EN 30993-6:1994
(Main)Biological evaluation of medical devices - Part 6: Tests for local effects after implantation (ISO 10993-6:1994)
Biological evaluation of medical devices - Part 6: Tests for local effects after implantation (ISO 10993-6:1994)
This part of ISO 10993 specifies test methods for the assessment of the local effects of an implant material on living tissue, at both the macroscopic and microscopic level. The test specimen is implanted into a site and tissue appropriate for evaluation of the biological safety of the material. The implant is not intended to be subjected to mechanical or functional loading. The local effects are evaluated by a comparison of the tissue response caused by a test specimen to that caused by materials used in medical devices whose clinical acceptability has been established. The test methods for local effects after implantation are used to assess subchronic effects (short-term, up to 12 weeks), or chronic effects (long-term, longer than 12 weeks).
Biologische Beurteilung von Medizinprodukten - Teil 6: Prüfungen auf lokale Effekte nach Implantationen (ISO 10993-6:1994)
Dieser Teil der ISO 10993 beschreibt Prüfungen auf die lokalen Effekte eines Implantatwerkstoffes auf lebendes Gewede, sowohl auf makroskopischer als auch auf mikroskopischer Ebene. Der Prüfkörper wird in eine Stelle und ein Gewebe implantiert, die geeignet sind, die biologische Sicherheit des Werkstoffes zu überprüfen. Es ist nicht beabsichtigt, das Implantat einer mechanischen oder funktionellen Belastung außusetzen.
Evaluation biologique des dispositifs médicaux - Partie 6: Essais concernant les effets locaux après implantation (ISO 10993-6:1994)
Biološko ovrednotenje medicinskih pripomočkov - 6. del: Preskusi, povezani z lokalnimi učinki po implantaciji (ISO 10993-6:1994)
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Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN 30993-6:2000
01-januar-2000
%LRORãNRRYUHGQRWHQMHPHGLFLQVNLKSULSRPRþNRYGHO 3UHVNXVLSRYH]DQL]
ORNDOQLPLXþLQNLSRLPSODQWDFLML ,62
Biological evaluation of medical devices - Part 6: Tests for local effects after implantation
(ISO 10993-6:1994)
Biologische Beurteilung von Medizinprodukten - Teil 6: Prüfungen auf lokale Effekte nach
Implantationen (ISO 10993-6:1994)
Evaluation biologique des dispositifs médicaux - Partie 6: Essais concernant les effets
locaux apres implantation (ISO 10993-6:1994)
Ta slovenski standard je istoveten z: EN 30993-6:1994
ICS:
11.100.20 %LRORãNRRYUHGQRWHQMH Biological evaluation of
PHGLFLQVNLKSULSRPRþNRY medical devices
SIST EN 30993-6:2000 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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INTERNATIONAL
IS0
STANDARD
10993-6
First edition
1994-07-l 5
Biological evaluation of medical devices -
Part 6:
Tests for local effects after implantation
haha tion biologique des dispositifs medicaux -
Partie 6: Essais concernan t /es effets locaux apr&s implantation
Reference number
IS0 10993-6:1994(E)
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IS0 10993=6:1994(E)
Contents
Page
1
1 Scope . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1
2 Normative references . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1
3 Common provisions for implantation test methods . . . . . . . . . . . . . . . . .
5
4 Test method for implantation in subcutaneous tissue . . . . . . . . . . . . .
5 Test method for implantation in muscle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
6 Test method for implantation in bone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Annexes
a
A Control materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
9
B Cylindrical specimen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
10
C Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
0 IS0 1994
All rights resewed. Unless otherwise specified, no part of this publication may be reproduced
or utilized in any form or by any means, electronic or mechanical, including photocopying and
microfilm, without permission in writing from the publisher.
International Organization for Standardization
Case Postale 56 l CH-1211 Geneve 20 l Switzerland
Printed in Switzerland
ii
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Foreword
IS0 (the International Organization for Standardization) is a worldwide
federation of national standards bodies (IS0 member bodies). The work
of preparing International Standards is normally carried out through IS0
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. IS0
collaborates closely with the International Electrotechnical Commission
(IEC) on all matters of electrotechnical standardization.
