ASTM F1439-03(2018)
(Guide)Standard Guide for Performance of Lifetime Bioassay for the Tumorigenic Potential of Implant Materials
Standard Guide for Performance of Lifetime Bioassay for the Tumorigenic Potential of Implant Materials
SIGNIFICANCE AND USE
4.1 This guide is not intended to specify the exact method of conducting a test for any particular material but only to present some of the criteria that should be considered in method design and possible problems that could lead to misleading results. In the development of the actual test protocol, it is recommended that recognized tumorigenesis bioassay procedures be consulted.
4.2 The recommendations given in this guide may not be appropriate for all applications or types of implant materials. These recommendations should be utilized by experienced testing personnel in conjunction with other pertinent information and the requirements of the specific material application.
SCOPE
1.1 This guide is intended to assist the biomaterials testing laboratory in the conduct and evaluation of tumorigenicity tests to evaluate the potential for new materials to evoke a neoplastic response. The procedure is generally reserved only for those materials which have not previously been used for human implantation for a significant period of time.
1.2 Assessment of tumorigenicity is one of several procedures employed in determining the biological response to a material as recommended in Practice F748. It is assumed that the investigator has already determined that this type of testing is necessary for a particular material before consulting this guide. The recommendations of Practice F748 should be considered before a study is commenced.
1.3 Whenever possible, it is recommended that a battery of genotoxicity procedures be initiated and proposed as an alternative to an in-vivo tumorigenicity bioassay. Genotoxicity assays may also be considered as initial screening procedures due to the sensitivity of the assays, the significant reduction in time to gain valuable data, and the desire to reduce the use of animals for testing. Genotoxicity assays that may be considered are outlined in Guides E1262, E1263, E1280, and E2186, and Practices E1397 and E1398. Additionally, other genotoxicity testing which might be considered (but which do not yet have ASTM test methods) include Salmonella/Mammalian-Microsomal Plate Incorporation Mutagenicity Assay, In Vivo Cytogenetics Bone Marrow Chromosomal Damage Assay, BALB/3T3 Morphological Transformation of Mouse Embryo Cells, and the Mouse Micronucleus Assay. The investigator is advised to consider carefully the appropriateness of a particular method for his application after a review of the published literature.
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
General Information
Relations
Standards Content (Sample)
This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: F1439 − 03 (Reapproved 2018)
Standard Guide for
Performance of Lifetime Bioassay for the Tumorigenic
Potential of Implant Materials
This standard is issued under the fixed designation F1439; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
1.1 This guide is intended to assist the biomaterials testing
1.5 This international standard was developed in accor-
laboratoryintheconductandevaluationoftumorigenicitytests
dance with internationally recognized principles on standard-
toevaluatethepotentialfornewmaterialstoevokeaneoplastic
ization established in the Decision on Principles for the
response. The procedure is generally reserved only for those
Development of International Standards, Guides and Recom-
materials which have not previously been used for human
mendations issued by the World Trade Organization Technical
implantation for a significant period of time.
Barriers to Trade (TBT) Committee.
1.2 Assessment of tumorigenicity is one of several proce-
dures employed in determining the biological response to a
2. Referenced Documents
material as recommended in Practice F748. It is assumed that
2.1 ASTM Standards:
the investigator has already determined that this type of testing
E1262 Guide for Performance of Chinese Hamster Ovary
is necessary for a particular material before consulting this
Cell/Hypoxanthine Guanine Phosphoribosyl Transferase
guide. The recommendations of Practice F748 should be
Gene Mutation Assay
considered before a study is commenced.
E1263 Guide for Conduct of Micronucleus Assays in Mam-
1.3 Whenever possible, it is recommended that a battery of
malian Bone Marrow Erythrocytes (Withdrawn 2014)
genotoxicity procedures be initiated and proposed as an alter-
E1280 Guide for Performing the Mouse Lymphoma Assay
native to an in-vivo tumorigenicity bioassay. Genotoxicity
for Mammalian Cell Mutagenicity (Withdrawn 2014)
assays may also be considered as initial screening procedures
E1397 Practice for In Vitro Rat Hepatocyte DNA Repair
due to the sensitivity of the assays, the significant reduction in
Assay (Withdrawn 2013)
time to gain valuable data, and the desire to reduce the use of
E1398 Practice for In Vivo Rat Hepatocyte DNA Repair
animals for testing. Genotoxicity assays that may be consid-
Assay (Withdrawn 2013)
ered are outlined in Guides E1262, E1263, E1280, and E2186,
E2186 Guide for Determining DNA Single-Strand Damage
and Practices E1397 and E1398. Additionally, other genotox-
in Eukaryotic Cells Using the Comet Assay
icity testing which might be considered (but which do not yet
F748 PracticeforSelectingGenericBiologicalTestMethods
have ASTM test methods) include Salmonella/Mammalian-
for Materials and Devices
Microsomal Plate Incorporation Mutagenicity Assay, In Vivo
2.2 Other Documents:
Cytogenetics Bone Marrow Chromosomal Damage Assay,
National Toxicology Program General Statement of Work
BALB/3T3 Morphological Transformation of Mouse Embryo
fortheConductofToxicityandCarcinogenicityStudiesin
Cells, and the Mouse Micronucleus Assay. The investigator is 4
Laboratory Animals
advisedtoconsidercarefullytheappropriatenessofaparticular
OECD Guidelines for Testing of Chemicals: Guideline 451,
method for his application after a review of the published
literature.
