ISO 18189:2016
(Main)Ophthalmic optics — Contact lenses and contact lens care products — Cytotoxicity testing of contact lenses in combination with lens care solution to evaluate lens/solution interactions
Ophthalmic optics — Contact lenses and contact lens care products — Cytotoxicity testing of contact lenses in combination with lens care solution to evaluate lens/solution interactions
Iso 18189:2016 describes an in vitro test method to assess the potential cytotoxic effects that may arise due to interaction of contact lenses with contact lens care solutions. NOTE The potential of a contact lens or a contact lens care solution to cause cytotoxicity by itself can be evaluated in accordance with general guidance in ISO 10993‑5.
Optique ophtalmique — Lentilles de contact et produits d'entretien des lentilles de contact — Essais de cytotoxicité des lentilles de contact en association avec une solution d'entretien des lentilles de contact pour évaluer les interactions solution/lentille
General Information
Buy Standard
Standards Content (Sample)
INTERNATIONAL ISO
STANDARD 18189
First edition
2016-06-01
Ophthalmic optics — Contact lenses
and contact lens care products —
Cytotoxicity testing of contact lenses
in combination with lens care solution
to evaluate lens/solution interactions
Optique ophtalmique — Lentilles de contact et produits d’entretien
des lentilles de contact — Essais de cytotoxicité des lentilles de
contact en association avec une solution d’entretien des lentilles de
contact pour évaluer les interactions solution/lentille
Reference number
ISO 18189:2016(E)
©
ISO 2016
---------------------- Page: 1 ----------------------
ISO 18189:2016(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2016, Published in Switzerland
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written 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
Ch. de Blandonnet 8 • CP 401
CH-1214 Vernier, Geneva, Switzerland
Tel. +41 22 749 01 11
Fax +41 22 749 09 47
copyright@iso.org
www.iso.org
ii © ISO 2016 – All rights reserved
---------------------- Page: 2 ----------------------
ISO 18189:2016(E)
Contents Page
Foreword .iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 1
5 Direct contact cytotoxicity test for lens/lens care solution combination .1
5.1 General . 1
5.2 Experimental procedure . 2
5.2.1 Basic procedure . 2
5.2.2 Material . 2
5.2.3 Preparation of test sample . 3
5.2.4 Methods . 3
6 Assessment of results . 5
7 Test report . 5
Annex A (normative) Measurement of zone of cell lysis for the direct contact cytotoxicity
test method for testing contact lens in combination with lens care solution .7
Bibliography . 9
© ISO 2016 – All rights reserved iii
---------------------- Page: 3 ----------------------
ISO 18189:2016(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out
through ISO technical committees. Each member body interested in a subject for which a technical
committee has been established has the right to be represented on that committee. International
organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.
ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of
electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www.iso.org/patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation on the meaning of ISO specific terms and expressions related to conformity assessment,
as well as information about ISO’s adherence to the World Trade Organization (WTO) principles in the
Technical Barriers to Trade (TBT) see the following URL: www.iso.org/iso/foreword.html.
The committee responsible for this document is ISO/TC 172, Optics and photonics, Subcommittee SC 7,
Ophthalmic optics and instruments.
iv © ISO 2016 – All rights reserved
---------------------- Page: 4 ----------------------
INTERNATIONAL STANDARD ISO 18189:2016(E)
Ophthalmic optics — Contact lenses and contact lens
care products — Cytotoxicity testing of contact lenses
in combination with lens care solution to evaluate lens/
solution interactions
1 Scope
This International Standard describes an in vitro test method to assess the potential cytotoxic effects
that may arise due to interaction of contact lenses with contact lens care solutions.
NOTE The potential of a contact lens or a contact lens care solution to cause cytotoxicity by itself can be
evaluated in accordance with general guidance in ISO 10993-5.
2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated
references, the latest edition of the referenced document (including any amendments) applies.
ISO 18369-1, Ophthalmic optics — Contact lenses — Part 1: Vocabulary, classification system and
recommendations for labelling specifications
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 18369-1 and the following apply.
3.1
room temperature
temperature defined as 18 °C to 25 °C
4 Principle
The chemicals in a lens care solution can cause cytotoxic effects by direct contact with ocular tissues
or by indirect contact through contact lenses. Uptake of the care product preservative or other
solution ingredients by the lens and subsequent release of these chemicals in the ocular environment
can compromise ocular biocompatibility. The potential interactions between a lens care product and
various contact lens materials should be taken into account in designing the tests to fully evaluate the
cytotoxicity potential of a new contact lens or a lens care product.
5 Direct contact cytotoxicity test for lens/lens care solution combination
5.1 General
The following protocol describes the test method for evaluating potential cytotoxic effects of contact
lenses exposed to contact lens care solution. The cytotoxicity can result from contact lens/lens care
solution interactions.
With the exception of daily disposable contact lenses, the potential interaction of a new contact lens
with marketed representative multipurpose solutions to produce cytotoxicity shall be evaluated.
