SECTION

II.3

Biological Testing of
Biomaterials
CHAPTER II.3.1 HOW WELL WILL IT
WORK? INTRODUCTION TO TESTING
BIOMATERIALS
Buddy D. Ratner
Professor, Bioengineering and Chemical Engineering, Director of University
of Washington Engineered Biomaterials (UWEB), Seattle, WA, USA

When an engineer designs a suspension bridge over

a river, he or she will start with a precise engineering
drawing and then, based on well-established mathematical models, assess the strength of materials and corrosion
resistance needed to ensure the structure will stand for
some anticipated lifetime, e.g., 100 years. The strength
of materials and corrosion resistance are compiled in
widely accepted handbooks containing data obtained
through rigorous testing. The engineer can confidently
design the bridge from computer models and handbook
data. The bridge will stand and function appropriately
for 100 years. Moreover, such a bridge manufactured
according to a given set of design criteria and specifications should perform approximately the same whether it
crosses a river in Seattle, Birmingham or Boston.
Now, consider an engineer who sets out to design a
heart assist device or a hip prosthesis. This engineer will
find the basic mechanical property data relevant to the
materials used in handbooks, but little on specific biointeractions, biodurability, and the implications of these
biological reactions for the ultimate performance of the
complex device. Because of the limited amounts of validated and qualified data on how a given material will
perform in the challenging invivo environment, biomaterials scientists devise tests to observe the performance
of materials (and devices), and assess potential biointeractions. These tests are often (but not always) focused on
the mechanics and biology relevant to the specific application (orthopedic, cardiovascular, urological, etc.), and
they do not consider the potential for different circumstances and possibly different biomaterialstissue interactions in different patients. This section of the textbook
focuses on useful tools and ideas for testing biomaterials.
After this introduction, the section leads off with a
chapter titled The Concept and Assessment of Biocompatibility (Chapter II.3.2). Biocompatibility is possibly

the most central concept in understanding biomaterials;

and you cannot test for biocompatibility until you understand what it is. Chapter II.3.3 (In Vitro Assessment of
Cell and Tissue Compatibility,) and Chapter II.3.4 (In
Vivo Assessment of Tissue Compatibility) provide fine
overviews of thinking circa early 21st century on testing for biocompatibility, primarily focused on toxicology
(invitro) and the foreign-body reaction (invivo), and parallel the thinking by most national regulatory agencies (for
example, the US Food and Drugs Administration, FDA).
So, why do we need a chapter on the concept of biocompatibility? There are new developments that will probably
change the definition of biocompatibility. For example,
Chapter II.3.2 discusses a material (with no toxic leachables) fabricated as a solid film that heals with the classic
foreign-body reaction, and the same material fabricated as
a precise, porous structure that heals in a vascularized, afibrotic manner. Todays definitions and regulatory climate
are not structured to appreciate the significance of these
differences. Furthermore, there are other strategies to shift
the healing of todays biocompatible biomaterials to a
more tissue-integrated, vascularized reconstruction. To
prepare this third edition of the Biomaterials Science textbook for changes that will occur in the next few years, we
introduce the issues and concerns stimulating new thinking on biocompatibility in Chapter II.3.2. Chapter II.2.2
(Inflammation, Wound Healing, and the Foreign-Body
Response) provides essential background to understand
the material in Chapters II.3.2, II.3.3, and II.3.4.
Chapter II.3.5, Evaluation of BloodMaterials Interactions, looks at another face of the biological reaction
to synthetic materials. Millions of medical devices are
interfaced with the bloodstream each year. The reaction
of blood with biomaterials is strikingly different from
the reactions seen in soft tissue and bone in both kinetics (rapid) and outcomes (clot). Although blood compatibility assessment goes back to the 1930s, to this day
we have no widely accepted list of biomaterials that
are blood-compatible. Indeed, there is little agreement
about the structural/chemical/molecular characteristics
of a material that engender blood compatibility. Why
this lack of clarity and consensus? Chapter II.3.5 introduces the issues in blood compatibility assessment (the
nature of blood, the impact of flow, and the nature of
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SECTION II.3 Biological Testing of Biomaterials

