FprEN ISO 10993-10

Biological evaluation of medical devices - Part 10: Tests for skin sensitization (ISO 10993-10:2021)

This document specifies the procedure for the assessment of medical devices and their constituent materials with regard to their potential to induce skin sensitization.
This document includes:
—    details of in vivo skin sensitization test procedures;
—    key factors for the interpretation of the results.
NOTE      Instructions for the preparation of materials specifically in relation to the above tests are given in Annex A.

Biologische Beurteilung von Medizinprodukten - Teil 10: Prüfungen auf Hautsensibilisierung (ISO 10993-10:2021)

Dieses Dokument legt das Verfahren für die Beurteilung von Medizinprodukten und ihren Bestandteilen hinsichtlich ihres Potentials, eine Hautsensibilisierung auszulösen, fest.
Dieses Dokument enthält:
   Einzelheiten zur Durchführung von In-vivo-Hautsensibilisierungsprüfungen;
   Schlüsselfaktoren für die Interpretation der Ergebnisse.
ANMERKUNG   Anhang A enthält Anweisungen für die spezielle Vorbereitung von Materialien im Zusammenhang mit den vorstehend angeführten Prüfungen.

Évaluation biologique des dispositifs médicaux - Partie 10: Essais de sensibilisation cutanée (ISO 10993-10:2021)

Le présent document spécifie le mode opératoire pour l'évaluation du potentiel des dispositifs médicaux et de leurs matériaux constitutifs à provoquer une sensibilisation cutanée.
Le présent document comprend:
—    des informations détaillées relatives aux modes opératoires d'essai de sensibilisation cutanée in vivo;
—    les facteurs clés pour l'interprétation des résultats.
NOTE      Des instructions pour la préparation des matériaux spécifiquement pour les essais ci-dessus sont données dans l'Annexe A.

Biološko ovrednotenje medicinskih pripomočkov - 10. del: Preskusi preobčutljivosti kože (ISO/FDIS 10993-10:2021)

General Information

• Status: Not Published
• Publication Date: 07-Feb-2023
• ICS: 11.100.20 - Biological evaluation of medical devices
• Technical Committee: CEN/TC 206 - Biocompatibility of medical and dental materials and devices
• Drafting Committee: CEN/TC 206 - Biocompatibility of medical and dental materials and devices
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 08-Feb-2023
• Completion Date: 08-Feb-2023
• Directive: 2007/47/EC - Directive 2007/47/EC of the European Parliament and of the council of 5 September 2007 amending Council Directive 90/385/EEC on the approximation of the laws of the Member States relating to active implantable medical devices, Council Directive 93/42/EEC concerning medical devices and Directive 98/8/EC concerning the placing of biocidal products on the market
• Directive: 2017/745 - Regulation (EU) 2017/745 of the European Parliament and of the Council of 5 April 2017 on medical devices, amending Directive 2001/83/EC, Regulation (EC) No 178/2002 and Regulation (EC) No 1223/2009 and repealing Council Directives 90/385/EEC and 93/42/EEC
• Directive: 90/385/EEC - Council Directive 90/385/EEC of 20 June 1990 on the approximation of the laws of the Member States relating to active implantable medical devices
• Directive: 93/42/EEC - Council Directive 93/42/EEC of 14 June 1993 concerning medical devices
• Mandate: M/565 - Medical devices and in vitro diagnostic medical devices in support of Regulation (EU) 2017/745 of the European Parliament and of the Council and Regulation (EU) 2017/746
• Mandate: M/575 - Medical devices in support of Regulation (EU) 2017/745 and in vitro diagnostic medical devices in support of Regulation (EU) 2017/746
• Mandate: M/BC/CEN/89/9 - Joint Standardization request to CEN/CENELEC concerning harmonized standards relating to horizontal aspects in the field of medical devices
• Ref Project: kSIST FprEN ISO 10993-10:2021 - Biological evaluation of medical devices - Part 10: Tests for skin sensitization (ISO/FDIS 10993-10:2021)

Relations

• Replaces: EN ISO 10993-10:2010 - Biological evaluation of medical devices - Part 10: Tests for irritation and skin sensitization (ISO 10993-10:2010)
• Effective Date: 19-Jan-2023

