Nanotechnology & Medicine - ianano.org Nanobio - Nanomedicine.pdf · Dendrimer/microbicide Topical...
Transcript of Nanotechnology & Medicine - ianano.org Nanobio - Nanomedicine.pdf · Dendrimer/microbicide Topical...
Nanotechnology & Medicine:
An Introduction
Lloyd L. TranDirector
California Institute of Nanotechnology
Professional Development Training in Nanotechnology © California Institute of Nanotechnology
Section 3.16 Nanobio - Nanomedicine © 2009 California Institute of Nanotechnology & International Association of Nanotechnology Certified Nanotech & Clean Tech Professional
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Nanobiotechnology is the branch of nanotechnology focusing on the biological and biochemical applications or uses.
The term bionanotechnology is often used interchangeably with nanobiotechnology.
Bionanotechnology normally refers to the biological or life science aspects in the design, fabrications, production and applications of nanostructures or nanodevices.
Nanobiotechnology
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Nanomedicine is the medical application of nanotechnology.
Nanomedicine focuses on the state-of-the-art research topics such as : treatment of diseases using nanoscale formulation of drugs and biologics, targeted drug delivery systems, biosensors, nanotoxicity
In April 2006, the journal Nature Materials estimated that 130 nanotech-based drugs and delivery systems were being developed worldwide.
Nanomedicine
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Single-Walled Tube
(SWNT)
Diameter ~ 0.5-2 nm5 nm
Multi-Walled Tube
(MWNT)
Diameter ~ 10 - 50 nm
Applications of Carbon Nanotubes for Biodetection
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Molecular Perfection: FULLERENE
• The Strongest fiber that will ever be made.
• Electrical Conductivity of Copper or Silicon.
• Thermal Conductivity of Diamond.
• The Chemistry of Carbon.
• The size and perfection of DNA.
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Nanomechanics for Biomolecular Recognition
Scanning electron micrograph of a microfabricated cantilever array. Eight cantilevers with dimension of 500 um x 80 um x 7 um.
Concentris, GmbH Instrument Prototype Available
Nanomechanics+Biochemical Surface Functionalization (Coating)
9Ref: Roszek- Medical Nanotechnology
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Nanotechnology based Drug
Delivery Systems
Polymeric biodegradable nanoparticles
Ceramic (inorganic) nanoparticles
Polymeric micelles (amphililic copolymers)
Liposomes
Dendrimers
Nanocrystals (Quantum dots)
Magnetic nanoparticles (iron oxide for MRI)
Ref: Sahoo and Labhasetwar , DDT, 2003
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Product Type of Nanoparticles Indication Particle Size
Magenvist Gadolinum Dimeglumine MRI contrast agent < 1nm
Feridex Superparamagnetic Iron Oxide MRI contrast agent 120- 180 nm
Rapamune Nano-crystal/ Sirolimus Immunosuppressant 100-180 nm
Emend Nano-crystal/Aprepitant Anti-emitic 100-1000 nm
TriCor Nanocrystal/Fenofibrate Hypolipidemic
Doxil Liposome/Doxorubicin Antineoplastic ~100 nm
Abraxane Albumin-coated nanoparticles Antineoplastic ~130 nm
Megace ES Nanocrystal/MegesterolAcetate
Appetite enhancer
AmBisome Liposome/Ampotericin Antifugal
Diprivan Albumin-coated nanoparticles Anesthetic
Definity Lipid coated gas particles Echography contrast agent
Currently Marketed Prescription Drugs with Nanoparticles
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Product Type of nanoparticle or drug Indication FDA Company
AmBisome® Liposomal/amphotericin B Fungal infections Approved8/11/1997 FDA50740
Gilead Sciences
Diprivan® Liposomal/propofol Anesthetic Approved10/2/1989FDA19627
Zeneca Pharmaceuticals
Doxil® PEGylated liposome/doxorubicin hydrochloride
Ovarian cancer Approved 11/17/1995FDA50718
OrthoBiotech
VivaGel® Dendrimer/microbicide Topical microbicide for HIV
FDA Fast Track (Phase II) StarPharma
Definity® Nanolipid/perflutren Echocardiogram contrast agent
Approved 7/31/2001FDA21064
ImaRx/BMS
Abraxane® Nanoparticulate albumin/paclitaxel Various cancers Approved 1/7/2005 FDA21660
American Pharmaceutical Partners
Cyclosert® Cyclodextrin nanoparticle Solid tumors Phase I Insert TherapeuticsRapamune® Nanocrystal/sirolimus Immunosuppressant for