1.4 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
responsibility of the user of this standard to establish appro-
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book ofASTM
Standards volume information, refer to the standard’s Document Summary page on
This guide is under the jurisdiction of ASTM Committee F04 on Medical and the ASTM website.
Surgical Materials and Devices and is the direct responsibility of Subcommittee The last approved version of this historical standard is referenced on
F04.16 on Biocompatibility Test Methods. www.astm.org.
Current edition approved Feb. 1, 2018. Published April 2018. Originally Available from National Institute of Environmental Health Sciences, 111 T. W.
approved in 1992. Last previous edition approved in 2013 as F1439 – 03 (2013). Alexander Drive, Research Triangle Park, NC, August 1988. http://
DOI: 10.1520/F1439-03R18. www.niehs.nih.gov/.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
F1439 − 03 (2018)
Carcinogenicity Studies recommendedtheuseofLong-EvansorWistarRatsbecauseof
OECD Guidelines for Testing of Chemicals: Guideline 453, the difficulty of achieving a two-year lifespan for Fischer and
Combined Chronic Toxicity/Carcinogenicity Studies Sprague-Dawley rats.
Good Laboratory Practice for Nonclinical Laboratory Stud-
5.2 The currently accepted level of testing in a particular
ies
site of implantation or medical specialty should be carefully
researched and regulatory requirements determined before a
3. Terminology
study design is finalized to ensure acceptability of the final
3.1 Definitions of Terms Specific to This Standard:
results.
3.1.1 carcinogenic—a substance is considered to be carci-
5.3 The appropriate choice of male or female animals or a
nogenicifitcanbeshowntobecausallyrelatedtoanincreased
combination should be carefully considered in light of the
incidence of malignant neoplastic formation.
particular material and application being investigated. If the
3.1.2 maximum implantable dose—the maximum weight or
device will ultimately be used only in the male or female, only
volume of the test article which can be reasonably implanted
one sex may need to be evaluated. Otherwise, both sexes
into the test site taking into account the gross distention of
should be used.
tissue which can occur and its possible effects on test results.
5.4 The decision to use other species for study should be
3.1.3 mutagenic—a substance is said to be mutagenic if it
carefully documented in terms of a clear need. The use of
induces alterations in the genetic code of the cell.
species which have not previously been used may reduce the
3.1.4 tumorigenic—a substance is said to be tumorigenic if amount of comparative data available on control animals.
it can be shown to be causally related to an increased incidence Typical tumor rates for hamsters, rats, and mice have been
of neoplastic formation whether malignant or benign. tabulated and are available in Refs. (1, 2, 3).
6. Selection of Size and Form of Implant
4. Significance and Use
6.1 Tumorigenicity bioassays have traditionally been per-
4.1 Thisguideisnotintendedtospecifytheexactmethodof
formed using chemical substances as the challenge. The
conducting a test for any particular material but only to present
evaluation of implant materials requires that solid material be
someofthecriteriathatshouldbeconsideredinmethoddesign
implanted in some form. It is important to realize that the
and possible problems that could lead to misleading results. In
down-sized implants necessary for use in animals will have a
the development of the actual test protocol, it is recommended
greatersurfaceareatovolumeratio,andthisdifferencemustbe
that recognized tumorigenesis bioassay procedures be con-
considered in experimental design.
sulted.
6.2 It may be important to determine the site of administra-
4.2 The recommendations given in this guide may not be
tion of the test material that is most appropriate to the end use
appropriate for all applications or types of implant materials.
before determining implant size. The site of implantation
These recommendations should be utilized by experienced
testing personnel in conjunction with other pertinent informa- should be the paravertebral muscle unless the size of the
implant causes this site to be unacceptable. Alternatively, the
tion and the requirements of the specific material application.
site of implantation should mimic the anticipated end use, if
5. Choice of Animal Model possible. Where a specific material may be utilized in more
than one type of device, multiple sites of administration should
5.1 These types of bioassays for chemical substances have
be considered if different types of tissue will be contacted. (For
traditionally been performed in mice or rats, or both, because
instance, materials that may be in contact with bone or
of their small size, relative cost factors, and lifespan. For the
implanted into internal organ tissue might be tested in both
testing of biomaterials, mice are not recommended because the
tissues.)
small animal size is not conducive to the placement of solid
implants. The investigator should seriously consider the use of 6.3 It should be recognized that the response of the test
one of the traditional models in order to draw upon the animal to an extract of a material may not fully represent the
extensive information available about typical tumor formation response that might be seen if the material itself were to be
rates and sites in control animals. The National Toxicology implanted. In general, an extract should not be used as a
Program recommends the use of Fischer 344 (F344/N) rats. substitute for the actual material of interest.