Standards Content (Sample)

SLOVENSKI STANDARD
oSIST prEN ISO 10993-10:2020
01-april-2020
Biološko ovrednotenje medicinskih pripomočkov - 10. del: Preskusi
preobčutljivosti kože (ISO/DIS 10993-10:2020)

Biological evaluation of medical devices - Part 10: Tests for skin sensitization (ISO/DIS

10993-10:2020)
Biologische Beurteilung von Medizinprodukten - Teil 10: Prüfungen auf
Hautsensibilisierung (ISO/DIS 10993-10:2020)

Évaluation biologique des dispositifs médicaux -- Partie 10: Essais de sensibilisation

cutanée (ISO/DIS 10993-10:2020)
Ta slovenski standard je istoveten z: prEN ISO 10993-10
ICS:
11.100.20 Biološko ovrednotenje Biological evaluation of
medicinskih pripomočkov medical devices
oSIST prEN ISO 10993-10:2020 en

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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oSIST prEN ISO 10993-10:2020
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oSIST prEN ISO 10993-10:2020
DRAFT INTERNATIONAL STANDARD
ISO/DIS 10993-10
ISO/TC 194 Secretariat: DIN
Voting begins on: Voting terminates on:
2020-02-14 2020-05-08
Biological evaluation of medical devices —
Part 10:
Tests for skin sensitization
Évaluation biologique des dispositifs médicaux —
Partie 10: Essais de sensibilisation cutanée
ICS: 11.100.20
THIS DOCUMENT IS A DRAFT CIRCULATED
This document is circulated as received from the committee secretariat.
FOR COMMENT AND APPROVAL. IT IS
THEREFORE SUBJECT TO CHANGE AND MAY
NOT BE REFERRED TO AS AN INTERNATIONAL
STANDARD UNTIL PUBLISHED AS SUCH.
IN ADDITION TO THEIR EVALUATION AS
ISO/CEN PARALLEL PROCESSING
BEING ACCEPTABLE FOR INDUSTRIAL,
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
STANDARDS MAY ON OCCASION HAVE TO
BE CONSIDERED IN THE LIGHT OF THEIR
POTENTIAL TO BECOME STANDARDS TO
WHICH REFERENCE MAY BE MADE IN
Reference number
NATIONAL REGULATIONS.
ISO/DIS 10993-10:2020(E)
RECIPIENTS OF THIS DRAFT ARE INVITED
TO SUBMIT, WITH THEIR COMMENTS,
NOTIFICATION OF ANY RELEVANT PATENT
RIGHTS OF WHICH THEY ARE AWARE AND TO
PROVIDE SUPPORTING DOCUMENTATION. ISO 2020
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oSIST prEN ISO 10993-10:2020
ISO/DIS 10993-10:2020(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2020

All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may

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ii © ISO 2020 – All rights reserved
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oSIST prEN ISO 10993-10:2020
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Contents Page

Foreword ..........................................................................................................................................................................................................................................v

Introduction ................................................................................................................................................................................................................................vi

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 1

4 General principles — Step-wise approach ................................................................................................................................ 3

5 Pretest considerations ................................................................................................................................................................................... 4

5.1 General ........................................................................................................................................................................................................... 4

5.2 Types of material................................................................................................................................................................................... 4

5.2.1 Initial considerations ................................................................................................................................................... 4

5.2.2 Ceramics, metals and alloys ................................................................................................................................... 4

5.2.3 Polymers ................................................................................................................................................................................. 4

5.2.4 Biologically derived materials .............................................................................................................................. 4

5.3 Information on chemical composition ............................................................................................................................... 5

5.3.1 General...................................................................................................................................................................................... 5

5.3.2 Existing data sources ................................................................................................................................................... 5

6 Skin sensitization tests .................................................................................................................................................................................. 5

6.1 Choice of test methods ..................................................................................................................................................................... 5

6.2 Murine Local Lymph Node Assay (LLNA) ........................................................................................................................ 6

6.2.1 Principle .................................................................................................................................................................................. 6

6.2.2 Test sample preparation............................................................................................................................................ 6

6.2.3 Animals and husbandry ............................................................................................................................................. 7