kidney transplantsApproved 9/15/1999 FDA21083
Elan/Wyeth
Emend® Nanocrystal/aprepitant Nausea Approved 3/27/2003FDA21549
Elan/Merck
TriCor® Nanocrystal/fenofibrate Reduces cholesterol Approved 11/5/2004FDA19304
Elan/Abbott
Megace ES® Nanocrystal/megestrol acetate Breast cancer Approved 7/5/2005 FDA21778
Elan/Par Pharmaceutical Companies
INGN-401 Liposomal/FUS1 Lung cancer Phase I IntrogenCombidex® Iron oxide Tumor imaging Phase III Advanced MagneticsAurimune® Colloidal gold/TNF Solid tumors Phase I CytImmune SciencesSGT-53 Liposome/p53 gene Solid tumors Phase I SynerGene Therapeutics
US FDA-approved Nanomedicines and Nanomedicines in Clinical Trials
Ref: Nanomedicine. 2007;2(6):789-803. ©2007 Future Medicine Ltd
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Paclitaxel is one of the best antineoplastic drugs found in nature.
Paclitaxel s highly hydrophobic with water solubility less than 0.03 mg/L. Its current clinical dosage from Taxol® is formulated in Cremophor EL, which has been found to be responsible for serious side effects including hypersensitivity reactions, nephrotoxicity, neurotoxicity and cardiotoxicity.
Paclitaxel
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New formulation of Paclitaxel contained in poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs)
Nanoparticles Paclitaxel can be are prepared by the solvent extraction/evaporation method with d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) as a novel emulsifier.
Nanoparticles Formulation of Paclitaxel was found to be 1.37 times more effective than Taxol® after 24 hour culture at 500 ng/ml Paclitaxel concentration.
The drug loaded PLGA nanoparticles can be used either for local delivery by balloon catheter or for development of a novel type of cardiovascular stent which is coated by nanoparticle formulation instead of the pristine drug.
Paclitaxel Nanoparticles
Paclitaxel Nanoparticles
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Paclitaxel Nanoparticles
Photomicrograph shows paclitaxel in albumin nanoparticles seen under electron microscope at x39.000 magnification. Round albumin nanoparticles are arranged on formvar carbon—coated grid of electron microscope. Arrows indicate nanoparticles
(Damascelli, B. et al. Am. J. Roentgenol. 2003;181:253-260 )
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Why prefer the nanoparticle formulation is preferred over bulk formulation?
The reason is that pure paclitaxel is not bioadhesive to the cell membrane due to its poor pharmaceutical properties, including the MDR (multiple drug resiatence) effect.
Clinical research has demonstrated that nanoparticles are more adhesive to, and thus easier to be taken than the microparticle formulation and the free drug itself.
Abraxane (paclitaxel protein-bound particles for injectable suspension), a next generation taxane, and the first in a new class of albumin-bound nanotechnology, was approved for the treatment of metastatic breast cancer.
( Ref: US Oncology)
Paclitaxel Nanoparticles
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Self-regulated Therapeutic Agent Delivery System and Method
Patent # 5,019,723
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Electromechanical Infusion Pump
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NanoBio Membrane Drug Delivery System
NanoBio Membrane
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NanoBio Membrane Drug Delivery System
The patented NanoBio Drug Delivery System is based on the advanced development of the novel nanostructure bio-electric membrane for drug delivery and diagnostic applications.
(Ref: Neurobiomed – Lloyd (Loi) Tran)
50 nm pore size
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NanoBio Drug Delivery
System
• NanoBio Membrane™ properties make it an ideal
drug delivery platform, with high efficiency/capacity of drug loading-up to 97%
• Safe with bio-compatible NanoBio membrane
• Totally needle-free delivery system for injectable drug formulation.
• Heat and radiation stable for ease of gamma radiation
• Ensure aseptic system with built-in nanoscale membrane (50 nanometers) eliminating particulates, bacteria, and endotoxin.
