Use of transgenic zebrafish as a tool to study nanomaterial inflammogenicity
Lead Research Organisation:
Heriot-Watt University
Department Name: Sch of Engineering and Physical Science
Abstract
This project will use zebrafish to assess the inflammatory responses induced by nanomaterials (NMs) in order to identify if zebrafish are a uesful (alternative) tool to investigate NM mediated inflammatory responses (e.g. neutrophil accumulation). A panel of NMs will be tested to identify whether the physico-chemical properties of NMs influence their pathogenicity. This will require that the physico-chemical properties of the NMs under investigation are characterised thoroughly (e.g. using AFM, TEM, SEM, ICP-MS). Where possible state of the art microscopy will be used to visualise and track immune cell movement in zebrafish models, and to quantify their accumulation. Of benefit, is that the ability of some of these NMs to induce pulmonary inflammation has already been tested in previous in vivo (rodent) studies. This will allow us to identify whether zebrafish are an appropriate alternative to rodent testing.
Publications
Johnston HJ
(2018)
Adoption of in vitro systems and zebrafish embryos as alternative models for reducing rodent use in assessments of immunological and oxidative stress responses to nanomaterials.
in Critical reviews in toxicology
Johnston H
(2023)
Application of transgenic zebrafish for investigating inflammatory responses to nanomaterials: Recommendations for new users
in F1000Research
Gillies S
(2022)
Transgenic zebrafish larvae as a non-rodent alternative model to assess pro-inflammatory (neutrophil) responses to nanomaterials.
in Nanotoxicology
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| EP/N509474/1 | 01/10/2016 | 30/09/2021 | |||
| 1813003 | Studentship | EP/N509474/1 | 01/10/2016 | 31/03/2020 | Suzanne Gillies |
| Description | Developing the transgenic zebrafish larvae as a model organism for nanotoxicology and investigating the use of alternative in vitro assays using human cells (primary neutrophils) and cell lines (HL-60 neutrophil-like cells) |
| Exploitation Route | A protocol has been developed which utilises transgenic zebrafish larvae from 3dpf to 5dpf to assess neutrophil (inflammatory) responses to nanomaterials, either by microinjection or by tail fin wound assay. Further work is required to establish the optimal injection sites in larvae, otic vesicle is possible but is associated with high mortality. The tail fin wound should be further optimised as this assay has the potential to aid the investigation of the mechanism of nanomaterial toxicity and uptake/fate of nanomaterials, especially in other reporter strains (e.g. oxidative stress line). |
| Sectors | Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology |
| Description | Air Pollution in Thailand and its detrimental health impacts |
| Amount | £78,978 (GBP) |
| Organisation | Government of Scotland |
| Department | Scottish Funding Council |
| Sector | Public |
| Country | United Kingdom |
| Start | 01/2020 |
| End | 05/2021 |
| Description | Investigation of the toxicity of Particulate Matter Emitted from Biomass Burning in Thailand |
| Organisation | Australian National University (ANU) |
| Country | Australia |
| Sector | Academic/University |
| PI Contribution | There are uncertainties regarding the effects of haze pollution on human health in Thailand. The haze pollution is caused by biomass burning (e.g burning of agricultural land), which releases particles into the air. It is known that similar combustion derived particles that are emitted from other sources (e.g. traffic) are harmful, but it is unknown whether particles emitted from biomass burning have the same detrimental impacts on health. This knowledge gap urgently needs to be addressed to inform the implementation of appropriate, evidence-based policy interventions by regional and national Governments that protect public health and improve the welfare of the population. Therefore our research project aims to investigate the toxicity of particulate matter (PM) emitted from biomass burning in Thailand and compare this to the toxicity of PM emitted from other sources (e.g. traffic). HWU is responsible for assessing the toxicity of the PM samples (using in vitro and zebrafish embryo models) and will employ the methods that were developed in our NC3Rs project to perform the toxicity assessment. |
| Collaborator Contribution | Collaborators in Thailand and Australia will collect PM samples. HWU will assess the toxicity of the PM samples using in vitro and zebrafish models using the methods developed in the NC3Rs project. The IOM will compare the findings obtained from the alternative models to existing epidemiological data to identify if these models make a good prediction of toxicity. |
| Impact | The protocols developed in the NC3Rs project to assess inflammatory responses in transgenic zebrafish have been employed in our new project to assess the toxicity of PM samples from Thailand. HWU has also, for the first time, used zebrafish models to assess the developmental toxicity of air pollution particles. This new project may therefore also provide evidence that zebrafish and in vitro models may be suitable to assess the toxicity of particles (for a range of endpoints). The new project is extremely multidisciplinary involving toxicologists, exposure scientists, and epidemiologists. We are also engaging with interested stakeholders (e.g. policy makers in the Thai Government) to ensure they are kept abreast of project outcomes and will arrange a stakeholder event later this year to disseminate project findings. Our international, multidisciplinary team will investigate how harmful different sources of pollution are in Thailand to inform the introduction of interventions to protect the health of vulnerable communities. We will share expertise between research groups and supporting the training and education of early career researchers. |