6.2.4 Test procedure ................................................................................................................................................................... 7

6.2.5 Treatment groups ........................................................................................................................................................... 8

6.2.6 Determination of cellular proliferation and tissue preparation ............................................. 8

6.2.7 Results and interpretation ...................................................................................................................................... 9

6.2.8 Test report ............................................................................................................................................................................. 9

6.3 Guinea-pig assays for the detection of skin sensitization .................................................................................. 9

6.3.1 Principle .................................................................................................................................................................................. 9

6.3.2 Choice of test sample concentrations ............................................................................................................ 9

6.3.3 Induction..............................................................................................................................................................................10

6.3.4 Challenge .............................................................................................................................................................................10

6.4 Important factors affecting the outcome of the test ............................................................................................10

6.5 Guinea-pig maximization test (GPMT) ............................................................................................................................11

6.5.1 Principle ...............................................................................................................................................................................11

6.5.2 Test sample preparation.........................................................................................................................................11

6.5.3 Animals and husbandry ..........................................................................................................................................11

6.5.4 Test procedure ................................................................................................................................................................11

6.5.5 Observation of animals ........................................................................................................................................... .13

6.5.6 Evaluation of results ..................................................................................................................................................14

6.5.7 Test report ..........................................................................................................................................................................14

6.6 Closed-patch test (Buehler test) ...........................................................................................................................................14

6.6.1 Principle ...............................................................................................................................................................................14

6.6.2 Test sample preparation.........................................................................................................................................14

6.6.3 Animals and husbandry ..........................................................................................................................................14

6.6.4 Test procedure ................................................................................................................................................................15

6.6.5 Observation of animals ........................................................................................................................................... .16

6.6.6 Evaluation of results ..................................................................................................................................................16

6.6.7 Test report ..........................................................................................................................................................................16

7 Key factors in interpretation of test results ..........................................................................................................................17

Annex A (normative) Preparation of materials for sensitization testing....................................................................18

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Annex B (informative) Method for the preparation of extracts from polymeric test materials ..........20

Annex C (informative) in vitro tests for skin sensitization .........................................................................................................23

Annex D (informative) Background information .................................................................................................................................36

Annex E (informative) Reference to the general safety and performance requirements ..........................39

Annex ZA (informative) Relationship between this European Standard and the essential

requirements of Directive 93/42/EEC [OJ L 169] aimed to be covered ....................................................41

Annex ZB (informative) Relationship between this European Standard and the essential

requirements of Directive 90/385/EEC [OJ L 189] aimed to be covered ................................................43

Annex ZC (informative) Relationship between this European standard and the General

Safety and Performance requirements of Regulation (EU) 2017/745 aimed to be

covered ......... ................................................................................................................................................................................................................44

Bibliography .............................................................................................................................................................................................................................46

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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 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,.

This fourth edition cancels and replaces the third edition (ISO 10993-10:2010), which has been

technically revised.
The main changes compared to the previous edition are as follows:
— This document now contains a description of skin sensitization testing only.
— Annex C on alternative test methods for skin sensitization has been updated.
— The testing for irritation is now described in ISO 10993-23
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.
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oSIST prEN ISO 10993-10:2020
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Introduction

This document assesses possible contact hazards from chemicals released from medical devices, which

may produce skin sensitization.

Some materials that are included in medical devices have been tested, and their skin sensitization

potential has been documented. Especially for dental material sensitizing properties were reported see

Reference [50]. Other materials and their chemical components have not been tested and may induce

adverse effects when in contact with human tissue. The manufacturer is thus obliged to evaluate each

device for potential adverse effects prior to marketing.

Traditionally, small animal tests are performed prior to testing on humans to help predict human

response. Since 2015, several in chemico and in vitro assays have been validated and OECD test

[72][76][99]

guidelines released to assess the skin sentization potential of chemicals. These test methods

developed to address a specific key event may not be sufficient to conclude on the presence or absence

of skin sensitisation potential of chemicals and should be considered in the context of integrated

approaches such as IATA, combining them with other complementary information. It should be noted

that the in vitro and in chemico tests for skin sensitization have so far been validated only for neat

chemicals and not for medical device. In order to apply these assays for the testing of skin sensitization

potential of medical devices, further validation for this specific area is essential. . Where appropriate,

the preliminary use of in vitro methods is encouraged for screening purposes prior to animal testing. In

order to reduce the number of animals used, this document presents a step-wise approach, with review

and analysis of test results at each stage. It is intended that, for regulatory submission, sensitization

study/studies be conducted using GLP/ISO 17025 as applicable to the respective country. and comply

with regulations related to animal welfare. Statistical analysis of data is recommended and should be

used whenever appropriate.

