prEN ISO 10993-3
(Main)Biological evaluation of medical devices - Part 3: Evaluation of genotoxicity, carcinogenicity, reproductive toxicity, and developmental toxicity (ISO/DIS 10993-3:2025)
Biological evaluation of medical devices - Part 3: Evaluation of genotoxicity, carcinogenicity, reproductive toxicity, and developmental toxicity (ISO/DIS 10993-3:2025)
ISO 10993-3:2014 specifies strategies for risk estimation and selection of hazard identification tests, with respect to the possibility of the following potentially irreversible biological effects arising as a result of exposure to medical devices:
genotoxicity;
carcinogenicity;
reproductive and developmental toxicity.
ISO 10993-3:2014 is applicable when the need to evaluate a medical device for potential genotoxicity, carcinogenicity, or reproductive toxicity has been established.
Biologische Beurteilung von Medizinprodukten - Teil 3: Bewertung der Genotoxizität, Karzinogenität, Reproduktionstoxizität und Entwicklungstoxizität (ISO/DIS 10993-3:2025)
Évaluation biologique des dispositifs médicaux - Partie 3: Évaluation de la génotoxicité, de la cancérogénicité, de la toxicité sur la reproduction et le développement (ISO/DIS 10993-3:2025)
L'ISO 10993-3:2014 spécifie les stratégies pour l'estimation des risques, le choix des essais d'identification des risques et la gestion des risques, en fonction du risque d'apparition des effets biologiques potentiellement irréversibles suivants résultant de l'exposition à des dispositifs médicaux:
génotoxicité;
cancérogénicité;
toxicité sur la reproduction et le développement.
L'ISO 10993-3:2014 est applicable lorsque le besoin d'évaluer un dispositif médical dont le risque de génotoxicité, de cancérogénicité ou de toxicité sur la reproduction a été identifié.
Biološko ovrednotenje medicinskih pripomočkov - 3. del: Ovrednotenje genske toksičnosti, kancerogenosti, toksičnosti za razmnoževanje in toksičnosti za razvoj (ISO/DIS 10993-3:2025)
General Information
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Standards Content (Sample)
SLOVENSKI STANDARD
01-junij-2025
Biološko ovrednotenje medicinskih pripomočkov - 3. del: Ovrednotenje genske
toksičnosti, kancerogenosti, toksičnosti za razmnoževanje in toksičnosti za razvoj
(ISO/DIS 10993-3:2025)
Biological evaluation of medical devices - Part 3: Evaluation of genotoxicity,
carcinogenicity, reproductive toxicity, and developmental toxicity (ISO/DIS 10993-3:2025)
Biologische Beurteilung von Medizinprodukten - Teil 3: Bewertung der Genotoxizität,
Karzinogenität, Reproduktionstoxizität und Entwicklungstoxizität (ISO/DIS 10993-3:2025)
Évaluation biologique des dispositifs médicaux - Partie 3: Évaluation de la génotoxicité,
de la cancérogénicité, de la toxicité sur la reproduction et le développement (ISO/DIS
10993-3:2025)
Ta slovenski standard je istoveten z: prEN ISO 10993-3
ICS:
11.100.20 Biološko ovrednotenje Biological evaluation of
medicinskih pripomočkov medical devices
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
DRAFT
International
Standard
ISO/DIS 10993-3
ISO/TC 194
Biological evaluation of medical
Secretariat: DIN
devices —
Voting begins on:
Part 3: 2025-04-04
Evaluation of genotoxicity,
Voting terminates on:
2025-06-27
carcinogenicity, reproductive
toxicity, and developmental toxicity
ICS: 11.100.20
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ISO/DIS 10993-3:2025(en)
DRAFT
ISO/DIS 10993-3:2025(en)
International
Standard
ISO/DIS 10993-3
ISO/TC 194
Biological evaluation of medical
Secretariat: DIN
devices —
Voting begins on:
Part 3:
Evaluation of genotoxicity,
Voting terminates on:
carcinogenicity, reproductive
toxicity, and developmental toxicity
ICS: 11.100.20
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Published in Switzerland Reference number
ISO/DIS 10993-3:2025(en)
ii
ISO/DIS 10993-3:2025(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Assessment strategies . . 3
4.1 General .3
4.2 Other considerations .3
5 Genotoxicity assessment . 3
5.1 General .3
5.2 Testing strategy .4
5.2.1 General .4
