SIST EN ISO 10993-6:2026

Biological evaluation of medical devices - Part 6: Tests for local effects after implantation (ISO 10993-6:2026)

This document specifies requirements for implantation test methods for preclinical assessment of the local effects after implantation of medical devices or materials intended for use in medical devices. This document is applicable to the evaluation of local tissue responses from medical devices that are intended to be used where skin or mucosal tissue is breached, when required.
This document is applicable to medical device or materials that require implantation evaluation and can be solid or non-solid (such as porous materials, liquids, gels, pastes, powders, and particulates), absorbable, degradable, non- absorbable, or can be tissue-engineered medical products (TEMPs).
These implantation tests are not intended to evaluate or determine the performance of the test sample in terms of mechanical loading or functional performance. This document also does not provide guidance on methods and study design to satisfy requirements for systemic toxicity, carcinogenicity, teratogenicity or mutagenicity. However, the study designs can be modified to also assess other biological effects.

Biologische Beurteilung von Medizinprodukten - Teil 6: Prüfungen auf lokale Effekte nach Implantationen (ISO 10993-6:2026)

Dieses Dokument legt Anforderungen für Implantationsprüfverfahren fest zur vorklinischen Beurteilung lokaler Effekte nach der Implantation von Medizinprodukten oder von Materialien, die zur Verwendung in Medizinprodukten vorgesehen sind. Um lokale Gewebereaktionen auf Medizinprodukte zu bewerten, die für einen Einsatz vorgesehen sind, bei dem die Haut oder Schleimhaut unterbrochen wird, ist dieses Dokument bei Bedarf anwendbar.
Dieses Dokument ist anwendbar auf Medizinprodukte oder Materialien, die eine Implantationsprüfung erfordern und die fest oder nicht fest (beispielsweise poröse Materialien, Flüssigkeiten, Gels, Pasten, Puder und Partikel) sein können, die resorbierbar, abbaufähig oder nicht resorbierbar sein können oder die durch Tissue Engineering hergestellte medizinische Produkte (TEMPs) sein können.
Diese Implantationsprüfungen sind nicht dazu vorgesehen, die Leistung des Prüfmusters im Sinne der mechanischen Belastbarkeit oder funktionellen Leistung zu bewerten oder zu ermitteln. Dieses Dokument stellt auch keinen Leitfaden zu Verfahren oder zum Studienaufbau bereit, um Anforderungen bezüglich systemischer Toxizität, Karzinogenität, Teratogenität oder Mutagenität zu erfüllen. Die Studienaufbauten können jedoch modifiziert werden, um auch andere biologische Effekte zu bewerten.

Évaluation biologique des dispositifs médicaux - Partie 6: Essais concernant les effets locaux après implantation (ISO 10993-6:2026)

Le présent document spécifie les exigences relatives aux méthodes d'essai d'implantation pour l'évaluation préclinique des effets locaux après l'implantation de dispositifs médicaux ou de matériaux destinés à être utilisés dans des dispositifs médicaux. Le présent document s'applique à l'évaluation des réponses tissulaires locales à des dispositifs médicaux destinés à être utilisés en cas de lésion de la peau ou des muqueuses, si nécessaire.
Le présent document s'applique aux dispositifs médicaux/matériaux qui nécessitent une évaluation de l'implantation et qui peuvent être solides ou non solides (tels que des matériaux poreux, des liquides, des gels, des pâtes, des poudres et des particules), absorbables, dégradables ou non absorbables, ou qui peuvent être des produits médicaux issus de l’ingénierie tissulaire (TEMPs).
Ces essais d'implantation ne sont pas destinés à évaluer ou à déterminer la performance de l'échantillon d'essai en termes de charge mécanique ou de performance fonctionnelle. Le présent document ne fournit pas non plus de recommandations sur les méthodes et la conception des études permettant de satisfaire aux exigences relatives à la toxicité systémique, à la cancérogénicité, à la tératogénicité ou à la mutagénicité. Cependant, les conceptions d’étude peuvent être modifiées afin d'évaluer d'autres effets biologiques.

