Standard Guide for Characterization of Type I Collagen as Starting Material for Surgical Implants and Substrates for Tissue Engineered Medical Products (TEMPs)

SCOPE
1.1 This guide for characterizing collagen-containing biomaterials is intended to provide characteristics, properties, and test methods for use by producers, manufacturers, and researchers to more clearly identify the specific collagen materials used. With greater than 20 types of collagen and the different properties of each, a single document would be cumbersome. This guide will focus on the characterization of Type I collagen, which is the most abundant collagen in mammals, especially in skin and bone. Collagen isolated from these sources may contain other types of collagen, for example, Type III and Type V. This guide does not provide specific parameters for any collagen product or mix of products or the acceptability of those products for the intended use. The collagen may be from any source, including, but not limited to animal or cadaveric sources, human cell culture, or recombinant sources. The biological, immunological, or toxicological properties of the collagen may vary depending on the source material. The properties of the collagen prepared from each of the above sources must be thoroughly investigated, as the changes in the collagen properties as a function of source materials is not thoroughly understood. This guide is intended to focus on purified Type I collagen as a starting material for surgical implants and substrates for Tissue Engineered Medical Products (TEMPs); some methods may not be applicable for gelatin nor for tissue implants. This guide may serve as a template for characterization of other types of collagen.
1.2 The biological response to collagen in soft tissue has been well documented by a history of clinical use (,) and laboratory studies (3,4,5,2.1). Biocompatibility and appropriateness of use for a specific application(s) is the responsibility of the device manufacturer.
The following precautionary caveat pertains only to the test method portion, Section 5, of this guide. 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 and health practices and determine the applicability of regulatory requirements prior to use.

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ASTM F2212-02(2007) - Standard Guide for Characterization of Type I Collagen as Starting Material for Surgical Implants and Substrates for Tissue Engineered Medical Products (TEMPs)
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Designation:F2212–02 (Reapproved 2007)
Standard Guide for
Characterization of Type I Collagen as Starting Material for
Surgical Implants and Substrates for Tissue Engineered
Medical Products (TEMPs)
This standard is issued under the fixed designation F 2212; 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 (e) indicates an editorial change since the last revision or reapproval.
INTRODUCTION
Collagen-based medical devices are becoming more prevalent especially in the area of soft tissue
augmentation. The use of collagen in surgery dates back to the late 1800s, with the use of catgut
sutures, human cadaveric skin, and fascia. More recently, collagen has been used in hemostatic
sponges, dermal equivalents, injectables for soft tissue augmentation, as a matrix for cell-based
products and as a vehicle for drug delivery. It is because of the versatility of collagen in medical
applications that specific characterizations should be performed as a way to compare materials.
1. Scope 1.2 The biological response to collagen in soft tissue has
been well documented by a history of clinical use (1,2) and
1.1 This guide for characterizing collagen-containing bio-
laboratory studies (3,4,5,21). Biocompatibility and appropri-
materials is intended to provide characteristics, properties, and
ateness of use for a specific application(s) is the responsibility
test methods for use by producers, manufacturers, and re-
of the device manufacturer.
searchers to more clearly identify the specific collagen mate-
1.3 The following precautionary caveat pertains only to the
rials used. With greater than 20 types of collagen and the
test method portion, Section 5, of this guide. This standard
different properties of each, a single document would be
does not purport to address all of the safety concerns, if any,
cumbersome. This guide will focus on the characterization of
associated with its use. It is the responsibility of the user of this
Type I collagen, which is the most abundant collagen in
standard to establish appropriate safety and health practices
mammals, especially in skin and bone. Collagen isolated from
and determine the applicability of regulatory requirements
thesesourcesmaycontainothertypesofcollagen,forexample,
prior to use.