5.2.2 Test battery .4
5.2.3 In vivo genotoxicity testing .5
5.2.4 Follow-up evaluation .5
5.3 Test sample preparation .5
6 Carcinogenicity Assessments . 6
6.1 General .6
6.2 Evaluation strategy .6
6.2.1 General .6
6.2.2 Genotoxic carcinogens .6
6.2.3 Non-genotoxic carcinogens .7
7 Evaluation of reproductive and developmental toxicity . 7
7.1 General .7
7.2 Evaluation methods .8
8 Test report . 9
Annex A (informative) Test sample preparation procedure for genotoxicity testing .10
Annex B (normative) Tests for genotoxicity .16
Annex C (informative) Carcinogenicity Tests in animals .26
Annex D (informative) Genotoxicity assessment of nanomaterials .29
Annex E (informative) Recipe for S9 mix .30
Annex F (informative) Endocrine Disruptors .31
Annex G (informative) Evaluation of reproductive and developmental toxicity .32
Annex ZA (informative) Relationship between this European standard and the General Safety
and Performance Requirements of Regulation (EU) 2017/745 aimed to be covered .36
Bibliography .40
iii
ISO/DIS 10993-3:2025(en)
Foreword
ISO (the International Organization for Standardization) and IEC (the International Electrotechnical
Commission) form the specialized system for worldwide standardization. National bodies that are
members of ISO or IEC participate in the development of International Standards through technical
committees established by the respective organization to deal with particular fields of technical activity.
ISO and IEC technical committees collaborate in fields of mutual interest. Other international organizations,
governmental and non-governmental, in liaison with ISO and IEC, also take part in the work. In the field of
information technology, ISO and IEC have established a joint technical committee, ISO/IEC JTC 1.
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 voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO's adherence to the World
Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following URL:
www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 194, Biological and clinical evaluation of medical
devices, in collaboration with the European Committee for Standardization (CEN) Technical Committee
CEN/TC 206, Biocompatibility of medical and dental materials and devices, in accordance with the Agreement
on technical cooperation between ISO and CEN (Vienna Agreement).
This fourth edition of ISO 10993-3 cancels and replaces the third edition (ISO 10993-3:2014), which has
been technically revised.
The main changes compared to the previous edition are as follows:
— Deletion of the Annex on Cellular Transformation;
— Restructure of Annex A on Guidance on selecting an appropriate test sample preparation procedure for
genotoxicity testing;
— Addition of Annex C on in vivo carcinogenicity assays that includes the Annex on solid state carcinogenesis;
— Addition of Annex D on Genotoxicity Assessment of Nanomaterials;
— Expanded Annex B on genotoxicity test methods;
— Addition of Annex E on Recipe for S9 mix;
— Addition of Annex G on the evaluation of reproductive and developmental toxicity;
— Update of normative references and bibliography.
A list of all parts in the ISO 10993 series can be found on the ISO website.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html
iv
ISO/DIS 10993-3:2025(en)
Introduction
The basis for biological evaluation of medical devices is often empirical and driven by relevant concerns for
human safety. The risk of serious and irreversible effects, such as cancer or second generation abnormalities,
is of particular public concern. It is inherent in the provision of safe medical devices that such risks be
minimized to the greatest extent feasible. The assessment of mutagenic, carcinogenic, reproductive, and
developmental toxicity hazards is an essential component of the control of these risks. Not all test methods
for the assessment of genotoxicity, carcinogenicity, reproductive toxicity, and developmental toxicity are
equally well developed, nor is their validity well established for the testing of medical devices.
