Mechanisms of bioreactivity of engineered nanoparticles with pulmonary gas exchange barrier

Lead Research Organisation: Imperial College London
Department Name: National Heart and Lung Institute

Abstract

Nanotechnology is a rapidly emerging area of science involving synthesis of engineered nanoparticles that are less that the width of a hair (ie less than 100 nanometres in size). Nanoparticles are processed to form larger products which are not harmful; they are used in engineering, medicine, cosmetics and many other day-to-day products. Nanoparticles have unique properties which make them very attractive and useful in producing new products; however, the properties that make them unique may unfortunately have health effects. We know very little about the hazards or risks associated with exposure to these materials because they are so new. As nanotechnology is moving so fast it is important to establish whether nanoparticles are likely to be toxic or to have unwanted health or environmental effects. If we know this we can protect people and the environment from exposure. We know that when susceptible individuals are exposed to high levels of air pollution which contains nano-sized particles they may develop heart and lung problems and have to go to hospital and might even die. This suggests that inhalation of very small, nanosized engineered particles might cause heart and lung problems, although they have very different properties to air pollution particles. The aim is to discover whether inhaled engineered nanoparticles are harmful and whether they cause breathing or heart problems if they reach the delicate air sacs of the lung. We will use cells that line human lungs and the blood vessels of the lung. These cells will be exposed to the nanoparticles to discover whether the they are toxic and if so, how toxic and why? Do the exposed cells release molecules that might cause inflammation or blood clotting? We want to know what properties of the particles make them dangerous - are they sharp, or do they pass into the cells, or between the cells or are any particular surface chemicals responsible for nanoparticle toxicity? We will watch nanoparticles under a unique microscope to see how they alter cell behaviour. We hope to learn whether inhaled, engineered nanoparticles are likely to be harmful. If they are, is it possible to make particles that are not harmful? This information will allow us to provide guidelines on the possible risks of inhaling engineered nanoparticles, as well as preventing exposure, particularly at work, where levels may be high.

Technical Summary

There has been a rapid emergence and expansion of nanotechnology - hundreds of products utilise nanoparticles. Unfortunately, the properties that make nanomaterials so attractive for use eg in electronics, cosmetics, textiles and medicine may have negative health effects. There are serious, urgent concerns that research on possible toxicity and risk of nano- sized materials to human health is insufficient compared to their current rate of production and use; recent reports describe the situation as urgent. Epidemiological studies of particulate air pollution suggest that inhalation of high levels of nano-sized particles causes increased cardiorespiratory morbidity and mortality, particularly in those with existing disease. This may be because the alveolar units are easily accessible by inhaled nano-sized particles ( 100nm), and delivery of large numbers (but low mass) of nanosized particles, with a large reactive surface area, to the alveolar epithelial-endothelial barrier of the lung may elicit pulmonary inflammation, release of pro-thrombotic factors and facilitate particle translocation into the vasculature. We hypothesise that the degree of bioreactivity and translocation of inhaled nanoparticles at the alveolar epithelial-endothelial blood-gas barrier depends on the physicochemical properties of the particles and nature/response of the target cell. In this cross-disciplinary study, materials scientists, a lung cell biologist, a cardiovascular cell biologist and biophysicist will collaborate to address this hypothesis. Materials scientists will synthesise and fully characterise a series of designer nanoparticles initially using silver and titanium dioxide, as well as carbon nanotubes. These particles have been chosen because their production and use is increasing fast; other particles may be used depending on the early findings. The chemists can strictly control nanoparticle synthesis and functionalisation for in vitro studies. Primary human pulmonary alveolar epithelial cells, capillary endothelial cells as well as monocyte-derived macrophages and dendritic cells will be exposed to the particles in both monoculture and co-culture to represent the alveolar-endothelial membrane in vivo. Effects will be assessed using conventional toxicology as well as determining more subtle molecular-biochemical changes, mechanisms of particle internalisation and intracellular localisation. Living cells will be examined at the level of the electron microscope by scanning ion conductance microscopy and scanning surface confocal microscopy to discover the exact features controlling particle uptake and how particles might affect the integrity of the epithelial-endothelial barrier. Any effects of test particles will be related to their physicochemistry and functionalisation to establish possible patterns or trends that predict reactivity or non-reactivity which will help in determining risk factors.

Publications

10 25 50
 
Description National Nanotoxicology Research Committee
Geographic Reach National 
Policy Influence Type Participation in a national consultation
 
Description CUPIDO: Cardio Ultraefficient nanoParticles for Inhalation of Drug prOducts
Amount € 6,500,000 (EUR)
Funding ID 720834 
Organisation European Commission H2020 
Sector Public
Country Belgium
Start 02/2017 
End 01/2021
 
Description European Respiratory Society Long Term In vitro study of the bioreactivitry of diesel exhaust particles at the human alveolar gas-blood barrier.
Amount € 37,000 (EUR)
Funding ID •European Respiratory Society Long Term Research Fellowship - 37,200 Euro - Research fellow (April 2016 - March 2017) In vitro study of the bioreactivitry of diesel exhaust particles at the human alveolar gas-blood barrier. 
Organisation European Respiratory Society (ERS) 
Sector Charity/Non Profit
Country European Union (EU)
Start 04/2016 
End 03/2017
 
