BIOCOMPATIBILITY TESTING AT SGS...different from both USP and ISO tests. SGS highly recommends...

BIOCOMPATIBILITY TESTING AT SGS For over thirty years, SGS has conducted biocompatibility testing for the medical device and pharmaceutical industries. Our facility in NJ includes a 10,000 square foot vivarium which is registered and inspected by the USDA and US FDA. With ISO 9001 certified operations, SGS is certain to meet your quality and regulatory requirements.

Our experienced staff can help you design a cost-effective safety test program for your product. We provide next-day quotes on most biocompatibility testing projects. And we are dedicated to providing you with clear, well-written reports and prompt, personalized service. Please call Business Development at (888) 747 8782 to discuss your testing requirements, or visit our website at www.us.sgs.com/lifescience.

SGS�s testing capabilities for medical device companies include the following:

Biocompatibility Tests

! Cytotoxicity ! Genotoxicity ! Subchronic Toxicity ! Sensitization ! Implantation ! Systemic Toxicity ! Irritation ! Hemocompatibility

QA/QC Testing

! Bioburden ! Biological Indicator Tests ! Bacterial Endotoxin (LAL) ! AAMI/ISO Dose Audits ! Environmental Monitoring

Validation Support

! AAMI/ISO Sterilization Validation ! Cleaning, Disinfection, ! Accelerated Aging and ! Reusable Device and Sterilization Validation Expiration ! Package Integrity Testing

Extractable Material Characterization

! USP Physiochemical Tests ! Sterilant Residues ! Total Organic Carbon (TOC) � Plastics or Elastomeric Closures ! AA, IR, GC, HPLC ! Organic Solvent Residues

United States Pharmacopeia (USP) – an independent, not-for-profifit, nongovernmental organization to improve global health through public standards and related programs.

SGS L i f e Sc ienc e Se rv i c es

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ASSESSING BIOCOMPATIBILITY A GUIDE FOR MEDICAL DEVICE MANUFACTURERS

TABLE OF CONTENTSBiocompatibility Testing at SGS.......................................................................................................................................................................................1 Introduction to Biocompatibility Testing.....................................................................................................................................................................3

What is Device Biocompatibility?.................................................................................................................................................................................3 What are the FDA and EU/ISO Requirements for Biocompatibility Testing? ..............................................................................................................3 Do I Need Biocompatibility Data? ................................................................................................................................................................................4 How Do I Determine which Tests I Need?...................................................................................................................................................................4 Should I Test Device Materials, or only a Composite of the Finished Device? ...........................................................................................................5 Is GLP Treatment Required for Biocompatibility Testing?...........................................................................................................................................5

The Biocompatibility Planning Tool .............................................................................................................................................................................6 All About Extracts.............................................................................................................................................................................................................7 Sample Preparation .........................................................................................................................................................................................................8 Formulas for Surface Area Calculation ............................................................................................................................................................................8 ISO 10993 - Listing of Individual Parts.............................................................................................................................................................................9 Device Categories – Definitions & Examples.................................................................................................................................................................10 Non-Contact Devices .....................................................................................................................................................................................................10 ISO Materials Biocompatibility Matrix........................................................................................................................................................................11 Test Turnaround Time and Sample Requirements...................................................................................................................................................12 Biological Test Methods .................................................................................................................................................................................................13

Cytotoxicity (Tissue Culture)......................................................................................................................................................................................13 Sensitization Assays..................................................................................................................................................................................................13 Irritation Tests ............................................................................................................................................................................................................14 Acute Systemic Toxicity.............................................................................................................................................................................................14 Subchronic Toxicity....................................................................................................................................................................................................14 Genotoxicity ...............................................................................................................................................................................................................15 Implantation Tests .....................................................................................................................................................................................................15 Hemocompatibility .....................................................................................................................................................................................................16 Carcinogenesis Bioassay ..........................................................................................................................................................................................17 Reproductive and Developmental Toxicity ................................................................................................................................................................17 Pharmacokinetics ......................................................................................................................................................................................................17 Preclinical Safety Testing ..........................................................................................................................................................................................18 Histopathology Services ............................................................................................................................................................................................18

Analytical Testing of Biomaterials ..................................................................................................................................................................................19 Material Characterization ...............................................................................................................................................................................................20 SGS’s Commitment to Excellence .................................................................................................................................................................................21

Quality Systems.........................................................................................................................................................................................................21 cGMP Compliance.....................................................................................................................................................................................................21 GLP Compliance........................................................................................................................................................................................................21 Accreditations ............................................................................................................................................................................................................22

References.....................................................................................................................................................................................................................22 Contact Information........................................................................................................................................................................................................22

REQUEST FOR TESTING � Please photocopy the form at the end of this booklet when submitting samples for testing.

To view this booklet online, go to www.us.sgs.com/publications_us Rev. 1.1, 05-07

As s es s ing B ioc ompa t i b i l i t y

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INTRODUCTION TO BIOCOMPATIBILITY TESTING By: SGS Life Science Services

WHAT IS DEVICE BIOCOMPATIBILITY?

The word biocompatibility refers to the interaction between a medical device and the tissues and physiological systems of the patient treated with the device. An evaluation of biocompatibility is one part of the overall safety assessment of a device. Biocompatibility of devices is investigated using analytical chemistry, in vitro tests, and animal models. The biocompatibility of a device depends on several factors, including:

• the chemical and physical nature of its component materials • the types of patient tissue that will be exposed to the device • the duration of that exposure.

While in use, substances may leach off of a medical device into adjacent tissue. Some leachables or extractables are not biologically safe. Many of the various biocompatibility tests include sample preparation steps designed to rinse the leachables and extractables form the device. The extract is then tested. The actual device material may be used in some other tests.

Of course, the primary purpose of a device biocompatibility assessment is to protect patient safety. Manufacturers should consider both corporate regulatory goals and compliance risks and potential future product liability exposure in planning a biocompatibility testing program. Ultimately, evaluating the biocompatibility of a device is a risk assessment exercise. There is no risk-free device or device material. The goal of device designers is to minimize risk while maximizing benefit to patients.

WHAT ARE THE FDA AND EU/ISO REQUIREMENTS FOR BIOCOMPATIBILITY TESTING?

The best starting point for understanding biocompatibility requirements is ISO Standard 10993, Biological Evaluation of Medical Devices. Part 1 of the standard is the Guidance on Selection of Tests, Part 2 covers animal welfare requirements, and Parts 3 through 19 are guidelines for specific test procedures or other testing-related issues. (A list of the individual sections of ISO 10993 can be found on page 9.)

Testing strategies that comply with the ISO 10993 family of documents are acceptable in Europe and Asia. In 1995, FDA issued a Blue Book Memorandum G95-1, in which it substantially adopted the ISO guideline. However, in some areas FDA�s testing requirements go beyond those of ISO.

The specific ISO test procedures vary slightly from the USP procedures historically used for FDA submissions. The ISO procedures tend to be more stringent, so companies planning to register their product in both Europe and the U.S. should follow ISO test methods. FDA requirements should be verified since additional testing may be needed. Japanese procedures for sample preparation and testing are slightly different from both USP and ISO tests.

SGS highly recommends discussing your proposed biocompatibility testing plan with an FDA reviewer before initiating testing

The FDA G95-1 has been replaced by "Use of International Standard ISO-10993, ‘Biological Evaluation of Medical Devices - Part 1: Evaluation and Testing.’”


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DO I NEED BIOCOMPATIBILITY DATA?

Biocompatibility data of one kind or another is almost always required for devices that have significant tissue contact. Refer to the chart from ISO 10993-1 (page 11) to help determine if your device needs biocompatibility testing.

Most commonly, companies arrange for their own biocompatibility studies. You may be able to reduce the amount of testing you will need on a specific device if you have some or all of the following types of biocompatibility data.

1. Data from previous submissions � If data is available from a previous submission, consider the following points as you apply it to your current device. You will need to perform confirmatory testing if there are significant changes in any of these areas.

a. Materials selection b. Manufacturing processes c. Chemical composition of materials d. Nature of patient contact e. Sterilization methods

2. Data from suppliers of materials or components � If vendor data is used, manufacturers should obtain copies of the original study reports. It is important that the laboratory that generated the reports had an experienced staff and a strong track record of cGMP/GLP compliance. Usually manufacturers should perform some confirmatory testing of their own (e.g. cytotoxicity and hemocompatibility studies) to support vendor data. Note that molding, fabrication, gluing, welding, assembly and sterilization can affect the biocompatibility of a medical device.

3. Analytical data � Manufacturers may use analytical data to help demonstrate that a device has a low overall risk or a low risk of producing a given biological effect. Section 18 of ISO Standard 10993, Chemical Characterization of Materials, gives some guidance on this process. (See also pages 19 -20.)