Draft International Standards adopted by the technical committees are
circulated to the member bodies for voting. Publication as an International
Standard requires approval by at least 75 % of the member bodies casting
a vote.
International Standard IS0 10993-6 was prepared by Technical Committee
lSO/TC 194, Biological evaluation of medical devices.
IS0 10993 consists of the following parts, under the general title Biological
evaluation of medical devices:
- Part 1: Guidance on selection of tests
- Part 2: Animal welfare requirements
- Part 3: Tests for genotoxicity, carcinogenicity and reproductive
toxicity
- Part 4: Selection of tests for interactions with blood
- Part 5: Tests for cytotoxicity: in vitro methods
- Part 6: Tests for local effects after implantation
- Part 7: Ethylene oxide steriiiza tion residuals
- Part 9: Degradation of materials related to biological testing
[Technical Report]
- Part 10: Tests for irritation and sensitization
- Part 11: Tests for systemic toxicity
- Part 12: Sample preparation and reference materials
- Part 13: ldentifica tion and quantification of degradation products
from polymers
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0 IS0
IS0 10993=6:1994(E)
- Part 14: lden tifica tion and quantification of degradation products
from ceramics
- Part 15: lden tifica tion and quantification of degradation products
from coated and uncoated metals and alloys
- Part 16: General guidance on toxicokinetic study design for degra-
dation products and leachables from medical devices
- Part 17: Glutaraldehyde and industrially
formaldehyde residues
sterilized medical devices
Future parts will deal with other relevant aspects of biological testing.
Annexes A, B and C of this part of IS0 10993 are for information only.
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Introduction
This International Standard gives methods of biological testing of medical
and dental materials and devices, and their evaluation in regard to their
biocompatibility.
IS0 10993-l offers a guide for selection of methods for biological testing.
The intention is to reduce animal tests to the justifiable minimum (see
IS0 10993-2). A search of the literature precedes any testing, as data
concerning the biological safety of the candidate material could be avail-
able.
The test methods described in this part of IS0 10993 are based on es-
tablished implantation tests. This part of IS0 10993 describes animal tests
for the study of local effects after implantation. The use of in vivo im-
plantation techniques for characterizing the biological response of tissues
to materials allows for the assessment of such materials not achieved by
other procedures.
These test methods may not be appropriate for all types of medical de-
vices. The user is cautioned to consider the appropriateness of the
method in view of the materials being tested, their potential applications,
and the recommendations contained in IS0 10993-l.
lSO/TC 194 appreciates any information for the further development of
this part of IS0 10993.
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IS0 10993=6:1994(E)
INTERNATIONAL STANDARD 0 IS0
Biological evaluation of medical devices -
Part 6:
Tests for local effects after implantation
IS0 10993-l : 1992, Biological evaluation of medical
1 Scope
devices - Part 1: Guidance on selection of tests.
This part of IS0 10993 specifies test methods for the
IS0 10993-2: 1992, Biological evaluation of medical
assessment of the local effects of an implant material
- Part 2: Animal welfare requirements.
devices
on living tissue, at both the macroscopic and micro-
scopic level.
3 Common provisions for implantation
The test specimen is implanted into a site and tissue
test methods
appropriate for evaluation of the biological safety of
the material. The implant is not intended to be sub-
jected to mechanical or functional loading. The local
3.1 General
effects are evaluated by a comparison of the tissue
response caused by a test specimen to that caused
For the purposes of this part of IS0 10993, the defi-
by materials used in medical devices whose clinical
nitions given in IS0 10993-l and IS0 10993-2 apply.
acceptability has been established.
The provisions in this clause shall apply to the test
The test methods for local effects after implantation
methods described in clauses 4 to 6.
are used to assess subchronic effects (short-term, up
It is important that the researcher plans the study in
to 12 weeks), or chronic effects (long-term, longer
such detail that the maximum of information can be
than 12 weeks).
extracted from the use of each animal (see
IS0 10993-2).
