Journal of Biomechanics 35 (2002) 401–414

ESB Keynote LectureFDublin 2000

Why mechanobiology?
A survey article
Marjolein C.H. van der Meulena,b, Rik Huiskesc,*
a
Sibley School of Mechanical & Aerospace Engineering, Cornell University, Ithaca, NY 14853, USA
b
Department of Biomedical Mechanics and Materials, The Hospital for Special Surgery, New York, NY 10021, USA
c
Department of Biomedical Engineering, Eindhoven University of Technology, Wh 4.131, PO Box 513, 5600 MB Eindhoven, Netherlands
Accepted 5 September 2001

Abstract

The central paradigm of skeletal mechanobiology is that mechanical forces modulate morphological and structural fitness of the
skeletal tissuesFbone, cartilage, ligament and tendon. Traditionally, skeletal biomechanics has focussed on how these tissues
perform the structural and locomotory functions of the vertebrate skeleton. In mechanobiology the central question is how these
same load-bearing tissues are produced, maintained and adapted by cells as an active response to biophysical stimuli in their
environment. The idea that ‘form follows function’ is not new, but we now believe that the scientific community has the knowledge
and tools to prove, understand and use functional adaptation to benefit medicine and human health. In this Survey Article the
philosophy and progress of skeletal mechanobiology are discussed. The revival of this science, with roots dating back to the 19th
Century, is now driven by new developments in cellular, molecular and computational technologies. These developments are still in
an early stage of application, but if modern mechanobiology fulfills the promises of its ambitions, the results will bring great benefits
to tissue engineering and to the treatment and prevention of skeletal conditions such as congenital deformities, osteoporosis,
osteoarthritis and bone fractures. r 2002 Elsevier Science Ltd. All rights reserved.

Keywords: Tissue differentiation; Bone adaptation; Computer simulation; Tissue engineering; Bone diseases

1. Introduction alization, chondrocyte apoptosis and bone modeling

Skeletal growth and development is an intricately
choreographed process involving many cell and tissue
types (Fig. 1). During growth in the embryo, the long
bones initially form as mesenchymal condensations
which chondrify to become cartilaginous anlagen. As
development progresses, the cartilage cellsFchondro-
cytesFhypertrophy and their extracellular matrix
mineralizes, forming the primary ossification center.
The chondrocytes die by apoptosis and the mineralized
cartilage is modeled to bone by the coordinated activity
of bone forming cells, osteoblasts, and bone resorbing
cells, osteoclasts. Osteoblasts that continue to produce
extracellular matrix and become trapped within the
matrix develop into mature bone cells, osteocytes. The
cycle of chondrocyte hypertrophy, extracellular miner-
*Corresponding author. Tel.: +31-40-247-4060; fax: +31-40-244-
7355.
E-mail address: r.huiskes@tue.nl (R. Huiskes).
proceeds spatially from the mid-diaphysis towards
the epiphyses. As the primary center progresses axially,
in some bones a secondary center forms, which
mineralizes through the same endochondral process.
The growth plate is where the two growth fronts meet.
Longitudinal growth at the growth plate occurs through
chondrocyte division and hypertrophy, and continues
until skeletal maturity. Concurrently, radial diaphyseal
growth occurs through direct apposition of bone by
osteoblasts on the periosteal surface and resorption by
osteoclasts on the endosteal surface. In parallel with the
formation of the mineralized skeleton, layers of cartilage
at the joint surfaces develop into articular cartilage and
fibrous anlagen develop into tendons and ligaments.
Joints form at the interzones between the cartilage
anlagen.
The premise of mechanobiology is that these biolo-
gical processes are regulated by signals to cells generated
by mechanical loading, a concept dating back to Roux
(1881). The relevant questions include how external and

0021-9290/02/$ - see front matter r 2002 Elsevier Science Ltd. All rights reserved.
PII: S 0 0 2 1 - 9 2 9 0 ( 0 1 ) 0 0 1 8 4 - 1
402 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414