Significant issues with test sample size and preparation, scientific understanding of disease processes
and test validation can be cited as limitations of available methods. Since the previous document revision,
many genotoxicity test methods have been updated with revised OECD guidelines. However, these generally
provide clearer recommendations but little alteration in test methods. Scientifically sound alternatives to
the proposed testing can be acceptable insofar as they address relevant matters of safety assessment.
In the selection of tests needed to evaluate a particular medical device, a careful assessment of expected
human uses and potential interactions with biological systems is important, particularly in such areas as
reproductive and developmental toxicology.
This document presents test methods and evaluation strategies for the identification of specific biological
harms. Testing is not always necessary or helpful in managing toxicological risks associated with exposure
to medical device materials but, where it is appropriate, it is important that maximum test sensitivity is
achieved.
In view of the multitude of possible outcomes and the importance of factors such as extent of exposure,
species differences and mechanical or physical considerations, risk assessment is typically performed on a
case-by-case basis. Suggestions for risk consideration and integration of ISO 10993-17 and -18 are provided.
v
DRAFT International Standard ISO/DIS 10993-3:2025(en)
Biological evaluation of medical devices —
Part 3:
Evaluation of genotoxicity, carcinogenicity, reproductive
toxicity, and developmental toxicity
1 Scope
This document specifies strategies for risk estimation and selection of biological harms identification tests
with respect to the possibility of the following potentially irreversible biological effects arising as a result of
exposure to medical devices:
— genotoxicity;
— carcinogenicity;
— reproductive and developmental toxicity.
This document is applicable when the need to evaluate a medical device for potential genotoxicity,
carcinogenicity, reproductive toxicity, and developmental toxicity has been established.
NOTE Guidance on selection of relevant endpoints is provided in ISO 10993-1.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content constitutes
requirements of this document. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
ISO 10993-1:2018, Biological evaluation of medical devices — Part 1: Requirements and general principles for
the evaluation of biological safety within a risk management process
ISO 10993-2:2022, Biological evaluation of medical devices — Part 2: Animal welfare requirements
ISO 10993-12:2021, Biological evaluation of medical devices — Part 12: Sample preparation and reference
materials
ISO 10993-16:2017, Biological evaluation of medical devices — Part 16: Toxicokinetic study design for
degradation products and leachables
ISO 10993-17, Biological evaluation of medical devices — Part 17: Toxicological risk assessment of medical
device constituents
ISO 10993-18, Biological evaluation of medical devices — Part 18: Chemical characterization of medical device
materials within a risk management process
ISO 14971:2019, Medical devices — Application of risk management to medical devices
OECD 414, OECD Guideline for the Testing of Chemicals — Prenatal Development Toxicity Study
OECD 421, OECD Guideline for the Testing of Chemicals — Reproduction/Developmental Toxicity Screening Test
OECD 422, OECD Guideline for the Testing of Chemicals — Combined repeated dose toxicity study with the
reproduction/ developmental toxicity screening test
ISO/DIS 10993-3:2025(en)
OECD 443, OECD Guideline for the Testing of Chemicals — Extended One-Generation Reproductive Toxicity Study
OECD 471, OECD Guideline for the Testing of Chemicals — Bacterial Reverse Mutation Test
OECD 473, OECD Guideline for the Testing of Chemicals — In vitro Mammalian Chromosome Aberration Test
OECD 474, OECD Guideline for the Testing of Chemicals — Mammalian Erythrocyte Micronucleus Test
OECD 475, OECD Guideline for the Testing of Chemicals — Mammalian Bone Marrow Chromosomal Aberration Test
OECD 487, OECD Guideline for the Testing of Chemicals — In vitro Mammalian Cell Micronucleus Test
OECD 488, OECD Guideline for the Testing of Chemicals — Transgenic Rodent Somatic and Germ Cell Gene
Mutation Assays
OECD 489, OECD Guideline for the Testing of Chemicals — In Vivo Mammalian Alkaline Comet Assay
OECD 490, OECD Guideline for the Testing of Chemicals — In vitro Mammalian Cell Gene Mutation Tests Using
