ISO/TR 10993-33:2015

Biological evaluation of medical devices — Part 33: Guidance on tests to evaluate genotoxicity — Supplement to ISO 10993-3

There are differences between the views of regulatory bodies on the subject of genotoxicity testing. The purpose of this ISO/TR 10993-33: 2015 is to provide background information to facilitate the selection of tests and guidance on the performance of tests.

Évaluation biologique des dispositifs médicaux — Partie 33: Directives sur les essais pour évaluer la génotoxicité — Supplément à l'ISO 10993-3

General Information

• Status: Published
• Publication Date: 05-Mar-2015
• ICS: 11.100.20 - Biological evaluation of medical devices
• Technical Committee: ISO/TC 194 - Biological and clinical evaluation of medical devices
• Drafting Committee: ISO/TC 194/WG 6 - Mutagenicity, carcinogenicity and reproductive toxicity
• Current Stage: 6060 - International Standard published
• Due Date: 08-Oct-2016
• Completion Date: 06-Mar-2015

Standards Content (Sample)

TECHNICAL ISO/TR
REPORT 10993-33
First edition
2015-03-01
Biological evaluation of medical
devices —
Part 33:
Guidance on tests to evaluate
genotoxicity — Supplement to ISO
10993-3
Évaluation biologique des dispositifs médicaux —
Partie 33: Directives sur les essais pour évaluer la génotoxicité —
Supplément à l’ISO 10993-3
Reference number
ISO/TR 10993-33:2015(E)
©
ISO 2015

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ISO/TR 10993-33:2015(E)

COPYRIGHT PROTECTED DOCUMENT
© ISO 2015
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ii © ISO 2015 – All rights reserved

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ISO/TR 10993-33:2015(E)

Contents Page
Foreword .vi
Introduction .viii
1 Scope . 1
2 Selection of tests . 1
3 Recommended tests . 1
4 Use of in vitro tests to detect genotoxicity. 2
5 Use of in vivo tests to detect genotoxicity . 2
6 Bacterial reverse mutation assay . 3
6.1 General . 3
6.2 Preparations . 3
6.2.1 Bacteria . 3
6.2.2 Medium . 4
6.2.3 Metabolic activation . 4
6.2.4 Test sample preparation. 4
6.3 Test conditions . 4
6.3.1 Solvents . 4
6.3.2 Exposure concentrations . 5
6.3.3 Controls . 6
6.4 Procedure . 7
6.4.1 Treatment with test sample . 7
6.4.2 Incubation . 7
6.4.3 Data collection . 7
6.5 Data and reporting . 8
6.5.1 Treatment of results. 8
6.5.2 Evaluation and interpretation of results . 8
6.5.3 Criteria for a valid test . 8
6.5.4 Test report . 9
7 In vitro mammalian chromosome aberration test .11
7.1 General .11
7.2 Preparations .11
7.2.1 Cells .11
7.2.2 Media and culture conditions .11
7.2.3 Preparation of cultures . .11
7.2.4 Metabolic activation .11
7.2.5 Test sample preparation.12
7.3 Test conditions .12
7.3.1 Solvents .12
7.3.2 Exposure concentrations .12
7.3.3 Controls .13
7.4 Procedure .14
7.4.1 Treatment with test sample or extract and harvest time .14
7.4.2 Chromosome preparation .14
7.4.3 Analysis .14
7.5 Data and reporting .15
7.5.1 Treatment of results.15
7.5.2 Evaluation and interpretation of results .15
7.5.3 Test report .15
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ISO/TR 10993-33:2015(E)

Contents Page
8 In vitro mammalian micronucleus test .17
8.1 General .17
8.2 Preparations .18
8.2.1 Cells .18
8.2.2 Media and culture conditions .18
8.2.3 Preparation of cultures . .18
8.2.4 Metabolic activation .18
8.2.5 Use of cytoB as a cytokinesis blocker .18
8.2.6 Test sample preparation.19
8.3 Test conditions .19
8.3.1 Solvents .19
8.3.2 Exposure concentrations .19
8.3.3 Controls .20
8.4 Procedure .21
8.4.1 Treatment with test sample or extract and harvest time .21
8.4.2 Cell harvest and slide preparation . .21
8.4.3 Analysis .22
8.5 Data and reporting .22
8.5.1 Treatment of results.22
8.5.2 Evaluation and interpretation of results .22
8.5.3 Test report .23
9 In vitro mammalian cell gene mutation test using mouse lymphoma (L5178Y) cells .25
9.1 General .25
9.2 Preparations .25
9.2.1 Cells .25
9.2.2 Media and culture conditions .25
9.2.3 Preparation of cultures . .25
9.2.4 Metabolic activation .25
9.2.5 Test sample preparations .26
9.3 Test conditions .26
9.3.1 Solvent/vehicle .26
9.3.2 Exposure concentrations .26
9.3.3 Controls .27
9.4 Procedure .28
9.4.1 General.28
9.4.2 Treatment with test sample .29
9.4.3 Measurement of survival, viability and mutant frequency.29
9.5 Data and reporting .29
9.5.1 Treatment of results.29
9.5.2 Evaluation and interpretation of results .30
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ISO/TR 10993-33:2015(E)

