide
variety of environmental contaminants, including asbestos, lead, radon, and vermiculite insulation.
Weis completed his Ph.D. in medical physiology and toxicology at Michigan State University in 1987, and was
awarded two NIH postdoctoral fellowship positions at the University of Virginia School of Medicine, Department of
Physiology and Biophysics.
Scott McNeil, Ph.D.
Dr. McNeil serves as the Director of the Nanotechnology
Characterization Laboratory (NCL) for SAIC-Frederick and
Frederick National Laboratory for Cancer Research, where he
coordinates preclinical characterization of nanotech cancer
therapeutics and diagnostics. At the NCL, Dr. McNeil leads a team
of scientists responsible for testing candidate nanotech drugs and
diagnostics, evaluating safety and efficacy, and assisting with
product development -- from discovery-level, through scale-up and
into clinical trials. NCL has assisted in characterization and
evaluation of more than 300 nanotechnology products, several of
which are now in human clinical trials. Dr. McNeil is a member of
several working groups on nanomedicine, environmental health
and safety, and other nanotechnology issues. He is an invited speaker to numerous
nanotechnology-related conferences and has several patents pending related to
nanotechnology and biotechnology. He is also a Vice President of SAIC-Frederick.
Prior to establishing the NCL, he served as a Senior Scientist in the Nanotech Initiatives
Division at SAIC where he transitioned basic nanotechnology research to government and
commercial markets. He advises industry and State and US Governments on the development
of nanotechnology and is a member of several governmental and industrial working groups
related to nanotechnology policy, standardization and commercialization. Dr. McNeil's
professional career includes tenure as an Army Officer, with tours as Chief of Biochemistry at
Tripler Army Medical Center, and as a Combat Arms officer during the Gulf War. He received
his bachelor's degree in chemistry from Portland State University and his doctorate in cell
biology from Oregon Health Sciences University.
Jennifer Sass, Ph.D.
Jennifer Sass is a Senior Scientist in NRDC's Health and
Environment program (since 2001), and a Professorial
Lecturer at George Washington University. She reviews
the science underpinning the regulation of toxic chemicals,
and advocates for health-protective regulations consistent
with environmental laws. She holds a doctoral degree in
Cell Biology from University of Saskatchewan, Canada,
and a post-doctoral certificate in Environmental Toxicology
from the University of Maryland.
Student Speakers
Student Speaker 1
Title: Use of automated high content fluorescence imaging and in silico models for in vitro assessment of
nanomaterial toxicity in Balb/c3T3 cells.
Authors: Georgina Harris1, Taina Palosaari2, Milena Mennecozzi2, David Asturiol2, Jean-Michel Gineste2, Luis
Saavedra2, Roman Liska2, Anne Milcamps2, Maurice Whelan2
1
Center for Alternatives to Animal Testing (CAAT), Johns Hopkins Bloomberg School of Public Health,
Baltimore, MD, USA. 2JRC - European Commission, Institute for Health and Consumer Protection, Systems
Toxicology Unit, Via E. Fermi 2749 TP202, I-21027 Ispra (VA), Italy
There are an increasing number of studies using in vitro approaches to determine the possible cellular effects of
nanomaterials. We propose automated high content fluorescence-based imaging combined with in silico models,
as a useful quantitative tool to screen nanomaterials for their adverse toxicological effects on cells in culture.
Additionally, the technical challenges and pitfalls associated with automated high throughput/content in vitro
nanotoxicity testing are considered. Over 20 nanomaterials (including titanium oxides, zinc oxides, silicon
dioxides, iron oxides, carbon nano-tubes) were tested in a series of assays, measuring three endpoints relevant for
safety assessment, namely, cell viability, reactive oxygen species production and DNA double-strand breaks. In
silico analysis was performed to compare the toxicity of the nanomaterials relative to physico-chemical
characteristics. The reliability and relevance of the results obtained is discussed in the context of the potential of
this approach to advance safety assessment science in the support of regulatory decision making specific to
nanomaterials.
Student Speaker 2
Title: Short- and Long-Term Effects of Commercially Available Gold Nanoparticles in Rodents
Authors: Javiera Bahamonde,* Bonnie Brenseke,*† Matthew Chan,* and M. Renee Prater*§
*Virginia Tech, Blacksburg, VA; †Campbell University School of Osteopathic Medicine, Buies Creek, NC;
§Edward Via College of Osteopathic Medicine, Blacksburg, VA
Gold nanoparticles (GNPs) are being intensely investigated for their potential use in biomedical applications.
Nanotoxicity studies are urgently needed to validate their safety in clinical practice. The objective of this research
was to assess the acute and chronic effects of a single exposure to commercially available GNPs in rats and mice.
Gold nanoparticles were purchased and independently characterized. Animals received either GNPs (1000 mg/kg)
or phosphate buffered saline intravenously. Subsets of animals were euthanized 1, 7, 14, 21, 28 days or 20 weeks
post-exposure and samples were collected. The physicochemical properties of the purchased GNPs were in good
agreement with the information provided by the supplier. Important differences in GNP-induced immune
responses were identified when comparing mice and rats. Liver microgranulomas and a transient increase in
serum levels of interleukin-18 were observed in GNP-exposed mice. No such alterations were found in rats.