| Start Year | 2020 |
| Description | Investigation of the toxicity of Particulate Matter Emitted from Biomass Burning in Thailand |
| Organisation | Chiang Mai University |
| Country | Thailand |
| Sector | Academic/University |
| PI Contribution | There are uncertainties regarding the effects of haze pollution on human health in Thailand. The haze pollution is caused by biomass burning (e.g burning of agricultural land), which releases particles into the air. It is known that similar combustion derived particles that are emitted from other sources (e.g. traffic) are harmful, but it is unknown whether particles emitted from biomass burning have the same detrimental impacts on health. This knowledge gap urgently needs to be addressed to inform the implementation of appropriate, evidence-based policy interventions by regional and national Governments that protect public health and improve the welfare of the population. Therefore our research project aims to investigate the toxicity of particulate matter (PM) emitted from biomass burning in Thailand and compare this to the toxicity of PM emitted from other sources (e.g. traffic). HWU is responsible for assessing the toxicity of the PM samples (using in vitro and zebrafish embryo models) and will employ the methods that were developed in our NC3Rs project to perform the toxicity assessment. |
| Collaborator Contribution | Collaborators in Thailand and Australia will collect PM samples. HWU will assess the toxicity of the PM samples using in vitro and zebrafish models using the methods developed in the NC3Rs project. The IOM will compare the findings obtained from the alternative models to existing epidemiological data to identify if these models make a good prediction of toxicity. |
| Impact | The protocols developed in the NC3Rs project to assess inflammatory responses in transgenic zebrafish have been employed in our new project to assess the toxicity of PM samples from Thailand. HWU has also, for the first time, used zebrafish models to assess the developmental toxicity of air pollution particles. This new project may therefore also provide evidence that zebrafish and in vitro models may be suitable to assess the toxicity of particles (for a range of endpoints). The new project is extremely multidisciplinary involving toxicologists, exposure scientists, and epidemiologists. We are also engaging with interested stakeholders (e.g. policy makers in the Thai Government) to ensure they are kept abreast of project outcomes and will arrange a stakeholder event later this year to disseminate project findings. Our international, multidisciplinary team will investigate how harmful different sources of pollution are in Thailand to inform the introduction of interventions to protect the health of vulnerable communities. We will share expertise between research groups and supporting the training and education of early career researchers. |
| Start Year | 2020 |
| Description | Investigation of the toxicity of Particulate Matter Emitted from Biomass Burning in Thailand |
| Organisation | Institute of Occupational Medicine (IOM) |
| Country | United Kingdom |
| Sector | Charity/Non Profit |
| PI Contribution | There are uncertainties regarding the effects of haze pollution on human health in Thailand. The haze pollution is caused by biomass burning (e.g burning of agricultural land), which releases particles into the air. It is known that similar combustion derived particles that are emitted from other sources (e.g. traffic) are harmful, but it is unknown whether particles emitted from biomass burning have the same detrimental impacts on health. This knowledge gap urgently needs to be addressed to inform the implementation of appropriate, evidence-based policy interventions by regional and national Governments that protect public health and improve the welfare of the population. Therefore our research project aims to investigate the toxicity of particulate matter (PM) emitted from biomass burning in Thailand and compare this to the toxicity of PM emitted from other sources (e.g. traffic). HWU is responsible for assessing the toxicity of the PM samples (using in vitro and zebrafish embryo models) and will employ the methods that were developed in our NC3Rs project to perform the toxicity assessment. |
| Collaborator Contribution | Collaborators in Thailand and Australia will collect PM samples. HWU will assess the toxicity of the PM samples using in vitro and zebrafish models using the methods developed in the NC3Rs project. The IOM will compare the findings obtained from the alternative models to existing epidemiological data to identify if these models make a good prediction of toxicity. |
| Impact | The protocols developed in the NC3Rs project to assess inflammatory responses in transgenic zebrafish have been employed in our new project to assess the toxicity of PM samples from Thailand. HWU has also, for the first time, used zebrafish models to assess the developmental toxicity of air pollution particles. This new project may therefore also provide evidence that zebrafish and in vitro models may be suitable to assess the toxicity of particles (for a range of endpoints). The new project is extremely multidisciplinary involving toxicologists, exposure scientists, and epidemiologists. We are also engaging with interested stakeholders (e.g. policy makers in the Thai Government) to ensure they are kept abreast of project outcomes and will arrange a stakeholder event later this year to disseminate project findings. Our international, multidisciplinary team will investigate how harmful different sources of pollution are in Thailand to inform the introduction of interventions to protect the health of vulnerable communities. We will share expertise between research groups and supporting the training and education of early career researchers. |