3.2 Preparation of specimens for
implantation
2 Normative references
3.2.1 Solid specimens (excluding powders)
The following standards contain provisions which,
Physical characteristics (that is form, density, hard-
through reference in this text, constitute provisions
ness, surface finish) can influence the character of the
of this part of IS0 ‘iO993. At the time of publication,
tissue response to the test material.
the editions indicated were valid. All standards are
subject to revision, and parties to agreements based
Each implant shall be manufactured, processed,
on this part of IS0 10993 are encouraged to investi-
cleaned of contaminants and sterilized by the method
gate the possibility of applying the most recent edi-
intended for the final product.
tions of the standards indicated below. Members of
IEC and IS0 maintain registers of currently valid
After final preparation and sterilization, the implant
International Standards. specimens shall be handled in such a way as to en-
1
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IS0 10993=6:1994(E) 0 IS0
sure that they are not scratched, damaged or con-
3.3 Animals, tissues, test periods, surgery,
taminated in any way prior to or during insertion.
postoperative care, euthanasia
3.3.1 Animals and tissues
3.2.2 Non-solid specimens (including powders)
Animal husbandry shall be in accordance with
IS0 10993-2 and/or national regulatory requirements
Non-solid specimens may be liquids, pastes and
for laboratory animals.
particulates, as distinct from the materials covered in
3.2.1. The components may be mixed before use (e.g.
Select an animal species with due consideration of
bone cements, dental materials), and set after varying
the size of the implant test specimens, the intended
time periods.
duration of the test in relation to the expected life-
span of the animals, as well as the recognized species
The materials may be contained in tubes for the pur-
differences in biological response in both hard and
pose of testing for local effects after implantation.
soft tissues.
Polyethylene (PE), polypropylene (PP), or polytetra-
fluoroethylene (PTFE) tubes are commonly used for
For short-term testing in subcutaneous tissue and
this purpose.
muscle, animals such as mice, rats, guinea-pigs and
rabbits are commonly used. Select one species
Prior to test the tubes shall be rinsed with
among these.
70 % (VW) ethanol and distilled water and sterilized
by autoclaving or other appropriate methods relevant
For long-term testing in subcutaneous tissue, muscle
for clinical applications. Materials tested in their
and bone, animals such as rats, guinea-pigs, rabbits,
freshly mixed state shall be tested for microbiological
dogs, sheep, goats, pigs and other animals with a
contamination.
relatively long life expectancy are suitable. Select one
species among these.
Prepare the test material according to the manufac-
turer’s instructions and insert the material into the
The specimens of test and control materials shall be
tube until level with the top. Exercise the utmost care
implanted under the same conditions in the same
to prevent contamination of the outer surface of the
species of the same age, sex and strain in corre-
tube by the test material. Avoid entrapment of air in
sponding anatomical sites. The number and size of
the tube and ensure that the end surfaces of the in-
implants inserted in an animal depends on the size of
serted material in the tube and the tube ends are
the species and the anatomical location.
smooth.
NOTE 1 PE tubes may be deformed by autoclaving. It is 3.3.2 Test periods
difficult to section PTFE tubes in the microtome, and sub-
stitution by PE or PP tubes of the same dimensions may
The local tissue response to implanted materials is
be preferable when the tubes are to remain in the tissue
assessed in short-term tests up to 12 weeks and in
blocks during sectioning.
long-term tests exceeding 12 weeks.
Test periods are chosen to ascertain that a steady
state has been reached with respect to biological re-
3.2.3 Control specimens
sponse. The local biological response to implanted
materials depends both on the properties of the ma-
The size, shape, and especially the surface condition
terials and on the trauma of surgery. The tissue con-
of the control(s) shall be as similar to that of the im-
figuration found in the vicinity of an implant changes
plant test specimens as is practically possible. When
with the time elapsed after surgery. Usually, at one
the test material is contained in a tube, the control
week observation periods, a high cell activity is found,
shall be a rod of the same material as the tube and
followed by a transitional stage. In muscle and
with the same diameter as the outer diameter of the
connective tissue, depending on the species, a steady
tube. The control specimens shall be handled, cleaned
state is seen in the cell population after 9 to 12
and sterilized in such a manner as to maintain them
weeks. Implantation in bone tissues may need longer
as acceptable and well characterized controls.
obsewation periods.
Selection of control material(s) should be based on
Test periods shall be selected from those specified in
their established use in clinical applications similar to
table 1 for short-term implantation, or from table 2 for
those proposed for the candidate test material and is
long-term implantation.
not restricted to those indicated in annex A and C.l.