Higher accumulation of GNPs in spleen and longer fecal excretion were observed in rats. In the long-term,
exposure to GNPs incited chronic inflammation in mice, with persistent microgranulomas in liver, spleen, and
lymph nodes, as well as further increased serum levels of interleukin-18. Impairment of body weight gain was
also observed in the GNP-exposed group. In conclusion, GNPs can incite a robust macrophage response in mice.
However, considering the mildness of the toxic effects identified despite the high dose selected for the study,
GNPs continue to have great potential for biomedical uses.
Student Speaker 3
Title: Toxicity and allergy responses in mice following pulmonary exposure to nanoparticle silver
Authors: CE McLoughlin1, S. Anderson1, K. Anderson1, D. Schwegler-Berry1, BT Chen1, JR Roberts1,
1
NIOSH, Morgantown, WV
Expansive commercial use of silver nanoparticles (AgNP) raises the concern of effects following respiratory
exposure in manufacturing workers. Previous work on AgNP has shown dose-dependent lung toxicity with
inflammation and alterations in lung immune parameters in rodents. The goal of the current study is to
characterize effects of AgNP for potential modulation of respiratory allergy in an ovalbumin (OVA)-induced
allergy model in BALB/c mice. Range-finding (RF) studies were conducted in mice exposed to physiological
dispersion medium (DM), 6.1g (LO), 18.2g (ME), or 73g (HI) AgNP. 20 nm diameter AgNP with 0.3% wt
polyvinylpovidone coating (NanoAmor, Inc.), were suspended and sonicated before pharyngeal aspiration (PA)
on day 0. Airway hyperreactivity was measured as PenH. Whole lung lavage (WLL) fluid and cells, lymph node
lymphocytes (LN), and serum were collected for analysis of parameters of lung injury, inflammation,
immunophenotyping, and allergic response. Results indicated a dose-dependent injury and inflammation by day
10 which began to resolve by day 29 with no changes in PenH with AgNP alone. For the allergy model, animals
received i.p. injections of OVA + aluminum hydroxide gel (alum) during the sensitization phase on days 1 and 10.
To elicit an OVA-specific response, 2 PA challenges of OVA were given on days 19 and 29. The second
challenge was given immediately prior to PenH measurement. PenH, WLL and LN cell number were increased in
OVA animals, and the responses were enhanced by both AgME and AgHI exposure. Results indicate potential for
AgNP to exacerbate allergic response in the lung.
Student Speaker 4
Title: Laser 3D printing with nanoscale resolution: improving biocompatibility and mitigating toxicity from
photoinitiators.
Authors: Peter Petrochenko1, 2, Roger Narayan2, Peter Goering1, Aleksandr Ovsianikov3
1
Office of Science and Engineering Laboratories, Center for Devices and Radiological Health, USFDA, Silver
Spring, Maryland, USA. 2 Joint Dept of Biomedical Engineering, University of North Carolina at Chapel Hill,
NC, USA. 3 Institute of Materials Science and Technology, Vienna University of Technology (TU Wien),
Favoritenstrasse 9-11, Vienna, Austria
Three-dimensional (3D) structural features of a scaffold critically influence cellular functions and the overall
performance of a scaffold material. Recent developments in laser-assisted 3D printing using two-photon
polymerization (2PP) have allowed for previously unattainable submicron resolution and opened a number of
possibilities for creating 3D cell scaffolds. However, a significant barrier to using 2PP for biological applications
exists due to the toxic nature of photoinitiators required for radical polymerization using light energy. In this
study we demonstrate two main approaches for creating nanotextured porous 3D scaffolds. The first involves
printing the scaffold beforehand, removing residual toxic substances and seeding cells afterwards. The second,
more complex approach, involves using lower laser energy and trapping cells directly in a collagen matrix. Our
results from the first method indicate that stable scaffolds with porosities of over 60% can be custom printed to fit
standard well plates. Cells grown on 3D scaffolds exhibit increased growth and proliferation compared to smooth
2D scaffold controls. Scaffolds additionally adsorb larger amounts of proteins due to their larger surface area and
allow cells to attach in multiple planes and completely infiltrate the porous scaffolds. Preliminary results from the
second approach show some dead cells in encapsulated regions. In order to improve cytocompatibility 3 possible
antioxidants: Trolox (water-soluble vitamin E), vitamin C, and glutathione; and collagenase are used to mitigate
the toxic effects of photoinitiators. Preliminary improved cytotoxicity and encapsulation results are demonstrated.
2PP is shown to be a promising technique for fabricating custom 3D scaffolds from a computer model with
potential for in situ cell encapsulation.
Questions?
Contact Susan Laessig, NCAC-SOT Councilor-Student Liaison (202-564-5232, Laessig.susan@epa.gov)
or Mark Miller, NCAC-SOT Vice-President (919-541-7758; mark.miller2@nih.gov)
NCAC-SOT Website: http://www.toxicology.org/isot/rc/ncac/index.asp