the biomaterial), and illustrates how simple assessment

schemes can lead to erroneous conclusions. Accepted
blood compatibility tests are introduced but, more
importantly, the chapter provides guidelines for drawing conclusions from the results of these tests. Some
thoughts on which materials might be blood compatible
are offered in this chapter.
Chapter II.3.6 and Chapter II.3.7 address the use of
animals in biomaterials, and in medical device research.
Animal implants (invivo assessment) are usually required
for preclinical testing of medical devices. Without animal
testing, it is less likely that the research we perform will
transition from laboratory bench to patient. The use of
animals in testing is fraught with ethical issues respect
for the animal lives to be sacrificed and minimization
of pain to the animal (see Chapter III.2.7). Also, a key
issue is how significant are the results obtained in animals for understanding the performance of the medical
device in humans? Humans are different genetically,
biochemically, and biomechanically from animals. Also,
humans are different genetically, biochemically, and biomechanically from each other, whereas animal models
are genetically homogeneous. Can predictive models
be developed? The answer is, in most cases, yes, but
not with some forethought and planning. Without careful consideration and understanding of specific animal
models, animal lives will be sacrificed and research funds

CHAPTER II.3.2 THE CONCEPT AND

ASSESSMENT OF BIOCOMPATIBILITY
Buddy D. Ratner1 and Frederick J. Schoen2
1Professor, Bioengineering and Chemical Engineering,
Director of University of Washington Engineered Biomaterials
(UWEB), Seattle, WA, USA
2Professor of Pathology and Health Sciences and Technology (HST),
Harvard Medical School, Executive Vice Chairman, Department of
Pathology, Brigham and Womens Hospital, Boston, MA, USA

What do we mean when we say a biomaterial is

biocompatible?
Is biocompatibility yes or no, or is there a continuum

of biocompatibilities ranging from good to bad?

How can we measure biocompatibility?
How do we improve or enhance the biocompatibility
of a biomaterial?
The idea of biocompatibility is central to what makes
a material a biomaterial. Also, new developments and
concepts in cell biology are shifting thinking about biocompatibility, and this chapter offers an opportunity of
a glimpse into the future of biocompatible biomaterials.
Biocompatibility is so central to what makes a material a biomaterial, that a focused discussion of the subject is justified.

wasted, since the results obtained may not be useful for

the human device. The biomechanics issue is particularly
challenging. Almost any orthopedic device (spine, joints)
will be subjected to different mechanical forces in the
skeletal system of an animal walking on four legs versus
an upright, bipedal human. Other considerations are: (1)
should we go directly from invitro testing to testing in
human subjects (see Chapter III.2.7); and (2) what about
developing devices for veterinary use (for example, about
5000 intraocular lenses are implanted in dogs each year)?
Biomaterials researchers are largely dependent on animal
models, and understanding them is essential Chapters
II.3.6 and II.3.7 provide a start in appreciating the complex issues surrounding animal models.
The section on testing concludes with a chapter on
the basics of microscopy (Chapter II.3.8). Microscopic
examination of tissues and implants is a key element in
most biomaterials testing, invitro and invivo. To visually observe the cellular and tissue reactions at the interface between a biomaterial and a living system provides
clues as to the functionality and prospects for success
of a biomaterial or medical device. The range of microscopic techniques, and the resolution and information
content of these methods, is now in a period of evolutionary development, so this chapter primarily addresses
the basics for observing biomaterials and bioreactions at
the micron scale and the nanoscale.

This chapter clarifies some of the issues in biocompatibility, and also raises questions that will likely
impact the field in the coming years. In contrast to
empirical approaches and practical considerations
focused solely about the safety of implanted devices
(for example, toxicology, the state of the art today),
modern cell and molecular biology ideas may give us a
useful theory of biocompatibility with quantifiable
parameters, testable hypotheses, and validated engineering rules.

BIOCOMPATIBILITY TODAY
We start with an overview of the state of the art in biocompatibility today, i.e., in the first decade in the 21st
century.
A definition for biocompatibility, widely used in the
biomaterials/medical device community, has been presented previously in the textbook and is repeated here:
the ability of a material to perform with an
appropriate host response in a specific application
(Williams, 1987).
This definition, though accurate and quite useful in
the design, development, and application of biomaterials in medicine, nevertheless offers no insights into