(Ref: Neurobiomed – Lloyd Tran)
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Scanning electron micrograph from the Cornell Nanofabrication Facility shows rat hippocampal neurons cultured on a silicon pillar structure. Photo credit: Andrea M. P. Turner, Stephen W. P.
Turner, Natalie Dowell, Jim Turner, William Shain and Harold Craighead.
Nanoscience experiment of Neurons
Nanobiotechnology studies into the effect of surface topography on the attachment and growth of central nervous system could lead to improved devices for restoring neural function in injured patients.
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Quantum Dots
• . Quantum Dots: semiconductor nanocrystals exhibiting fluorescence and can be used as alternatives to organic dyes for biological labeling
• This size leads to a quantum confinement effect, which endows nanocrystals with unique optical and electronic properties.
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Quantum Dots
• Qdots are generally composed of atoms from groups II–VI or III–V of the periodic table
• Qdots have size-tunable emission (from the ultraviolet to the infrared), narrow spectral line widths , high luminescence, continuous absorption profiles, and stability against photobleaching
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Schematic representation of qdot targeting. Intravenous delivery of qdots into specific tissues of the mouse.
(Left ) Design of peptide-coated qdots. (Right ) Qdots were coated with either peptides only or with peptides and PEG.
PEG helps the qdots maintain solubility in aqueous solvents and minimize nonspecific binding.
Akerman & Chan et al (2002), PNAS .
Targeted Qdots to tumor endothelium in vivo
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Subcellular localization of QDS. PEG-QDs were conjugated to localization sequence peptides, which permit active transport to the nucleus or mitochondria. Here, fluorescence and phase contrast micrographs of a HeLa cell 24 hr after co-injection of NLS-QDs with 70kDa rhodamine dextran control.
Ref: Derfus et al (2004). Adv. Mat.
Subcellular localization of QDS
Nanotechnology and
Tissue Engineering
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Avg. Implant Lifetime ≈ 12-15 years
(Dowson D. Proceedings of the Institution of Mechanical Engineers. Part H- Journal of Engineering
in Medicine 2001; 215(4): 335-358. )
Age of Patients Receiving Total Hip Replacement Surgery
Over 6566%
45-6524%
Under 4510%
17.9 15.9 19.2
0
10
20
Years
At age 65 males
(age 65)
females
(age 65)
Life Expectancy
http://www.aaos.org/wordhtml/press/joinrepl.htm; http://www.aaos.org/wordhtml/press/hip_knee.htm; Minino AM, MPH, and Smith BL. National Vital Statistics Reports 2001; 49(12); http://www.cdc.gov/nchs/fastats/lifexpec.htm; http://www.oxmed.com/docs/datafiles/hip%20replacement%20cemented%20or%20uncemented.html
25% failure rate for dental implants after 15 years
Many patients receiving the implants are 35-54 years old
Current Orthopedic Implant Failures
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Cell Adhesion
F
A
IL
U
R
E
!!
Fibrous Tissue Encapsulation
Direct Bone Apposition
Inflammation
Wear Particles
Interfacial Stresses and Strains
Microdamage to Surrounding Bone
Loosening
Pain
Osteolysis
Anderson JM et al. Host reactions to biomaterials and their evaluation. In: Biomaterials
science: An introduction to materials in medicine. San Diego: Academic Press, Inc., 1996.
p. 165-214.
Dowson D. Proceedings of the Institution of Mechanical Engineers. Part H- Journal of
Engineering in Medicine 2001; 215(4): 335-358.
Huiskes R and Boeklagen R. Biomat 1989; 22: 793-804.
http://www.dental-implants.com/implant_systems/system_types.html
Current Orthopedic Implant Failures
More Fully
Integrated Interface
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Bone is a Nanophase Material
Cancellous
Bone
Osteocl
ast
Osteobla
stOsteocy
te
Cancellous Bone
Capillar
y Capillar
yOsteoc
yte
Lamella
Collagen Fibers: composed of Type I
collagen which is a triple helix 300 nm in
length; 0.5 nm in width; and periodicity of
67 nm
Hydroxyapatite Crystals:
less than 50 nm in length
and 5 nm in diameter
Compact
Bone
Redrawn and adapted from Fung Biomechanics: Mechanical Properties of Living Tissue, Springer-Verlag, New York, 1993 and Keaveny and Hayes, Bone 7:285, 1993.