| Start Year | 2020 |
| Description | Investigation of the toxicity of Particulate Matter Emitted from Biomass Burning in Thailand |
| Organisation | Mahidol University |
| Country | Thailand |
| Sector | Academic/University |
| PI Contribution | There are uncertainties regarding the effects of haze pollution on human health in Thailand. The haze pollution is caused by biomass burning (e.g burning of agricultural land), which releases particles into the air. It is known that similar combustion derived particles that are emitted from other sources (e.g. traffic) are harmful, but it is unknown whether particles emitted from biomass burning have the same detrimental impacts on health. This knowledge gap urgently needs to be addressed to inform the implementation of appropriate, evidence-based policy interventions by regional and national Governments that protect public health and improve the welfare of the population. Therefore our research project aims to investigate the toxicity of particulate matter (PM) emitted from biomass burning in Thailand and compare this to the toxicity of PM emitted from other sources (e.g. traffic). HWU is responsible for assessing the toxicity of the PM samples (using in vitro and zebrafish embryo models) and will employ the methods that were developed in our NC3Rs project to perform the toxicity assessment. |
| Collaborator Contribution | Collaborators in Thailand and Australia will collect PM samples. HWU will assess the toxicity of the PM samples using in vitro and zebrafish models using the methods developed in the NC3Rs project. The IOM will compare the findings obtained from the alternative models to existing epidemiological data to identify if these models make a good prediction of toxicity. |
| Impact | The protocols developed in the NC3Rs project to assess inflammatory responses in transgenic zebrafish have been employed in our new project to assess the toxicity of PM samples from Thailand. HWU has also, for the first time, used zebrafish models to assess the developmental toxicity of air pollution particles. This new project may therefore also provide evidence that zebrafish and in vitro models may be suitable to assess the toxicity of particles (for a range of endpoints). The new project is extremely multidisciplinary involving toxicologists, exposure scientists, and epidemiologists. We are also engaging with interested stakeholders (e.g. policy makers in the Thai Government) to ensure they are kept abreast of project outcomes and will arrange a stakeholder event later this year to disseminate project findings. Our international, multidisciplinary team will investigate how harmful different sources of pollution are in Thailand to inform the introduction of interventions to protect the health of vulnerable communities. We will share expertise between research groups and supporting the training and education of early career researchers. |
| Start Year | 2020 |
| Description | Brightest Watts - practical demonstration of alternative models to schoolchildren (2019) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Schoolchildren participated in practical work during 2, 2.5 hour sessions as part of the HWU Brightest Watts summer week which allowed them to gain experience of working with in vitro and zebrafish models. This was also an opportunity to educate schoolchildren about the need to find alternatives to animal testing. |
| Year(s) Of Engagement Activity | 2019 |
| Description | HWU Seminars (2020) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Other audiences |
| Results and Impact | Seminars were given to academics and postgraduate students in the School of Engineering and Physical Sciences at HWU to disseminate project findings. |
| Year(s) Of Engagement Activity | 2020 |
| Description | Invited Talk at the Edinburgh Science Festival (Apr 2019) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | H Johnston was invited to give a talk as part of the Edinburgh Science Festival. As part of the talk she discussed what alternative models are available to assess nanomaterial toxicity A member of the audience contacted H Johnston after the event to ask that she spoke to the U3A (University for the Third Age) |
| Year(s) Of Engagement Activity | 2019 |
| Description | Invited talk at the NC3Rs PhD studentship Information Day (2019) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Other audiences |
| Results and Impact | An overview of the research activities of the project was provided to academics hoping to apply for NC3Rs PhD studentships - this will hopefully promote the more widespread use of the model. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Oral Presentation at SETAC EUROPE (May 2020) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | Talk entitled | Exploring the use of zebrafish embryo models as alternatives to rodent testing for investigating inflammatory responses to nanomaterials (NMs) given at the SETAC EUROPE Virtual conference in May 2020 After this conference the organisers of the SETAC North America conference contacted the research team at HWU to request that we provide an oral presentation at this work. This demonstrate interest of the wider scientific community in the research that has been performed as part of the NC3Rs projec. |
| Year(s) Of Engagement Activity | 2020 |
| Description | Oral Presentation at SETAC NORTH AMERICA (NOV 2020) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | Talk entitled Exploring the use of zebrafish embryo models as alternatives to rodent testing for investigating inflammatory responses to nanomaterials (NMs) provided at the conference |