However, other readily available species and strains may also
6.4 The physical form of the test material should be repre-
be acceptable for the performance of these studies. Other rat
sentative of that intended for use in human patients and should
species which have been recommended include Sprague-
consider potential material debris, if appropriate. The investi-
Dawley, Long-Evans, and Wistar. Some investigators have
gator should be aware that tests have shown (4) that powdered
polymeric materials may not elicit a tumorigenic response
subcutaneously even when prepared from polymers that do
Available from Organization for Economic Cooperation and Development,
induce tumors when implanted in the form of a film. The
2001 L Street, N.W., Suite 650, Washington, D.C. 20036-4922. http://
www.oecd.org/washington/contact.htm.
Available from 21 CFR, Part 58, U.S. Government Printing Office Superinten-
dent of Documents, 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC The boldface numbers in parentheses refer to the list of references at the end of
20401, http://www.access.gpo.gov. this guide.
F1439 − 03 (2018)
impact of physical form and surface properties on tumorigen- each group is adequate for statistical and regulatory purposes
esis must be carefully considered, in making decisions about before proceeding. In order to ensure valid data analysis, the
the physical form of the implants (5, 6, 7, 8, 9, 10). animals should be randomly assigned to control and experi-
mentalgroups.Considerationsspecifictotheparticularimplant
6.5 Researchers have found that the aspect ratio (length/
applicationormedicalspecialtymaymandateagreaternumber
diameter) of fiber materials may play a role in the tumorigen-
of animals in each group. Additional animals in interim
esis of a particular material (11, 12). When new fibrous
sacrifice groups or satellite groups may be added.
materials are being tested, the actual fiber length to be
anticipated in practice should be studied. If fragmentation can
8.2 The number of test animals in each group shall be
be anticipated or is a worse case possibility, an attempt should
determined based upon a sound statistical analysis of the
be made to document a clinically relevant fiber length.
scientific questions to be addressed by the study. This analysis
should take into account predicted survival rates (if available)
6.6 The material to be tested should originate from
for the species being used as well as being consistent with
sample(s) representative of all processing including surface
responsible use of experimental animals. If a statistically valid
finishing,passivation,andsterilizationorotherfinalprocessing
experimentcanbeperformedwithfewerthantheusualnumber
that will occur to a finished device.
of animals per group, that fact should be documented and the
6.7 Dosage:
study design should proceed accordingly.
6.7.1 Inmostmaterials,theratiobetweenthesurfaceareaof
the implant and the body weight of the animal or person will
9. Duration of Study
have an effect on the amount of extractable substances (if any)
9.1 Recommended durations for evaluation of tumorigenic-
which leach out of the material. The total weight or volume of
ity in rats is two years.
material used in each animal should be in excess of the
anticipated dosages to be seen in clinical practice when
9.2 Depending upon the material being evaluated, the early
calculated based upon the ratio of surface area of sample to
results may suggest that the study can be terminated earlier
body weight of the animal. Consideration should be given to
than two years without compromising the validity of the study.
using the maximum implantable dose as the dosage or as one
Examples might include studies in which a significantly
of multiple dosage levels. For the special case of degradable
increased rate of tumor formation or toxicity is being seen in
materials, the sample size should be calculated based on the
the test animals or in one or more dosage groups.
ratio of sample weight to animal body weight.
9.3 Attheterminationofthestudy,amajorityoftheanimals
6.7.2 Whenever possible, more than one exposure level
in each group should have survived for euthanasia or been
should be considered to evaluate a dose-response effect.
terminated early for study-related reasons such as increased
7. Choice of Control tumor incidence, spontaneous tumors, or toxicity of the test
article. It is expected that a minimum of 50 % of the animals
7.1 Control groups for this type of study will usually consist
per sex and per group should survive until final study termi-
of identical animals that have not received an implant of the
nation barring the above reasons. Moreover, the number of
test material but have been subjected to the remainder of the
survivors or study-related terminations should be sufficient for
surgical procedures. Additional groups such as housing (ani-
detection of effects at the p < 0.05 level of significance. If
mals which receive no treatment but are housed
...
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