Non-biologically Inspired
Surface Roughness of Conventional Implants
Conventional (Rolled) Ti Sheet: ASTM grain size number, 7.5; ave. grain
diameter, 50 µm; bar = 100 µm.
• It is believed that one reason why current orthopedic implants only have a 15 year lifetime isdue to non-biologically-inspiredsurface roughness.
• Such surface roughness does not promote sufficient new bone growthfor long term implant integration into surrounding bone.
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Bar = 1mm
(Goodman S.L. et al., Biomaterials. 1996 Nov;17(21):2087-95.
Cast Replica of Vascular Tissue Demonstrating Nanometer
Roughness *
• Due to the presence of numerous nano-structures (i.e., proteins) in the body, cells are accustomed to interacting with surfaces that have a large degree of nanometer roughness.
• Despite this fact, current syntheticmaterials used as tissue engineeringscaffolds possess conventional surface features only.
Nanophase Materials
for Tissue Engineering
Ways to Synthesize Nanophase
Materials
There are many techniques to synthesize nanophase materials(or nano-structured surface roughness):
• Physical Vapor Synthesis• Electro-explosion,• Chemical Vapor Deposition (CVD),• Sol-gel, • Nanolithography, • Chemical Etching, and• etc.
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Physical Vapor Synthesis was used:– Arc energy applied to solid metal which creates a vapor at high temperature.– A reactant gas is added and cooled at a controlled rate.– The vapor condenses to form nanoparticles with a defined crystalline.
Nanospherical Ceramic Synthesis
From T. J. Webster, in Advances in Biochemical Engineering/Biotechnology(K. Lee and D.L. Kaplan, editors), Springer-Verlag, in press, 2005. 35
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Health & Safety Issues relating to Nanomaterials
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Nanoparticle Safety
What are the nanoparticle’s pathways inside the human body?
How long do the nanoparticles remain in the tissues and how are they cleared?
What effects do nanoparticles have on cellular and tissue functions?
Can nanoparticles gain access to the systemic circulation from dermal exposure? If nanoparticles enter skin cells, is there an effect on cellular functions?
What are unanticipated reactions in vivo?
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Single Wall Carbon Nanotube contains Redox-Active Iron
Source: Valerie E. Kagan, Dept. Environmental & Occupational Health, University of Pittsburgh
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Source: Valerie E. Kagan, Dept. Environmental & Occupational Health, University of Pittsburgh
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1. Researchers in the University of Texas found that carbon nanotubes squirted into the trachea of mice caused serious inflammation of the lungs and granulomas(tumour-like nodules of bloated white blood cells in the lining of the lungs), and five of the nine mice treated with the higher dose died.
2. In a similar experiment carried out at the National Institute of Occupational Safety and Health (NIOSH) in Morgantown, West Virginia, researchers not only found granulomas in the lungs, but also damage to mitochondrial DNA in the heart and the aortic artery, and substantial oxidative damage, both foreshadowing atherosclerosis.
3. In another similar experiment in Tottori University, Japan, researchers showed that within a minute of contacting the mice’s tiniest airways, carbon nanotubes began to burrow through gaps between the surface lining cells and into the blood capillaries, where the negatively charged nanoparticles latched onto the normally positively charged red blood cells surface, thereby potentially causing the red blood cells to clump and the blood to clot.
Examples of Nanotoxicity
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4. Researchers from the University of Rochester, New York, reported anincreased susceptibility to clotting in rabbits that had inhaled buckyballs.
5. Buckballs present in water at 0.5 parts per million were taken up by largemouth bass, which suffered severe brain damage 48 hours later, the extent of damage being 17 times greater than that seen in controls.
6. Carbon Nanotubes in the lungs are translocated to the circulatory system and from there throughout the body, accumulating in the liver, spleen, and bone marrow. Carbon Nanotubes inhaled through the nose and air passages are translocated to the brain through the olfactory nerves, and accumulate in the brain.
Examples of Nanotoxicity
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7. Nanoparticles can enter the body through the skin; and quantum dots injected into the skin accumulate in lymph nodes with potential effects on the immune system [5].
8. Quantum dots consisting of a core of fluorescent cadmium selenide, touted as a non-invasive way to image internal body tissues, break down in the body, releasing cadmium, a toxic heavy metal [6].