Development of bone

A B C D E F
Embryonic Postnatal

Fig. 1. Schematic illustration of the development of long bones from embryonic to mature (figures not to scale). (A) Cartilaginous anlage with
chondrocytes; (B) Chondrocytes in the center swell; (C) Cartilage mineralization occurs around hypertrophic chondrocytes, proliferating flattened
cells develop; (D) Blood vessels penetrate the tissue, mineralized tissue is resorbed, bone formation takes place, and longitudinal growth commences;
(E) Secondary ossification centers develop and growth plates remain in between; (F) The bone is mature, the growth plates are closed; courtesy of Dr.
E. Tanck (2001).

muscle loads are transferred to the tissues, how the cells
sense these loads, and how the signals are translated into
the cascade of biochemical reactions to produce cell
expression or differentiation. Ultimately, we want to
predict growth and differentiation in quantitative terms,
based on a given force exerted on a given tissue matrix
populated by cells.
While understanding the mechanobiology of bone
development and growth can be relevant for the
prevention and treatment of congenital deformities,
there is also another motive for its study. The processes
of bone development continue in the mature organism
during bone regeneration in fracture healing and during
skeletal adaptation to implants. These involve tissue
differentiation from granulation through fibrous con-
nective tissue and cartilage to bone. The differentiation
pathway from granulation tissue to cartilage is a gradual
process that can be influenced over time. Mineralization,
on the other hand, is a ‘catastrophic event’, occurring
almost instantly and irreversibly. This event transforms
the mechanical and morphological status of the tissue
dramatically and completely, not a single chondrocyte
remains afterwards. After ossification, bone differentia-
tion continues within the tissue. Osteoclasts and
osteoblasts now build the characteristic microstructures
of cortical or cancellous bone tissue. This process of
structural modulation and growth is called modeling
(Frost, 1987). Mechanics also modulates the bone
modeling process, as evident from the trabecular
patterns that line up with external forces, suggesting
an optimized structure of minimal weight and maximal
strength (Wolff, 1892).
After the tissue has matured, bone is continually
maintained by osteoclastic resorption and subsequent
osteoblastic formation in a process called remodeling
(Frost, 1987). One purpose of this process is the removal
of microcracks and microdamage (Verborgt et al., 2000),
hence the term ‘maintenance’. Osteoclasts are believed
to be attracted by microcracks, through the apoptosis of
osteocytes (Noble et al., 1997). At maturity, bone
modeling continues due to variations in physical
activity; exercise enhances bone mass, while inactivity
reduces it. In the elderly osteoporosis can develop, the
etiology of which is also thought to be affected by
mechanobiology. And finally, the development of
osteoarthritis in the elderly may be related to mechanical
loading as well.
To understand how a mechanical stimulus produces a
biological signal for cells to differentiate or adapt a
tissue, we need to understand the relevant stimuli, the
signal transduction pathways and the response processes
(Duncan and Turner, 1995). Here, we described some
experimental and analytical approaches to address these
questions, with examples from work examining skeletal
adaptation and differentiation.
As in many sciences, the integration of experimental
and analytical models is critical to gaining an under-
standing of the skeletal response to mechanical factors.
Experiments provide insights and measurements, which
can then be interpreted within the context of analytical
 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414
frameworks. Analytical simulations permit investigation
of possible explanations that require in vivo validation
and will suggest further experimental investigations.
These investigations are greatly impacted by recent
technological advances in imaging, computational me-
chanics, genetics and molecular biology. The integration
of these techniques will provide many important insights
into skeletal development and diseases.
2. Experimental mechanobiology
Adaptation experiments examining skeletal mechan-
obiology have been performed for more than a century
and have examined the influence of both increasing and
decreasing the habitual loads applied to the skeleton.
Yet, despite the multitude of studies that have been
completed, there are still many unanswered questions
regarding the skeletal response to normal, perturbed and
pathological mechanical loading. Some of the difficulties