4. Clinical data � Clinical data can be used to satisfy some biological effects categories from the ISO 10993-1 test selection matrix. The data may come from clinical trials of the device in question, or from clinical experience with predicate devices or devices containing similar components or materials.

HOW DO I DETERMINE WHICH TESTS I NEED?

The core of the ISO Standard is confirmation of the suitability of device material for its intended application. The first step in this process is chemical characterization of device components. See page 19 for specifics of such a program.

Biological testing is probably the most critical step in a biocompatibility evaluation. The ISO materials biocompatibility matrix (page 11) categorizes devices based on the type and duration of body contact. It also presents a list of potential biological effects. For each device category, certain effects must be considered and addressed in the regulatory submission for that device. ISO 10993-1 does not prescribe a specific battery of tests for any particular medical device. Rather, it provides a framework that can be used to design a biocompatibility testing program.

Device designers should generally consult with an experienced device toxicologist and their clinical investigators to determine how best to meet the requirements of the materials biocompatibility matrix. For each biological effect category, the rationale for the testing strategy should be documented. This is especially true when a manufacturer decides not to perform testing for an effect specified by the matrix for their category of devices.

cGMP - current good manufacturing practice GLP - good laboratory practice


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SHOULD I TEST DEVICE MATERIALS, OR ONLY A COMPOSITE OF THE FINISHED DEVICE?

As a manufacturer, you should gather safety data on every component and material used in a device. In addition, you should definitely conduct testing on the finished device as specified by ISO 10993-1. Generally, the best approach is to:

1. assemble vendor data on candidate materials

2. conduct analytical and vitro screening of materials

3. conduct confirmatory testing on a composite sample from the finished device.

There is a risk in testing the finished device without developing data on component materials. If an adverse result occurs, it can be difficult to track down the component that is causing the problem. You may end up delaying your regulatory submission while you repeat testing on the individual components.

Screening device materials minimizes this risk. The initial chemical characterization should detect leachable materials that could compromise device safety. Inexpensive non-animal studies (such as cytotoxicity and hemocompatibility tests) provide an additional screen for material safety. Material screening tests also help insure that you will not be forced to redesign your device due to biocompatibility test failures. Many manufacturers assemble data on a library of qualified materials used in their products.

Some test procedures do not lend themselves to testing of composite samples. Due to physical limitations, agar overlay or direct contact cytotoxicity tests and implant studies require separate testing of each device component.

For all biocompatibility studies, test samples should be sterilized using the same method as will be used for the finished device.

IS GLP TREATMENT REQUIRED FOR BIOCOMPATIBILITY TESTING?

As a general rule, all biocompatibility testing should be performed in compliance with Good Laboratory Practice (GLP) regulations (FDA or OECD).

GLP regulations apply to biological safety studies conducted in support of regulatory submissions. They govern all phases of testing, including preparation and approval of study protocols, monitoring tests in progress, and issuance of final reports, as well as facility and study management and the role of the Quality Assurance Unit.

GLP treatment is explicitly required for IDE and PMA submissions. FDA reviewers say they strongly prefer GLP treatment for studies supporting 510(k)s. For European submissions, ISO 10993-1 seems to require GLP treatment, but the wording is somewhat ambiguous. In practice, studies are usually not rejected for lack of GLP treatment.

Manufacturers of device components and materials should have their biocompatibility studies done per GLP so that their clients can use the data in any type of regulatory submission.


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THE BIOCOMPATIBILITY PLANNING TOOL Device companies spend a tremendous amount of time, money and energy developing and implementing biocompatibility testing programs. We have developed this Biocompatibility Planning Tool to guide you through the basic concepts of device testing and to help manufacturers select testing procedures to comply with current regulatory requirements.

The chart below gives you an overview of the process. Follow the page references to get more detail on each specific topic. For information on materials characterization and analytical testing of devices, see page 19-20.

THE BIOCOMPATIBILITY PLANNING TOOL

Refer to the Materials Biocompatibility Matrix on page 11 to determine which biological test categories and procedures are appropriate for your device. Pages 13-18 discuss specific test procedures.

More information on extracts and sample preparation is found on pages 7 and 8. Page 8 shows formulas used to calculate the surface area of your device.

Test

Pla

nnin

g Te

stin

g

Device category definitions and examples of devices that fall into each category are given on page 10. See page 10 for definitions of the three device duration categories.

What is the device category and body contact duration?

Alternative sources for biocomp data are presented on page 4.

Test

ing

See page 12 for sample requirements for each test. Or, contact SGS for a price quote which will specify sample requirements for each test procedure.

GLP treatment is required for certain types of regulatory submissions. More information can be found on page 5.

Please call Business Development at (888) 747 8782.

Sign and return the quote acceptance form, complete and submit the Request for Testing form on pages 23-24, and send sterilized samples to SGS.

What biocompatibility data is already available?

How do I select tests from the Biocompatibility Matrix?

How do I decide on extracting media and conditions?

How do I determine test sample requirements?

Should the studies be performed following Good Laboratory Procedures (GLP) regulations?

How do I determine testing costs?

How do I initiate testing?


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ALL ABOUT EXTRACTS

Medical device biocompatibility problems are most often caused by toxins that leach out of the device into the surrounding tissues or body fluids. So in the laboratory, extracts of device materials are often used in assessing biocompatibility. These extracts are generally prepared using exaggerated conditions of time and temperature to allow a margin of safety over normal physiological conditions.

Analytical extraction studies allow the chemist to identify and quantitate specific leachable substances. This data can in turn help the device toxicologist or risk assessor determine the worst case scenario for patient exposure and the risk to patient health.

Extracts are also used in many of the biological tests specified by ISO 10993. Table 1 at the bottom of this page lists the most commonly used extracting media. For most devices, only saline and vegetable oil extracts are needed.

Extracts are selected on the basis of the biological environment in which the test material is to be used. A saline (SCI) extract approximates the aqueous, hydrophilic fluids in the body. It also permits the use of extreme temperatures in preparing the extracts, thus simulating certain sterilization conditions.

Tissue culture media may even more closely approximate aqueous body fluids, but cannot be used for high temperature extractions. Vegetable oils are non-polar, hydrophobic solvents and simulate the lipid fluids in the body. For technical reasons, DMSO extracts are often used in certain genotoxicity and sensitization tests. Two other common extracting media � Alcohol in SCI and PEG � should be used only if they approximate the solvent properties of drugs or other materials that will contact the device during its normal use.

Extraction conditions (temperature and time) should be at least as extreme as any conditions the device or material will encounter during sterilization or clinical use. Generally, you will want to choose the highest extraction temperature that does not melt or fuse the material or cause chemical changes. To provide some margin of safety for use conditions, SGS recommends an extraction condition of at least 50°C for 72 hours. For devices that are susceptible to heat, an extraction condition of 37°C for 72 hours may be acceptable. Table 2 lists common extraction conditions.

TABLE 1: EXTRACTING MEDIA Sodium Chloride for Injection, USP (SCI)

Vegetable Oil

1:20 Alcohol in SCI

Polyethylene Glycol 400 (PEG)

DMSO

Clinically Relevant Solvents

TABLE 2: EXTRACTION CONDITIONS 37°C for 24 hours

37°C for 72 hours

50°C for 72 hours

70°C for 24 hours

121°C for 1 hour

Other Conditions (justification required)

Dimethyl sulfoxide (DMSO) is an organosulfur compound with the formula (CH3)2SO.


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SAMPLE PREPARATION

The simplest method for determining the surface area of a device is usually to use the CAD program from the design engineering group. Typically the surface area can be calculated with a just a few keystrokes. Alternatively, you can calculate the surface area using the equations below. Or you can submit a sample device and/or an engineering drawing to SGS, and our staff will perform the calculations.

Typically, the standard surface area of your device is used to determine the volume of extract needed for each test performed. This area includes the combined area of both sides of the device but excludes indeterminate surface irregularities. If the surface area cannot be determined due to the configuration of the device, a mass/volume of extracting fluid can be used. In either case, the device is cut into small pieces before extraction to enhance exposure to the extracting media. In some cases, it is not appropriate to cut the device; such devices are tested intact.

The table on page 12 lists the amount of sample required for many procedures. Generally, we recommend using the ratio of sample to extracting media specified in ISO 19993-12 (i.e. either 6 cm²/mL or 3 cm²/mL, depending on the thickness of the test material). For some types of materials, the ratio used for USP Elastomeric Closures for Injections (1.25 cm² per mL) is preferred.

FORMULAS FOR SURFACE AREA CALCULATION Device Shape Formula Square or Rectangle A = L x W

Hollow Cylinder A = (ID + OD) π x L

Disk A (one side) = π r2

Ellipse A = (π x X x Y)/4

Regular Polygon A = (b x h x n)/2

Solid Cylinder (including ends) A = (OD x π x L) + (2 π r2)

Triangle A = (b x h)/2

Sphere A = 4 x π r2

Trapezoid A = (h x [p + q])/2

Circular Ring 4 π2Rrrc

Legend

A = surface area

OD = outer diameter

W = width

RR = ring radius (circular ring)

X, Y = longest and shortest distances through the center of an ellipse

h = height

p, q = length of the parallel sides of a trapezoid

ro = ½ OD

ID = inner diameter

L = length

R = radius

rc = cross section radius (circular ring)

π = 3.14

b = base length

n = number of sides of a polygon

ri = ½ ID


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ISO 10993 - BIOLOGICAL EVALUATION OF MEDICAL DEVICES LISTING OF INDIVIDUAL PARTS

Part Topic

1 Evaluation and Testing

2 Animal Welfare Requirements

3 Tests for Genotoxicty, Carcinogenicity, and Reproductive Toxicity

4 Selection of Tests for Interactions with Blood

5 Tests for Cytotoxicity � In Vitro Methods

6 Tests for Local Effects after Implantation

7 Ethylene Oxide Sterilization Residuals

8 Selection and Qualification of Reference Materials for Biological Test

9 Framework for Identification & Quantification of Potential Degradation Products

10 Test for Irritation and Sensitization

11 Test for Systemic Toxicity

12 Sample Preparation and Reference Materials

13 Identification and Quantification of Degradation Products from Polymers

14 Identification and Quantification of Degradation Products from Ceramics

15 Identification and Quantification of Degradation Products from Coated and Uncoated Metals and Alloys

16 Toxicokinetic Study Design for Degradation Products and Leachables

17 Establishment of Allowable Limits for Leachable Substances

18 Chemical Characterization of Materials*

19 Physicochemical, Mechanical and Morphological Characterization (Draft)

20 Principles and Methods for Immunotoxicology Testing of Medical Devices (Draft)

* ANSI/AAMI (The United States ISO Member Body) is considering a version of this document for use in the U.S.


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DEVICE CATEGORIES – DEFINITIONS & EXAMPLES

Device Categories Examples

Skin Devices that contact intact skin surfaces only. Examples include electrodes, external prostheses, fixation tapes, compression bandages and monitors of various types.

Mucous membrane

Devices communicating with intact mucosal membranes. Examples include contact lenses, urinary catheters, intravaginal and intraintestinal devices (stomach tubes, sigmoidoscopes, colonoscopes, gastroscopes), endotracheal tubes, bronchoscopes, dental prostheses, orthodontic devices and IUD�s.

Surface Device

Breached or compromised surfaces

Devices that contact breached or otherwise compromised external body surfaces. Examples include ulcer, burn and granulation tissue dressings or healing devices and occlusive patches.

Blood path indirect

Devices that contact the blood path at one point and serve as a conduit for entry into the vascular system. Examples include solution administration sets, extension sets, transfer sets, and blood administration sets.

Tissue/bone/dentin communicating

Devices communicating with tissue, bone, and pulp/dentin system. Examples include laparoscopes, arthroscopes, draining systems, dental cements, dental filling materials and skin staples. This category also includes devices which contact internal tissues (rather than blood contact devices). Examples include many surgical instruments and accessories.

External Communicatin

g Device

Circulating blood

Devices that contact circulating blood. Examples include intravascular catheters, temporary pacemaker electrodes, oxygenators, extracorporeal oxygenator tubing and accessories, hemoadsorbents and immunoabsorbents.

Tissue/bone

Devices principally contacting bone. Examples include orthopedic pins, plates, replacement joints, bone prostheses, cements and intraosseous devices. Devices principally contacting tissue and tissues fluid. Examples include pacemakers, drug supply devices, neuromuscular sensors and stimulators, replacement tendons, breast implants, artificial larynxes, subperiosteal implants and ligation clips.

Implant Device

Blood

Devices principally contacting blood. Examples include pacemaker electrodes, artificial arteriovenous fistulae, heart valves, vascular grafts and stents, internal drug delivery catheters, and ventricular assist devices.

NON-CONTACT DEVICES

These are devices that do not contact the patient's body directly or indirectly. Examples include in vitro diagnostic devices. Regulatory agencies rarely require biocompatibility testing for such devices.

Intrauterine Device (IU): birth control

***


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ISO MATERIALS BIOCOMPATIBILITY MATRIX Medical Device Categorization by

Contact Duration Biological Effect

Initial Evaluation Tests Supplementary Evaluation Tests

Nature of Body Contact

Category Contact

A - Limited (< 24 hours) B - Prolonged (24 hours-30 days) C - Permanent (> 30 days)

Cyt

otox

icity

Sen

sitiz

atio

n

Irrita

tion

or In

tracu

tane

ous

Rea

ctiv

ity

Sys

tem

ic T

oxic

ity (a

cute

)

Sub

acut

e an

d S

ubch

roni

c To

xici

ty

Gen

otox

icity

Impl

anta

tion

Hem

ocom

patib

ility

Chr

onic

Tox

icity

Car

cino

geni

city

Rep

rodu

ctiv

e/D

evel

opm

enta

l!

Bio

degr

adat

ion!

A ! ! ! B ! ! ! Skin

C ! ! ! A ! ! !

B ! ! ! F F F Mucosal Membrane

C ! ! ! F ! ! F F

A ! ! ! F

B ! ! ! F F F

Surface Device

Breached or Compromised

Surface C ! ! ! F ! ! F F

A ! ! ! ! ! B ! ! ! ! F ! Blood Path, Indirect

C ! ! F ! ! ! F ! ! ! A ! ! ! F B ! ! ! ! ! ! ! Tissue/Bone/Dentin"

C ! ! ! ! ! ! ! ! ! A ! ! ! ! F# ! B ! ! ! ! ! ! ! !

External Communicating

Device

Circulating Blood

C ! ! ! ! ! ! ! ! ! !

A ! ! ! F B ! ! ! ! ! ! ! Tissue/Bone

C ! ! ! ! ! ! ! ! ! A ! ! ! ! ! ! ! B ! ! ! ! ! ! ! !

Implant Device

Blood

C ! ! ! ! ! ! ! ! ! !

This table is only a framework for the development of an assessment program for your device and is not a checklist.

� = ISO Evaluation Tests for Consideration Note" Tissue includes tissue fluids and subcutaneous spaces

F = Additional Tests which may be required Note# For all devices used in extracorporeal circuits by FDA for US submissions Note! Depends on specific nature of the device and its component

materials

Consult with the FDA before performing any biocompatibility testing if you are submitting an IDE or you have a device/drug combination.

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TEST TURNAROUND TIME AND SAMPLE REQUIREMENTS

SAMPLE AMOUNT" TURN AROUND REQUIREMENT TEST NAME Surface Area (cm2) Weight (gram or mL) (in weeks)

USP Agar Overlay 1 cm2 x 2 pieces USP MEM Elution 1 cm2 x 2 pieces USP Direct Contact 60# ISO Agar Overlay 1 cm2 x 3 pieces ISO MEM Elution 1 cm2 x 3 pieces

Cytotoxicity

ISO Direct Contact 60#

2 2 � 3

Maximization Test 240# 16 8 Sensitization Closed Patch Test 1 in2 x 130 pieces 60-80 8 � 9

USP Intracutaneous Test 60# 4 ISO Intracutaneous Test 60# 4

3

ISO Dermal Irritation 60# 10 FHSA Primary Skin Irritation NA 10

3 � 4

ISO Ocular Irritation 60# 10 FHSA Primary Eye Irritation NA 10

3 � 4

Irritation

Mucous Membrane Irritation 60# Varies Varies

USP Systemic Injection Test 60# 4 3 Systemic Toxicity

Material Mediated Pyrogen Test 10 devices/540 cm2 4 2 - 3

Intraperitoneal Test 12 devices 55 6 � 7 Intravenous Test Implant Tests

12 devices $ Varies

Subchronic (14 � 180 Days)

Other Procedures Varies Varies Varies Ames Test 120# 8 6

Mouse Micronucleus Assay 120# 8 11 Genotoxicity Chromosomal Aberration Test 120# 8 12 Implantation Test Acute 7 Day Subchronic 14 � 180 Day Chronic > 180 Day

12 strips 1 x 10 mm

3

7 � 28 54

Implantation

� Histopathology NA 3 � 4 Hemolysis � Direct Contact (duplicate) NA 4 2 Hemolysis � Direct Contact (triplicate) 3 devices 6 2 Hemolysis � Sample Extract (duplicate) 120# NA 2 Hemolysis � Sample Extract (triplicate) 3 devices NA 2

In Vitro Platelet Aggregation Assay 240# 10 4 - 6 In Vitro Hemocompatibility Assay 150# 10 4 - 6 Partial Thromboplastin Time (PTT), Prothrombin Time (PT) 60# 4 4 - 6

Hemocompatibility

Complement Activation 120# 2 4 - 7 Long Term Implant Inquire Inquire

Chronic Lifetime Toxicity Inquire Inquire Carcinogenesis Lifetime Toxicity Inquire Inquire

USP Physicochemical Tests 720 NA 2 Infrared Scan 5 cm x 1 cm (min.) NA 2 Analytical Tests Other Procedures Inquire Inquire

" - Sample requirements based on surface area calculations. The weight of the device may be used if the surface area cannot be calculated. # - Double these amounts for materials < 0.5 mm in thickness ! - Depends on duration of implant $ - 15 strips per time point, each strip 1 mm x 10 mm, sample should be supplied by Sponsor in specified size, separately packaged

and sterilized, ends should be smooth and rounded

***


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BIOLOGICAL TEST METHODS The following pages describe some of the specific procedures recommended for biocompatibility testing. This listing does not imply that all procedures are necessary for any given device, nor does it indicate that these are the only available tests.

CYTOTOXICITY (TISSUE CULTURE)

Cell culture assays are used to assess the biocompatibility of a material or extract through the use of isolated cells in vitro. These techniques are useful in evaluating the toxicity or irritancy potential of materials and chemicals. They provide an excellent way to screen materials prior to in vivo tests.

There are three cytotoxicity tests commonly used for medical devices. The Direct Contact procedure is recommended for low density materials, such as contact lens polymers. In this method, a piece of test material is placed directly onto cells growing on culture medium. The cells are then incubated. During incubation, leachable chemicals in the test material can diffuse into the culture medium and contact the cell layer. Reactivity of the test sample is indicated by malformation, degeneration and lysis of cells around the test material.

The Agar Diffusion assay is appropriate for high density materials, such as elastomeric closures. In this method, a thin layer of nutrient-supplemented agar is placed over the cultured cells. The test material (or an extract of the test material dried on filter paper) is placed on top of the agar layer, and the cells are incubated. A zone of malformed, degenerative or lysed cells under and around the test material indicates cytotoxicity.

The MEM Elution assay uses different extracting media and extraction conditions to test devices according to actual use conditions or to exaggerate those conditions. Extracts can be titrated to yield a semi-quantitative measurement of cytotoxicity. After preparation, the extracts are transferred onto a layer of cells and incubated. Following incubation, the cells are examined microscopically for malformation, degeneration and lysis of the cells. (See page 7 for more information on the selection of extracting media and conditions.) At least one type of cytotoxicity test should be performed on each component of any device.

SENSITIZATION ASSAYS

Sensitization studies help to determine whether a material contains chemicals that cause adverse local or systemic effects after repeated or prolonged exposure. These allergic or hypersensitivity reactions involve immunologic mechanisms. Studies to determine sensitization potential may be performed using either specific chemicals from the test material, the test material itself, or most often, extracts of the test material. The Materials Biocompatibility Matrix recommends sensitization testing for all classes of medical devices.

The Guinea Pig Maximization Test (Magnusson-Kligman Method) is recommended for devices that will have externally communicating or internal contact with the body or body fluids. In this study, the test material is mixed with complete Freund�s adjuvant (CFA) to enhance the skin sensitization response.

The Closed Patch Test involves multiple topical doses and is recommended for devices that will contact only unbroken skin.

Eagle's minimal essential medium (MEM) is a synthetic cell culture medium developed by Harry Eagle in 1955/1959 that can be used to maintain cells in tissue culture.


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IRRITATION TESTS

These tests estimate the local irritation potential of devices, materials or extracts, using sites such as skin or mucous membranes, usually in an animal model. The route of exposure (skin, eye, mucosa) and duration of contact should be analogous to the anticipated clinical use of the device, but it is often prudent to exceed predicted exposure conditions somewhat to establish a margin of safety for patients.

In the Intracutaneous Test, extracts of the test material and blanks are injected intradermally. The injection sites are scored for erythema and edema (redness and swelling). This procedure is recommended for devices that will have externally communicating or internal contact with the body or body fluids. It reliably detects the potential for local irritation due to chemicals that may be extracted from a biomaterial.

The Primary Skin Irritation test should be considered for topical devices that have external contact with intact or breached skin. In this procedure, the test material or an extract is applied directly to intact and abraded sites on the skin of a rabbit. After a 24-hour exposure, the material is removed and the sites are scored for erythema and edema.

Mucous Membrane Irritation Tests are recommended for devices that will have externally communicating contact with intact natural channels or tissues. These studies often use extracts rather than the material itself. Some common procedures include vaginal and eye irritation studies. (See page 7 for more information on extracts.)

ACUTE SYSTEMIC TOXICITY

By using extracts of the device or device material, the Acute Systemic Toxicity test detects leachables that produce systemic (as opposed to local) toxic effects. The extracts of the test material and negative control blanks are injected into mice (intravenously or intraperitoneally, depending on the extracting media). The mice are observed for toxic signs just after injection and at four other time points. The Materials Biocompatibility Matrix recommends this test for all blood contact devices. It may also be appropriate for any other device that contacts internal tissues.

The Material Mediated Pyrogen test evaluates the potential of a material to cause a pyrogenic response, or fever, when introduced into the blood. Lot release testing for pyrogenicity is done in vitro using the bacterial endotoxin (LAL) test. It must be validated for each device or material. However, for assessing biocompatibility, the rabbit pyrogen test is preferred. The rabbit test, in addition to detecting bacterial endotoxins, is sensitive to material-mediated pyrogens that may be found in test materials or extracts.

SUBCHRONIC TOXICITY

Tests for subchronic toxicity are used to determine potentially harmful effects from longer-term or multiple exposures to test materials and/or extracts during a period of up to 10% of the total lifespan of the test animal (e.g. up to 90 days in rats). Actual use conditions of a medical device need to be taken into account when selecting an animal model for subchronic toxicity. Appropriate animal models are determined on a case-by-case basis.

Subchronic tests are required for all permanent devices and should be considered for those with prolonged contact with internal tissues.

induced fever


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GENOTOXICITY

Genotoxicity evaluations use a set of in vitro and in vivo tests to detect mutagens, substances that can directly or indirectly induce genetic damage directly through a variety of mechanisms. This damage can occur in either somatic or germline cells, increasing the risk of cancer or inheritable defects. A strong correlation exists between mutagenicity and carcinogenicity.

Genotoxic effects fall into one of three categories: point mutations along a strand of DNA, damage to the overall structure of the DNA, or damage to the structure of the chromosome (which contains the DNA). A variety of tests have been developed to determine if damage has occurred at any of these levels. These assays complement one another and are performed as a battery.

The most common test for mutagenicity, the Ames test, detects point mutations by employing several strains of the bacteria Salmonella typhimurium, which have been selected for their sensitivity to mutagens. The Mouse Lymphoma and the HGPRT assays are common procedures using mammalian cells to detect point mutations. The Mouse Lymphoma assay is also able to detect clastogenic lesions in genes (chromosome damage). Assays for DNA damage and repair include both in vitro and in vivo Unscheduled DNA Synthesis (UDS). Cytogenetic assays allow direct observation of chromosome damage. There are both in vitro and in vivo methods, including the Chromosomal Aberration and the Mouse Micronucleus assays.

ISO 10993-1 specifies an assessment of genotoxic potential for permanent devices and for those with prolonged contact (>24 hours) with internal tissues and blood. Extracorporeal devices with limited contact (<24 hours) may require a genotoxicity evaluation. Generally, devices with long-tem exposure require an Ames test and two in vivo methods, usually the Chromosomal Aberration and Mouse Micronucleus tests. Devices with less critical body contact may be able to be tested using only the Ames test.

When selecting a battery of genotoxicity tests, you should consider the requirements of the specific regulatory agency where your submission will be made. Because of the high cost of genotoxicity testing, SGS strongly recommends that you consult your FDA reviewer before you authorize testing.

IMPLANTATION TESTS

Implant studies are used to determine the biocompatibility of medical devices or biomaterials that directly contact living tissue other than skin (e.g. sutures, surgical ligating clips, implantable devices, etc.). These tests can evaluate devices, which, in clinical use, are intended to be implanted for either short-term or long-term periods. Implantation techniques may be used to evaluate both absorbable and non-absorbable materials. To provide a reasonable assessment of safety, the implant study should closely approximate the intended clinical use.

The dynamics of biochemical exchange and cellular and immunologic responses may be assessed in implantation studies, especially through the use of histopathology. Histopathological analysis of implant sites greatly increases the amount of information obtained from these studies.


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HEMOCOMPATIBILITY

Materials used in blood contacting devices (e.g. intravenous catheters, hemodialysis sets, blood transfusion sets, vascular prostheses) must be assessed for blood compatibility to establish their safety. In practice, all materials are to some degree incompatible with blood because they can either disrupt the blood cells (hemolysis) or activate the coagulation pathways (thrombogenicity) and/or the complement system.

The hemolysis assay is recommended for all devices or device materials except those which contact only intact skin or mucous membranes. This test measures the damage to red blood cells when they are exposed to materials or their extracts, and compares it to positive and negative controls.

DEVICES OR COMPONENTS WHICH CONTACT CIRCULATING BLOOD AND THE CATEGORIES OF APPROPRIATE TESTING — EXTERNAL COMMUNICATING DEVICES

Test Category

Device Examples

Thro

mbos

is

Coag

ulatio

n

Plate

lets

Hema

tolog

y

Comp

lemen

t

Syste

m

Atherectomy devices xa

Blood monitors x xa

Blood storage and administration equipment, Blood collection devices, Extension sets x x xa

Extracorporeal membrane oxygenator systems Haemodialysis/haemofiltration equipment, Percutaneous circulatory support devices

x x x x x

Catheters, guidewires, intravascular endoscopes, Intravascular ultrasound, laser systems, Retrograde coronary perfusion catheters.

x x xa

Cell savers x x xa

Devices for absorption of specific substances from blood x x x x

Donor and therapeutic apheresis equipment x x x x

a Hemolysis testing only

The complement system is a part of the immune system that enhances the ability of antibodies and phagocytic cells to clear microbes and damaged cells from an organism, promotes inflflammation, and attacks the pathogen's cell membrane.


P a g e 1 7

DEVICES OR COMPONENTS WHICH CONTACT CIRCULATING BLOOD AND THE CATEGORIES OF APPROPRIATE TESTING — IMPLANT DEVICES

Test Category

Device examples

Thro

mbo

sis

Coa

gula

tion

Pla

tele

ts

Hem

atol

ogy

Com

plem

ent

Sys

tem

Annuloplasty rings, mechanical heart valves x xa

Intra-aortic balloon pumps x x x x x

Total artificial hearts, ventricular-assist devices x x

Embolization devices xa

Endovascular grafts x xa

Implantable defibrillators and cardioverters x xa

Pacemaker leads x xa

Leukocyte removal filter x x xa

Prosthetic (synthetic) vascular grafts and patches,including arteriovenous shunts x xa

Stents x xa

Tissue heart valves x xa

Tissue vascular grafts and patches, including arteriovenous shunts x xa

Vena cava filters x xa a Hemolysis testing only

CARCINOGENESIS BIOASSAY

These assays are used to determine the tumorigenic potential of test materials and/or extracts from either a single or multiple exposures, over a period consisting of the total lifespan of the test system (e.g. two years for rat, 18 months for mouse, or seven years for dog).

Carcinogenicity testing of devices is expensive, highly problematic, and controversial. Manufacturers can almost always negotiate an alternative to full scale carcinogenicity testing of their devices.

REPRODUCTIVE AND DEVELOPMENTAL TOXICITY

These studies evaluate the potential effects of test materials and/or extracts on fertility, reproductive function, and prenatal and early postnatal development. They are often required for devices with permanent contact with internal tissues.

PHARMACOKINETICS

Pharmacokinetic or ADME (Absorption/Distribution/Metabolism/Excretion) studies are used to investigate the metabolic processes of absorption, distribution, biotransformation, and elimination of toxic leachables and potential degradation products from test materials and/or extracts. They are especially appropriate for bioabsorbable materials or for drug/device combinations. Our toxicology team is happy to work with you in setting up the appropriate PK or ADME study for your product.


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PRECLINICAL SAFETY TESTING

The objectives of preclinical safety studies are to define pharmacological and toxicological effects not only prior to initiation of human studies but throughout clinical development. Both in vitro and in vivo studies can contribute to this characterization.

HISTOPATHOLOGY SERVICES

Implant studies are often the most direct evaluation of device biocompatibility. The test material is placed in direct contact with living tissue. After an appropriate period, the implant site is recovered and examined microscopically for tissue reaction. The histopathologist can detect and describe many types of tissue and immune system reactions.

Similarly, in subchronic and chronic studies, various organs and tissues are harvested at necropsy and evaluated microscopically for toxic effects. Many of these studies also call for clinical chemistry analysis of specimens or serum samples form the test animals.


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MATERIALS CHARACTERIZATION AND ANALYTICAL TESTING OF BIOMATERIALS

Analytical procedures provide the initial means for investigating the biocompatibility of medical device materials. Knowledge of device materials and their propensity for releasing leachable matter will help manufacturers assess the risks of in vivo reactivity and preclude subsequent toxicology problems with finished devices.

Increasingly, FDA has been asking for analytical characterization of device materials and potential leachables per ISO 10993-17 and 10993-18. Many firms also use analytical procedures for routine QC of raw materials or finished products.

The degree of chemical characterization required should reflect the nature and duration of the clinical exposure and should be determined be based on the data necessary to evaluate the biological safety of the device. It will also depend on the nature of the materials used, e.g. liquids, gels, polymers, metals, ceramics, composites or biologically sourced material.

The following strategy is suggested as a sound program for chemical characterization of a device material:

1. Determine the qualitative composition of each device component or material. This information should be available from the material vendor, or it can be determined through laboratory testing. The list of constituents should include

a. the identity of the matrix (i.e. the major component such as the specific polymer, alloy, or metal) b. all plasticizers, colorants, anti-oxidants, fillers, etc. deliberately added during fabrication of the

material c. impurities such as unreacted monomers and oligomers d. manufacturing materials such as solvent residues, slip agents, and lubricants.

2. Estimate the potential for patient exposure for each item on the material constituent list. Use literature searches of toxicological databases to assess the likelihood of tissue reactivity. For potentially toxic constituents, design and conduct laboratory studies to determine the extractable levels of those constituents. Use exaggerated conditions of time and temperature, and consider appropriate detection limits. Additional studies may be needed to assess levels of extractables released in actual use conditions.

3. Data generated from this characterization process can be used to create a material data file. The information can then be used as a reference for continued testing of device materials to ensure consistency of future production lots. This may in turn reduce the need for routine biological testing.

Additional uses of analytical characterization data might include:

1. Use in an assessment of the overall biological safety of a medical device.

2. Measurement of the level of any leachable substance in a medical device in order to allow the assessment of compliance with the allowable limit derived for that substance from health based risk assessment.

3. Judging equivalence of a proposed material to a clinically established material.

4. Judging equivalence of a final device to a prototype device to check the relevance of data on the latter to be used to support the assessment of the former.

5. Screening of potential new materials for suitability in a medical device for a proposed clinical application.


P a g e 2 0

TRADITIONAL EXTRACTABLE MATERIAL CHARACTERIZATION

• USP Physicochemical Tests � Plastics

• USP Physicochemical Test Panel for Elastomeric Closures for Injections

• USP Polyethylene Containers Tests � Heavy Metals and Non-volatile Residues

• Indirect Food Additives and Polymers Extractables (21CFR Part 177)

• Sterilant Residues � Ethylene Oxide, Ethylene Chlorohydrin, Ethylene Glycol

TESTS PROCEDURES FOR EXTRACTABLE MATERIAL

• UV/Visible Spectroscopy

• Gas Chromatography

• Liquid Chromatography

• Infrared Spectroscopy (IR)

• Mass Spectrometry

• Residual Solvents

• Atomic Absorption Spectroscopy (AAS)

• Inductively-coupled Plasma Spectroscopy (ICP)

BULK MATERIAL CHARACTERIZATION

• Infrared Spectroscopy Analysis for Identity and Estimation of Gross Composition

o Reflectance Spectroscopy

o Transmission Spectroscopy

• Atomic Absorption Spectroscopy (AAS)

• Inductively-coupled Plasma Spectroscopy (ICP)

• Thermal Analysis

SURFACE CHARACTERIZATION

• IR Reflectance Spectroscopy

• Scanning Electron Microscopy (SEM)

• Energy-dispersive X-ray Analysis (EDX)


P a g e 2 1

SGS’S COMMITMENT TO EXCELLENCE Since 1878, quality testing services and client satisfaction have been our primary objectives in serving industry. During this period, SGS staff has increased to over 43,000 dedicated professionals. Operating 16 laboratories worldwide, SGS Life Science Services represents the largest network for GMP/GLP-compliant quality control testing, with unmatched geographic coverage serving local as well as global companies. With the addition of Northview Laboratories, SGS now has facilities in Northbrook, IL, Fairfield, NJ, and Mississauga, ON, expanding our ability to offer high quality analytical services to the pharmaceutical and medical device industry in North America. Our success grows from our commitment to the highest level of quality in all of our operations.

QUALITY SYSTEMS It is our commitment to:

• Measure our regulatory compliance by ensuring we meet or exceed all regulatory requirements and to ensure our core quality systems are operating properly.