Type III and Type V. This guide does not provide specific
parameters for any collagen product or mix of products or the
2. Referenced Documents
acceptability of those products for the intended use. The
2.1 ASTM Standards:
collagen may be from any source, including, but not limited to
E 1298 Guide for Determination of Purity, Impurities, and
animal or cadaveric sources, human cell culture, or recombi-
Contaminants in Biological Drug Products
nant sources. The biological, immunological, or toxicological
F 619 Practice for Extraction of Medical Plastics
properties of the collagen may vary depending on the source
F 720 Practice for Testing Guinea Pigs for Contact Aller-
material. The properties of the collagen prepared from each of
gens: Guinea Pig Maximization Test
the above sources must be thoroughly investigated, as the
F 748 Practice for Selecting Generic Biological Test Meth-
changes in the collagen properties as a function of source
ods for Materials and Devices
materials is not thoroughly understood. This guide is intended
F 749 Practice for Evaluating Material Extracts by Intracu-
to focus on purified Type I collagen as a starting material for
taneous Injection in the Rabbit
surgicalimplantsandsubstratesforTissueEngineeredMedical
F 756 Practice for Assessment of Hemolytic Properties of
Products (TEMPs); some methods may not be applicable for
Materials
gelatin nor for tissue implants. This guide may serve as a
template for characterization of other types of collagen.
The boldface numbers in parentheses refer to the list of references at the end of
This guide is under the jurisdiction of ASTM Committee F04 on Medical and this standard.
Surgical Materials and Devices and is the direct responsibility of Subcommittee For referenced ASTM standards, visit the ASTM website, www.astm.org, or
F04.42 on Biomaterials and Biomolecules for TEMPs. contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Current edition approved Feb. 1, 2007. Published February 2007. Originally Standards volume information, refer to the standard’s Document Summary page on
approved in 2002. Last previous edition approved in 2002 as F 2212 – 02. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
F2212–02 (2007)
F 763 Practice for Short-Term Screening of Implant Mate- United States Pharmacopeia (USP), Edition XXIV (24)
rials USP 24/NF 19 Viral Safety Evaluation of Biotechnology
F 813 PracticeforDirectContactCellCultureEvaluationof Products Derived from Cell Lines of Human or Animal
Materials for Medical Devices Origin
F 895 Test Method forAgar Diffusion Cell Culture Screen- 2.5 Code of Federal Regulations:
ing for Cytotoxicity Code of Federal Regulations, Title 21, Part 820
F 981 Practice for Assessment of Compatibility of Bioma- Federal Register Vol. 43, No. 141, Friday, July 21, 1978
terials for Surgical Implants with Respect to Effect of Human Cells, Tissues and Cellular and Tissue-Based Prod-
Materials on Muscle and Bone ucts, Establishment Registration and Listing. 21 CFR
F 1251 Terminology Relating to Polymeric Biomaterials in Parts 207, 807, and 1271
Medical and Surgical Devices Federal Register/Vol. 66, No. 13, Jan 19, 2001/Rules and
F 1439 Guide for Performance of Lifetime Bioassay for the Regulations, page 5447
Tumorigenic Potential of Implant Materials Suitability Determination for Donors of Human Cell and
F 1903 Practice for Testing For Biological Responses to Tissue-based Products, 21 CFR 1271 Part C, Proposed
Particles in vitro Rule
F 1904 Practice for Testing the Biological Responses to Current Good Tissue Practice for Manufacturers of Human
Particles in vivo Cellular and Tissue-Based Products, Inspection and En-
F 1905 Practice For Selecting Tests for Determining the forcement. Proposed Rule. Federal Register/Vol. 66, No.