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IS0 10993=6:1994(E)
0 IS0
be carried out under aseptic conditions and in a man-
ner that minimizes trauma at the implant site.
- Selection of test periods for
Table 1
short-term implantation in subcutaneous tissue
After surgery close the wound, using either wound
and muscle
clips or sutures, taking precautions to maintain aseptic
Implantation period conditions.
Species
weeks
3.3.4 Post-operative assessment
12
1 3 4 9
Obsewe each animal at appropriate intervals during
X X X
Mice
the test period and record any abnormal findings, in-
X X
Rats X
cluding local, systemic and behavioural abnormalities.
X X X
Guinea-pigs
X X
Rabbits X
3.3.5 Euthanasia
At the termination of the experimental period,
euthanize the animals with an overdose of anaesthetic
Table 2 - Selection of test periods for long-term
or by some other acceptable humane method (see
implantation in subcutaneous tissue, muscle and
IS0 10993-2).
bone
Species Implantation period
3.4 Evaluation of biological response
weeks
12 Evaluate the biological response by grading and
26 52 78 (104)
documenting the macroscopic and histopathological
Rats X
test responses as a function of time. Compare the
X responses to the test material and control material.
Guinea-pigs
Rabbits X
Carry out comparison of the control and the test im-
Dogs X
plants at equivalent locations relative to each implant
Sheep X
so that the effect of relative motion between the tis-
sue and implant is at a minimum (see note 2).
Goats X
Pigs X
NOTE 2 For a cylindrical specimen the region is midway
between its ends. With grooved cylindrical implants the
centre portions between the grooves as well as the flat top
Depending on the intended use of the test material,
end surfaces of the implant are suitable for evaluation.
not all implantation periods may be necessary (see
IS0 10993-l). An observation period of 104 weeks
For a non-solid or particulate material incorporated into
may be of interest in selected instances.
a tube, the area at the end of the tube is the only
available area for evaluation.
The number of implants per animal and the number
of animals per observation period are described in
clauses 4 to 6. A sufficient number of implants shall 3.4.1 Macroscopic assessment
be inserted to ensure that the final number of speci-
Examine each implant site with the aid of a low mag-
mens to be evaluated will give valid results.
nification lens. Record the nature and extent of any
tissue reaction observed.
3.3.3 Surgery
3.4.2 Preparation for histology - Implant
Anaesthetize the animals. Remove hair from the sur-
retrieval and specimen preparation
gical area by clipping, shaving or other mechanical
means. Wash the area with an antiseptic solution.
Excise the implant together with sufficient unaffected
Ensure that hair does not come in contact with the
surrounding tissue to enable evaluation of the local
implants or the wound surfaces. The specific insertion biological response. Process the excised tissue blocks
or implantation procedures are described in clauses 4
containing test or control implants for histopatho-
to 6.
logical and other studies as appropriate.
The surgical technique may profoundly influence the When conventional techniques are used, the tissue
result of any implantation procedure. The surgery shall envelope may be opened before or after exposure to
3
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IS0 10993-6: 1994(E)
a fixative and the condition of the implant surface and 3.5 Test report
tissue bed shall be reported. However, with this
technique the tissue layers closest to the implant are
3.51 Content of test report
usually destroyed.
The test report shall have sufficient detail to allow in-
When the implant/tissue interface is to be studied,
dependent assessment of the results. The report shall
embedding of the intact tissue envelope with the im-
include the items listed in 3.5.2 to 3.5.6.
plant in situ using hard plastics is preferred. Appropri-
ate sectioning or grinding techniques are employed
for the preparation of histological sections. It shall be
3.5.2 Implant specimens
demonstrated that the technique of embedding in
plastics does not markedly alter the interface tissue.
Description of test and control materials , material
condition, fabrication, surface condition, and the
shape and size of implants.
Report the rationale for selection o f control
material(s).
3.4.3 Histological assessment
The surface preparation of the specimens can affect
the tissue reaction. Therefore, the preparation pro-
The extent of response may be determined by
cedure should be noted in the report.
measurement of the distance from the
Report cleaning, handling and sterilization techniques
implant/tissue interface to unaffected areas with the
employed. If not done in-house, this information
characteristics of normal tissue and of normal
should be supplied b
...
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