Biološka ocena medicinskih pripomočkov - 6. del: Preskusi lokalnih učinkov po vsaditvi (ISO 10993-6:2026)

Ta dokument določa zahteve za metode testiranja implantacije za predklinično oceno lokalnih učinkov po implantaciji medicinskih pripomočkov ali materialov, namenjenih za uporabo v medicinskih pripomočkih. Ta dokument je uporaben za oceno lokalnih tkivnih odzivov medicinskih pripomočkov, ki so namenjeni uporabi tam, kjer je prekinjena koža ali sluznično tkivo, kadar je to potrebno.
Ta dokument je uporaben za medicinske pripomočke ali materiale, ki zahtevajo oceno implantacije in so lahko trdni ali netrdni (kot so porozni materiali, tekočine, geli, paste, praški in delci), absorbilni, razgradljivi, neabsorbilni ali pa so lahko tkivno inženirski medicinski izdelki (TEMPs).
Ti testi implantacije niso namenjeni ocenjevanju ali določanju zmogljivosti testnega vzorca v smislu mehanske obremenitve ali funkcionalne zmogljivosti. Ta dokument prav tako ne nudi smernic o metodah in zasnovi študij za izpolnjevanje zahtev glede sistemske toksičnosti, karcinogenosti, teratogenosti ali mutagenosti. Vendar pa se lahko zasnove študij prilagodijo tudi za oceno drugih bioloških učinkov.

General Information

• Status: Published
• Public Enquiry End Date: 30-Jul-2024
• Publication Date: 10-Jun-2026
• ICS: 11.100.20 - Biological evaluation of medical devices
• Technical Committee: VAZ - Healthcare
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 28-May-2026
• Due Date: 02-Aug-2026
• Completion Date: 11-Jun-2026
• Directive: Harmonized Standard2017/745 - 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
• 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/575 Amd 2 - Medical devices in support of Regulation (EU) 2017/745 and in vitro diagnostic medicial devices in support of Regulation (EU) 2017746
• Ref Project: EN ISO 10993-6:2026 - Biological evaluation of medical devices - Part 6: Tests for local effects after implantation (ISO 10993-6:2026)

Relations

• Replaces: SIST EN ISO 10993-6:2017 - Biological evaluation of medical devices - Part 6: Tests for local effects after implantation (ISO 10993-6:2016)
• Effective Date: 01-Jul-2026
• Referred By: SIST-TS CEN/TS 15277:2006 - Non-active surgical implants - Injectable implants
• Effective Date: 27-May-2026
• Referred By: SIST EN 60601-1:2007/A2:2021 - Medical electrical equipment - Part 1: General requirements for basic safety and essential performance (IEC 60601-1:2005/A2:2020)
• Effective Date: 27-May-2026
• Referred By: SIST EN 455-3:2007 - Medical gloves for single use - Part 3: Requirements and testing for biological evaluation
• Effective Date: 27-May-2026

Overview

SIST EN ISO 10993-6:2026 - Biological evaluation of medical devices - Part 6: Tests for local effects after implantation - provides internationally recognized requirements for evaluating the local biological effects of medical devices following implantation. Developed under the framework of ISO 10993, this standard is crucial for preclinical assessment to ensure safety and biocompatibility of medical devices or materials intended for use at locations where skin or mucosal tissue is breached.

This part of the ISO 10993 series applies to a wide range of device types, including solid, non-solid (porous, liquids, gels, pastes, powders, particulates), absorbable, degradable, non-absorbable, and tissue-engineered medical products (TEMPs). It is intended for use when direct contact with tissue necessitates assessment of the local biological response. Importantly, the standard outlines local tissue effects only, and does not address systemic toxicity, carcinogenicity, teratogenicity, or mutagenicity, although study designs may be adapted to examine those aspects.

Key Topics

• Scope of Application

  • Implantation test methods for devices/materials intended for use at breached tissue sites
  • Applicable to various forms and compositions of medical devices, including TEMPs
  • Focus on local effects: not designed to evaluate mechanical or functional device performance

• Study Design and Methodology

  • Emphasis on the 3 Rs principle (replace, reduce, refine animal use)
  • Adherence to animal welfare standards (ISO 10993-2)
  • Detailed preparation, sterilization, and handling procedures for test and control samples
  • Use of appropriate controls, either established biocompatible materials or compositionally similar products

• Test Methods and Assessment

  • Selection of relevant anatomical implantation site based on intended clinical use
  • Specification of sample size, test duration, and animal models
  • Guidelines for both absorbable and non-absorbable materials
  • Procedures for macroscopic and microscopic tissue evaluation
  • Recommendations for marking implantation sites and tracking degradation or absorption

• Reporting Requirements

  • Comprehensive documentation of study design, sample characteristics, animal care, and test results
  • Recording and justification of all controls, implantation periods, and observed effects

Applications

Implementation of SIST EN ISO 10993-6:2026 is critical throughout the medical device development lifecycle, especially for:

• Preclinical Safety Assessment:

  • Manufacturers use the standard to demonstrate the local biocompatibility of device materials before human clinical trials.
  • Data supports risk assessment and regulatory submissions for device approval in domestic and international markets.

• Design of Novel Devices:

  • Assists developers in selecting appropriate materials and design parameters to minimize adverse local tissue reactions.

• Regulatory Compliance:

  • Compliance with ISO 10993-6 is often required or expected by regulatory authorities, including those in the EU (Regulation (EU) 2017/745).

• Material Comparison and Improvement:

  • Enables comparative studies between new device materials and established controls with proven clinical safety.

Typical devices subject to these tests include sutures, meshes, bone substitutes, soft tissue fillers, and innovative tissue-engineered products, among others.

Related Standards

For comprehensive biological evaluation and regulatory compliance, SIST EN ISO 10993-6:2026 should be used alongside other standards in the ISO 10993 series, such as:

• ISO 10993-1: Biological evaluation - Part 1: Evaluation and testing within a risk management process
• ISO 10993-2: Animal welfare requirements
• ISO 10993-4: Selection of tests for interactions with blood
• ISO 10993-11: Tests for systemic toxicity
• ISO 10993-12: Sample preparation and reference materials
• ISO 10993-16: Toxicokinetic study design for degradation products and leachables

For specialized devices, reference may also be made to standards such as ISO 15798 (ocular devices), ISO 11979-5 (intraocular lenses), and ISO 7405 (dental materials).

Conclusion

Adhering to SIST EN ISO 10993-6:2026 ensures medical devices are evaluated for local tissue biocompatibility, supporting safe innovation and market entry. The standard is a vital tool for manufacturers, laboratories, and regulatory professionals committed to high standards of patient safety in medical device development.

Standards Content (Sample)

SLOVENSKI STANDARD
01-julij-2026
Nadomešča:
SIST EN ISO 10993-6:2017
Biološko ovrednotenje medicinskih pripomočkov - 6. del: Preskusi, povezani z
lokalnimi učinki po implantaciji (ISO 10993-6:2026)
Biological evaluation of medical devices - Part 6: Tests for local effects after implantation
(ISO 10993-6:2026)
Biologische Beurteilung von Medizinprodukten - Teil 6: Prüfungen auf lokale Effekte
nach Implantationen (ISO 10993-6:2026)
Évaluation biologique des dispositifs médicaux - Partie 6: Essais concernant les effets
locaux après implantation (ISO 10993-6:2026)
Ta slovenski standard je istoveten z: EN ISO 10993-6:2026
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.

EN ISO 10993-6
EUROPEAN STANDARD
NORME EUROPÉENNE
May 2026
EUROPÄISCHE NORM
ICS 11.100.20 Supersedes EN ISO 10993-6:2016
English Version
Biological evaluation of medical devices - Part 6: Tests for
local effects after implantation (ISO 10993-6:2026)
Évaluation biologique des dispositifs médicaux - Partie Biologische Beurteilung von Medizinprodukten - Teil 6:
6: Essais concernant les effets locaux après Prüfungen auf lokale Effekte nach Implantation (ISO
implantation (ISO 10993-6:2026) 10993-6:2026)
This European Standard was approved by CEN on 22 January 2026.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2026 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 10993-6:2026 E
worldwide for CEN national Members.

Contents Page
European foreword . 3
Annex ZA (informative) Relationship between this European Standard the General Safety
and Performance Requirements of Regulation (EU) 2017/745 aimed to be covered . 4

European foreword
This document (EN ISO 10993-6:2026) has been prepared by Technical Committee ISO/TC 194
"Biological and clinical evaluation of medical devices" in collaboration with Technical Committee
CEN/TC 206 “Biological and clinical evaluation of medical devices” the secretariat of which is held by
DIN.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by November 2026, and conflicting national standards
shall be withdrawn at the latest by November 2026.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document supersedes EN ISO 10993-6:2016.
This document has been prepared under a standardization request addressed to CEN by the European
Commission. The Standing Committee of the EFTA States subsequently approves these requests for its
Member States.
For the relationship with EU Legislation, see informative Annex ZA, which is an integral part of this
document.
Any feedback and questions on this document should be directed to the users’ national standards
body/national committee. A complete listing of these bodies can be found on the CEN website.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,
Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and the
United Kingdom.
Endorsement notice
The text of ISO 10993-6:2026 has been approved by CEN as EN ISO 10993-6:2026 without any
modification.
Annex ZA
(informative)
Relationship between this European Standard the General Safety and
Performance Requirements of Regulation (EU) 2017/745 aimed to be
covered
This European standard has been prepared under M/575 to provide one voluntary means of
conforming to the General Safety and Performance Requirements of Regulation (EU) 2017/745 of 5
April 2017 concerning medical devices [OJ L 117] and to system or process requirements including
those relating to quality management systems, risk management, post-market surveillance systems,
clinical investigations, clinical evaluation or post-market clinical follow-up.
Once this standard is cited in the Official Journal of the European Union under that Regulation,
compliance with the normative clauses of this standard given in Table ZA.1 and application of the
edition of the normatively referenced standards as given in Table ZA.2 confers, within the limits of the
scope of this standard, a presumption of conformity with the corresponding General Safety and
Performance Requirements of that Regulation, and associated EFTA Regulations.
Where a definition in this harmonised standard differs from a definition of the same term set out in
Regulation (EU) 2017/745, the differences shall be indicated in the Annex Z. For the purpose of using
this standard in support of the requirements set out in Regulation (EU) 2017/745, the definitions set
out in this Regulation prevail.
Where the European standard is an adoption of an International Standard, the scope of this document
can differ from the scope of the European Regulation that it supports. As the scope of the applicable
regulatory requirements differ from nation to nation and region to region the standard can only
support European regulatory requirements to the extent of the scope of the European Regulation for
medical devices ((EU) 2017/745).
NOTE 1 Where a reference from a clause of this standard to the risk management process is made, the risk
management process needs to be in compliance with Regulation (EU) 2017/745. This means that risks have to be
‘reduced as far as possible’, ‘reduced to the lowest possible level’, ‘reduced as far as possible and appropriate’,
‘removed or reduced as far as possible’, ‘eliminated or reduced as far as possible’, ’removed or minimized as far as
possible’, or ‘minimized’, according to the wording of the corresponding General Safety and Performance
Requirement.
NOTE 2 The manufacturer’s policy for determining acceptable risk must be in compliance with General Safety
and Performance Requirements 1, 2, 3, 4, 5, 8, 9, 10, 11, 14, 16, 17, 18, 19, 20, 21 and 22 of the Regulation.
NOTE 3 When a General Safety and Performance Requirement does not appear in Table ZA.1, it means that it is
not addressed by this European Standard.
Table ZA.1 — Correspondence between this European Standard and Annex I
of Regulation (EU) 2017/745 [OJ L 117] and to system or process requirements including those
relating to quality management systems, risk management, post-market surveillance systems, clinical
investigations, clinical evaluation or post-market clinical follow-up
General Safety and
Performance Clause(s)/subclause(s)
Requirements of of Remarks/Notes
Regulation this EN
(EU) 2017/745
GSPR 10.1 [(a), (b), (c), (d) and (g)] is partly
covered by this document, since the standard
does not provide requirements on design
and manufacture. However, this standard
provides a means to assess interactions of
medical devices with tissue after
4, 5 and 6, and Annex A,
implantation. Toxicity as systemic effect is
10.1 [(a), (b), (c), (d) and
Annex B, Annex C, Annex
covered only when the study is designed
(g)]
D
accordingly. Performance of the medical
device in terms of mechanical loading or
functional performance is not covered.
Flammability is not covered.
Selection and applicability of relevant Annex
should be performed according to 5.1.1.
GSPR 10.2 is partly covered by this
document, since the standard does not
provide requirements on design,
4, 5 and 6, and Annex A, manufacture and packaging. However, this
10.2 Annex B, Annex C, Annex standard provides a means to assess
D interactions of contaminants and residues
from medical devices with the tissue.
Selection and applicability of relevant Annex
should be performed according to 5.1.1.
10.4.1 (first paragraph) 4, 5 and 6, and Annex A, GSPR 10.4.1 (first paragraph) is partly
Annex B, Annex C, Annex covered by this document, since the standard
D does not provide requirements on design
and manufacture. However, this standard
provides a means to assess interactions of
contaminants and residues from medical
devices with the tissue. Selection and
applicability of relevant Annex should be
performed according to 5.1.1.
General Safety and
Performance Clause(s)/subclause(s)
Requirements of of Remarks/Notes
Regulation this EN
(EU) 2017/745
10.6 4, 5 and 6, and Annex A, GSPR 10.6 is partly covered by this
Annex B, Annex C, Annex document, since the standard does not
D provide requirements on design and
manufacture. However, this standard
provides a means to assess the properties of
particles which are or can be released into
the patient's body. No guidance on size
determination and nanomaterials is
provided.
Selection and applicability of relevant Annex
should be performed according to 5.1.1.
Table ZA.2 — Normative references from clause 2 of this document and their corresponding
European publications
Column 1 Column 2 Column 3 Column 4
Reference in International
Title Corresponding
Clause 2 Standard Edition
European Standard
Edition
ISO 10993-1 ISO 10993-1:2025 Biological evaluation of medical EN ISO 10993-1:2025
devices — Part 1: Requirements
and general principles for the
evaluation of biological safety
within a risk management process
ISO 10993-2 ISO 10993-2:2022 Biological evaluation of medical EN ISO 10993-2:2022
devices — Part 2: Animal welfare
requirements
ISO 10993-4 ISO 10993-4:2017 Biological evaluation of medical EN ISO 10993-4:2017
devices — Part 4: Selection of
ISO 10993- EN ISO 10993-
tests for interactions with blood
4:2017/Amd 1:2025 4:2017/A1:2025
ISO 10993-9 ISO 10993-9: 2019 Biological evaluation of medical EN ISO 10993-9:2021
devices — Part 9: Framework for
identification and quantification of
potential degradation products
ISO 10993-11 ISO 10993-11:2017 Biological evaluation of medical EN ISO 10993-11:2018
devices — Part 11: Tests for
systemic toxicity
ISO 10993-12:2021 ISO 10993-12:2021 Biological evaluation of medical EN ISO 10993-12:2021
devices — Part 12: Sample
ISO 10993- EN ISO 10993-
preparation and reference
12:2021/Amd 1:2025 12:2021/A1:2025
materials
ISO 10993-16 ISO 10993-16:2017 Biological evaluation of medical EN ISO 10993-16:2017
devices — Part 16: Toxicokinetic
Column 1 Column 2 Column 3 Column 4
Reference in International
Title Corresponding
Clause 2 Standard Edition
European Standard
Edition
study design for degradation
products and leachables
The documents listed in the Column 1 of Table ZA.2, in whole or in part, are normatively referenced in
this document, i.e. are indispensable for its application. The achievement of the presumption of
conformity is subject to the application of the edition of Standards as listed in Column 4 or, if no
European Standard Edition exists, the International Standard Edition given in Column 2 of Table ZA.2.
WARNING 1 — Presumption of conformity stays valid only as long as a reference to this European
standard is maintained in the list published in the Official Journal of the European Union. Users of this
standard should consult frequently the latest list published in the Official Journal of the European
Union.
WARNING 2 — Other Union legislation may be applicable to the product(s) falling within the scope of
this standard.
International
Standard
ISO 10993-6
Fourth edition
Biological evaluation of medical
2026-04
devices —
Part 6:
Tests for local effects after
implantation
Évaluation biologique des dispositifs médicaux —
Partie 6: Essais concernant les effets locaux après implantation
Reference number
ISO 10993-6:2026(en) © ISO 2026

ISO 10993-6:2026(en)
© ISO 2026
All rights reserved. Unless otherwise specified, or required in the context of its implementation, 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 permission. Permission can be requested from either ISO at the address below
or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii
ISO 10993-6:2026(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Common provisions for implantation test methods . 2
4.1 General .2
4.2 Preparation of samples for implantation .3
4.3 Selection of control samples .4
5 General aspects and requirements for implantation test . 4
5.1 Tissue and implantation site .4
5.2 Animal model .5
5.3 Test periods .6
5.4 Surgery and testing conditions .8
5.5 Evaluation .9
5.5.1 General .9
5.5.2 Macroscopic assessment .9
5.5.3 Implant retrieval and tissue sample collection .9
5.5.4 Microscopic assessment .10
5.5.5 Evaluation of responses .11
6 Test report .11
6.1 General .11
6.2 Test laboratory . 12
6.3 Implant samples . 12
6.4 Animals and implantation . 12
6.5 Retrieval and histological procedure . 12
6.6 Macroscopic and microscopic evaluation . 13
6.7 Final evaluation . 13
Annex A (normative) Test methods for implantation in subcutaneous tissue . 14
Annex B (normative) Test method for implantation in muscle .16
Annex C (normative) Test method for implantation in bone .18
Annex D (normative) Test method for implantation in neural tissue .21
Annex E (informative) Test methods for devices contacting peripheral nerve tissue .26
Annex F (informative) Examples of scoring systems used to support the evaluation of local
biological effects after implantation .30
Annex G (informative) Microscopic evaluation of tissue responses to implanted materials .33
Bibliography .38

iii
ISO 10993-6:2026(en)
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 document 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).
ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent
rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a)
patent(s) which may be required to implement this document. However, implementers are cautioned that
this may not represent the latest information, which may be obtained from the patent database available at
www.iso.org/patents. ISO shall not be held responsible for identifying any or all such patent rights.
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of 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 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, Biological and clinical evaluation of medical devices, in accordance with the Agreement on technical
cooperation between ISO and CEN (Vienna Agreement).
This fourth edition cancels and replaces the third edition (ISO 10993-6:2016), which has been technically
revised.
The main changes are as follows:
— new definitions for “comparative control”, “coupon”, “euthanasia”, “local effect”, “location marker”,
“steady-state” and “reference control” have been added to Clause 3;
— a new paragraph on the use of smaller compositionally representative samples or coupons has been
added to 4.2.2;
— a new subclause 4.3 “Selection of control materials” has been added;
— the discussion of assessment of lymph nodes for certain materials has been expanded;
— a new Annex E “Test methods for devices contacting peripheral nerve tissue” and Annex G “Microscopic
evaluation of tissue responses to implanted materials” have been added;
— tissue and pathological terminology has been updated throughout this document;
— bibliographical entries have been updated.
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 10993-6:2026(en)
Introduction
The objective of the implantation test methods is to characterize the local tissue response after implantation
of a medical device or material (test sample) including integration, degradation, or absorption in an
appropriate animal model.
The test sample is implanted into an anatomical site appropriate for the evaluation of the local effects of the
medical device (or portion of) in an animal.
The medical device or material local effects are evaluated by a comparison of the tissue response caused
by a test sample to that caused by comparative or reference control samples used in medical devices whose
clinical acceptability and biocompatibility characteristics have been established.
Careful study design can include other relevant biological effects to reduce the number of animals used to
evaluate safety and efficacy while accomplishing all study objectives. Additionally, a long-term systemic
toxicity study that is designed to incorporate the methods, biological effects, additional timepoints and
outcomes of implantation testing can satisfy the requirements of this document and ISO 10993-11.

v
International Standard ISO 10993-6:2026(en)
Biological evaluation of medical devices —
Part 6:
Tests for local effects after implantation
1 Scope
This document specifies requirements for implantation test methods for preclinical assessment of the local
effects after implantation of medical devices or materials intended for use in medical devices. This document
is applicable to the evaluation of local tissue responses from medical devices that are intended to be used
where skin or mucosal tissue is breached, when required.
This document is applicable to medical device or materials that require implantation evaluation and can be
solid or non-solid (such as porous materials, liquids, gels, pastes, powders, and particulates), absorbable,
degradable, non- absorbable, or can be tissue-engineered medical products (TEMPs).
These implantation tests are not intended to evaluate or determine the performance of the test sample in
terms of mechanical loading or functional performance. This document also does not provide guidance on
methods and study design to satisfy requirements for systemic toxicity, carcinogenicity, teratogenicity or
mutagenicity. However, the study designs can be modified to also assess other biological effects.
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, 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, Biological evaluation of medical devices — Part 2: Animal welfare requirements
ISO 10993-4, Biological evaluation of medical devices — Part 4: Selection of tests for interactions with blood
ISO 10993-9, Biological evaluation of medical devices — Part 9: Framework for identification and quantification
of potential degradation products
ISO 10993-12:2021, Biological evaluation of medical devices — Part 12: Sample preparation and reference
materials
ISO 10993-16, Biological evaluation of medical devices — Part 16: Toxicokinetic study design for degradation
products and leachables
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 10993-1, ISO 10993-2, ISO 10993-4,
ISO 10993-9, ISO 10993-12, ISO 10993-16 and the following apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/

ISO 10993-6:2026(en)
3.1
absorb
action of a non-endogenous (foreign) material or substance, or its decomposition products passing through
or being assimilated by cells and tissue over time
3.2
comparative control
medical device or material with a history of safe clinical use that is used as a point of comparison for a new
medical device seeking regulatory approval
3.3
coupon
multiple smaller compositionally representative portions of a more complex or larger device that together
contain all materials, surface finishes, and processing as the final, finished device
3.4
degradation product
intermediate or final substance which results from the physical, metabolic, or chemical decomposition of a
material or substance
[SOURCE: ISO/TS 37137-1:2021, 3.2, modified — “and/or” has been changed to “or” and “agent” has been
replaced with “substance”.]
3.5
degrade
physically, metabolically or chemically decompose a material or substance
[SOURCE: ISO/TS 37137-1:2021, 3.3, modified — “and/or” has been changed to “or”.]
3.6
euthanasia
humane killing of an animal by a method causing minimal physical and mental suffering
[SOURCE: ISO 10993-2:2022, 3.5]
3.7
local effect
tissue response to an implanted test sample that can be seen with gross pathology and histopathology at the
site of test sample administration or implantation
3.8
location marker
inert, non-absorbable material or process used to mark the location in tissue of an implanted material
3.9
reference control
material with known properties considered to be generally inert in terms of biological local tissue effects
and has been established in a historical basis within a pharmacopeia or similar standard
3.10
steady-state
biologically stable condition wherein change in the test system’s cellular activity, morphology or features is
no longer detected over a period of time
4 Common provisions for implantation test methods
4.1 General
The ISO 10993 series requires animal testing to be avoided unless the required data cannot be obtained by
other means. Prior to choosing to pursue implantation studies, consideration to the type of data required,

ISO 10993-6:2026(en)
the appropriate method and approach to implantation studies should be given to ensure that the data
collected covers the concerns for biological reactivity. It is important that the study be planned in sufficient
detail such that all relevant information can be extracted from the use of each animal and each study plan or
protocol according to ISO 10993-2, ISO 10993-11 and ISO 10993-16. The implantation tests shall be planned
and documented in the study documentation, including the information on the selected test methods, test
samples, test animals, implantation sites, test duration, measurement frequencies, assessment methods
and evaluation. When relevant vertical standards exist, refer to those as guidance for biological specific
evaluation (e.g. ISO 15798, ISO 11979-5, ISO 7405, ISO 14708 series, ISO 5840 series).
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 conform with the
requirements of ISO 10993-2. These studies shall be performed under Good Laboratory Practices or other
recognized, quality assurance systems. While pilot implant studies can or cannot be conducted under Good
Laboratory Practices, the test facility should have an adequate quality system.
The provisions of this clause shall apply to the test methods specified in Annex A, Annex B, Annex C and
Annex D.
4.2 Preparation of samples for implantation
4.2.1 Test and control samples shall be in a final finished state as described in ISO 10993-12 (unless used
in a pilot study). The implant size and shape shall be documented and justified. For certain devices, clinically
relevant dosing can be considered. A justification should be provided to demonstrate mimicking a clinically
relevant dose in the implantation study.
Test samples for various implant sites are described in Annex A, Annex B, Annex C and Annex D. Physical
properties (such as form, density, hardness, surface porosity and texture), geometrical characteristics (form,
shape and size) and composition can influence the character of the tissue response to the test material and
shall be recorded and taken into account when the response is characterized. Comparative control samples
should be matched as closely as reasonably possible for physical properties and geometrical (form and
shape) characteristics. Otherwise, a reference control (well characterized material with a well characterized
response) can be used, such as high density polyethylene (HDPE).
Additional control samples may be included in the study to help interpret tissue reactions [e.g. in case of
combination drugs, an additional control such as a placebo device (without drug) can be considered].
4.2.2 Each implant shall be manufactured, processed, cleaned of contaminants and sterilized by the
method intended for the final product and this shall be confirmed in the study documentation. After final
preparation and sterilization, the implant samples shall be handled aseptically and in such a way as to
ensure that they are not damaged or contaminated in any way prior to or during implantation.
The use of smaller test samples or test coupons that are consistent with the requirements as described in
ISO 10993-12 can be relevant if the medical device to be tested cannot be tested as is, due to size or complex
geometry. The test samples or test coupons shall contain all tissue-contacting materials and surface finishes
as in the final, finished medical device. For medical devices comprising two or more different materials,
implanted test samples should be of similar composition, surface finish and include each individual material
to the final product. With multiple materials, it can be necessary to implant several coupons together, each
containing a subset of device materials. The potential for synergies and interactions of different materials in
the final product should be considered in the choice of test sample.
NOTE Refer to the respective annexes for guidance on the number of test and control samples for each material
and implantation period.
4.2.3 For materials used as scaffolds for tissue-engineered medical products, it can be appropriate not
to use the final preparation pre-populated with cells or proteins, as the immune reaction of the animal
to human cellular or protein components of such products and the reaction of the cells to the animal can
interfere with the resulting local tissue response, making it difficult to interpret.

ISO 10993-6:2026(en)
NOTE 1 Implantation in an appropriate immune-deficient animal can be an option to avoid the xenograft responses,
if justified.
NOTE 2 Products with living cells are not considered medical devices in all jurisdictions.
4.2.4 For multicomponent material device designed to be cured prior to placement, the components
shall be mixed before use and allowed to set before implantation. However, materials that are designed to
polymerize in situ (e.g. bone cements, many dental materials, tissue sealants and glues) shall be introduced
in a manner such that in situ polymerization occurs. The procedure used shall be justified and documented.
4.3 Selection of control samples
The process of implanting any material in tissue induces some degree of cellular response. In the evaluation
of material local effects after implantation, the choice of control samples is important in determining
the acceptability of the observed tissue reaction to the test sample. Evaluation should be performed by
comparing the tissue reaction to that of a similar material (comparative control) whose clinical acceptability
and biocompatibility characteristics have been established. The geometrical and physical characteristics
such as shape, size, curvature and especially the surface condition (including porosity and texture) of the
control article(s), should be as similar to that of the implant test samples as is practical. Any deviations
shall be explained, justified and recorded. If no appropriate comparative control is available, a reference
control can be used. For further guidance, see ISO 10993-12. For example, reference materials as specified
in ISO 10993-12:2021, Table A.1 negative control column can provide a good control for solid, smooth,
non-absorbable materials. Conversely, the reaction can likely be different if it is used to compare against
a non-absorbable or degradable hernia mesh due to the differing physical form characteristics. Ideally, a
commercially available material should be used as a comparative control so the tissue reaction is similar
to what has been clinically proven. For non-absorbable medical devices or materials, a comparative control
shall be used made of a stable non-absorbable comparative material (see Annex A, Annex B, Annex C and
Annex D).
Since absorbable materials encounter changing tissue responses as degradation proceeds at rates that differ
based on composition, processing and sterilization techniques, similar absorbable comparative medical
device or material controls should be considered.
The absorption rate of the control material or medical device should be similar to the test material or medical
device (for more information, see ISO/TS 37137-1). Alternatively, a non-absorbable comparative control can
be used, if justified (e.g. there are no clinically relevant absorbable controls). If a comparative control is used,
typically reference materials as specified in ISO 10993-12:2021, Annex A are also implanted into the animal
for comparison and serving as a procedural control yielding a total of three separate articles included in the
study. For novel materials or medical devices, if a commercially available comparative material or medical
device does not exist, the choice of a control that is as close as possible is preferred. The nature of any
adverse inflammatory response or high reactivity rating or similar histopathological evaluation conclusion
should be discussed in the context of the control article chosen. The choice of control shall be documented
and justified. If no control or a sham control is used, this also shall be documented and justified.
5 General aspects and requirements for implantation test
5.1 Tissue and implantation site
5.1.1 The test sample shall be implanted into or onto the tissues most relevant to the intended clinical use
of the material. The justification for the choice of sample numbers, tissue, implantation sites and test period
shall be documented. Test methods for various implantation sites are given in Annex A, Annex B, Annex C
and Annex D. If other implantation sites are chosen, the general scientific principles behind the test methods
described in Annex A, Annex B, Annex C and Annex D shall still be adhered to and a justification be provided
for selecting an anatomical location that does not align with the use of the device.
NOTE For some devices, there are vertical standards prescribing specific implant studies to evaluate local tissue
responses, e.g. for intraocular lens implant (see ISO 11979-5) and dental devices (see ISO 7405). The studies described
in the standards can be used to satisfy the requirements in ISO 10993-6.

ISO 10993-6:2026(en)
5.1.2 For absorbable materials, the implantation site shall be marked in a manner suitable for identification
of the site at the end of the designated time periods, taking into consideration the growth and aging of the
implanted animal(s). The use of a non-invasive permanent skin marker or a template marking the placement
of the sample is recommended as a single application for short-term study intervals only. This method of
marking can come off as skin exfoliates and requires remarking as often as needed to maintain the marked
site. Additionally, in many laboratory animals, the loose highly mobile skin limits the precision of marking
sites that are deeper than skin. In most circumstances, a location marker comprising an appropriate non-
absorbable inert or biocompatible material (e.g. HDPE 1 mm by 2 mm by 5 mm, polypropylene suture, gold
band, clips) may be used to identify the location of the implant site. These location markers should be far
enough away from each implant site to not induce changes in the local tissue reaction to the implanted
material. If this is not possible, i.e. the marker of the implanted material is adjacent to or on the edge of the
implanted material, then the tissue reaction to this marker should not be included in the evaluation of the
tissue reaction to the implanted material.
5.1.3 A sham surgical procedure can be used to evaluate the impact of the procedure on the tissue involved;
in these cases, the specific justification shall be provided. If a sham surgical procedure is performed, the
same implantation procedure that is used for the test or control should be used for the sham procedure.
5.2 Animal model
5.2.1 All aspects of animal care and accommodation shall be performed in accordance with ISO 10993-2.
5.2.2 Animals are used to evaluate local effects following implantation and are described as animal
models in the context of this document. In general, small laboratory animals such as mice, rats, guinea-pigs
or rabbits are preferred. The use of larger animals such as dogs, sheep, goats or pigs may be justified based
upon special scientific considerations of the test sample under study, length of study in relation to animal
life expectancy, or if needed to accommodate implant size, with whole device testing or applicable sham
defect size.
5.2.3 Select an animal species in line with the principles set out in ISO
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