1、BSI Standards Publication Biological evaluation of medical devices Part 33: Guidance on tests to evaluate genotoxicity Supplement to ISO 10993-3 PD ISO/TR 10993-33:2015National foreword This Published Document is the UK implementation of ISO/TR 10993-33:2015. The UK participation in its preparation
2、was entrusted to Technical Committee CH/194, Biological evaluation of medical devices. A list of organizations represented on this committee can be obtained on request to its secretary. This publication does not purport to include all the necessary provisions of a contract. Users are responsible for
3、 its correct application. The British Standards Institution 2015. Published by BSI Standards Limited 2015 ISBN 978 0 580 84781 3 ICS 11.100.20 Compliance with a British Standard cannot confer immunity from legal obligations. This Published Document was published under the authority of the Stand- ard
4、s Policy and Strategy Committee on 31 March 2015. Amendments/corrigenda issued since publication Date Text affected PUBLISHED DOCUMENT PD ISO/TR 10993-33:2015 ISO 2015 Biological evaluation of medical devices Part 33: Guidance on tests to evaluate genotoxicity Supplement to ISO 10993-3 valuation bio
5、logique des dispositifs mdicaux Partie 33: Directives sur les essais pour valuer la gnotoxicit Supplment lISO 10993-3 TECHNICAL REPORT ISO/TR 10993-33 Reference number ISO/TR 10993-33:2015(E) First edition 2015-03-01 PD ISO/TR 10993-33:2015 ISO/TR 10993-33:2015(E)ii ISO 2015 All rights reserved COPY
6、RIGHT PROTECTED DOCUMENT ISO 2015 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior written permi
7、ssion. Permission can be requested from either ISO at the address below or ISOs member body in the country of the requester. ISO copyright office Case postale 56 CH-1211 Geneva 20 Tel. + 41 22 749 01 11 Fax + 41 22 749 09 47 E-mail copyrightiso.org Web www.iso.org Published in Switzerland PD ISO/TR
8、10993-33:2015 ISO/TR 10993-33:2015(E)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
9、.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 .
10、 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
11、 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 Ana
12、lysis .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 ISO 2015 All rights reserved iii Contents Page PD ISO/TR 10993-33:2015 ISO/TR 10993-33:2015(E)8 In vitro mammalian micronucleus test 17 8.1 General 17 8.2 Preparat
13、ions .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
14、 .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 c
15、ell 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 .2
16、6 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 iv
17、ISO 2015 All rights reserved Contents Page PD ISO/TR 10993-33:2015 ISO/TR 10993-33:2015(E)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
18、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 Bon
19、e 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
20、 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
21、.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 Bibliogra
22、phy .45 ISO 2015 All rights reserved v Contents Page PD ISO/TR 10993-33:2015 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
23、 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 w
24、ork. 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 di
25、fferent 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
26、 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/pa
27、tents). 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 ISOs adherence to the WTO prin
28、ciples 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
29、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
30、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: Sa
31、mple 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 a
32、nd alloys Part 16: Toxicokinetic study design for degradation products and leachables Part 17: Establishment of allowable limits for leachable substancesvi ISO 2015 All rights reserved PD ISO/TR 10993-33:2015 ISO/TR 10993-33:2015(E) Part 18: Chemical characterization of materials Part 19: Physico-ch
33、emical, 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)
34、 ISO 2015 All rights reserved vii PD ISO/TR 10993-33:2015 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
35、. Expression of gene mutations, large scale chromosomal damage, recombination, and numerical changes are generally considered to be essential for heritable effects and the multi-step carcinogenesis. A positive genotoxicity test provides an indication that further testing can be warranted to determin
36、e the carcinogenic potential of the compound. Because the relationship between exposure to particular chemicals and carcinogenesis is established for man, while a similar relationship has been difficult to prove for heritable diseases, genotoxicity tests have been used mainly for the prediction of c
37、arcinogenicity. Nevertheless, because germ line mutations are clearly associated with human disease, the suspicion that a compound can induce heritable effects is considered to be just as serious as the suspicion that a compound can induce cancer. In addition, the outcome of such tests can be valuab
38、le for the interpretation of carcinogenicity studies.viii ISO 2015 All rights reserved PD ISO/TR 10993-33:2015 Biological evaluation of medical devices Part 33: Guidance on tests to evaluate genotoxicity Supplement to ISO 10993-3 1 Scope There are differences between the views of regulatory bodies o
39、n the subject of genotoxicity testing. The purpose of this Technical Report is to provide background information to facilitate the selection of tests and guidance on the performance of tests. 2 Selection of tests Since chemicals can induce genetic damage by different mechanisms, a battery of tests s
40、ensitive to different types of genetic damage are thought to provide the best assurance for detecting genotoxic hazard. The tests selected usually include tests to detect point mutations and tests to detect chromosomal aberrations. Both bacterial cells and cultured mammalian cells are used to detect
41、 genotoxic agents. in vivo tests are sometimes incorporated into these test batteries. These tests are sometimes included in the initial test battery or are used to clarify results from in vitro tests, see Reference 13. 3 Recommended tests Although there are some variations in details, the same geno
42、toxicity tests are commonly recommended by most regulatory agencies. The following are commonly recommended tests: bacterial reverse mutation test (see OECD 471 1and Clause 6); in vitro mammalian chromosome aberration test (see OECD 473 2and Clause 7); in vitro mammalian micronucleus test (see OECD
43、487 6and Clause 8); in vitro mammalian cell gene mutation test using mouse lymphoma (L5178Y) cells (see OECD 475 4and Clause 9); in vivo mammalian erythrocyte micronucleus test (see OECD 474 3and Clause 10); in vivo chromosome aberration test (see OECD 475 5and Clause 11). For medical devices, a bat
44、tery of tests is commonly used for genotoxicity evaluations. The general strategy identified in ISO 10993-3 is as follows: a) test for gene mutations in bacteria. Bacterial Reverse Mutation Assay, OECD 471 1technically modified for medical devices to allow, for example, testing with extracts from de
45、vices (see Clause 6); and either b) an in vitro test with cytogenetic evaluation of chromosomal damage with mammalian cells, Chromosome aberration test, OECD 473 2technically modified for medical devices (see Clause 7), or c) an in vitro mouse lymphoma tk assay, OECD 476 5technically modified for me
46、dical devices (see Clause 8) including detection of small (slow growing) and large colonies, or TECHNICAL REPORT ISO/TR 10993-33:2015(E) ISO 2015 All rights reserved 1 PD ISO/TR 10993-33:2015 ISO/TR 10993-33:2015(E) d) an in vitro mammalian cell micronucleus test for chromosomal damage and aneugenic
47、ity, OECD 487 technically modified for medical devices, (see Clause 8). The International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) recommends a three-test battery described in the ICH S2(R1) Genotoxicity, which can be required for
48、medical devices by some regulatory authorities. 4 Use of in vitro tests to detect genotoxicity In vitro tests are commonly used for identifying the potential of chemicals to induce genotoxicity. Multiple tests are used because no single test detects all known genotoxins. Genotoxins often lead to dif
49、ferent effects (e. g. large scale or chromosomal damage vs. small scale damage or point mutations or different DNA sequence specificity). Also, the resulting genetic damage has differing susceptibility to DNA repair. The “ICH test battery” was developed to cast a wider net for detecting genotoxins. Although in vitro genotoxicity tests can be considered overly sensitive, these tests detect most rodent genotoxic carcinogens. Comparisons of the “scorecards
