ASTM_F_2148_-_13.pdf

Designation:F214813Standard Practice forEvaluation of Delayed Contact Hypersensitivity Using theMurine Local Lymph Node Assay(LLNA)1This standard is issued under the fixed designation F2148;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of last revision.A number in parentheses indicates the year of last reapproval.Asuperscript epsilon()indicates an editorial change since the last revision or reapproval.1.Scope1.1 This practice provides a methodology to use an in-situprocedure for the evaluation of delayed contact hypersensitiv-ity reactions.1.2 This practice is intended to provide an alternative to theuse of guinea pigs for evaluation of the ability of a devicematerial to stimulate delayed contact hypersensitivity reac-tions.This alternative is particularly applicable for materialsused in devices that contact only intact skin.However,theguinea pig maximization test is still the recommended methodwhen assessing the delayed hypersensitivity response to metalsor when testing substances that do not penetrate the skin but areused in devices that contact deep tissues or breached surfaces.This practice may be used for testing metals,with the excep-tion of nickel-containing metals,unless the unique physico-chemical properties of the materials may interfere with theability of LLNA to detect sensitizing substances.1.3 This practice consists of a protocol for assessing anincrease in lymphocyte proliferation within the nodes drainingthe site of administration on the ears of mice.1.4 The LLNA has been validated only for low-molecular-weight chemicals that can penetrate the skin.The absorbedchemical or metabolite must bind to macromolecules,such asproteins,to form immunogenic conjugates.1.5 This practice is one of several developed for theassessment of the biocompatibility of materials.Practice F748may provide guidance for the selection of appropriate methodsfor testing materials for a specific application.1.6 Identification of a supplier of materials or reagents is forthe convenience of the user and does not imply a single source.Appropriate materials and reagents may be obtained frommany commercial supply houses.1.7 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.1.8 This standard does not purport to address all of thesafety concerns,if any,associated with its use.It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2.Referenced Documents2.1 ASTM Standards:2F619 Practice for Extraction of Medical PlasticsF720 Practice for Testing Guinea Pigs for ContactAllergens:Guinea Pig Maximization TestF748 Practice for Selecting Generic Biological Test Methodsfor Materials and DevicesF750 Practice for Evaluating Material Extracts by SystemicInjection in the Mouse2.2 Other Documents:3ICCVAM NIH Publication No:99-4494 The Murine LocalLymph Node Assay,1999ICCVAM NIH Publication NO:11-7709 Usefulness andLimitations of the Murine Local Lymph Node Assay forPotency Categorization of Chemicals Causing AllergicContact Dermatitis in Humans3.Terminology3.1 Definitions:3.1.1 AOO,nacetone olive oil solution(4:1 v/v)is asuitable nonpolar solvent.3.1.2 aqueous solvent,nin this assay refers to the polarsolvent,saline.3.1.3 DMSO,ndimethylsulfoxide(nonaqueous,suitableorganic solvent).3.1.4 DNCB,n2,4-dinitrochlorobenzene.3.1.5 formalin,na110dilution of 37 to 39%formalde-hyde solution(formaldehyde)in PBS.1This practice is under the jurisdiction ofASTM Committee F04 on Medical andSurgical Materials and Devices and is the direct responsibility of SubcommitteeF04.16 on Biocompatibility Test Methods.Current edition approved June 1,2013.Published August 2013.Originallyapproved in 2001.Last previous edition approved in 2012 as F2148 07(2012).DOI:10.1520/F2148-13.2For referenced ASTM standards,visit the ASTM website,www.astm.org,orcontact ASTM Customer Service at serviceastm.org.For Annual Book of ASTMStandards volume information,refer to the standards Document Summary page onthe ASTM website.3Available from NICEATM,NIEHS,79 Alexander Dr.,Mail Drop EC-17,Research Triangle Park,NC 27709.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 3.1.6 ICCVAM,nInteragency Coordinating Committee onthe Validation of Alternative Methods.3.1.7 nonaqueous solvent,nin this assay refers to theorganic or nonpolar solvent,which shall be dimethylsulfoxide(DMSO)or acetone olive oil(AOO).3.1.8 PBS,nphosphate buffered saline,pH 7.2.3.1.9 positive control,na substance capable of consis-tently stimulating lymphocyte proliferation.3.1.10 saline,n0.9%sodium chloride(aqueous,polarsolvent).3.1.11 TCA,n5%trichloroacetic acid.3.1.12 tritiated thymidine,nH3methyl thymidine,specificactivity 2 Ci/mM(in PBS)I125IUDR-radioactive uridine.3.1.13 vehicle controls,nan aq