Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials

Abstract In vitro cytotoxicity testing is an indispensable part of the development of new biomaterials. However, the standard ISO 10993-5 enables variability in the testing conditions, which makes the results of the test incomparable. We studied the influence of media composition on the results of t...

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Autores principales:	Eva Jablonská, Jiří Kubásek, Dalibor Vojtěch, Tomáš Ruml, Jan Lipov
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3f0a2206c2984c4aa200af8d0e9f71f1
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spelling	oai:doaj.org-article:3f0a2206c2984c4aa200af8d0e9f71f12021-12-02T17:04:36ZTest conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials10.1038/s41598-021-85019-62045-2322https://doaj.org/article/3f0a2206c2984c4aa200af8d0e9f71f12021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85019-6https://doaj.org/toc/2045-2322Abstract In vitro cytotoxicity testing is an indispensable part of the development of new biomaterials. However, the standard ISO 10993-5 enables variability in the testing conditions, which makes the results of the test incomparable. We studied the influence of media composition on the results of the cytotoxicity test. Solutions of ZnCl2 served as simulated extracts and we also used extracts of three types of Zn-based and Mg-based degradable metals. We incubated the cells with the solutions prepared in two types of media with two concentrations of serum (5 and 10%). We compared the toxic effect of the extracts on L929 murine fibroblast-derived cell line, which is recommended by ISO standard and on “osteoblast-like cells” U-2 OS. We also compared two methods of exposition: solutions were added either to a sub-confluent layer or to the cell suspension. We evaluated the metabolic activity of the cells using the resazurin test. We found out that in vitro cytotoxicity is dramatically influenced by the concentration of serum and by the type of the medium as well as by the type of exposition and type of cells. Therefore, when performing in vitro cytotoxicity testing of biomaterials, the authors should carefully specify the conditions of the test and comparison of different studies should be carried out with caution.Eva JablonskáJiří KubásekDalibor VojtěchTomáš RumlJan LipovNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Eva Jablonská Jiří Kubásek Dalibor Vojtěch Tomáš Ruml Jan Lipov Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials
description	Abstract In vitro cytotoxicity testing is an indispensable part of the development of new biomaterials. However, the standard ISO 10993-5 enables variability in the testing conditions, which makes the results of the test incomparable. We studied the influence of media composition on the results of the cytotoxicity test. Solutions of ZnCl2 served as simulated extracts and we also used extracts of three types of Zn-based and Mg-based degradable metals. We incubated the cells with the solutions prepared in two types of media with two concentrations of serum (5 and 10%). We compared the toxic effect of the extracts on L929 murine fibroblast-derived cell line, which is recommended by ISO standard and on “osteoblast-like cells” U-2 OS. We also compared two methods of exposition: solutions were added either to a sub-confluent layer or to the cell suspension. We evaluated the metabolic activity of the cells using the resazurin test. We found out that in vitro cytotoxicity is dramatically influenced by the concentration of serum and by the type of the medium as well as by the type of exposition and type of cells. Therefore, when performing in vitro cytotoxicity testing of biomaterials, the authors should carefully specify the conditions of the test and comparison of different studies should be carried out with caution.
format	article
author	Eva Jablonská Jiří Kubásek Dalibor Vojtěch Tomáš Ruml Jan Lipov
author_facet	Eva Jablonská Jiří Kubásek Dalibor Vojtěch Tomáš Ruml Jan Lipov
author_sort	Eva Jablonská
title	Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials
title_short	Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials
title_full	Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials
title_fullStr	Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials
title_full_unstemmed	Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials
title_sort	test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/3f0a2206c2984c4aa200af8d0e9f71f1
work_keys_str_mv	AT evajablonska testconditionscansignificantlyaffecttheresultsofinvitrocytotoxicitytestingofdegradablemetallicbiomaterials AT jirikubasek testconditionscansignificantlyaffecttheresultsofinvitrocytotoxicitytestingofdegradablemetallicbiomaterials AT daliborvojtech testconditionscansignificantlyaffecttheresultsofinvitrocytotoxicitytestingofdegradablemetallicbiomaterials AT tomasruml testconditionscansignificantlyaffecttheresultsofinvitrocytotoxicitytestingofdegradablemetallicbiomaterials AT janlipov testconditionscansignificantlyaffecttheresultsofinvitrocytotoxicitytestingofdegradablemetallicbiomaterials
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Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials

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