| Year(s) Of Engagement Activity | 2020 |
| Description | Organisation of session on Alternatives to Animal Testing as part of the Health Innovation and Technologies Conference at HWU (2019) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Other audiences |
| Results and Impact | I (Helinor Johnston) arranged the Alternatives to Animal Testing session as part of the Health Innovation Technologies Conference at HWU. This allowed researchers working with non-rodent models to showcase their research to a range of interested stakeholders (e.g. academics, students, industry, policy makers, funders, patients, general public). |
| Year(s) Of Engagement Activity | 2019 |
| Description | Participation in Explorathon event in Edinburgh 2019 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | We had a stall at the Explorathon event in the botanical gardens in Edinburgh where we gave practical demonstrations of lab work (Related to the use of alternative models in research). Children (a range of ages), their parents and members of the public attended the event |
| Year(s) Of Engagement Activity | 2019 |
| Description | Poster Presentation at the First COST Action CA17140 training school "Cancer and Nanomedicine", Italy (2019) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | This training event allowed research activities to be communicated to other researchers working in the field of nanotoxicology and nanomedicine - this increased awareness of the alternative models employed in the project amongst the scientific community. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Poster presentation at 'The Nanosafety Forum for Young Scientists' (Malta, September 2018) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | A poster presentation which provided an overview of the research findings was given at The Nanosafety Forum for Young Scientists', (Malta) |
| Year(s) Of Engagement Activity | 2018 |
| Description | Poster presentation at 12th International Particle Toxicology Conference (Austria, 2019) |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Poster presentation at the 12th International Particle Toxicology Conference, Austria allowed research activities performed in the project to be communicated to the scientific community, and industry. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Poster presentation at the symposium "Fish and amphibian embryos as alternative models in toxicology and teratology" (Paris, December 2018) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | A poster was presented which provided an overview of the research findings of the project so far |
| Year(s) Of Engagement Activity | 2018 |
| Description | Poster presented at the 12th International Particle Toxicology Conference in Austria |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | I presented a poster at this conference |
| Year(s) Of Engagement Activity | 2019 |
| Description | Poster presented at the First COST Action CA17140 training school "Cancer and Nanomedicine" in Italy |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | I attended a training school which focused on nanomedicine safety assessment. I presented a poster of my research to share with the other participants of the school. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Presentation at the HWU Festival of Research (2019) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | I (Helinor Johnston) provided a presentation as part of the HWU Festival of Research in the session on Health Innovation and Technology. This talk discussed how alternative models could be used to a greater extent in biomedical research and presented data generated during the project. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Presentation to the U3A (University for the 3rd Age, 2019) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | A presentation entitled 'The benefits and limitations of employing alternative, non-rodent models to investigate the toxicity of nanomaterials' was given to attendees in order to provide an overview of how alternative, non rodent models can be employed in nanotoxicology testing. The participants requested that a lab tour was arranged so they could learn more about the alternative models. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Teaching of Undergraduate Students 2020-2021 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Undergraduate students |
| Results and Impact | Undergraduate students were educated about the 3Rs principles as part of their undergraduate Biological Sciences BSc degree. More specifically, lectures in several courses, for students at different stages of their degree, provided students with information on what the 3Rs principles are and examples of what alternatives to animal testing are available for biomedical research. In addition, H Johnston supervised several final year research project students who focused on using zebrafish or in vitro models for toxicology testing |
| Year(s) Of Engagement Activity | 2020,2021 |
| Description | Teaching of Undergraduate and Postgraduate Students (2019/2020) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Undergraduate students |
| Results and Impact | As part of several of my BSc and MSc courses (e.g. Molecular Toxicology, Pathobiology of Human Disease, Applied Studies in Human Health, Food Quality, Safety and Toxicity) I (Helinor Johnston) have given lectures on the 3Rs principles and use of alternative non-rodent models in biomedical research. This has given students a better theoretical understanding of the 3Rs principles and alternative models. I have also supervised final year BSc students who are conducting laboratory or desk based projects which employ alternative, non rodent models. |
| Year(s) Of Engagement Activity | 2019,2020 |