Propensity of Materials to Cause Immunotoxicity 5/January 8, 2001/Proposed Rules, pages 1552-1559
F 1906 Practice for Evaluation of Immune Responses In Guidance for Screening and Testing of Donors of Human
Biocompatibility Testing Using ELISATests, Lymphocyte Tissue Intended for Transplantation, Availability. Federal
Proliferation, and Cell Migration Register/Vol. 62, No. 145/July 29, 1997/NoticesDraft
F 1983 Practice for Assessment of Compatibility of Guidance for Preclinical and Clinical Investigations of
Absorbable/Resorbable Biomaterials for Implant Applica- Urethral BulkingAgents used in the Treatment of Urinary
tions Incontinence. November 29, 1995. (ODE/DRARD/
F 2148 Practice for Evaluation of Delayed Contact Hyper- ULDB), Document No. 850
sensitivity Using the Murine Local Lymph Node Assay Guidance for Industry and for FDA Reviewers, Medical
(LLNA) Devices Containing Materials Derived from Animal
2.2 ISO Standards: Sources(Exceptfor In VitroDiagnosticDevices),Novem-
ISO 10993-1 Biological Evaluation of Medical Devices— ber 6, 1998, U.S. Department of Health and Human
Part 1: Evaluation and Testing Services, Food and Drug Administration, Center for De-
ISO 10993-3—Part 3 Tests for Genotoxicity, Carcinogenic- vices and Radiological Health
ity and Reproductive Toxicity CFR 610.13(b), Rabbit Pyrogen Assay
ISO 10993-9—Part 9 Framework for Identification and 2.6 ICH Documents:
Quantification of Potential Degradation Products International Conference on Harmonization (1997) Guid-
ISO 10993-10 Biological Evaluation of Medical Devices— ance for Industry M3 Nonclinical Safety Studies for the
Part 10: Tests for Irritation and Delayed-Type Hypersen- Conduct of Human Clinical Trials for Pharmaceuticals 62
sitivity FR 62922
ISO 10993-17—Part 17 Methods for Establishment of Al- International Conference on Harmonization (1996) Guide-
lowable Limits for Leachable Substances Using Health- line for Industry S2A Specific Aspects of Regulatory
Based Risk Assessment Genotoxicity Tests for Pharmaceuticals. 61 FR 18199
ISO 13408-1: 1998 Aseptic Processing of Health Care International Conference on Harmonization (1997) Guid-
Products—Part 1: General Requirements ance for Industry S2B Genotoxicity: A Standard Battery
2.3 EN (European Norm) Documents: for Genotoxicity Testing of Pharmaceuticals 62 FR 62472
EN 12442-1 Animal Tissues and their Derivatives Utilized International Conference on Harmonization (1994) Guide-
in the Manufacture of Medical Devices—Part 1:Analysis line for Industry S5A Detection of Toxicity to Reproduc-
and Management of Risk tion for Medicinal Products. 59 FR 48746
EN 12442-2—Part 2 Controls on Sourcing, Collection and International Conference on Harmonization (1996) Guid-
Handling ance for Industry S5B Detection of Toxicity to Reproduc-
EN 12442-3—Part 3 Validation of the Elimination and/or tion for Medicinal Products: Addendum on Toxicity to
Inactivation of Virus and Transmissible Agents Male Fertility. 61 FR 15360
2.4 U. S. Pharmacopeia Documents: International Conference on Harmonization (1996) Guide-
line for Industry S1A The Need for Long-term Rodent
Available from International Organization for Standardization (ISO), 1 rue de
Varembé, Case postale 56, CH-1211, Geneva 20, Switzerland.
5 7
Available from European Committee for Standardization, CEN Management AvailablefromU.S.GovernmentPrintingOfficeSuperintendentofDocuments,
Centre 36, rue de Stassart B-1050 Brussels, Belgium. 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401.
6 8
Available from U.S. Pharmacopeia (USP), 12601Twinbrook Pkwy., Rockville, ICH Secretariat, c/o IFPMA, 30 rue de St-Jean, P.O. Box 758, 1211 Geneva 13,
MD 20852. Switzerland.
F2212–02 (2007)
Carcinogenicity Studies of Pharmaceuticals. 61 FR 8153 Devices—Requirements for Products Labeled “Sterile”
International Conference on Harmonization (1998) Guid- 2.9 Other References:
ance for Industry S1B Testing for Carcinogenicity of Draft Guidance for Preclinical and Clinical Investigations of
Pharmaceuticals. 63 FR 8983 UrethralBulkingAgentsUsedintheTreatmentofUrinary
International Conference on Harmonization (1995) Guide- Incontinence, November 29, 1995. (ODE/DRARD/
line for Industry S1C Dose Selection for Carcinogenicity ULDB), Document No. 850.