• SGS Life Science Services business is regulatory science. We use FDA current Good Manufacturing Practice (cGMP) regulations as our baseline level of compliance. When requested by the client, we adhere to Good Laboratory Practice (GLP) regulations. We also comply with the requirements of ISO 9001-2000 and for animal sciences, USDA. At SGS, the core quality systems are:

% Documentation Procedures % Standard Operation Procedures (SOPs) % Training Systems and Documents % Equipment Validation, Calibration, and Preventive Maintenance % Vendor Qualification and Monitoring % Quality Audits � Internal, Client, and Regulatory % Quality Objective Monitoring and Trending % Management Review Meetings

cGMP COMPLIANCE

SGS and all of its medical device and pharmaceutical clients are required to operate in accordance with cGMP (current Good Manufacturing Practices) regulations. SGS is committed to perform all testing in accordance with our understanding of the cGMPs. A comprehensive body of standard operating procedures covers all aspects of our laboratory operations. We are routinely inspected by the FDA and auditors from many of our clients.

SGS does not assume any responsibility for the appropriateness and/or regulatory acceptance of any client's testing program. It is the responsibility of each client to assess the testing and test validation requirements of their products and quality control systems. Our staff will endeavor to alert clients of testing programs that may need further consideration to determine conformance to cGMPs.

GLP COMPLIANCE

For most biocompatibility submissions, the FDA and EPA require that testing be performed in accordance with GLP (Good Laboratory Practice) regulations. It is the client's responsibility to determine when GLP treatment is required and to inform SGS in writing of this requirement at the time of sample submission. SGS will perform testing in accordance with GLPs when requested by the client. (There is an additional charge for GLP treatment.)


P a g e 2 2

ACCREDITATIONS

SGS has been ISO 9001 certified by the British Standards Institute and by National Quality Association. FDA registration numbers for SGS's facilities are available upon request. Please contact us if you need additional information. Our staff would be pleased to have you visit and audit our laboratories at your convenience.

REFERENCES

AAMI Standards and Recommended Practices, Volume 4: Biological Evaluation of Medical Devices, which includes AAMI/ANSI/ISO Standard 10993. (Annex B of 10993-1 is an extensive bibliography of U.S. and international reference documents.)

ASTM F-748-98, Practice for Selecting Generic Biological Test Methods for Materials and Devices

Biocompatibility Testing and Management, Nancy J. Stark; Clinical Design Group, Chicago, 1994

ISO Standard 10993, Biological Evaluation of Medical Devices - Parts 1 � 20

Guidelines for the Intraarticular Prosthetic Knee Ligament (FDA)

PTCA Catheter System Testing Guideline (FDA)

Safety Evaluation of Medical Devices, Shayne Cox Gad; Marcel Dekker, Inc., New York, 2002

USP <1031>, The Biocompatibility of Materials Used In Drug Containers, Medical Devices, and Implants

CONTACT INFORMATION

Business Development (888) 747 8782

SGS Life Science Services (973) 244 2435 75 Passaic Avenue Fairfield, NJ 07004 SGS Canada, Inc. (905) 890 4880 310 Brunel Road Mississauga, ON L4Z 2C2, CAN SGS Northview Laboratories (847) 564-8181 1880 Holste Road Northbrook, IL 60062

www.us.sgs.com/lifescience

REQUEST FOR TESTING: BIOCOMPATIBILITY, page 1 of 2

REPORT TO:

____________________________

____________________________

____________________________

____________________________

____________________________

INVOICE TO:

____________________________

____________________________

____________________________

____________________________

____________________________

PHONE NO. __________________

FAX NO. _____________________

P.O. NO._____________________

QUOTE NO. __________________

EMAIL _____________________________________________________

SEND LAB REPORT BY (Test fees include delivery by email, fax or mail. 2nd & 3rd means are $6 additional. FedEx and UPS at additional cost):

Email Fax Mail

FedEx – Acct. No. _______________ UPS – Acct. No.________________ Report cc to:___________________

SAMPLE IDENTIFICATION (Please use the exact wording as desired on final report): ___________________________________________________________________________________________________________________________

NO. OF SAMPLES _______________ SPLWT/VOL. ____________________ EXP. DATE______________________SAMPLE CODE _________________ LOT NO.________________________

SPECIAL HANDLING:___________________________________________________________________________________ HAZARDOUS TYPE OF HAZARD: _________________________________________________

NOT HAZARDOUS Please include MSDS if samples are hazardous.

List parts of test article to be tested: ________________________________________________________________________

Final intended use/application of test article: _________________________________________________________________STABILITY TESTING: N/A Completed To be completed by sponsor STERILITY TESTING: non-sterile sterile, method ________________CAN SAMPLE BE CUT? YES NO UNUSED SAMPLES: discard return to sponsor EXTRACTION CONDITIONS 121°C for 1 hour 70°C for 24 hours

50°C for 72 hours 37°C for 72 hours other____________________________SURFACE AREA IN cm2 , if known _____________________________________________ THICKNESS IN mm , if known_________________________________________________ SURFACE AREA CALC. COMPLETED BY: Sponsor Consultant

To Be Completed by SGS via CAD (technical) Drawing other____________________________

ISO 10993-12:2002(E) 10.3.2 The standard surface area can be used to determine the volume of extract needed. This area includes the combined area of both sides of the sample and excludes indeterminate surfaces irregularities. When the surface area cannot be determined due to the configuration of the sample, a mass/volume of extracting fluid shall be used.

Unusually complex surface area calculations that are to be performed by SGS will incur additional charges. Extraction of large samples may incur additional media charges.

Should these tests be performed according to GLP regulations?

YES NO

Most biocompatibility studies should be performed according to GLP regulations. It is the responsibility of the test sponsor to request GLP treatment. There is an additional charge for GLP treatment.

Date results of all tests needed:_____________________________________________

SPECIAL INSTRUCTIONS: ______________________________________________________________________________________________________________________________________________________________

Please continue on page 2

REQUEST FOR TESTING: BIOCOMPATIBILITY, page 2 of 2

TESTS REQUIRED: (continued from page 1)

CYTOTOXICITY: ISO 10993-5 ISO USP Please specify method:

Agar Diffusion MEM Elution (extract test) Direct Contact All cytotoxicity samples will be extracted and/or incubated at 37°C for 24 hours in MEM, unless otherwise specified by sponsor.

Other media:______________________ Other extraction and/or incubation period: ______________________________________SENSITIZATION: ISO 10993-10

Murine Local Lymph Node Assay (LLNA) Please select extraction media: Saline Acetone in Olive Oil DMSO Other____________________________

Maximization Test (ISO) Please select extraction media: Saline Vegetable Oil Other____________________________

Closed Patch Test (ISO)

IRRITATION: ISO 10993-10 ISO Intracutaneous USP Intracutaneous Please select extraction media: Saline Vegetable Oil Other____________________________

ISO Primary Eye FHSA Primary Eye Other guideline ____________________ ISO Primary Skin FHSA Primary Skin Other guideline ____________________

Mucous Membrane Test: Vaginal Rectal Penile Hamster Cheek Pouch SYTEMIC: ISO 10993-11

USP Systemic Toxicity Test Please select extraction media: Saline Vegetable Oil Other____________________________

Material Mediated Pyrogen Test (Saline Extract Only)

SUBACUTE: ISO 10993-11 Repeated Dose Toxicity Study Duration:

14 days 28 days Please select route of administration: Oral Dermal Intravenous Intraperitoneal Other____________________________

SUBCHRONIC/CHRONIC: ISO 10993-11 Repeated Dose Toxicity Study Duration: _________________ days

Please select route of administration: Oral Dermal Intravenous Intraperitoneal Other____________________________

GENOTOXICITY: ISO 10993-3 Ames Test Please select extraction media: Mouse Lymphoma Saline DMSO Chromosomal Aberration PEG 400 Other____________________________ Mouse Micronucleaus

IMPLANTATION: ISO 10993-6 ISO USP Histopathology

Subacute Duration – 7 days 14 days 30 days (Histopathology is included in all ISO implantation studies.) Subchronic Duration – 60 days 90 days

Chronic Duration – 180 days 365 days HEMOCOMPATIBLITY: ISO 10993-4

Hemolysis - ISO (triplicate) Direct Contact Hemolysis - USP (duplicate) Sample Extract Prothrombin Time (PT) Partial Thromboplastin Time (PTT) Complement Activation Platelet Aggregation Platelet Count Platelet Activation In vivo thrombogenicity (device must be tubular in nature) Other___________________________

OTHER TESTS/SPECIAL INSTRUCTIONS: _________________________________________________________________________________________________________________________________________________

TESTING AUTHORIZED BY:___________________________________ DATE:_______________________Form No. RFA 08-2.2© (please sign)

© ISO 2018

Biological evaluation of medical devices —Part 1: Evaluation and testing within a risk management processÉvaluation biologique des dispositifs médicaux —Partie 1: Évaluation et essais au sein d'un processus de gestion du risque

INTERNATIONAL STANDARD

ISO10993-1

Fifth edition2018-08

Reference numberISO 10993-1:2018(E)

Corrected version2018-10

Provläsningsexemplar / Preview

This is only the table of contents. The full PDF of ISO 10993-1 can be purchased from the ISO website for US$158:https://www.iso.org/standard/68936.html

APPENDIXISO 10993-1 Excerpt

ISO 10993-1:2018(E)

ii © ISO 2018 – All rights reserved

COPYRIGHT PROTECTED DOCUMENT

© ISO 2018All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of the requester.

