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Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™)
John W. Wills,
Nicole Hondow,
Adam David Thomas 
,
Katherine Chapman ,
David Fish,
Thierry Maffeis ,
Mark W. Penny,
Richard A. Brown,
Gareth Jenkins ,
Andy P. Brown,
Paul A. White,
Shareen Doak
,
Katherine Chapman ,
David Fish,
Thierry Maffeis ,
Mark W. Penny,
Richard A. Brown,
Gareth Jenkins ,
Andy P. Brown,
Paul A. White,
Shareen Doak
Particle and Fibre Toxicology, Volume: 13, Issue: 1
Swansea University Authors:
Adam David Thomas , Katherine Chapman , Thierry Maffeis , Gareth Jenkins , Shareen Doak
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DOI (Published version): 10.1186/s12989-016-0161-5
Abstract
BackgroundThe rapid production and incorporation of engineered nanomaterials into consumer products alongside research suggesting nanomaterials can cause cell death and DNA damage (genotoxicity) makes in vitro assays desirable for nanosafety screening. However, conflicting outcomes are often observe...
| Published in: | Particle and Fibre Toxicology |
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| ISSN: | 1743-8977 1743-8977 |
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2016
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| URI: | https://cronfa.swan.ac.uk/Record/cronfa29852 |
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<?xml version="1.0"?><rfc1807><datestamp>2021-01-14T13:03:19.3060942</datestamp><bib-version>v2</bib-version><id>29852</id><entry>2016-09-09</entry><title>Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™)</title><swanseaauthors><author><sid>fab7e04239bbf899b6a51a97334e91b1</sid><ORCID>NULL</ORCID><firstname>Adam David</firstname><surname>Thomas</surname><name>Adam David Thomas</name><active>true</active><ethesisStudent>true</ethesisStudent></author><author><sid>19e7d85eec17117858d867ec0c9f575e</sid><ORCID>0000-0001-6668-0705</ORCID><firstname>Katherine</firstname><surname>Chapman</surname><name>Katherine Chapman</name><active>true</active><ethesisStudent>false</ethesisStudent></author><author><sid>992eb4cb18b61c0cd3da6e0215ac787c</sid><ORCID>0000-0003-2357-0092</ORCID><firstname>Thierry</firstname><surname>Maffeis</surname><name>Thierry Maffeis</name><active>true</active><ethesisStudent>false</ethesisStudent></author><author><sid>a44095d26187304e903da7ca778697b6</sid><ORCID>0000-0002-5437-8389</ORCID><firstname>Gareth</firstname><surname>Jenkins</surname><name>Gareth Jenkins</name><active>true</active><ethesisStudent>false</ethesisStudent></author><author><sid>8f70286908f67238a527a98cbf66d387</sid><ORCID>0000-0002-6753-1987</ORCID><firstname>Shareen</firstname><surname>Doak</surname><name>Shareen Doak</name><active>true</active><ethesisStudent>false</ethesisStudent></author></swanseaauthors><date>2016-09-09</date><abstract>BackgroundThe rapid production and incorporation of engineered nanomaterials into consumer products alongside research suggesting nanomaterials can cause cell death and DNA damage (genotoxicity) makes in vitro assays desirable for nanosafety screening. However, conflicting outcomes are often observed when in vitro and in vivo study results are compared, suggesting more physiologically representative in vitro models are required to minimise reliance on animal testing.MethodBASF Levasil® silica nanoparticles (16 and 85 nm) were used to adapt the 3D reconstructed skin micronucleus (RSMN) assay for nanomaterials administered topically or into the growth medium. 3D dose-responses were compared to a 2D micronucleus assay using monocultured human B cells (TK6) after standardising dose between 2D / 3D assays by total nanoparticle mass to cell number. Cryogenic vitrification, scanning electron microscopy and dynamic light scattering techniques were applied to characterise in-medium and air-liquid interface exposures. Advanced transmission electron microscopy imaging modes (high angle annular dark field) and X-ray spectrometry were used to define nanoparticle penetration / cellular uptake in the intact 3D models and 2D monocultured cells.ResultsFor all 2D exposures, significant (p < 0.002) increases in genotoxicity were observed (≥100 μg/mL) alongside cell viability decreases (p < 0.015) at doses ≥200 μg/mL (16 nm-SiO2) and ≥100 μg/mL (85 nm-SiO2). In contrast, 2D-equivalent exposures to the 3D models (≤300 μg/mL) caused no significant DNA damage or impact on cell viability. Further increasing dose to the 3D models led to probable air-liquid interface suffocation. Nanoparticle penetration / cell uptake analysis revealed no exposure to the live cells of the 3D model occurred due to the protective nature of the skin model’s 3D cellular microarchitecture (topical exposures) and confounding barrier effects of the collagen cell attachment layer (in-medium exposures). 