arise from the design of experiments, others in the
interpretation of data, and many from the inability to
compare across different species, ages, bones, loading
protocols, and so forth. Many of these issues arise from
the inherent challenge of in vivo experiments. An
introduction to in vivo bone mechanobiology experi-
ments, what is known from these experiments and the
open questions are presented below. Emphasis is placed
on bone adaptation models due to the greater presence
in the literature. However, analogous questions can be
posed for skeletal tissue differentiation and the adapta-
tion of other musculoskeletal tissues in response to
mechanical loading. Underlying this discussion is the
fundamental goal of understanding skeletal mechano-
transduction, which can be considered to consist of a
mechanical signal, response pathway and cellular
adaptive response.
2.1. Skeletal tissue differentiation
Fracture healing is a natural skeletal tissue differ-
entiation process. A well-defined temporal-spatial tissue
generation and replacement pathway is present normally
during secondary fracture healing. Three phases follow
bone injury: an initial inflammatory phase, a tissue
formation phase and finally tissue remodeling. Biophy-
sical stimuli are integral to the healing process. This can
already be appreciated by comparing primary fracture
healing to secondary healing. Primary healing is
completely stable and forms bone directly (Einhorn,
1998). Secondary healing involves lower fragment
stability and larger gap sizes and heals through
successive tissue formation and replacement (Einhorn,
1998). Experiments have demonstrated a significant role
of mechanical forces in the process of secondary fracture
healing. When compared to rigid fixation, fractures
403
subjected to cyclic loading after initial healing demon-
strate significantly greater increases in stiffness and
produce larger calluses (Goodship and Kenwright,
1985). Cyclic compression produces stronger, but more
compliant, healing fractures than static compression
(Panjabi et al., 1979). Excessive strains inhibit the
normal healing process, producing less bone tissue and
more initial inflammatory tissue (Augat et al., 1998).
However, growth factors and other chemical stimuli
likely play a more critical regulatory role than mechan-
ical stimuli, confounding these purely mechanobiologi-
cal models. Understanding the interaction between
growth factor expression and mechanical stimuli
may provide significant insights into the healing
process. Computer models and simulations can aid our
understanding of these fracture experiments (Blenman
!
et al., 1989; Bailon-Plaza and van der Meulen, 2001;
Lacroix et al., 2000). Distraction osteogenesis is a
similar process to fracture healing and can also be used
to study tissue differentiation (Carter et al., 1998; Tay
et al., 1998).
In vivo experiments have been used to examine both
tissue differentiation and adaptation to altered loading,
with the majority of experiments examining tissue
remodeling in response to mechanical stimuli. Skeletal
tissue differentiation experiments are complex to design;
however, historically many ‘natural’ examples have been
cited (Pauwels, 1941; Wolff, 1892). The most common
experimental design is a device that isolates or controls
the mechanical stimuli and then examines the tissues
that form (Goodman et al., 1994; Guldberg et al., 1997a;
Sballe et al., 1992; Tagil and Aspenberg, 1999). When
empty chambers are implanted into skeletal sites, new
tissue forms often in a process similar to fracture
healing. Bone can be formed both through intramem-
branous formation (Guldberg et al., 1997a) and
endochondral ossification (de Rooij et al., 2001). Often
fibrous tissue forms within the chambers under parti-
cular loading protocols (Goodman et al., 1994). Once
the process for a given model is established, cellular and
tissue differentiation can be examined in the absence of
loading or with the application of controlled micromo-
tion or loads in the device. These experiments demon-
strated the sensitivity of differentiating tissues to the
mechanical stimuli in their environment. With loading,
different tissues form (de Rooij et al., 2001) and the
morphology of the tissues also differs (Guldberg et al.,
1997a). While the intent is a simple loading situation,
this is difficult to realize in these in vivo devices due to
complex boundary conditions, tissue interfaces and
nonlinear behavior of the tissues, in addition to the
continually evolving nature of these conditions. Again,
mechanical-hypothesis-based theories can help explain
and interpret these tissue differentiation models, as
discussed below (Yuan et al., 2000; Prendergast et al.,
1997; