BS ISO 17853-2011 Wear of implant materials Polymer and metal wear particles Isolation and characterization《植入物材料的磨损 聚合物和金属磨损颗粒 离析和表征》.pdf

1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS ISO 17853:2011Wear of implant materials Polymer and metal wearparticles Isolation andcharacterizationBS ISO 17853:2011 BRITISH STANDARDNational forewordThis British Standard i

2、s the UK implementation of ISO 17853:2011. Itsupersedes BS ISO 17853:2010 which is withdrawn.The UK participation in its preparation was entrusted toTechnical Committee CH/150/4, Surgical Implants - Bone and JointReplacements.A list of organizations represented on this committee can beobtained on re

3、quest to its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. BSI 2011ISBN 978 0 580 72705 4ICS 11.040.40Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Sta

4、ndard was published under the authority of theStandards Policy and Strategy Committee on 31 March 2011.Amendments issued since publicationDate Text affectedBS ISO 17853:2011Reference numberISO 17853:2011(E)ISO 2011INTERNATIONAL STANDARD ISO17853Third edition2011-03-01Wear of implant materials Polyme

5、r and metal wear particles Isolation and characterization Usure des matriaux dimplant Particules dusure des polymres et des mtaux Isolation et caractrisation BS ISO 17853:2011ISO 17853:2011(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobes licensing policy, th

6、is file may be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In downloading this file, parties accept therein the responsibility of not infringing Adobes licensing policy. The ISO Central Secret

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9、n may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISOs member body in the country of the requester. ISO copyright office Case postale 56 CH-1211 Geneva 20

10、Tel. + 41 22 749 01 11 Fax + 41 22 749 09 47 E-mail copyrightiso.org Web www.iso.org Published in Switzerland ii ISO 2011 All rights reservedBS ISO 17853:2011ISO 17853:2011(E) ISO 2011 All rights reserved iiiContents Page Foreword iv Introduction.v 1 Scope1 2 Terms and definitions .1 3 Principle, re

11、agents and apparatus.1 3.1 Principle .1 3.2 Reagents 2 3.3 Apparatus.2 4 Methods of sampling and analysis of polymer and metal wear particles from tissue samples 3 4.1 Storage and preparation of samples.3 4.2 Procedure for polymer particle isolation 4 4.3 Procedure for metal particle isolation.5 4.4

12、 Collection of particles.6 4.5 Particle size and shape characterization 7 4.6 Particle identification 8 5 Methods of sampling and analysis of polymer and metal particles from joint simulator lubricants .9 5.1 General .9 5.2 Procedure for polymer materials For example UHMWPE and polyetheretherketone

13、(PEEK).9 5.3 Procedure for metal particles.10 5.4 Procedure for ceramic particles 13 6 Test report13 Bibliography15 BS ISO 17853:2011ISO 17853:2011(E) iv ISO 2011 All rights reservedForeword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodi

14、es (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

15、, 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. International Standards are drafted in accordance with the rules given in the IS

16、O/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. 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

17、member bodies casting a vote. 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. ISO 17853 was prepared by Technical Committee ISO/TC 150, Implants for

18、surgery, Subcommittee SC 4, Bone and joint replacements. This third edition cancels and replaces the second edition (ISO 17853:2010), of which it constitutes a minor revision. BS ISO 17853:2011ISO 17853:2011(E) ISO 2011 All rights reserved vIntroduction The biological responses to wear particles con

19、tribute to the failure of joint implants through bone resorption and consequent implant loosening. A standardized method of particle retrieval from the tissues followed by particle characterization is necessary to ensure that the investigations of wear particle effects have a uniform approach. The c

20、haracterization of the particles generated from implants in joint simulators also provides valuable information on the wear properties and performance of the implant being studied. In the protocols included in this International Standard, for isolation and characterization of particles from both tis

21、sues or test fluids from joint simulators, the particles are isolated and then dispersed using filtration or embedding in resin for scanning electron microscopy (SEM) or transmission electron microscopy (TEM) analysis. An alternative protocol for isolation and characterization of metal particles fro

22、m implants tested in joint simulators has recently been developed in which the particles are deposited on to wafers for SEM analysis, without filtration or embedding1. At the time of publication of this International Standard, this alternative method has not been tested for isolation and characteriz

23、ation of particles from tissues and no direct comparison between the different methods is available. Therefore, the latter method has not been included in detail. BS ISO 17853:2011BS ISO 17853:2011INTERNATIONAL STANDARD ISO 17853:2011(E) ISO 2011 All rights reserved 1Wear of implant materials Polyme

24、r and metal wear particles Isolation and characterization 1 Scope This International Standard specifies methods for sampling wear particles generated by joint replacement implants in humans and in joint simulators. It specifies the apparatus, reagents and test methods to isolate and characterize bot

25、h polymer and metal wear particles from samples of tissues excised from around the joint replacement implant, obtained at revision surgery or post mortem, and from samples of joint simulator test fluids. Some of these procedures could certainly be adapted for isolation and characterization of partic

26、les from human biological fluids (e.g. synovial fluid). The methods given in this International Standard do not quantify the level of wear the implant produces; neither do they determine the amount of wear from any particular surface. This International Standard does not cover the biological effects

27、 of wear particles or provide a method for evaluation of biological safety. 2 Terms and definitions For the purposes of this document, the following terms and definitions apply. 2.1 polymer wear particle particle generated from the wear of polymeric components of an implant 2.2 metal wear particle p

28、article and particulate corrosion product generated from the wear of metal components of an implant 2.3 ceramic wear particle particle generated from the wear of ceramic components of an implant 3 Principle, reagents and apparatus 3.1 Principle Particles of polymeric and metal wear are isolated from

29、 tissue samples and simulator lubricants by digestion. The yield of each particle species is then purified by eliminating any remaining organic debris. NOTE The methods involved in polymer and metal particle isolation are different and are described in 4.2 and 4.3, respectively. The particles are co

30、llected, and are characterized and counted (where applicable) using scanning electron microscopy (SEM) or transmission electron microscopy (TEM). BS ISO 17853:2011ISO 17853:2011(E) 2 ISO 2011 All rights reserved3.2 Reagents During the analysis, unless otherwise stated, use only reagents of recognize

31、d analytical grade and distilled water or water of equivalent purity. All reagent solutions shall be filtered through a filter of 0,2 m or smaller pore size prior to use, to avoid contamination of the sample by extraneous particles. 3.2.1 Absolute ethanol. 3.2.2 Acetone, 100 % or diluted with distil

32、led water, with a volume fraction of 80 % acetone. 3.2.3 Distilled water. 3.2.4 Fixative, e.g. formalin, diluted with distilled water, with a volume fraction of 10 % formalin. 3.2.5 Hydrochloric acid solution, HCl, c = 0,01 mol/l. 3.2.6 Isopropanol-water mixtures, = 0,96 g/cm3and = 0,90 g/cm3. 3.2.7

33、 Papain solution, at 4,8 U/1,5 ml of 250 mM sodium phosphate buffer containing 25 mM ethylenediaminetetraacetic acid solution (EDTA), pH 7,4. 3.2.8 Sodium phosphate buffer, at 250 mM containing 25 mM of EDTA, pH 7,4. 3.2.9 Proteinase K, 2 g/ml of 50 mM TRIS-hydrochloride (TRIS-HCl), pH 7,6. NOTE For

34、 particles isolated from joint simulator serum lubricant, the quantity should be adjusted depending on the serum percentage of the lubricant and initial serum volume from which particles are isolated. See 5.3.2. 3.2.10 Resin, epoxy, such as EMbed 812. 3.2.11 Sodium dodecyl sulfate (SDS), 2,5 g/100 m

35、l of distilled water or 3 g/100 ml of 80 % acetone. 3.2.12 Sodium hydroxide, NaOH, solutions and pellets, c = 5 M. 3.2.13 Sucrose solutions, = 1,35 g/cm3, 1,17 g/cm3, 1,08 g/cm3, 1,04 g/cm3and 1,02 g/cm3. 3.2.14 TRIS-hydrochloride buffer, TRIS-HCl, at 50 mM, pH 7,6. 3.3 Apparatus All apparatus shall

36、 be cleaned and triple rinsed with distilled water previously filtered through a filter of 0,2 m pore size (3.3.6) before use to remove any contaminating particles. 3.3.1 Aluminium stub. 3.3.2 Balance, with an accuracy of at least 0,1 mg. 3.3.3 Carbon stickers. 3.3.4 Centrifuge tubes, different size

37、s. 3.3.5 Centrifuge. 3.3.6 Filters, with a pore size of 0,2 m for filtering reagents and distilled water. BS ISO 17853:2011ISO 17853:2011(E) ISO 2011 All rights reserved 33.3.7 Filtration unit. 3.3.8 Formvar-coated copper grids, of 200 mesh size for TEM analysis. 3.3.9 Fourier Transform Infrared (FT

38、IR) Spectroscope. 3.3.10 Heating plate. 3.3.11 Lint-free cloth. 3.3.12 Pipettes, micropipettes and tips. 3.3.13 Polarizing light microscope. 3.3.14 Polycarbonate membrane filters, of pore sizes 10 m, 1 m, 0,1 m, 0,05 m and 0,015 m, for collecting particles. 3.3.15 Scanning electron microscope, SEM,

39、with an energy dispersive X-ray analysis (EDXA) module. 3.3.16 Sterile Petri dishes, with lids. 3.3.17 Syringe, with wide-bore needle. 3.3.18 Teflon-glass potter tissue grinder. 3.3.19 Transmission electron microscope, TEM, with an energy dispersive X-ray analysis (EDXA) module. 3.3.20 Ultrasonic ce

40、ll disrupter, equipped with a titanium microprobe. 3.3.21 Ultrasonic bath. 3.3.22 Water bath, agitating temperature controlled. 4 Methods of sampling and analysis of polymer and metal wear particles from tissue samples 4.1 Storage and preparation of samples Store the tissue at 70 C (or lower) in a f

41、reezer, or at room temperature in a fixative such as formalin (3.2.4), diluted with distilled water (3.2.3), with a volume fraction of 10 % formalin. Thaw the tissue, if applicable, and rinse it thoroughly in distilled water before continuing with the extraction method. Remove excess water from the

42、rinsed tissue by blotting with a lint-free cloth (3.3.11). Unfixed tissue should be handled under universal conditions. The nature of surgical instruments used for sample retrieval should be recorded in case of contamination. NOTE Sampling variability due to specimen origin can occur. BS ISO 17853:2

43、011ISO 17853:2011(E) 4 ISO 2011 All rights reserved4.2 Procedure for polymer particle isolation 4.2.1 Tissue digestion There are many published methods for polyethylene particle isolation from periprosthetic tissues. The methods presented here are based on those of Campbell et al.2, Tipper et al.3an

44、d Richards et al.4. Cut the tissue into smaller pieces using a scalpel and blade before digestion to speed up the digestion times. Extract the lipids from the minced tissue by placing into a 2:1 (volume ratio) chloroform:methanol mixture for 24 h or until the tissue sinks to the bottom of the contai

45、ner. Remove and rinse the tissue with PBS (3.2.8). Add 5 M NaOH (3.2.12) to the tissue (10 ml of 5 M NaOH to 1 g of tissue) and leave to digest for a minimum of 24 h in an agitating water bath (3.3.22) at 65 C. Digestion can be judged to be complete when no visible solid pieces of tissue remain in t

46、he suspension. 4.2.2 Purification of the polymer particle yield 4.2.2.1 General The polymer particles can be purified from the digested tissue in a number of ways. Use one of the methods described in 4.2.2.2 or 4.2.2.3. 4.2.2.2 Purification of polymer particles by high-speed centrifugation This meth

47、od enables all particle sizes to be collected from the nanometre-size range to several millimetres in length, enabling the total wear volume of particles to be isolated. Cool the digested tissue to 4 C. Add an equal volume of ice-cold absolute ethanol (3.2.1). At this point, salts might precipitate.

48、 If this is the case, add ultrapure water until the salt dissolves. Incubate the solution at 4 C overnight with stirring. Centrifuge the solution at 20 000 g for 2 h at 4 C. Decant the supernatant liquid into a clean tube (3.3.4) and dilute with 400 ml of ultrapure water prior to filtration. 4.2.2.3

49、 Purification of polymer particles by ultracentrifugation Place 2 ml of each sucrose solution (3.2.13) ( = 1,35 g/cm3, 1,17 g/cm3, 1,08 g/cm3, 1,04 g/cm3and 1,02 g/cm3) into centrifuge tubes (3.3.4) so that the tubes are roughly three-quarters full, and apply measured aliquots of the digested tissue suspension to the surface of the sucrose solution in each tube. Ultracentrifuge at 100 000 g for 3 h at 5 C. Carefully collect the top layer into a sterile tube and dilute with distilled water at 65 C t