Revisiting ISO 10993‐5 In Vitro Cytotoxicity Standard Tests for Evaluation of Extracellular Matrix‐Based Biomaterials.
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| Title: | Revisiting ISO 10993‐5 In Vitro Cytotoxicity Standard Tests for Evaluation of Extracellular Matrix‐Based Biomaterials. |
|---|---|
| Authors: | Ramakrishnan, Rashmi1,2 (AUTHOR), Daly, Andrew C.1,2 (AUTHOR) andrew.daly@universityofgalway.ie, Galli, Carlo (AUTHOR) carlo.galli@unipr.it |
| Source: | International Journal of Biomaterials. 5/30/2026, Vol. 2026, p1-27. 27p. |
| Subjects: | Extracellular matrix, International Organization for Standardization, Cytotoxins, Biocompatibility, Apoptosis, Metabolic profile tests, Cell survival, Toxicity testing |
| Abstract: | Cytotoxicity testing is a critical step in the preclinical evaluation of biomaterials and medical devices, with extracellular matrix (ECM)‐based biomaterials constituting a significant category. The current ISO 10993‐5:2009 standard, Biological Evaluation of Medical Devices‐Part 5: Tests for In Vitro Cytotoxicity, delineates four metabolic assays that primarily rely on colorimetric methods and colony formation. However, relying solely on colorimetric assays or colony formation fails to provide precise insights into cell function/activity and may yield false‐positive results, contributing to interlaboratory discrepancies. This study systematically evaluated ISO 10993‐5‐recommended assays for ECM‐based commercial products, specifically assessing the impact of key assay variables including cell types, contact mode (test extracts versus test material itself), and media components (with or without serum) on biological outcomes. These evaluations support the development of more accurate and robust test methods. While all four assays indicated the noncytotoxic nature of the test samples, metabolic activity readings varied substantially depending on the serum presence, cell types, and assay method employed. To address these limitations and achieve more precise insights into cellular activity, cell membrane integrity (live/dead staining), cell‐ECM attachment (actin cytoskeleton), proliferation (Ki67), and apoptosis (annexin V) were analyzed. Notably, despite observing increased metabolic activity (100%–150%) under serum‐free conditions measured using MTT and XTT assays, live/dead and actin staining showed no corresponding changes in cell viability or attachment, and Ki67 indicated only ∼15% proliferation. Annexin V staining was detected only in human primary dermal fibroblasts, highlighting their greater reliability over L929 cells for detecting apoptosis. These findings provide a valuable reference for researchers, regulatory bodies, and industry stakeholders in refining cytotoxicity testing protocols and guiding future ISO 10993‐5 revisions for more reliable assessment of biomaterials and medical devices. [ABSTRACT FROM AUTHOR] |
| Copyright of International Journal of Biomaterials is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
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| Header | DbId: egs DbLabel: Engineering Source An: 194204605 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: Revisiting ISO 10993‐5 In Vitro Cytotoxicity Standard Tests for Evaluation of Extracellular Matrix‐Based Biomaterials. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Ramakrishnan%2C+Rashmi%22">Ramakrishnan, Rashmi</searchLink><relatesTo>1,2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Daly%2C+Andrew+C%2E%22">Daly, Andrew C.</searchLink><relatesTo>1,2</relatesTo> (AUTHOR)<i> andrew.daly@universityofgalway.ie</i><br /><searchLink fieldCode="AR" term="%22Galli%2C+Carlo%22">Galli, Carlo</searchLink> (AUTHOR)<i> carlo.galli@unipr.it</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22International+Journal+of+Biomaterials%22">International Journal of Biomaterials</searchLink>. 5/30/2026, Vol. 2026, p1-27. 27p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Extracellular+matrix%22">Extracellular matrix</searchLink><br /><searchLink fieldCode="DE" term="%22International+Organization+for+Standardization%22">International Organization for Standardization</searchLink><br /><searchLink fieldCode="DE" term="%22Cytotoxins%22">Cytotoxins</searchLink><br /><searchLink fieldCode="DE" term="%22Biocompatibility%22">Biocompatibility</searchLink><br /><searchLink fieldCode="DE" term="%22Apoptosis%22">Apoptosis</searchLink><br /><searchLink fieldCode="DE" term="%22Metabolic+profile+tests%22">Metabolic profile tests</searchLink><br /><searchLink fieldCode="DE" term="%22Cell+survival%22">Cell survival</searchLink><br /><searchLink fieldCode="DE" term="%22Toxicity+testing%22">Toxicity testing</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Cytotoxicity testing is a critical step in the preclinical evaluation of biomaterials and medical devices, with extracellular matrix (ECM)‐based biomaterials constituting a significant category. The current ISO 10993‐5:2009 standard, Biological Evaluation of Medical Devices‐Part 5: Tests for In Vitro Cytotoxicity, delineates four metabolic assays that primarily rely on colorimetric methods and colony formation. However, relying solely on colorimetric assays or colony formation fails to provide precise insights into cell function/activity and may yield false‐positive results, contributing to interlaboratory discrepancies. This study systematically evaluated ISO 10993‐5‐recommended assays for ECM‐based commercial products, specifically assessing the impact of key assay variables including cell types, contact mode (test extracts versus test material itself), and media components (with or without serum) on biological outcomes. These evaluations support the development of more accurate and robust test methods. While all four assays indicated the noncytotoxic nature of the test samples, metabolic activity readings varied substantially depending on the serum presence, cell types, and assay method employed. To address these limitations and achieve more precise insights into cellular activity, cell membrane integrity (live/dead staining), cell‐ECM attachment (actin cytoskeleton), proliferation (Ki67), and apoptosis (annexin V) were analyzed. Notably, despite observing increased metabolic activity (100%–150%) under serum‐free conditions measured using MTT and XTT assays, live/dead and actin staining showed no corresponding changes in cell viability or attachment, and Ki67 indicated only ∼15% proliferation. Annexin V staining was detected only in human primary dermal fibroblasts, highlighting their greater reliability over L929 cells for detecting apoptosis. These findings provide a valuable reference for researchers, regulatory bodies, and industry stakeholders in refining cytotoxicity testing protocols and guiding future ISO 10993‐5 revisions for more reliable assessment of biomaterials and medical devices. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of International Journal of Biomaterials is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1155/ijbm/7395612 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 27 StartPage: 1 Subjects: – SubjectFull: Extracellular matrix Type: general – SubjectFull: International Organization for Standardization Type: general – SubjectFull: Cytotoxins Type: general – SubjectFull: Biocompatibility Type: general – SubjectFull: Apoptosis Type: general – SubjectFull: Metabolic profile tests Type: general – SubjectFull: Cell survival Type: general – SubjectFull: Toxicity testing Type: general Titles: – TitleFull: Revisiting ISO 10993‐5 In Vitro Cytotoxicity Standard Tests for Evaluation of Extracellular Matrix‐Based Biomaterials. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Ramakrishnan, Rashmi – PersonEntity: Name: NameFull: Daly, Andrew C. – PersonEntity: Name: NameFull: Galli, Carlo IsPartOfRelationships: – BibEntity: Dates: – D: 30 M: 05 Text: 5/30/2026 Type: published Y: 2026 Identifiers: – Type: issn-print Value: 16878787 Numbering: – Type: volume Value: 2026 Titles: – TitleFull: International Journal of Biomaterials Type: main |
| ResultId | 1 |