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Published in Switzerland


ISO 10993-1:2018(E)

Foreword ........................................................................................................................................................................................................................................ivIntroduction ................................................................................................................................................................................................................................vi1 Scope ................................................................................................................................................................................................................................. 12 Normative references ..................................................................................................................................................................................... 13 Termsanddefinitions ..................................................................................................................................................................................... 24 General principles applying to biological evaluation of medical devices ................................................. 55 Categorization of medical devices ...................................................................................................................................................... 9

5.1 General ........................................................................................................................................................................................................... 95.2 Categorization by nature of body contact ........................................................................................................................ 9

5.2.1 Non-contacting medical devices......................................................................................................................... 95.2.2 Surface-contacting medical devices .............................................................................................................105.2.3 Externally communicating medical devices ..........................................................................................105.2.4 Implant medical devices ......................................................................................................................................... 11

5.3 Categorization by duration of contact ............................................................................................................................. 115.3.1 Contact duration categories ................................................................................................................................ 115.3.2 Transitory-contacting medical devices .....................................................................................................115.3.3 Medical devices with multiple contact duration categories ....................................................11

6 Biological evaluation process ..............................................................................................................................................................126.1 Physical and chemical information for biological risk analysis .................................................................126.2 Gap analysis and selection of biological endpoints for assessment .......................................................126.3 Biological testing ................................................................................................................................................................................ 13

6.3.1 General................................................................................................................................................................................... 136.3.2 Testing for evaluation ...............................................................................................................................................14

7 Interpretation of biological evaluation data and overall biological risk assessment ...............18Annex A (informative) Endpoints to be addressed in a biological risk assessment.........................................20Annex B (informative) Guidance on the conduct of biological evaluation within a risk

management process ....................................................................................................................................................................................25Annex C (informative) Suggested procedure for literature review ...................................................................................38Bibliography .............................................................................................................................................................................................................................40

© ISO 2018 – All rights reserved iii

Contents Page


ISO 10993-1:2018(E)

Foreword

ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (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, 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.

The procedures used to develop this document and those intended for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/directives).

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. Details of any patent rights identified during the development of the document will be in the Introduction and/or on the ISO list of patent declarations received (see www .iso .org/patents).

Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.

For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and expressions related to conformity assessment, as well as information about ISO's adherence to the World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following URL: www .iso .org/iso/foreword .html.

This document was prepared by Technical Committee ISO/TC 194, Biological and clinical evaluation of medical devices.

This fifth edition cancels and replaces the fourth edition (ISO 10993-1:2009), which has been technically revised. It also incorporates the Technical Corrigendum ISO 10993-1:2009/Cor.1:2010.

The main changes compared to the previous edition are as follows:

a) revised Annex A “Endpoints to be addressed in a biological risk assessment” with new columns for “physical and/or chemical information” and “material mediated pyrogenicity” as well as columns for “chronic toxicity,” “carcinogenicity,” “reproductive/developmental toxicity,” and “degradation” which now indicates “endpoints” to be considered with “E” (instead of “tests” to be conducted with an “X”);

b) replaced Annex B “Guidance on the risk management process” with “Guidance on the conduct of biological evaluation within a risk management process” (formerly ISO TR 15499);

c) additional definitions for terms used throughout the ISO 10993 series of standards;

d) additional information on the evaluation of “Non-contacting medical devices” and new information on the evaluation of “Transitory-contacting medical devices”;

e) additional information on the evaluation of nanomaterials, and absorbable materials;

f) additional reference to ISO 18562 (all parts) for “Biocompatibility evaluation of breathing gas pathways in healthcare applications”;

g) significant editing changes throughout the document;

A list of all parts in the ISO 10993 series can be found on the ISO website.

This corrected version of ISO 10993-1:2018 incorporates the following correction.

iv © ISO 2018 – All rights reserved


https://www.iso.org/directives-and-policies.html

https://www.iso.org/iso-standards-and-patents.html

https://www.iso.org/foreword-supplementary-information.html

ISO 10993-1:2018(E)

Introduction

The primary aim of this document is the protection of humans from potential biological risks arising from the use of medical devices. It is compiled from numerous International and national standards and guidelines concerning the biological evaluation of medical devices. It is intended to describe the biological evaluation of medical devices within a risk management process, as part of the overall evaluation and development of each medical device. This approach combines the review and evaluation of existing data from all sources with, where necessary, the selection and application of additional tests, thus enabling a full evaluation to be made of the biological responses to each medical device, relevant to its safety in use. The term “medical device” is wide-ranging and, at one extreme, consists of a single material, which can exist in more than one physical form, and at the other extreme, of a medical device consisting of numerous components made of more than one material.

This document addresses the determination of the biological response to medical devices, mostly in a general way, rather than in a specific device-type situation. Thus, for a complete biological evaluation, it classifies medical devices according to the nature and duration of their anticipated contact with human tissues when in use and indicates, in a matrix, the biological endpoints that are thought to be relevant in the consideration of each medical device category. See also 3.14, Note 1 to entry.

The range of biological hazards is wide and complex. The biological response to a constituent material alone cannot be considered in isolation from the overall medical device design. Thus, in designing a medical device, the choice of the best material with respect to its biocompatibility might result in a less functional medical device, biocompatibility being only one of a number of characteristics to be considered in making that choice. Where a material is intended to interact with tissue in order to perform its function, the biological evaluation needs to address this.

Biological responses that are regarded as adverse, caused by a material in one application, might not be regarded as such in a different situation. Biological testing is based upon, among other things, in vitro and ex vivo test methods and upon animal models, so that the anticipated behaviour when a medical device is used in humans can be judged only with caution, as it cannot be unequivocally concluded that the same biological response will also occur in this species. In addition, differences in the manner of response to the same material among individuals indicate that some patients can have adverse reactions, even to well-established materials.

The primary role of this document is to serve as a framework in which to plan a biological evaluation. A secondary role is to utilize scientific advances in our understanding of basic mechanisms, to minimize the number and exposure of test animals by giving preference to in vitro models and to chemical, physical, morphological, and topographical characterization testing, in situations where these methods yield equally relevant information to that obtained from in vivo models.

It is not intended that this document provide a rigid set of test methods, including pass/fail criteria, as this might result in either an unnecessary constraint on the development and use of novel medical devices, or a false sense of security in the general use of medical devices. Where a particular application warrants it, experts in the product or in the area of application concerned can choose to establish specific tests and criteria, described in a product-specific vertical standard.

ISO 10993 series is intended for use by professionals, appropriately qualified by training and experience, who are able to interpret its requirements and judge the outcome of the evaluation for each medical device, taking into consideration all the factors relevant to the medical device, its intended use and the current knowledge of the medical device provided by review of the scientific literature and previous clinical experience.

Informative Annex A contains a table that is generally helpful in identifying endpoints recommended in the biocompatibility evaluation of medical devices, according to their category of body contact and duration of clinical exposure. Informative Annex B contains guidance for the application of the risk management process to medical devices which encompasses biological evaluation.

vi © ISO 2018 – All rights reserved


INTERNATIONAL STANDARD ISO 10993-1:2018(E)

Biological evaluation of medical devices —

Part 1: Evaluation and testing within a risk management process

1 Scope

This document specifies:

— the general principles governing the biological evaluation of medical devices within a risk management process;

— the general categorization of medical devices based on the nature and duration of their contact with the body;

— the evaluation of existing relevant data from all sources;

— the identification of gaps in the available data set on the basis of a risk analysis;

— the identification of additional data sets necessary to analyse the biological safety of the medical device;

— the assessment of the biological safety of the medical device.

This document applies to evaluation of materials and medical devices that are expected to have direct or indirect contact with:

— the patient's body during intended use;

— the user’s body, if the medical device is intended for protection (e.g., surgical gloves, masks and others).

This document is applicable to biological evaluation of all types of medical devices including active, non-active, implantable and non-implantable medical devices.

This document also gives guidelines for the assessment of biological hazards arising from:

— risks, such as changes to the medical device over time, as a part of the overall biological safety assessment;

— breakage of a medical device or medical device component which exposes body tissue to new or novel materials.