Description FP7 Programme
Amount £437,500 (GBP)
Organisation European Commission 
Department Seventh Framework Programme (FP7)
Sector Public
Country European Union (EU)
Start 11/2008 
End 10/2012
 
Description Immortalisation of human lung alveolar epithelium and study of metal oxides in vitro
Amount € 56,000 (EUR)
Organisation Government of Spain 
Sector Public
Country Spain
Start 09/2015 
End 11/2017
 
Description MRC-HPA Center PhD studentship
Amount £80,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 09/2011 
End 08/2014
 
Description NE/H012893/1 Risk Assessment for Manufactured Nanoparticles Used in Consumer Products (RAMNUC) (£1.4m shared between T Lee, P Elliott and T Tetley
Amount £1,200,000 (GBP)
Funding ID NE/H012893/1 
Organisation Natural Environment Research Council 
Sector Public
Country United Kingdom
Start 05/2010 
End 04/2014
 
Description Nanoparticles effects on cell membranes and function - programme grant/Naional Institute for Environmentyal Health, US
Amount £1,200,000 (GBP)
Funding ID NIEHS No. U19ES019536 
Organisation National Institutes of Health (NIH) 
Sector Public
Country United States
Start 05/2010 
End 04/2015
 
Description Nanoparticles, sensory irritation and the lung: environmental influences on biological processes.
Amount £80,000 (GBP)
Funding ID 1243159 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2012 
End 09/2015
 
Description PhD Studentship
Amount £140,000 (GBP)
Organisation Unilever 
Sector Private
Country United Kingdom
Start 10/2010 
End 09/2013
 
Description PhD studentship
Amount £80,000 (GBP)
Organisation Public Health England 
Sector Public
Country United Kingdom
Start 10/2011 
End 01/2015
 
Description PhD studentship
Amount £80,000 (GBP)
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 01/2012 
End 01/2015
 
Description PhD studentship National Heart & Lung Institute
Amount £60,000 (GBP)
Organisation Imperial College London 
Department National Heart & Lung Institute (NHLI)
Sector Academic/University
Country United Kingdom
Start 10/2010 
End 09/2013
 
Description RCUK Fellowship
Amount £125,000 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start  
 
Description Royal Brompton NHS Trust PhD studentship
Amount £80,000 (GBP)
Organisation Royal Brompton & Harefield NHS Foundation Trust 
Sector Public
Country United Kingdom
Start 09/2010 
End 08/2013
 
Description Travel award
Amount £340 (GBP)
Organisation European Respiratory Society (ERS) 
Sector Charity/Non Profit
Country European Union (EU)
Start 09/2009 
End 09/2010
 
Description Travel award
Amount £750 (GBP)
Organisation British Lung Foundation (BLF) 
Sector Charity/Non Profit
Country United Kingdom
Start 05/2009 
End 05/2009
 
Description Travel award
Amount £1,000 (GBP)
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom
Start 05/2007 
End 05/2007
 
Description Travel award NanImpactNet EU
Amount £400 (GBP)
Organisation European Commission 
Sector Public
Country European Union (EU)
Start  
 
Description Travel award NanoImpact EU
Amount £400 (GBP)
Organisation European Commission 
Sector Public
Country European Union (EU)
Start  
 
Description Travel award NanoImpactnet EU
Amount £400 (GBP)
Organisation European Commission 
Sector Public
Country European Union (EU)
Start  
 
Title In vitro models of human respiratory unit 
Description The work in this project was the start of examining exactly how inhaled nanomaterials would interact with the alveolar epithelium and translocate into the pulmonary interstitium or possibly the vasculature. Very exact mechanisms have been identified, including punching holes into the epithelium and active processes involving clathrin. a small percentage (3%) of particles have been shown to translocate to the basal compartment. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2012 
Provided To Others? Yes  
Impact This research tool allows for very accurate determinstion of the uptake and translocation of nanosized objects through human respiratory epithelium. 
URL http://pubs.acs.org/doi/abs/10.1021/nn505399e
 
Description Exposure of experimental animals to aerosolised nanoparticulates 
Organisation Public Health England
Department Centre for Radiation, Chemicals and Environmental Hazards (CRCE)
Country United Kingdom 
Sector Public 
PI Contribution We performed cellular/molecular analyses. The material scientists associated with this research provided the nanoparticles.
Collaborator Contribution The agency has uniqure inhalation facilities which have been established by experts in aerosol science who set up the facility, advised on exposure protocols and performed the exposures and lung function studies. Although there was no direct funding through this collaboration, sections of two grants, RAMNUC and RESAC, included in vivo exposures to be carried out at the HPA which we believe proved attractive to the reviewers.
Impact This collaboration involved aerosol scientists, physicists, pathologists, cell biologists, material scientists and chemists.
Start Year 2009
 
Description Identification of mechanisms of platelet activation by nano-particle pollutants in the UK 
Organisation Imperial College London
Country United Kingdom 
Sector Academic/University 
PI Contribution Lung cell biology, knowledge of role of inhaled particulate air pollution in cardiorespiratory disease, working with nanoparticles
Collaborator Contribution Knowledge of the cardiovascular system and measurement of platelet activation
Impact Three publications and still collaborating.
Start Year 2011
 
Description NanoRetox programme 
Organisation Natural History Museum
Country United Kingdom 
Sector Public 
PI Contribution Our expertise with primary human lung cells and our long experience studying environmental toxicants in experimental models in vitro and in vivo.
Collaborator Contribution We have set up NaNoRISK, Nanotoxicology Research In South Kensington. This facilitates collaborative, multidisciplinary research between the instutions and has provided a platform from which to obtain funding.
Impact We have, between us, obtained over ten million pounds worth of funding from the UK, EU and US agencies. Those involving me have been listed. These studies involve cell biologists, materials scientists, chemists, physicists, aerosol scientists, clinicians, epidemiologists.
Start Year 2006
 
Description Nanoparticles, sensory irritation and the lung: environmental influences on biological processes 
Organisation Imperial College London
Department National Heart & Lung Institute (NHLI)
Country United Kingdom 
Sector Academic/University 
PI Contribution I am providing the nanoparticle expertise and analytical techniques
Collaborator Contribution One partner works on the vagus nerve model and cough, the other is an expert in nanomaterials and imaging.
Impact Featured in News and on TV. Brings together a materials scientist, a pharmacologist and lung cell biologist
Start Year 2012
 
Description Respiratory effects of silver and carbon nanomaterials (RESAC) 
Organisation University of Southern California
Country United States 
Sector Academic/University 
PI Contribution I am the UK PI on this study and the working hypothesis was proposed by me. I co-ordinated all the UK applicants and integrated the core research (materials scientists) with the 3 research WPs. I was responsible for writing 2 of the 3 WPs. We have set up unique in vitro models of the human lung, we can establish well-characterised in vivo models to study lung surfactant metabolism, we work closely with the material scientists and use unique imaging methods to study particle-cell interactions.
Collaborator Contribution The collaborators have experience in modelling, in lung surfactant biology and in oxidative stress that will add to the research programme in my laboratory.
Impact This collaboration was funded end of September 2010. We worked with modellers, epidemiologists, experts in lung surfactant biology, immunology and oxidative stress to complement the imperial College expertise in cell biology, in vivo models of lung disease and materials science. There are still one or two papers to be written
Start Year 2010
 
Description Risk Assessment for Manufactured Nanoparticles Used in Consumer Products (RAMNUC) 
Organisation Rutgers University
Country United States 
Sector Academic/University 
PI Contribution I am a co-PI of the grant. I brought together all the PIs from the UK consortium (material scientists, the collaborators at the HPA inhalation facility, the in vivo scientist) and wrote a significant portion of the work packages for the in vitro and in vivo studies. I will be the PI for the in vitro work package (WP1), reflecting my group's expertise with primary human lung cells in vitro and models to assessnanoparticle reactivity.
Collaborator Contribution We have links with modellers to investigate the possible environmental and other risks associated with exposure to ZnO and Ag nanomaterials that are being used in commercial products. Specifally to study nanoparticles' life cycle: (1) sources, (2) environmental fate and transport, (3) concentrations in environmental media, (4) human exposure, (5) dose at target organs and bioavailability, (6) toxicological effects, and (7) ecological impacts. In addition, access to immune in vitro models of human monocytes and macrophages to investigate susceptibility to TB in the presence of nanoparticles. The combined expertise allows us to examine and understand better the impact of these nanopartoicles throughout their lifespan and therefore to predict hazard and risk.
Impact We started this research this year and there aree no outputs/outcomes as yet. The research is multidisciplinary, involving cell biologists, clinicians, epidemiologists, modellers, aerosol scientists and material scientists.
Start Year 2009
 
Description The cytoxicity of carbon nanostructures to lung epithelial cells: The effect of aspect ratio and surface functionalisation 
Organisation Imperial College London
Department Faculty of Engineering
Country United Kingdom 
Sector Academic/University 
PI Contribution I am the lung cell biologist, providing expertise with human lung cell models in vitro, knowledge of the lung and analysis of the exposure models
Collaborator Contribution The Materials scientists are synthesising the carbon nanotubes in specific formats. The interaction of these nanomaterials with the lung cells are being studied using state of the art electron microscopy (Titan) and other analytical techniques
Impact I am a cell biologist, one partner is a chemist, the other is a materials scientist.
Start Year 2011
 
Description Breathing Easy 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Schools
Results and Impact The lecture theatre at the Royal Instuitute was full (200?) of schoolchildren between 8 and 12 years. They were very excited at some of the demonstrations and asked lots of questions. They alos came to talk to me afterwards, as did their parents/teachers..

I did not get any feedback; perhaps the RI did.
Year(s) Of Engagement Activity 2008