2D monocultured cells meanwhile showed extensive internalisation of both silica particles causing (geno)toxicity.ConclusionsThe results establish the importance of tissue microarchitecture in defining nanomaterial exposure, and suggest 3D in vitro models could play a role in bridging the gap between in vitro and in vivo outcomes in nanotoxicology. Robust exposure characterisation and uptake assessment methods (as demonstrated) are essential to interpret nano(geno)toxicity studies successfully.</abstract><type>Journal Article</type><journal>Particle and Fibre Toxicology</journal><volume>13</volume><journalNumber>1</journalNumber><paginationStart/><paginationEnd/><publisher/><placeOfPublication/><isbnPrint/><isbnElectronic/><issnPrint>1743-8977</issnPrint><issnElectronic>1743-8977</issnElectronic><keywords>3D cell culture, Silica, Genotoxicity, Nanotoxicology, Physico-chemical characterisation, Nanoparticles,Reconstructed skin, RSMN, Micronucleus assay, Air-liquid interface</keywords><publishedDay>9</publishedDay><publishedMonth>9</publishedMonth><publishedYear>2016</publishedYear><publishedDate>2016-09-09</publishedDate><doi>10.1186/s12989-016-0161-5</doi><url/><notes/><college>COLLEGE NANME</college><department>Swansea University Medical School</department><CollegeCode>COLLEGE CODE</CollegeCode><institution>Swansea University</institution><degreesponsorsfunders>EPSRC (EP/H008683/1), NC3Rs (Jenkins.G.10-07-2009)</degreesponsorsfunders><apcterm/><lastEdited>2021-01-14T13:03:19.3060942</lastEdited><Created>2016-09-09T12:51:00.4552435</Created><path><level id="1">Faculty of Science and Engineering</level><level id="2">School of Engineering and Applied Sciences - Uncategorised</level></path><authors><author><firstname>John W.</firstname><surname>Wills</surname><order>1</order></author><author><firstname>Nicole</firstname><surname>Hondow</surname><order>2</order></author><author><firstname>Adam David</firstname><surname>Thomas</surname><orcid>NULL</orcid><order>3</order></author><author><firstname>Katherine</firstname><surname>Chapman</surname><orcid>0000-0001-6668-0705</orcid><order>4</order></author><author><firstname>David</firstname><surname>Fish</surname><order>5</order></author><author><firstname>Thierry</firstname><surname>Maffeis</surname><orcid>0000-0003-2357-0092</orcid><order>6</order></author><author><firstname>Mark W.</firstname><surname>Penny</surname><order>7</order></author><author><firstname>Richard A.</firstname><surname>Brown</surname><order>8</order></author><author><firstname>Gareth</firstname><surname>Jenkins</surname><orcid>0000-0002-5437-8389</orcid><order>9</order></author><author><firstname>Andy P.</firstname><surname>Brown</surname><order>10</order></author><author><firstname>Paul A.</firstname><surname>White</surname><order>11</order></author><author><firstname>Shareen</firstname><surname>Doak</surname><orcid>0000-0002-6753-1987</orcid><order>12</order></author></authors><documents><document><filename>0029852-16092016163446.pdf</filename><originalFilename>GeneticToxicityDoak.pdf</originalFilename><uploaded>2016-09-16T16:34:46.1630000</uploaded><type>Output</type><contentLength>4173315</contentLength><contentType>application/pdf</contentType><version>Version of Record</version><cronfaStatus>true</cronfaStatus><documentNotes>Released under the terms of a Creative Commons Attribution 4.0 International License (CC-BY).</documentNotes><copyrightCorrect>true</copyrightCorrect><language>eng</language><licence>http://creativecommons.org/licenses/by/4.0/</licence></document></documents><OutputDurs/></rfc1807> |
| spelling |