9. Lawrence Berkeley National Lab team working with laboratory-grown cells showed that carbon nanotubes specifically activate "cell suicide genes.
Examples of Nanotoxicity
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Nano-Toxicology study
• bioavailability
• bioaccumulation
• bio-interaction
• pathogenicity
• toxic levels: ppm, ppb
• acute vs chronic
• in vitro vs. in vivo
• by-products, metabolites, degradation products
• exposure route: external, ingestion and inhalation,
• animal toxicity vs. human toxicity
• susceptibility: children vs. adult exposure
• short-term and long term toxicity
Etc…
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Nanoparticle Characterization :
1. What are the forms in which particles are presented to host, cells and organelles?
2. What are residual solvents, processing variables, impurities and excipients?
3. What are validated assays to detect and quantify nanoparticles in in vitro and in vivo?
4. How do we determine long and short-term stability of nanomaterials?
5. How do you analyze the degradation of nanoparticles
6. How to maintain quality assurance in scaling-up to mass production.
7. What are the reference materials and manufacturing standardization
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Nanoparticle Safety
What are the nanoparticle’ pathways inside the human body?
How long do the nanoparticles remain in the tissues and how are they cleared?
What effects do nanoparticles have on cellular and tissue functions?
Can nanoparticles gain access to the systemic circulation from dermal exposure? If nanoparticles enter skin cells, is there an effect on cellular functions?
What are unanticipated reactions in vivo?
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Toxic Substances Control Act15 U.S.C. s/s 2601 et seq. (1976)
The Toxic Substances Control Act (TSCA) of 1976 was enacted by Congress to give EPA the ability to track the 75,000 industrial chemicals currently produced or imported into the United States.
EPA repeatedly screens these chemicals and can require reporting or testing of those that may pose an environmental or human-health hazard.
EPA can ban the manufacture and import of those chemicals that pose an unreasonable risk.
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European Union’s REACH
Registration Evaluation Authorization of Chemicals
New chemicals. A new chemical is defined as one that does not appear on the EINECS (European Inventory of Existing Commercial Substances) inventory. When a new chemical is produced, before introduction to the market, the producer of that chemical is required to conduct testing, and in the meantime take such precautions as are practicable. The level of testing required is determined by the mass produced, with the lowest mass trigger currently set at 10kg per annum.
Only changes in chemical structure constitute a new substance, whereas changes in form (for example size or shape) do not. An exception is made for polymers: those produced entirely from EINECS listed monomers are exempt from notification.
Mass (tonnage) triggers: Essentially, the more of an existing substance that is produced, the more data on its properties are required by regulators.
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Sources: Friends of the Earth
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CTA Petitions on FDA's Failure to Regulate Health Threats from NanomaterialsMay 16, 2006
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Legal Petition to the FDA (continued)
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Legal Petition to the FDA (continued)
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A case study: Asbestos
Scanning electron micrograph of asbestiform amphibole from a former vermiculite mining site near Libby, Montana. Source:
U.S. Geological Survey and U.S. Environmental Protection Agency, Region 8, Denver, Colorado.
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Asbestos: over the past century has caused millions of deaths and asbestos toll exceeds 100,000 per year around the world
http://www.ohiotoxicmold.com/images/IAQ%20Images/exposure.gif
Health Effects of Asbestos
After asbestos are inhaled, they can remain and accumulate in the lungs. Asbestos can cause lung cancer, mesothelioma (a cancer of the chest and abdominal linings), and asbestosis (irreversible lung scarring that can be fatal). Symptoms of these diseases do not show up until many years after exposure began.
Argentina
Australia
Austria
Belgium
Bulgaria
Chile
Crotia
Cyprus
Czech Republic
Denmark
Egypt
Estonia
Finland
France
Gabon
Germany
Greece
Honduras
Hungary
Iceland
Ireland
Italy
Japan
Korea
Kuwait
Latvia
Lithuania
Luxembourg
Malta
Netherlands
New Zealand
New Caledonia
Norway
Poland
Portugal*
Saudi Arabia
Seychelles
Slovakia
Slovenia
Spain
Sweden
Switzerland
United Kingdom
Uruguay
South Africa
(Source: compiled by Laurie Kazan-Allen)
CURRENT ASBESTOS BANS AND RESTRICTIONS
(Revised April 2008)
The USA, Canada and Russia are 3 major developed countries that have not totally banned Asbestos
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• In 1979 EPA issued a notice of its intent to regulate asbestos under the authority of Section 6 of the Toxic Substances Control Act (TSCA).