Studies of Pharmaceuticals. 60 FR 11278
3. Terminology
International Conference on Harmonization (1997)
S1C(R) Guidance for IndustryAddendum to Dose Selec-
3.1 Definitions:
tion for Carcinogenicity Studies of Pharmaceuticals: Ad- 3.1.1 adventitious agents—an unintentionally introduced
dition of a Limit Dose and Related Notes. 62 FR 64259
microbiologicalorotherinfectiouscontaminant.Intheproduc-
International Conference on Harmonization (ICH) Q1A tionofTEMPs,theseagentsmaybeunintentionallyintroduced
ICH HarmonizedTripartiteGuidanceforStabilityTesting
into the process stream or the final product, or both.
of New Drug Substances and Products (September 23, 3.1.2 biocompatibility—a material may be considered bio-
1994)
compatible if the materials perform with an appropriate host
U.S. Food and Drug Administration (FDA and Committee
response in a specific application (22).
for Proprietary Medicinal Products (CPMP), 1998 Inter-
3.1.3 collagen—Type I collagen is a member of a family of
national Conference on Harmonization (ICH), Quality of
structural proteins found in animals. Type I collagen is part of
Biotechnological Products: Viral Safety Evaluation of the fibrillar group of collagens. It derives from the COL1A1
Biotechnology Products Derived from Cell Lines of Hu-
and COL1A2 genes, which express the alpha chains of the
man or Animal Origin, Consensus Guideline ICH Viral collagen. All collagens have a unique triple helical structure
Safety Document: Step 5
configuration of three polypeptide units known as alpha-
2.7 FDA Documents: chains. Proper alignment of the alpha chains of the collagen
FDA Guideline on Validation of the Limulus Amebocyte
molecule requires a highly complex enzymatic and chemical
Test as an End-Product Endotoxin Test for Human and interaction in vivo. As such, preparation of the collagen by
AnimalParenteralDrugs,BiologicalProductsandHealth-
alternate methods may result in improperly aligned alpha
care Products, DHHS, December 1987 chains and, putatively, increase the immunogenicity of the
U.S. Food and Drug Administration (FDA) Center for
collagen. Collagen is high in glycine, L-alanine, L-proline, and
Biologics Evaluation and Research (CBER), 1993 Points 4-hydroxyproline, low in sulfur, and contains no L-tryptophan.
to Consider in the Characterization of Cell Lines Used to
Natural, fibrillar Type I collagen is normally soluble in dilute
produce Biologicals acids and alkalis. When heated (for example, above approxi-
U.S. Food and Drug Administration (FDA) Center for
mately 40°C), collagen is denatured to single alpha chains
Biologics Evaluation and Research (CBER), 1997 Points (gelatin). At each end of the chains are short non-helical
toConsiderintheManufactureandTestingofMonoclonal
domains called telopeptides, which are removed in some
Antibody Products for Human Use, 94D-0259
collagen preparations. Through non-covalent interactions with
FDA Interim Guidance for Human and Veterinary Drug
sites on adjacent helixes, fibrillogenesis is achieved. Subse-
Products and Biologicals, Kinetic LAL techniques,
quently, non-reducible cross-links are formed. Type I collagen
DHHS, July 15, 1991 can be associated with Type III and Type V collagen and also
2.8 AAMI Documents:
with the other non-collagenous proteins like elastin and other
ANSI/AAMI/ISO 11737-1: 1995 Sterilization of Medical structural molecules like glycosaminoglycans and complex
Devices—Microbiological Methods—Part 1: Estimation
lipoproteins and glycoproteins.
of Bioburden on Product 3.1.4 degradation—change in chemical, physical, or mo-
ANSI/AAMI/ISO 11737-2: 1998 Sterilization of Medical
lecular structure or appearance (that is, gross morphology) of
Devices—Microbiological Methods—Part 2:Tests of Ste- material.
rilityPerformedintheValidationofaSterilizationProcess
3.1.5 endotoxin—a high molecular weight lipopolysaccha-
AAMI TIR No. 19-1998 Guidance for ANSI/AAMI/ISO ride (LPS) complex associated with the cell wall of gram-
10993-7: 1995, Biological Evaluation of Medical
negative bacteria that is pyrogenic in humans. Though endot-
Devices—Part 7: Ethylene Oxide Sterilization Residuals o
...

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