2021-01-14T13:03:19.3060942 v2 29852 2016-09-09 Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™) fab7e04239bbf899b6a51a97334e91b1 NULL Adam David Thomas Adam David Thomas true true 19e7d85eec17117858d867ec0c9f575e 0000-0001-6668-0705 Katherine Chapman Katherine Chapman true false 992eb4cb18b61c0cd3da6e0215ac787c 0000-0003-2357-0092 Thierry Maffeis Thierry Maffeis true false a44095d26187304e903da7ca778697b6 0000-0002-5437-8389 Gareth Jenkins Gareth Jenkins true false 8f70286908f67238a527a98cbf66d387 0000-0002-6753-1987 Shareen Doak Shareen Doak true false 2016-09-09 BackgroundThe rapid production and incorporation of engineered nanomaterials into consumer products alongside research suggesting nanomaterials can cause cell death and DNA damage (genotoxicity) makes in vitro assays desirable for nanosafety screening. However, conflicting outcomes are often observed when in vitro and in vivo study results are compared, suggesting more physiologically representative in vitro models are required to minimise reliance on animal testing.MethodBASF Levasil® silica nanoparticles (16 and 85 nm) were used to adapt the 3D reconstructed skin micronucleus (RSMN) assay for nanomaterials administered topically or into the growth medium. 3D dose-responses were compared to a 2D micronucleus assay using monocultured human B cells (TK6) after standardising dose between 2D / 3D assays by total nanoparticle mass to cell number. Cryogenic vitrification, scanning electron microscopy and dynamic light scattering techniques were applied to characterise in-medium and air-liquid interface exposures. Advanced transmission electron microscopy imaging modes (high angle annular dark field) and X-ray spectrometry were used to define nanoparticle penetration / cellular uptake in the intact 3D models and 2D monocultured cells.ResultsFor all 2D exposures, significant (p < 0.002) increases in genotoxicity were observed (≥100 μg/mL) alongside cell viability decreases (p < 0.015) at doses ≥200 μg/mL (16 nm-SiO2) and ≥100 μg/mL (85 nm-SiO2). In contrast, 2D-equivalent exposures to the 3D models (≤300 μg/mL) caused no significant DNA damage or impact on cell viability. Further increasing dose to the 3D models led to probable air-liquid interface suffocation. Nanoparticle penetration / cell uptake analysis revealed no exposure to the live cells of the 3D model occurred due to the protective nature of the skin model’s 3D cellular microarchitecture (topical exposures) and confounding barrier effects of the collagen cell attachment layer (in-medium exposures). 2D monocultured cells meanwhile showed extensive internalisation of both silica particles causing (geno)toxicity.ConclusionsThe results establish the importance of tissue microarchitecture in defining nanomaterial exposure, and suggest 3D in vitro models could play a role in bridging the gap between in vitro and in vivo outcomes in nanotoxicology. Robust exposure characterisation and uptake assessment methods (as demonstrated) are essential to interpret nano(geno)toxicity studies successfully. Journal Article Particle and Fibre Toxicology 13 1 1743-8977 1743-8977 3D cell culture, Silica, Genotoxicity, Nanotoxicology, Physico-chemical characterisation, Nanoparticles,Reconstructed skin, RSMN, Micronucleus assay, Air-liquid interface 9 9 2016 2016-09-09 10.1186/s12989-016-0161-5 COLLEGE NANME Swansea University Medical School COLLEGE CODE Swansea University EPSRC (EP/H008683/1), NC3Rs (Jenkins.G.10-07-2009) 2021-01-14T13:03:19.3060942 2016-09-09T12:51:00.4552435 Faculty of Science and Engineering School of Engineering and Applied Sciences - Uncategorised John W. Wills 1 Nicole Hondow 2 Adam David Thomas NULL 3 Katherine Chapman 0000-0001-6668-0705 4 David Fish 5 Thierry Maffeis 0000-0003-2357-0092 6 Mark W. Penny 7 Richard A. Brown 8 Gareth Jenkins 0000-0002-5437-8389 9 Andy P. Brown 10 Paul A. White 11 Shareen Doak 0000-0002-6753-1987 12 0029852-16092016163446.pdf GeneticToxicityDoak.pdf 2016-09-16T16:34:46.1630000 Output 4173315 application/pdf Version of Record true Released under the terms of a Creative Commons Attribution 4.0 International License (CC-BY). true eng http://creativecommons.org/licenses/by/4.0/ |
| title |
Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™) |
| spellingShingle |
Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™) Adam David Thomas Katherine Chapman Thierry Maffeis Gareth Jenkins Shareen Doak |
| title_short |
Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™) |
| title_full |
Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™) |
| title_fullStr |
Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™) |
| title_full_unstemmed |
Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™) |
| title_sort |
Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™) |
| author_id_str_mv |
fab7e04239bbf899b6a51a97334e91b1 19e7d85eec17117858d867ec0c9f575e 992eb4cb18b61c0cd3da6e0215ac787c a44095d26187304e903da7ca778697b6 8f70286908f67238a527a98cbf66d387 |
| author_id_fullname_str_mv |
fab7e04239bbf899b6a51a97334e91b1_***_Adam David Thomas 19e7d85eec17117858d867ec0c9f575e_***_Katherine Chapman 992eb4cb18b61c0cd3da6e0215ac787c_***_Thierry Maffeis a44095d26187304e903da7ca778697b6_***_Gareth Jenkins 8f70286908f67238a527a98cbf66d387_***_Shareen Doak |
| author |
Adam David Thomas Katherine Chapman Thierry Maffeis Gareth Jenkins Shareen Doak |
| author2 |
John W. Wills Nicole Hondow Adam David Thomas Katherine Chapman David Fish Thierry Maffeis Mark W. Penny Richard A. Brown Gareth Jenkins Andy P. Brown Paul A. White Shareen Doak |
| format |
Journal article |
| container_title |
Particle and Fibre Toxicology |
| container_volume |
13 |
| container_issue |
1 |
| publishDate |
2016 |
| institution |
Swansea University |
| issn |
1743-8977 1743-8977 |
| doi_str_mv |
10.1186/s12989-016-0161-5 |
| college_str |
Faculty of Science and Engineering |
| hierarchytype |
|
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facultyofscienceandengineering |
| hierarchy_top_title |
Faculty of Science and Engineering |
| hierarchy_parent_id |
facultyofscienceandengineering |
| hierarchy_parent_title |
Faculty of Science and Engineering |
| department_str |
School of Engineering and Applied Sciences - Uncategorised{{{_:::_}}}Faculty of Science and Engineering{{{_:::_}}}School of Engineering and Applied Sciences - Uncategorised |
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| description |
BackgroundThe rapid production and incorporation of engineered nanomaterials into consumer products alongside research suggesting nanomaterials can cause cell death and DNA damage (genotoxicity) makes in vitro assays desirable for nanosafety screening. However, conflicting outcomes are often observed when in vitro and in vivo study results are compared, suggesting more physiologically representative in vitro models are required to minimise reliance on animal testing.MethodBASF Levasil® silica nanoparticles (16 and 85 nm) were used to adapt the 3D reconstructed skin micronucleus (RSMN) assay for nanomaterials administered topically or into the growth medium. 3D dose-responses were compared to a 2D micronucleus assay using monocultured human B cells (TK6) after standardising dose between 2D / 3D assays by total nanoparticle mass to cell number. Cryogenic vitrification, scanning electron microscopy and dynamic light scattering techniques were applied to characterise in-medium and air-liquid interface exposures. Advanced transmission electron microscopy imaging modes (high angle annular dark field) and X-ray spectrometry were used to define nanoparticle penetration / cellular uptake in the intact 3D models and 2D monocultured cells.ResultsFor all 2D exposures, significant (p < 0.002) increases in genotoxicity were observed (≥100 μg/mL) alongside cell viability decreases (p < 0.015) at doses ≥200 μg/mL (16 nm-SiO2) and ≥100 μg/mL (85 nm-SiO2). In contrast, 2D-equivalent exposures to the 3D models (≤300 μg/mL) caused no significant DNA damage or impact on cell viability. Further increasing dose to the 3D models led to probable air-liquid interface suffocation. Nanoparticle penetration / cell uptake analysis revealed no exposure to the live cells of the 3D model occurred due to the protective nature of the skin model’s 3D cellular microarchitecture (topical exposures) and confounding barrier effects of the collagen cell attachment layer (in-medium exposures). 2D monocultured cells meanwhile showed extensive internalisation of both silica particles causing (geno)toxicity.ConclusionsThe results establish the importance of tissue microarchitecture in defining nanomaterial exposure, and suggest 3D in vitro models could play a role in bridging the gap between in vitro and in vivo outcomes in nanotoxicology. Robust exposure characterisation and uptake assessment methods (as demonstrated) are essential to interpret nano(geno)toxicity studies successfully. |
| published_date |
2016-09-09T03:36:23Z |
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1763751580812705792 |
| score |
11.037056 |