Nanoparticle Exposure NASAs LongTerm Space Travel Jacqueline Jordan, Ph'D' Clayton State University

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Nanoparticle Exposure NASAs Long-Term Space
TravelJacqueline Jordan, Ph.D.Clayton State
University
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Overview

• What is nanotechnology?
• General information about nanotoxicity
• NASA concerns and current research

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What is nanotechnology?

• Defined by National Nanotechnology Initiative of
  National Science Foundation (NSF)
• Nanotechnology is the understanding and control
  of matter at dimensions of roughly 1 to 100
  nanometers to produce new structures, materials,
  and devices

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Terminology

• Nanometer (nm) one billionth of a meter, which
  is 10 -9, anything smaller than 100 nanometer
• For example, a single human hair is about
  80,000nm wide a red blood cell is about 7,000nm
  wide and a water molecule is approximately 0.3nm
  wide
• Nanoscale the size range from 100nm down to 0.2
  nm

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Richard Smalley

• 1996 Nobel Prize in Chemistry
• Fullerenes (buckyballs)
• Spherical cages of 60 carbon atoms arranged as 20
  hexagons and 12 pentagons

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Uses of nanomaterials

• Carbon nanotubes
• Tensile strength fifty times that of stainless
  steel.
• Surface to volume ratio
• Zyvexs NanoSolve Technology (www.zyvex.com)
• Quantum dot
• Fluorescent tags for Biological Research
• DNA, Protein, and Lipids

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Uses of nanomaterials in medicine

• Cancer Research

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Other uses in everyday use materials

• Cosmetics
• Toothpaste
• Food
• Detergents
• Bullet-proof vest

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Vanderbilt Engineering To Lead New Defense
Nanotechnology Program
Nashville TN (SPX) Jun 25, 2004The Vanderbilt
School of Engineering will lead a new 2.4
million multi-institutional nanotechnology
program funded by the U.S. Army Research
Laboratory to develop radically improved
electronics, sensors, energy-conversion devices
and other critical defense systems. The
Advanced Carbon Nanotechnology Research Program
will explore various nanostructures of carbon,
including diamond, at the molecular level to
develop next-generation materials that can be
used in a wide range of defense devices and
systems.
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Nanoparticles are being introduced at a rapid pace

• Commercial products and potential medical
  applications
• If controlled, this technology has great promise.
• If not controlled, harm may be caused to human
  health and the environment
• Various debilitating diseases
• CANCER

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NIOSH Strategic goals for the emerging field of
nanotechnology

• Understand and prevent work-related injuries and
  illnesses possibly caused by nanomaterials
• Promote healthy workplaces through interventions,
  recommendations, and capacity building
• Enhance global workplace safety and health
  through international collaboration on
  nanotechnology

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Airborne Particulate Matter

• Dust, dirt, soot, smoke, etc.
• Natural sources- pollen, dust, volcanic eruptions
• Combustion processes fossil fuels
• Solid particles or liquid droplets
• Many different sizes and shapes
• Over 10,000 people in US die each year from the
  inhalation of air pollution

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Examples of Airborne dust found in workplace

• Minerals dust free crystalline silica, coal,
  cement dust
• Metallic dust lead, cadmium, nickel, and
  beryllium dusts
• Organic and vegetable dusts flour, wood,
  cotton, pollens
• Biohazard molds and spores

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Respiratory System

• Respiratory system
• Provides for gas exchange intake of O2 and
  elimination of CO2. What would happen if you
  couldnt breath?
• Other functions include regulating blood pH,
  sense of smell, filtration of inspired air,
  sounds, exhale heat.

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Particle size (EPA classification)

• Larger than 10 microns is normally trapped by the
  nasal cavity
• Particles smaller than 10 microns in diameter can
  reach the human lungs
• Particles smaller than 1 micron travel to the
  delicate alveolar region

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New EPA Classification of Respiratory Particles
Course Particles (2.5 10 µm)
red blood cells (8 µm)
Cells
Fine Particles (?2.5 µm)
1µm
mitochondria (1 µm)
Organelles
Nasal cavity
Ultrafines (nano) (?0.1 µm)
0.1µm
DNA (.070 µm)
Macromolecules
bronchus
trachea
lungs
diaphragm
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General function of the Respiratory system
Pictures taken from Tortora and
Grabowski Principles of Anatomy and Physiology,
2003

• Nasal hairs and mucosa (mucus)
• Filters out dust particles and bacteria.
• (also regulates the temperature
• and moisten air)
• Trachea, bronchi and bronchioles
• Lined with ciliated mucus membrane
• Contain cilia that beat rhythmically to
• Sweep debris out of the lungs