the Thymidine Kinase Gene
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 10993-1, ISO 10993-12 and the
following apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
1) ISO Online browsing platform: available at https:// www .iso .org/ obp
2) IEC Electropedia: available at https:// www .electropedia .org/
3.1
carcinogenicity test
test to determine cancer outcomes after long term or life-span exposure of animals
3.2
developmental toxicity test
test to determine adverse effects on the developing organism during prenatal development, or postnatally
to the time of sexual maturation
3.3
genotoxicity test
test using cells, bacteria, yeast, fungi, animals, or other biological systems to determine whether gene
mutations, chromosomal alterations, or other genetic changes are caused by the test sample
3.4
reproductive toxicity test
test to evaluate the potential effects of test samples on male or female reproductive function during any
stage of development or fertility
3.5
sham-incubated vehicle
vehicle control exposed to the same extraction time and temperature as the test sample extract
3.6
threshold of toxicological concern TTC
level of exposure for constituents, below which there would be no appreciable risk to human health
Note 1 to entry: The constituent is defined as a chemical or compound present in or on a finished medical device
or material(s) of construction. Constituents may be intentionally present (e.g., an additive such as an antioxidant) or
unintentionally present (e.g., an impurity).
[SOURCE: ISO/TS 21726:2019 3.5, modified — Note 1 to entry has been added.]
ISO/DIS 10993-3:2025(en)
4 Assessment strategies
4.1 General
ISO 10993-1 indicates circumstances where the potential for genotoxicity, carcinogenicity, reproductive
toxicity, and developmental toxicity are relevant biological harms for consideration in an overall biological
safety evaluation. Evaluation of the biological harms shall be conducted with consideration of the following
factors:
— whether assessment is necessary based on patient population (e.g. not in the case of <6 months life
expectancy); and the anticipated impact of test results on risk management judgements,
— whether assessment is necessary based on the worst-case quantity, type of contact and duration of
exposure,
NOTE Quantity can be influenced by device size and number used per patient.
— an analysis of the chemical constituents of the device material(s), including manufacturing process
residues and degradation products or metabolites, to identify causes of concern. The identification can
be based on chemical-specific toxicity data if available; otherwise, structure-activity relationships or
previous demonstration of relevant toxicity in the chemical class can be used,
— existing information relevant to the genotoxicity, carcinogenicity, reproductive toxicity, and
developmental toxicity evaluation of the medical device, and
— previous use of equivalent (see ISO 10993-1:2018, 4.1) materials and processing in relevant applications.
The decision to waive in vitro genotoxicity testing shall be justified. The decision to conduct in vivo testing
shall be justified.
4.2 Other considerations
Toxicity evaluation can be warranted for additional states of the device, such as wear debris generated from
the device, absorbable materials, or materials that cure in situ (e.g. cements, adhesives, and pre-polymer
mixtures) unless toxicological risk assessment based on material information determines no cause for
concern. For guidance on sample preparation for in situ curing devices see ISO 10993-12.
NOTE Material information can include chemical composition, degradation products, debris size and morphology,
pre-polymerization, and post-polymerization products etc.
If exposure to constituents identified in accordance with ISO 10993-18 is determined to be within acceptable
levels based on toxicological risk assessment conducted in accordance with ISO 10993-17, no additional
testing needs to be performed to address the relevant risks.
5 Genotoxicity assessment
5.1 General
Assessment of genotoxic risk should be made:
a) by toxicological risk assessment based on chemical characterization in accordance with ISO 10993-17
and ISO 10993-18 or;
b) by performing genotoxicity testing in the standard test battery in accordance with 5.2.2.
In certain cases (e.g. a material which has not been used in a legally marketed device), both toxicological risk
assessment and genotoxicity testing can be appropriate.