© ISO 2016 – All rights reserved 1
---------------------- Page: 5 ----------------------
ISO 18189:2016(E)
For evaluating a new contact lens care solution, the potential interaction of new contact lens care
solution with representative contact lenses to produce cytotoxicity shall be evaluated.
5.2 Experimental procedure
5.2.1 Basic procedure
The test contact lens is incubated in ~10 ml of contact lens care solution in a sterile compatible container
2+
for 24 h ± 2 h at room temperature. Similarly, a Dulbecco’s Phosphate Buffered Saline with Ca and
2+
Mg (DBPS)-treated control lens (“Lens Control”) is prepared by incubating the contact lens in ~10 ml
of DPBS in the same type of container for 24 h ± 2 h at room temperature.
For the purpose of this International Standard, a compatible container refers to a container in which
there is little to no uptake of the disinfecting agent and/or preservative. Rinsing of the container with
the contact lens care product may be used to reduce uptake by the container.
Following the 24 h ± 2 h soak period, the lenses may be cut in a pinwheel fashion (3 to 4 cuts
approximately 1/3 to 1/2 into the lens) and immediately used for cytotoxicity testing. If the lens is
not cut, it shall be placed on the cells in a concave manner. Each lens is placed in the centre on the cell
surface in a 60 mm diameter tissue culture plate containing subconfluent monolayer of L-929 cells in
1,6 ml Minimal Essential Medium (MEM) supplemented with 5 % fetal bovine serum (FBS).
Similarly, negative and positive controls are placed in the designated 60 mm diameter tissue culture
plates containing subconfluent monolayer of L-929 cells in 1,6 ml MEM supplemented with 5 % FBS.
The tissue culture plates are incubated at 37 °C ± 1 °C in 5 % ± 1 % CO for 24 h ± 2 h.
2
Following incubation, the lenses and the controls are removed from each plate and the cells are stained
with Trypan Blue to facilitate observation of dead or damaged cells. The cytotoxicity is assessed by
evaluating the cells macroscopically and microscopically (100×) for any abnormal cell morphology and
lysis around the test article and controls to determine the zone of lysis (if any).
5.2.2 Material
5.2.2.1 Cell line
L-929 cells [NCTC clone 929: CCL 1, American Type Culture Collection (ATCC), Manassas, VA, USA;
ECACC No. 88102702 or equivalent, European Collection of Cell Cultures, Salisbury, Wiltshire SP4 0JG,
UK]. Cell cultures shall be free of mycoplasma.
The passage number of the cells for testing should be 10 – 30.
5.2.2.2 Technical equipment
5.2.2.2.1 Incubator, 37 °C ± 1 °C, humidified, 5 % ± 1 % CO /air.
2
5.2.2.2.2 Laminar flow cabinet, standard: “biological hazard”.
5.2.2.2.3 Water bath, 37 °C.
5.2.2.2.4 Inverse phase contrast microscope.
5.2.2.2.5 Laboratory burner.
5.2.2.2.6 Centrifuge.
2 © ISO 2016 – All rights reserved
---------------------- Page: 6 ----------------------
ISO 18189:2016(E)
5.2.2.2.7 Laboratory balance.
5.2.2.2.8 Cell counter or hemocytometer.
5.2.2.2.9 Tissue culture flasks and 60 mm diameter tissue culture plat
...
DRAFT INTERNATIONAL STANDARD
ISO/DIS 18189
ISO/TC 172/SC 7 Secretariat: DIN
Voting begins on: Voting terminates on:
2015-08-17 2015-11-17
Ophthalmic optics — Contact lenses and contact lens care
products — Cytotoxicity testing of contact lenses and
contact lens care solutions
Titre manque
ICS: 11.040.70
THIS DOCUMENT IS A DRAFT CIRCULATED
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
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 18189:2015(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 2015
---------------------- Page: 1 ----------------------
ISO/DIS 18189:2015(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2015, Published in Switzerland
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written 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
Ch. de Blandonnet 8 • CP 401
CH-1214 Vernier, Geneva, Switzerland
Tel. +41 22 749 01 11
Fax +41 22 749 09 47
copyright@iso.org
www.iso.org
ii © ISO 2015 – All rights reserved
---------------------- Page: 2 ----------------------
ISO/DIS 18189
Contents Page
Foreword . iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle. 1
5 Direct contact cytotoxicity test for lens-lens care solution combination . 1
5.1 General . 1
5.2 Experimental procedure . 2
5.2.1 Basic procedure . 2
5.2.2 Material . 2
5.2.3 Preparation of test sample . 3
5.2.4 Methods . 3
6 Assessment of results . 5
7 Test report . 6
Annex A (normative) Measurement of zone of cell lysis for the direct contact cytotoxicity test
method for testing contact lens in combination with lens care solution . 7
A.1 Plate layout . 7
A.2 Grades 0 to 2 . 7
A.3 Grades 3 and 4 . 8
Bibliography . 9
© ISO 2015 – All rights reserved iii
---------------------- Page: 3 ----------------------
ISO/DIS 18189
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. 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. 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.
The committee responsible for this document is ISO/TC 172, Optics and photonics, Subcommittee SC 7,
Ophthalmic optics and instruments.
iv © ISO 2015 – All rights reserved
---------------------- Page: 4 ----------------------
DRAFT INTERNATIONAL STANDARD ISO/DIS 18189
Ophthalmic optics — Contact lenses and lens care products —
Cytotoxicity testing of contact lens in combination with lens
care solution to evaluate lens - solution interactions
1 Scope
This International Standard describes an in vitro test method to assess the potential cytotoxic effects that may
arise due to interaction of contact lenses with contact lens care solutions.
NOTE The potential of a contact lens or a contact lens care solution to cause cytotoxicity by itself can be evaluated in
accordance with general guidance in ISO 10993-5.
2 Normative references
The following referenced documents are indispensable for the application 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 18369-1, Ophthalmic optics — Contact lenses — Part 1: Vocabulary, classification system and
recommendations for labelling specifications
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 18369-1 and the following apply.
3.1
room temperature
temperature defined as 18 °C to 25 °C
4 Principle
The chemicals in a lens care solution can cause cytotoxic effects by direct contact with ocular tissues or by
indirect contact through contact lenses. Uptake of the care product preservative or other solution ingredients
by the lens and subsequent release of these chemicals in the ocular environment can compromise ocular
biocompatibility. The potential interactions between a lens care product and various contact lens materials
should be taken into account in designing the tests to fully evaluate the cytotoxicity potential of a new contact
lens or a lens care product.
5 Direct contact cytotoxicity test for lens-lens care solution combination
5.1 General
The following protocol describes the test method for evaluating potential cytotoxic effects of contact lenses
exposed to contact lens care solution. The cytotoxicity can result from contact lens – lens care solution
interactions.
With the exception of daily disposable contact lenses, the potential interaction of a new contact lens with
marketed representative multipurpose solutions to produce cytotoxicity shall be evaluated.
© ISO 2015 – All rights reserved
1
---------------------- Page: 5 ----------------------
ISO/DIS 18189
For evaluating a new contact lens care solution, the potential interaction of new contact lens care solution with
representative conventional and silicone hydrogel lenses to produce cytotoxicity shall be evaluated.
5.2 Experimental procedure
5.2.1 Basic procedure
The test contact lens is incubated in ~10 ml of contact lens care solution in a sterile compatible container for
2+
24 h ± 2 h at room temperature. Similarly, a DPBS (Dulbecco’s Phosphate Buffered Saline with Ca and
2+
Mg )-treated control lens (“Lens Control”) is prepared by incubating the contact lens in ~10 ml of DPBS in the
same type of container for 24 h ± 2 h at room temperature.
For the purpose of this document, a compatible container refers to a container in which there is little to no
uptake of the disinfecting agent and/or preservative. Rinsing of the container with the contact lens care
product may be used to reduce uptake by the container.
Following the 24 h ± 2 h soak period, the lenses may be cut in a pinwheel fashion (3 to 4 cuts approximately
1/3 to 1/2 into the lens) and immediately used for cytotoxicity testing. If the lens is not cut, it shall be placed on
the cells in a concave manner. Each lens is placed in the centre on the cell surface in a 60 mm diameter
tissue culture plate containing subconfluent monolayer of L-929 cells in 1,6 ml Minimal Essential Medium
supplemented with 5 % serum (MEM).
Similarly, negative and positive controls are placed in the designated 60 mm diameter tissue culture plates
containing subconfluent monolayer of L-929 cells in 1,6 ml Minimal Essential Medium supplemented with 5 %
serum (MEM).
The tissue culture plates are incubated at 37 °C ± 1 °C in 5 % ± 1 % CO for 24 h ± 2 h.
2
Following incubation, the lenses and the controls are removed from each plate and the cells are stained with
Trypan Blue to facilitate observation of dead or damaged cells. The cytotoxicity is assessed by evaluating the
cells macroscopically and microscopically (100×) for any abnormal cell morphology and lysis around the test
article and controls to determine the zone of lysis (if any).
5.2.2 Material
5.2.2.1 Cell line
L-929 cells (NCTC clone 929: CCL 1, American Type Culture Collection [ATCC], Manassas, VA, USA;
ECACC No. 88102702, European Collection of Cell Cultures, Salisbury, Wiltshire SP4 0JG, UK). Cell cultures
shall be free of mycoplasma.
The passage number of the cells for testing should be 10 – 30.
5.2.2.2 Technical equipment
5.2.2.2.1 Incubator, 37 °C ± 1 °C, humidified, 5 % CO /air.
2
5.2.2.2.2 Laminar flow cabinet, standard: “biological hazard”.
5.2.2.2.3 Water bath, 37 °C.
5.2.2.2.4 Inverse phase contrast microscope.
5.2.2.2.5 Laboratory burner.
5.2.2.2.6 Centrifuge.
5.2.2.2.7 Laboratory balance.
© ISO 2015 – All rights reserved
2
---------------------- Page: 6 ----------------------
ISO/DIS 18189
5.2.2.2.8 Cell counter or hemocytometer.
5.2.2.2.9 Tissue culture flasks and 60 mm diameter tissu
...
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.