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Site of gas exchange Respiratory Membrane
Oxygen passes through the thin walls of the
alveoli into the bloodstream. Carbon dioxide, a
waste product, passes from the bloodstream into
the alveoli and then is exhaled. Alveolar
macrophages help remove particles from this
delicate area
Pictures taken from Tortora and Grabowski
Principles of Anatomy and Physiology, 2003
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Clearance of particles form respiratory tract

• Mucociliary clearance trachea and bronchi, down
  to the terminal bronchioles
• Swallowed, coughed out, sneezing
• Phagocytosis epithelium of alveolar region is
  not ciliated. Particles are removed by phagocytic
  cells (alveolar macrophages)
• What happens if particles are not removed from
  the alveolar region?

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Several things can occur if they travel to deep
lung

• Remain in pulmonary space
• Enter the lymphatic space or bloodstream
  (interstitial)
• Or kill the macrophage cells (release hydrolytic
  enzymes)
• Damage delicate respiratory region and initiate
  an inflammatory response

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Tissue Repair

• Release of chemical mediators attract other
  macrophages, neutrophils and stimulate fibroblast
  cells
• Rapid division of fibroblast cells to repair
  damage
• Fibrosis (scar tissue) that makes the lungs stiff
  and less compliable

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Pneumoconiosis

• General terms used for lung disease caused by
  inhalation of mineral dust
• Pulmonary fibrosis or malignancy
• Industrial diseases
• Big ones Coal Workers Lung, Asbestosis, and
  Silicosis

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Silicosis

• Oldest form of pneumoconiosis
• Inhalation of free crystalline silica
• Mining, quarrying of granite and sandstone, stone
  masonry, sandblasting
• Pulmonary silicotic nodules (1-3 microns in
  diameter)
• Interstitial fibrosis and silicotuberculosis

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Coal Workers Pneumoconiosis (CWP)

• Black lung disease
• Inhalation of black, carbonaceous pigment
• Carbon is minimally fibrogenic, but often
  combined with irritant gases and minerals.
• Fibrotic lesions that make the lungs stiff.
• Result in respiratory distress and decreased
  oxygen in tissues

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X-Ray and Fibrotic Nodule of CWP
Taken from Silicosis and Coal Workers
Pneumoconiosis, Vicent Castranova and Val
Vallyathan, Environ. Health, Vol 108, 2000
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Asbestosis

• Refers to a family of fiber types linked to tumor
  formation, Approximately 4 in 10,000 people
• Takes commonly 20 years to manifest itself
• Macrophages attempt to ingest and clear fibers
  and activate fibrogenic mediators as a repair
  response
• Generalized pulmonary inflammation and
  interstitial fibrosis
• Risk of malignancy is related to exposure to long
  fibers (gt10 microns)- Mesothelioma

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Exact mechanism of toxicity is being investigated

• Alveolar macrophage clearance of particles from
  deep lung is overwhelmed
• In vivo ( animal testing or human studies) under
  the whole body response
• In vitro (under the mechanism of toxicity)
  outside the body in test tubes, glass or plastic
  support

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In Vitro Studies examining the mechanisms of dust
toxicity

• Castranova, et. al. Particles and the airway
  basic biological mechansims of pulmonary
  pathogenicity, Appl. Occup, Environ Hyg. Vol 13,
  1998
• Schins, et al. Mechanisms and mediators in coal
  dust induced toxicity. Ann Occup Hyg. Vol. 43,
  1998
• Renwick, et al. Increased inflammation and
  altered macrophage chemotatic response caused by
  two ultrafine particle types (titanium dioxide
  and carbon black). Occupational and Environmental
  Medicine. Vol 61, 2004
• Fubini, B. The surface chemistry of crushed
  quartz in relation to its pathogenicity. Org.
  Chem Acta Vol. 138, 1987

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New studies of ultrafines

• Higher deposition pattern
• Harder to clear
• Avoid normal clearance mechanism and move to
  interstitial space
• Not all are inflammatory or tumorgenic
• Surface properties are important in causing
  disease- oxidative stress

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NASA has near-term plans to return humans to the
Moon, 2015
http//www.astrosurf.com/cidadao/camedia.htm
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Exploration of the Lunar Surface
Credit NASA Photo
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Mining lunar soil to study the properties of
these materials
Credit NASA Photo
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Astronauts from the Apollo 17 mission complained
of air in LM smelling of gun powder. Symptoms
of congestion and fever
Credit NASA
Dusty spacesuits
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Close-up Of Apollo 11 Lunar Dust
Terry Slezak With Lunar Dust
NASA Photo
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What is Lunar Dust?