• Asbestos producers and the Canadian government began to pressure the Reagan Administration to halt EPA's efforts. Canada took a particular interest in the matter because 95 percent of the 85,000 tons of asbestos used in the US came from Canada, primarily Quebec.
• In 1989, after conducting a ten year study, EPA announced that it would phase out and ban virtually all products containing asbestos.
• EPA's stated rationale for the ban was simple: "asbestos is a human carcinogen and is one of the most
hazardous substances to which humans are exposed in both occupational and non-occupational settings."
• Proposals to ban asbestos in the United States were announced on June 18, 2002 by Senator Patty Murray
• Asbestos product manufacturers and industry organizations swiftly filed a lawsuit challenging the ban's validity under TSCA in Corrosion Proof Fittings v. EPA.
• The Fifth Circuit vacated the ban, finding that EPA failed to present "substantial evidence" to justify the ban under TSCA.
• The administration of George Bush chose not to appeal the decision to the Supreme Court.
• Many Americans are unaware that the use of asbestos is still permitted in the USA.
Effort to ban asbestos in the USA
Bruce Vento Ban Asbestos and Prevent Mesothelioma Act of 2007 - Amends the Toxic Substances Control Act to require the Director of the National Institute for Occupational Safety and Health to conduct studies and report on the health effects of non-asbestiform minerals and elongated mineral particles.
Requires the Administrator of the Environmental Protection Agency (EPA) to establish a plan to: (1) increase awareness of the dangers posed by asbestos-containing materials and products and contaminants in homes and workplaces and by asbestos-related diseases; (2) provide information to, and encourage participation in research and treatment endeavors by, asbestos-related disease patients and their families and front-line health care providers; and (3) encourage health care providers and researchers to provide to patients and their families information relating to research, diagnostic, and clinical treatments relating to asbestos.
Requires the Administrator to promulgate regulations that prohibit the importing, manufacturing, processing, or distributing of asbestos-containing materials, subject to limited exemption upon petition and specified exemptions sought by the Department of Defense (DOD) and the National Aeronautics and Space Administration (NASA). States that such prohibitions do not apply to specified diaphragm electrolysis installations.
Requires the disposal of asbestos-containing materials within two years. Exempts products that are no longer in the stream of commerce or that are in the possession of an end user.
Amends the Public Health Service Act to: (1) direct the Secretary of Health and Human Services to expand and coordinate research programs on diseases caused by asbestos exposure, particularly mesothelioma, asbestosis, and pleural injuries; (2) provide for establishment of a national clearinghouse for data and specimens relating to asbestos-related diseases; (3) require the Director of the National Institutes of Health (NIH) to establish an asbestos-related disease research and treatment network; and (4) direct the Secretary to support research on mesothelioma and other asbestos-related diseases that is directly relevant to the health of the Armed Forces.
Bruce Vento Ban Asbestos and Prevent Mesothelioma Act of 2007August 2, 2007
(Sources: http://www.govtrack.us/congress/bill.xpd?bill=h110-3339&tab=summary)
It is difficult to assess the risk factors of
nanomaterials without internationally
adopted standards
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Challenges in developing
international standards for Nanoparticle Toxicology
Communication across multi-disciplines
National and international geo-political differences
Intellectual Property protection
Confidential business information
Lack of funding for this long term endeavor
Concern about speeding up government regulation and public policy
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Conclusion
Nanomedicine is one of the most promising new frontiers of medical research
Study on nanomaterial toxicology is the key for the advancement of nanomedicine
International nomenclature and standards are urgently needed
Health safety of nanoparticles needed to be addressed
Pro-active risk management to prevent abuse and unintended misuse
Nanobiotechnology potentially offers breakthrough solutions for medical diagnostics,
biosensor and treatment of diseases such as cancer, neurological disorders.
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Thank you
Lloyd L. TranDirector
California Institute of [email protected]
www.cinano.comTel. 408-280-6266
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