If the device is made with and releases nanomaterials, additional evaluation shall be considered.
NOTE See ISO/TR 10993-22 and Annex D for more information.
ISO/DIS 10993-3:2025(en)
If the device contains identified genotoxicants which are not easily detected using non-targeted analytical
approaches described in ISO 10993-18 (e.g. glutaraldehyde, formaldehyde), targeted analysis of genotoxic
residuals shall be conducted. Where compositional profiling or analytical chemistry reveals the presence of
chemical constituents with inadequate genotoxicity data and where QSAR (quantitative structure activity
relationship) information is inconclusive (see ISO 10993-17), testing of individual chemicals or device
components containing the chemical should be considered. In either case the test sample shall be justified.
5.2 Testing strategy
5.2.1 General
No single test is capable of detecting all genotoxic agents. Therefore, the usual approach is to conduct a
battery of in vitro and under certain circumstances also in vivo tests.
Genotoxicity tests are designed to detect the two major classes of genetic damage:
— gene mutations (DNA single base pair changes, small deletions or additions), and
— Chromosomal damage (structural aberrations such as translocations, small or large deletions and
insertions, and gain or loss of whole chromosomes(aneuploidy) or parts of chromosomes).
NOTE Bacterial reverse mutation assays have been shown to detect relevant small-scale mutational changes
produced by the majority of genotoxic carcinogens detected by rodent bioassays.
5.2.2 Test battery
When genotoxicity testing is performed, it shall be conducted in accordance with Annex B and the test
battery shall include:
1) a test for gene mutations in bacteria (see OECD 471), modified for medical devices, see clause B.2, and
2) one of the following three validated in vitro mammalian cell tests:
a) an in vitro test with cytogenetic evaluation of chromosomal damage with mammalian cells (see
OECD 473), modified for medical devices, see clause B.3 and B.4.1.2, or
+/-
b) an in vitro mouse lymphoma tk assay using L5178Y cells (see OECD 490), modified for medical
devices, including detection of small (slow growing) and large colonies, see clause B.3 and B.4.1.1, or
c) an in vitro mammalian cell micronucleus test for chromosomal damage and aneugenicity (see OECD
487), modified for medical devices, see clause B.3 and B.4.1.3.
NOTE 1 The OECD 471 can be inappropriate for nanomaterials, see Annex D.
Tests shall be performed in the presence and absence of metabolic activation. Metabolic activation is
achieved by addition of a co-factor-supplemented post-mitochondrial fraction (S9).
NOTE 2 An example for a S9 recipe is given in Annex E.
In vitro mammalian tests shall include both a short-term exposure with and without S9 and a long-term
exposure without S9.
+/-
Results from both chromosomal aberration assays and the in vitro mouse lymphoma tk assay (MLA) have
a relatively high level of congruence for compounds that are regarded as genotoxic but yield negative results
in the bacterial reverse mutation assay. The MLA detects the broadest set of genetic damage, including both
small-scale and large-scale genetic damage. The micronucleus assay detects chromosomal damage in the
form of whole chromosome loss or chromosomal fragments and uniquely detects aneuploidy. The three tests
are equally acceptable as an in vitro mammalian genotoxicity test (see OECD 473, OECD 487 and OECD 490).
ISO/DIS 10993-3:2025(en)
5.2.3 In vivo genotoxicity testing
1) In vivo genotoxicity tests are not part of the recommended genotoxicity test battery because they are
relatively insensitive and not likely to detect the concentrations of substances generally found in medical
[52][53]
devices or in medical device extracts. However, under certain circumstances, an in vivo test can
be informative. These circumstances can include assessment of nanomaterial or other particulates
suspected of sequestration in particular organs or tissues (see Annex D, [2] and ISO/TR 10993-22); or
2) when additional relevant factors e.g. toxico-kinetics (see ISO 10993-16) or organ-specific toxicity need
to be considered.