Other parts of ISO 10993 cover specific aspects of biological assessments and related tests. Device-specific or product standards address mechanical testing.

This document excludes hazards related to bacteria, moulds, yeasts, viruses, transmissible spongiform encephalopathy (TSE) agents and other pathogens.

2 Normative references

The following documents are referred to in the text in such a way that some or all of their content constitutes requirements of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.

ISO 10993-2:2006, Biological evaluation of medical devices — Part 2: Animal welfare requirements

© ISO 2018 – All rights reserved 1


ISO 10993-1:2018(E)

ISO 10993-3, Biological evaluation of medical devices — Part 3: Tests for genotoxicity, carcinogenicity and reproductive toxicity

ISO 10993-4, Biological evaluation of medical devices — Part 4: Selection of tests for interactions with blood

ISO 10993-5, Biological evaluation of medical devices — Part 5: Tests for in vitro cytotoxicity

ISO 10993-6, Biological evaluation of medical devices — Part 6: Tests for local effects after implantation

ISO 10993-7, Biological evaluation of medical devices — Part 7: Ethylene oxide sterilization residuals

ISO 10993-9, Biological evaluation of medical devices — Part 9: Framework for identification and quantification of potential degradation products

ISO 10993-10, Biological evaluation of medical devices — Part 10: Tests for irritation and skin sensitization

ISO 10993-11:2017, Biological evaluation of medical devices — Part 11: Tests for systemic toxicity

ISO 10993-12, Biological evaluation of medical devices — Part 12: Sample preparation and reference materials

ISO 10993-13, Biological evaluation of medical devices — Part 13: Identification and quantification of degradation products from polymeric medical devices

ISO 10993-14, Biological evaluation of medical devices — Part 14: Identification and quantification of degradation products from ceramics

ISO 10993-15, Biological evaluation of medical devices — Part 15: Identification and quantification of degradation products from metals and alloys

ISO 10993-16, Biological evaluation of medical devices — Part 16: Toxicokinetic study design for degradation products and leachables

ISO 10993-17, Biological evaluation of medical devices — Part 17: Establishment of allowable limits for leachable substances

ISO 10993-18, Biological evaluation of medical devices — Part 18: Chemical characterization of materials

ISO/TS 10993-20, Biological evaluation of medical devices — Part 20: Principles and methods for immunotoxicology testing of medical devices

ISO 14971:2007, Medical devices — Application of risk management to medical devices

3 Termsanddefinitions

For the purposes of this document, the following terms and definitions apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— IEC Electropedia: available at http: //www .electropedia .org/

— ISO Online browsing platform: available at https: //www .iso .org/obp

3.1biocompatibilityability of a medical device (3.14) or material (3.12) to perform with an appropriate host response in a specific application

3.2biological riskcombination of the probability of harm to health occurring as a result of adverse reactions associated with medical device (3.14) or material (3.12) interactions, and the severity of that harm

2 © ISO 2018 – All rights reserved


http://www.electropedia.org/

https://www.iso.org/obp

ISO 10993-1:2018(E)

3.3biological safetyfreedom from unacceptable biological risk (3.2) in the context of the intended use

3.4chemical constituentany synthetic or natural substance that is used in a process for manufacturing materials (3.12) and/or medical devices (3.14), including the base material(s), additives (antioxidants, UV stabilizers, color additives, dyes, etc.), and processing aids (solvents, lubricants, antifoaming agents, etc.)

3.5data setinformation, such as physical and/or chemical characterization, toxicity data, etc. from a variety of sources necessary to characterize the biological response to a medical device

3.6direct contactmedical device (3.14) or medical device component that comes into physical contact with body tissue

3.7externally communicating medical devicemedical device (3.14) or medical device component that is partially or wholly located outside the body but has either direct or indirect contact with the internal body fluids and/or tissues

3.8finalproductmedical device (3.14) or medical device component that has been subjected to all manufacturing processes for the “to be marketed” medical device including packaging and if applicable, sterilization

3.9geometrydeviceconfigurationshape and relative arrangement of the parts of the medical device (3.14)

3.10implantmedical device (3.14) which is intended to be totally introduced into the human body or to replace an epithelial surface or the surface of the eye by means of clinical intervention and which is intended to remain in place after the procedure

3.11indirect contactmedical device (3.14) or medical device component through which a fluid or gas passes, prior to the fluid or gas coming into physical contact with body tissue (in this case the medical device or medical device component itself does not physically contact body tissue)

3.12materialsynthetic or natural polymer, metal or alloy, ceramic, or composite, including tissue rendered non-viable, used as a medical device (3.14) or any part thereof

3.13material characterizationbroad and general process of collecting existing information about a material’s chemistry, structure and other properties, and if appropriate, new data, to facilitate the evaluation of these properties

3.14medical deviceany instrument, apparatus, implement, machine, appliance, implant, reagent for in vitro use, software, material (3.12) or other similar or related article, intended by the manufacturer to be used, alone or in combination, for human beings, for one or more of the specific medical purpose(s) of:



ISO 10993-1:2018(E)

— diagnosis, prevention, monitoring, treatment or alleviation of disease;

— diagnosis, monitoring, treatment, alleviation of or compensation for an injury;

— investigation, replacement, modification, or support of the anatomy or of a physiological process;

— supporting or sustaining life;

— control of conception;

— disinfection of medical devices;

— providing information by means of in vitro examination of specimens derived from the human body;

and does not achieve its primary intended action by pharmacological, immunological or metabolic means, in or on the human body, but which may be assisted in its intended function by such means.Medical devices include dental devices.

Note 1 to entry: Products which may be considered to be medical devices in some jurisdictions but not in others include:

— disinfection substances;

— aids for persons with disabilities;

— devices incorporating animal and/or human tissues;

— devices for in vitro fertilization or assisted reproduction technologies;

[SOURCE: GHTF/SG1/N071: 2012, 5.1 modified to clarify that dental devices are included]

3.15nanomaterialmaterial (3.12) with any external dimension in the nanoscale or having internal structure or surface structure in the nanoscale

[SOURCE: ISO/TR 10993-22:2017, 3. 7, modified — Notes to entry have been deleted.]

3.16non-contactingindicates that the medical device (3.14) or medical device component has neither direct nor indirect contact with body tissues

3.17physical and chemical informationknowledge regarding formulation, manufacturing processes, geometric and physical properties and type of body contact and clinical use that is used to determine whether any additional biological or material characterization testing is needed

3.18risk analysissystematic use of available information to identify hazards and to estimate the risk

[SOURCE: ISO 14971:2007, 2.17, modified— The Note has been deleted.]

3.19risk assessmentoverall process comprising a risk analysis (3.18) and a risk evaluation (3.20)

[SOURCE: ISO 14971:2007, 2.18]

4 © ISO 2018 – All rights reserved


ISO 10993-1:2018(E)

3.20risk evaluationprocess of comparing the estimated risk against given risk criteria to determine the acceptability of the risk

[SOURCE: ISO 14971:2007, 2.21]

3.21risk managementsystematic application of management policies, procedures and practices to the tasks of analysing, evaluating, controlling and monitoring risk

[SOURCE: ISO 14971:2007, 2.22]

3.22toxiccapable of causing an adverse biological response

3.23toxicological hazardpotential for a chemical substance or material (3.12) to cause an adverse biological reaction, taking into account the nature of the reaction and the dose required to elicit it

3.24toxicological riskprobability of a specified degree of an adverse reaction occurring in response to a specified level of exposure

3.25toxicological thresholdlimit, such as a tolerable intake (TI), tolerable exposure (TE), allowable limit (AL) value, or Threshold of Toxicological Concern (TTC) below which adverse effects are not expected for relevant biological endpoints

3.26transitory contactmedical device (3.14) or medical device component that has a very brief duration of contact with body tissue

4 General principles applying to biological evaluation of medical devices

4.1 The biological evaluation of any material or medical device intended for use in humans shall form part of a structured biological evaluation plan within a risk management process in accordance with ISO 14971:2007, Annex I, as given in Figure 1 of this document. This risk management process involves identification of biological hazards, estimation of the associated biological risks, and determination of their acceptability. Annex B provides guidance on this process. The biological evaluation shall be planned, carried out, and documented by knowledgeable and experienced professionals.

The risk management plan should identify aspects of the biological evaluation requiring specific technical competencies and shall identify the person(s) responsible for the biological evaluation.

The evaluation shall include documented, informed consideration of advantages/disadvantages and relevance of:

a) medical device configuration (e.g. size, geometry, surface properties) and a listing of a medical device’s materials of construction (qualitative) and where necessary, the proportion and amount (mass) of each material in the medical device (quantitative);

b) the physical and chemical characteristics of the various materials of construction and their composition;



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