This document is intended for use by professionals, appropriately qualified by training and experience,

who are able to interpret its requirements and judge the outcomes of the evaluation for each medical

device, taking into consideration all the factors relevant to the device, its intended use and the current

knowledge of the medical device provided by review of the scientific literature and previous clinical

experience.

The tests included in this document are important tools for the development of safe products, provided

that these are executed and interpreted by trained personnel.

This document is based on numerous standards and guidelines, including OECD Guidelines,

U.S. Pharmacopoeia and the European Pharmacopoeia. It is intended to be the basic document for the

selection and conduct of tests enabling evaluation of dermal sensitization responses relevant to the

safety of medical materials and devices.
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oSIST prEN ISO 10993-10:2020
DRAFT INTERNATIONAL STANDARD ISO/DIS 10993-10:2020(E)
Biological evaluation of medical devices —
Part 10:
Tests for skin sensitization
1 Scope

This document specifies the procedure for the assessment of medical devices and their constituent

materials with regard to their potential to induce skin sensitization.
This document includes:
— details of in vivo sensitization test procedures;
— key factors for the interpretation of the results.

NOTE Instructions for the preparation of materials specifically in relation to the above tests are given in

Annex A.
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: Evaluation and testing within a risk

management process

ISO 10993-2, Biological evaluation of medical devices — Part 2: Animal welfare requirements

ISO 10993-9, Biological evaluation of medical devices — Part 9: Framework for identification and

quantification of potential degradation products

ISO 10993-12, Biological evaluation of medical devices — Part 12: Sample preparation and reference

materials

ISO 10993-13, Biological evaluation of medical devices — Part 13: Identification and quantification of

degradation products from polymeric medical devices

ISO 10993-14, Biological evaluation of medical devices — Part 14: Identification and quantification of

degradation products from ceramics

ISO 10993-15, Biological evaluation of medical devices — Part 15: Identification and quantification of

degradation products from metals and alloys

ISO 10993-18, Biological evaluation of medical devices — Part 18: Chemical characterization of materials

ISO 14155, Clinical investigation of medical devices for human subjects — Good clinical practice

3 Terms and definitions

For the purposes of this document, the terms and definitions given in ISO 10993-1 and the following apply.

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ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— IEC Electropedia: available at http:// www .electropedia .org/
— ISO Online browsing platform: available at http:// www .iso .org/ obp
3.1
allergen
sensitizer

substance or material that is capable of inducing a specific hypersensitivity reaction upon repeated

contact with that substance or material
3.2
blank

extraction vehicle not containing the test material, retained in a vessel identical to that which holds

the test material and subjected to identical conditions to which the test material is subjected during its

extraction

Note 1 to entry: The purpose of the blank control is to evaluate possible confounding effects due to the extraction

vessel, vehicle and extraction process.
3.3
challenge
elicitation

process following the induction phase, in which the immunological effects of subsequent exposures in

an individual to the inducing material are examined
3.4
erythema
reddening of the skin or mucous membrane
3.5
extract
liquid that results from extraction of the test sample or control
[SOURCE: ISO 10993-12:2012]
3.6
induction

process that leads to the de novo generation of an enhanced state of immunological activity in an

individual, after initial exposure to a specific material
3.7
irritant
agent that produces irritation (see 3.9)
3.8
irritation

localized non-specific inflammatory response to single, repeated or continuous application of a

substance/material

Note 1 to entry: Skin irritation is a reversible reaction and is mainly characterized by local erythema (redness),

swelling, itching, peeling, cracking, scaling etcetera of the skin.
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3.9
negative control

well-characterized material or substance that, when evaluated by a specific test method, demonstrates

the suitability of the procedure to yield a reproducible, appropriately negative, non-reactive or minimal

response in the test system
[SOURCE: SOURCE; ISO 10993-12:2012]

Note 1 to entry: In practice, negative controls include blanks, vehicles/solvents and reference materials.