In vivo tests are valid only with data demonstrating the test sample reached the target cells, organs, or
tissues. Mutational analysis in target organs of rats or mice can be determined using one of several rodent
transgenic mutation assays (see OECD 488); chromosomal damage can be assessed in the erythrocyte
micronucleus assay (see OECD 474), the in vivo comet assay (see OECD 489), or the in vivo chromosomal
aberration assay (see OECD 475). See B.5 for additional considerations for in vivo assays.
All animal studies shall be performed in a facility approved by a nationally recognized organization and
in accordance with all appropriate regulations dealing with laboratory animal welfare to comply with the
requirements of ISO 10993-2.
5.2.4 Follow-up evaluation
If genotoxicity tests are performed in accordance with 5.2.2 and if the in vitro tests are negative, the test
article can be considered non-genotoxic and further testing is not necessary.
When any single in vitro test is positive, then the following options shall be considered:
a) Identification of genotoxic constituents such as impurities or residues by chemical characterization
(information gathering or analytical chemistry testing) or other testing, with appropriate steps taken
to eliminate or manage any identified risks.
b) If confounding factors are suspected to have caused the positive response in an initial genotoxicity test,
the dosing and/or test conditions may be altered.
c) Presume that a genotoxicity risk exists and manage the risk in accordance with ISO 14971. Equivocal
results (i.e. elevated results that don’t meet criteria for a positive result), can require repeating tests
with a modified protocol such as adjusting dilution of the extract or the test chemical in the test system.
5.3 Test sample preparation
Test samples should be extracted in vehicles compatible with the test system (see Annex A). Use of extraction
vehicles that are incompatible with the test system require additional manipulation (e.g. solvent removal)
that can alter the composition of the extract and, therefore, affect the results of the test. Use of extraction
vehicles that are incompatible with the test system shall be justified.
Extraction shall be performed in accordance with ISO 10993-12 unless otherwise justified.
Tests can be performed on solutions (e.g. soluble or liquid devices), suspensions (e.g. Method A in Annex A),
extracts (e.g. Method B in Annex A) of the finished product, device components or individual chemicals
from the medical device. If individual chemicals are tested, then the appropriate OECD guidelines shall be
followed.
NOTE Some authorities having jurisdictions prefer Method C in Annex A to be used.
ISO/DIS 10993-3:2025(en)
6 Carcinogenicity Assessments
6.1 General
The objective of carcinogenicity studies is to determine tumorigenic potential in animals and to evaluate the
relevant risk to patients. Mechanisms of carcinogenesis can be genotoxic or non-genotoxic. Medical devices
shall be evaluated for both types of carcinogenicity, preferably by alternative approaches, such as chemical
characterization and toxicological risk assessment, that do not use in vivo testing.
Animal testing for carcinogenicity is technically challenging. It requires implantation of many medical
devices or representative samples of the medical device or the application of concentrated medical device
extracts to achieve the elevated doses necessary for a valid test in a reasonable number of animals. For these
reasons animal testing is generally not justified when the risks can be adequately assessed by other means.
NOTE For additional information on animal carcinogenicity testing, see Annex C.
Where genotoxicity of a medical device is established, additional evaluation of carcinogenic risk shall be
performed (e.g. ISO 10993-17). In this situation, additional factors such as genotoxicity results, history of
clinical use, and the indication for use should be considered in determining the risk. It can be informative to
identify the genotoxic constituent(s) and focus the evaluation on that chemical.
6.2 Evaluation strategy
6.2.1 General
Carcinogenicity should be evaluated initially using chemical characterization in accordance with
ISO 10993-18 and toxicological risk assessment in accordance with ISO 10993-17. In cases where devices
or materials are not readily amenable to analytical chemistry testing and adequate material information
is available along with genotoxicity testing data on the final finished device, it is always preferable to use
chemical information and toxicological risk assessment to determine potential carcinogenicity rather than
performing an animal carcinogenicity study.