Contents Page
10 In vivo mammalian erythrocyte micronucleus test .32
10.1 General .32
10.2 Preparations .33
10.2.1 Selection of animal species .33
10.2.2 Housing and feeding conditions .33
10.2.3 Preparation of the animals .33
10.2.4 Test sample preparation.33
10.3 Test conditions .34
10.3.1 Solvent/vehicle .34
10.3.2 Controls .34
10.4 Procedure .34
10.4.1 Number and sex of animals .34
10.4.2 Treatment schedule . .34
10.4.3 Limit test .35
10.4.4 Dose levels .35
10.4.5 Routes of administration and doses levels .36
10.4.6 Bone marrow/blood preparation .36
10.4.7 Analysis .36
10.5 Data and reporting .37
10.5.1 Evaluation of results .37
10.5.2 Evaluation and interpretation of results .37
10.5.3 Test report .37
11 Chromosome aberration test (in vivo) .39
11.1 General .39
11.2 Preparations .39
11.2.1 Selection of animal species .39
11.2.2 Housing and feeding conditions .39
11.2.3 Preparation of the animals .39
11.2.4 Test sample preparation.39
11.3 Test conditions .40
11.3.1 Solvent/vehicle .40
11.3.2 Controls .40
11.4 Procedure .40
11.4.1 Number and sex of animals .40
11.4.2 Treatment schedule . .40
11.4.3 Dose levels .41
11.4.4 Limit test .41
11.4.5 Dose levels and routes of exposure.41
11.4.6 Bone marrow collection and preparation of slides .42
11.4.7 Analysis of Metaphase Cells .42
11.5 Data and reporting .42
11.5.1 Treatment of results.42
11.5.2 Evaluation and interpretation of results .42
11.5.3 Test report .43
Bibliography .45
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ISO/TR 10993-33:2015(E)

Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out
through ISO technical committees. Each member body interested in a subject for which a technical
committee has been established has the right to be represented on that committee. International
organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.
ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of
electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any
patent rights identified during the development of the document will be in the Introduction and/or on
the ISO list of patent declarations received (see www.iso.org/patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation on the meaning of ISO specific terms and expressions related to conformity
assessment, as well as information about ISO’s adherence to the WTO principles in the Technical Barriers
to Trade (TBT), see the following URL: Foreword — Supplementary information.
The committee responsible for this document is ISO/TC 194, Biological and clinical evaluation of
medical devices.
ISO 10993 consists of the following parts, under the general title Biological evaluation of medical devices:
— Part 1: Evaluation and testing within a risk management process
— Part 2: Animal welfare requirements
— Part 3: Tests for genotoxicity, carcinogenicity and reproductive toxicity
— Part 4: Selection of tests for interactions with blood
— Part 5: Tests for in vitro cytotoxicity
— Part 6: Tests for local effects after implantation
— Part 7: Ethylene oxide sterilization residuals
— Part 9: Framework for identification and quantification of potential degradation products
— Part 10: Tests for irritation and delayed-type hypersensitivity
— Part 11: Tests for systemic toxicity
— Part 12: Sample preparation and reference materials
— Part 13: Identification and quantification of degradation products from polymeric medical devices
— Part 14: Identification and quantification of degradation products from ceramics
— Part 15: Identification and quantification of degradation products from metals and alloys
— Part 16: Toxicokinetic study design for degradation products and leachables
— Part 17: Establishment of allowable limits for leachable substances
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ISO/TR 10993-33:2015(E)

— Part 18: Chemical characterization of materials
— Part 19: Physico-chemical, morphological and topographical characterization of materials (Technical
specification)
— Part 20: Principles and methods for immunotoxicology testing of medical devices (Technical specification)
— Part 33: Guidance on tests to evaluate genotoxicity - Supplement to ISO 10993-3 (Technical Report)
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ISO/TR 10993-33:2015(E)

Introduction
Genotoxicity tests are designed to detect compounds which induce genetic damage directly or
indirectly by various mechanisms. These tests should enable hazard identification with respect to
genetic damages. Expression of gene mutations, large scale chromosomal damage, recombination, and
numerical changes are generally considered to be essential for heritable effects and the mul
...