3.10
oedema
swelling due to abnormal infiltration of fluid into the tissues
3.11
positive control

well-characterized material or substance that, when evaluated by a specific test method, demonstrates

the suitability of the test system to yield a reproducible, appropriately positive or reactive response in

the test system
3.12
skin sensitization
allergic contact dermatitis
immunologically mediated cutaneous reaction to a substance

Note 1 to entry: In the human, the responses can be characterized by pruritis, erythema, oedema, papules,

vesicles, bullae or a combination of these. In other species the reactions can differ and only erythema and oedema

can be seen.
3.13
test material

material, device, device portion or component thereof that is sampled for biological or chemical testing

3.14
test sample

material, device, device portion, component, extract or portion thereof that is subjected to biological or

chemical testing or evaluation
3.15
vehicle

liquid used to moisten, dilute, suspend, extract or dissolve the test substance/material

4 General principles — Step-wise approach

The available methods for testing sensitization were developed specifically to detect skin sensitization

potential. Other types of adverse effects are generally not predicted by these tests. For medical devices

that are used as implants or external communicating devices, intradermal testing is more relevant in

approaching the application.

This document requires a step-wise approach, which shall include one or more of the following:

a) characterization of test material, involving chemical characterization and analysis of the test

sample according to the general principles described in ISO 10993-9, ISO 10993-13, ISO 10993-14,

ISO 10993-15, ISO 10993-17 and ISO 10993-18;

b) literature review, including an evaluation of chemical and physical properties, and information on

the sensitization potential of any product constituent as well as structurally-related chemicals and

materials;

c) in accordance with ISO 10993-2, in vitro tests in preference to in vivo tests shall be considered, and

replacement of the latter as new in vitro tests are scientifically validated and become reasonably

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and practicably available. There are currently a number of internationally validated and accepted

in vitro tests to detect the skin sentization potential of chemicals; However, none of these tests has

been qualified for use with medical devices.
d) in vivo animal tests

NOTE 1 Sensitization potential of medical device extracts can at the moment only be determined by an

in vivo assay. This can be accomplished by using the local lymph node assay (LLNA) in mice, the occluded

patch test in guinea-pigs or the guinea-pig maximization test (GPMT).

NOTE 2 In vivo animal tests are appropriate when test materials cannot be characterized and risk

assessments cannot be undertaken using information obtained by the means set out in a), b) and c).

e) Clinical studies, in accordance with ISO 14155 and ethics principles, shall not be performed before

the results of the other evaluations in a) to e) are known.
5 Pretest considerations
5.1 General

It is important to emphasise that pretest considerations may result in the conclusion that testing for

sensitization is not necessary.
The requirements given in ISO 10993-1:2018, Clause 5, and the following apply.

Non-sterile samples shall be investigated by topical investigation only, as the possibility of microbial

contamination of the test sample could confound the final assay interpretation. In cases where

the sterility of a test sample cannot be guaranteed, but the sample is still considered to be non-

contaminated, intradermal administration may be justified.
5.2 Types of material
5.2.1 Initial considerations

It shall be taken into consideration that during manufacture and assembly of medical devices, additional

chemical components may be used as processing aids, e.g. lubricants or mould-release agents. In addition

to the chemical components of the starting material and manufacturing process aids, adhesive/solvent

residues from assembly and also sterilant residues or reaction products resulting from the sterilization

process may be present in a finished product. Whether these components pose a health hazard/risk

depends on the leaching or degradation characteristics of the finished products. These components

shall be taken into account for their potential sensitization activity.
5.2.2 Ceramics, metals and alloys

These materials are normally less complex than polymers and biologically derived materials in terms of

the number of chemical constituents.
5.2.3 Polymers

These materials are normally chemically more complex than those in 5.2.2 in terms of composition. A

number of reaction products/impurities/additives/residual catalyst can be present and the degree or

extent of polymerization can vary.
5.2.4 Biologically derived materials

These materials are inherently complex in their composition. They often also contain process

residues, e.g. cross-linke
...