NOTE 1 Some authorities having jurisdictions can require consideration of carcinogenicity testing for novel
materials (e.g. when data are not available to provide an adequate assessment).
If the results of toxicological risk assessment indicate no unacceptable risk of carcinogenicity from the
device, then no further assessment is required.
Carcinogenicity testing shall not be performed when risks can be adequately assessed or managed
without generating new carcinogenicity test data or when the benefit/risk analysis obviates the need for
a carcinogenicity assessment. If the toxicological risk assessment demonstrates an unacceptable risk of
carcinogenicity from the device, the risk shall be managed accordingly (see also ISO 14971). Alternatively,
if the device contains a chemical with inadequate toxicological data, biological testing shall be justified on
the basis of a need for additional information. The assessment should address both genotoxic (see 6.2.2)
and non-genotoxic carcinogenicity (see 6.2.3) on the individual chemical identified in the toxicological risk
assessment.
NOTE 2 The history of human clinical use can be considered in the risk assessment. The clinical evidence for
carcinogenicity risk is generally not monitored over an adequate timeframe and this aspect of device safety can be
unknown.
6.2.2 Genotoxic carcinogens
The risk from genotoxic carcinogens can be established by genotoxicity testing or by toxicological risk
assessment. Testing will require a battery of tests (see 5.2.2).
Where genotoxicity of a medical device is established, additional evaluation of carcinogenic risk shall be
performed in accordance with ISO 10993-17. In these situations, additional factors such as genotoxicity
results, history of clinical use, and the indication for use should be considered in determining the risk
ISO/DIS 10993-3:2025(en)
assessment. It can be informative to identify the genotoxic constituent and focus the evaluation on that
chemical.
NOTE While genotoxic carcinogenicity involves non-threshold mechanisms, there are exposure levels below
which impact on lifetime cancer risk is considered acceptable (usually less than 1 in 100 000 additional lifetime
incidence).
6.2.3 Non-genotoxic carcinogens
Non-genotoxic carcinogens do not cause direct DNA damage but induce cancer via other mechanisms (see
[29], [30] and [31]). These carcinogens predominantly induce their effects (leading to cancer) upon repeated
exposure leading to prolonged interference with homeostatic processes that lead to irreversible and
uncontrolled cellular proliferation. The prediction of carcinogenic potential of non-genotoxic chemicals is
complicated by:
— the diversity of modes of action;
— the tissue and species specificity; and
— the absence of currently validated tests methods.
While a reliable evaluation is to conduct a rodent carcinogenicity test (see [30], [32], [33] and [34]), there
are many practical challenges (see 6.1). Therefore, toxicological risk assessment in accordance with
ISO 10993-17 should be used.
If the substance is not genotoxic and carcinogenicity data is not available, then a tolerable intake based on
subchronic, or chronic systemic toxicity data can be useful for evaluating the risk. If there is insufficient data
to set a TI, then the TTC described in ISO/TS 21726 may be used to estimate the risk. Recent publications
indicate that the TTCs based on Cramer Class are protective for non-genotoxic carcinogens (see [37]).
[36]
NOTE 1 Non-genotoxic carcinogens have thresholds, below which the risk of cancer is unlikely (ECHA 2017 ).
NOTE 2 Approaches for prediction of non-genotoxic carcinogenicity are under active investigation (e.g., [33] and [35]).
NOTE 3 Examples for non-genotoxic carcinogens can be found in (see [37], supplementary material).
7 Evaluation of reproductive and developmental toxicity
7.1 General
Reproductive toxicity evaluation shall be considered for devices intended for use in individuals of
reproductive age under any of the following conditions:
— devices have prolonged or long-term direct or indirect contact with reproductive organs or gametes;
— devices have prolonged or long-term in vitro contact with gametes/ reproductive tissue;
— devices have prolonged or long-term use and contain known or presumed reproductive toxicants
(materials with a known adverse effect on reproduction);
— devices are absorbable or cure in situ, and are in close proximity to the reproductive organs, regardless
of duration of contact; or
— materials which have not been used in a legally marketed device, for a similar intended purpose.