• Similar to Volcanic Ash (meteorite impacts)
• Diverse Size Distribution
• Nominal geometric diameter below 20 µm in
  diameter (Dust)
• SiO2 (44.72) and Al2O3 (14.86)
• Properties Magnetic, Potentially surface
  active, glassy

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Major Chemical Elements found in Lunar Dust
SiO2silicon dioxide FeO ferric
oxide Al2O3aluminum oxide CaOcalcium
oxide MgOmagnesium oxide TiO2titanium
oxide Othernumerous element including
sodium, potassium chromium, etc.
McKay, et al. 1991. Lunar Regolith in the Lunar
Sourcebook A Users Guide to Moon, pg. 285-356.
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Effects of Lunar Dust

• Lunar Dust can cause problems for human
  exploration activities
• Equipment degradation and failure
• Potential process contaminant for In Situ
  Resource Utilization (ISRU)
• Potential effects on human health due to
  inhalation of these fine materials
• What are the longer term problems?

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Research at NASA Glenn

• In vitro (outside the body)
• Using cells grown in culture
• Primary respiratory cells
• Alveolar Macrophages and Type II epithelial cells
• Exposure to dust particles (including lunar dust
  stimulants) and examine the cellular response

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Experimental Design

• Compare the in vitro response of nanoparticles
  and components of lunar dust
• Important parameters
• Cellular uptake of nanoparticles
• Changes in cellular morphology
• Cellular toxicity

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Materials and Methods

• Test Materials
• Fluorescent latex beads (0.1 and 0.5 ?m)
• Aluminum oxide particles (0.7?m) and silica
  particles (gt1.6 ?m)
• Cells
• Alveolar macrophages (RAW 264.7)
• Type II epithelial cells (A549)
• LDH
• Standard cytotoxicity assay

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CELL CULTURE
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Alveolar Macrophages
American Type Cultures Collections, 2006
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EPITHELIAL CELLS
American Type Culture Collections, 2006
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A. Alveolar Macrophages
B. Type II epithelial cells

• Fluorescence microscopic images of untreated RAW
  264.7 cells
• and untreated A549 cells (B)
• Note Nucleus is stained blue and the cytoplasm
  is stained red.
• Total magnification 400X

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A. RAW 0.1 ?m
B. RAW 0.5 ?m
Fluorescence microscopic image of RAW 264.7
(alveolar macrophage) cells exposed to ultrafine
and fine polystyrene fluorescent beads for 6
hours Note Fluorescent beads appear blue and
cytoplasm stained red. Total magnification
400X.
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A.
A549 cells exposed to 0.1 ?m for 6 h
B.
C.
A549 cells exposed to 0.5 ?m beads (6 h)
A549 cells exposed 0.1?m beads
for 24 h
Fluorescence microscopic image of A549 cells
exposed to ultrafine and fine polystyrene
fluorescent beads. Mag400 x
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RAW 264.7


A549
Percent uptake of fluorescent polystyrene beads
by respiratory cells in vitro. RAW and A549
cells were incubated at 37?C with 0.1 ?m or 0.5
?m beads for 6 hrs. indicates significantly
different from control (t-test).
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A. Raw control
B. 1.0 mg/ml Al203
C. 1.0 mg/ml SiO2


D. A549 control
.
.
E. 1.0 mg/ml Al203
F. 1.0 mg/ml SiO2


Phase contrast microscopy. 6 h exp. Mag400x


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A.
B.
A549 cells
RAW 264.7 cells


RAW 264.7 cells
C.
A549 cells



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Summary

• Respiratory cells were cultured as the in vitro
  model
• We determined the phagocytic activity of both
  cell types
• We investigated the cellular response of cells to
  SiO2 and Al2O3
• This study suggests differential cellular
  toxicity associated with exposure to ultrafine
  and fine particles

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Future studies

• Changes in inflammatory mediators
• Effects on normal cellular responses
• Cell division
• Apoptosis
• Animal studies

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AcknowledgementsNASA Scientists
David Fischer, Ph.D. Optical Scientist
Paul Greenberg Mechanical Engineer
Ashley Verhoff NASA Coop UC Aerospace
Engineer/ Astronaunt
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Acknowledgements

• Amber Cardell- graduated last year
• Jewels Morgan- current research student
• CSU faculty and staff

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