NOTE 1 Material means material of construction, intended additives, or process residuals, or any combination
of the three.
ISO/DIS 10993-3:2025(en)
NOTE 2 Similar use means the nature of contact category and contact-duration category are the same; and the
device risk level is considered the same or lower compared to the legally marketed device. In addition to contact
category and contact-duration, additional factors to consider in the determination of the device risk level include: the
material type and quantity, the material constituent composition, device contact frequency, nature/degree of tissue
contact with the device, and user population.
Developmental toxicity evaluation shall be considered for:
— devices with prolonged or long-term direct or indirect contact with embryos or fetuses; and
— devices for prolonged or long-term use on individuals in early childhood and contain known or presumed
developmental toxicants [e.g. endocrine disruptors (see Annex F) and lead]. Potential exposure from a
medical device contacting lactating women should also be considered.
— devices are absorbable or cure in situ, and are in close proximity to the reproductive organs, regardless
of duration of contact; or
— materials which have not been used in a legally marketed device, for an equivalent intended purpose.
7.2 Evaluation methods
Assessment of reproductive and developmental risk should be made:
a) by toxicological risk assessment based on chemical characterization in accordance with ISO 10993-17
and ISO 10993-18 or;
b) by performing developmental and reproductive testing as described in Annex G.
Current biological methods are for single substances and are not validated for mixtures (e.g. medical
device extracts). Evaluation for medical devices shall first consider a toxicological risk assessment focusing
on reproductive and developmental toxicity in accordance with ISO 10993-17. Exposure estimates can be
determined through chemical characterization outlined in ISO 10993-18.
Exposure estimates can be improved with simulated-use extraction studies. If it is determined that the
amount of a reproductive and developmental toxicant in the device is above acceptable limits, the presence
of the toxicant shall be carefully considered for risk control where possible and in the benefit-risk analysis in
the (overall) risk management file in accordance with ISO 14971:2019, 7.4.
The following factors should be considered in the toxicological risk assessment:
— an assessment of reproductive potential of the subject population (devices used in early adulthood, or
reproductive age-individuals);
— a specific assessment of reproductive toxicity data, if available;
— the potential for exposure of reproductive tissues, the developing offspring, or nursing child, including
available chemical characterization information in accordance with ISO 10993-18 and available
toxicokinetic studies in accordance with ISO 10993-16;
— device physical characteristics;
— available device genotoxicity information;
— history of clinical use;
— indication for use; and
— the need for additional information relevant to the reproductive and developmental toxicity risk arising
from use of the device (see ISO 10993-1).
NOTE For more information see Annex G.
When the remaining risk is unacceptable due to a lack of robust experimental data, individual chemical
[38]
substances can be tested with in vitro or in vivo biological test methods (e.g. ICH S5 ) (see clause G.2).
ISO/DIS 10993-3:2025(en)
More information regarding test selection is given in ICH S5 and OECD 43. For example, testing an individual
chemical in a developmental toxicity study would yield an effect level or no effect level that could help refine
the risk assessment.
8 Test report
If relevant, the test report shall include at least the following details:
a) description of test sample (e.g. identifier, material composition, processing, size and weight) including
its intended use;
b) description and rationale/justification of test methods, test conditions, test materials, test dose and test
procedures;
c) description of analytical methods including quantification limits;
d) statement of compliance to appropriate current/valid best laboratory/quality practices, for example
Good Laboratory Practices (GLP) or ISO/IEC 17025, where applicable;
e) description of S9 mix (including manufacturers as applicable);
f) test results including summary;
g) statistical methods, if applicable;
h) interpretation and discussion of results;
i) further details as specified in the relevant OECD guidelines (OECD 414, OECD 421, OECD 422 and OECD
443) or Annex G;
j) name and certifications of the testing la
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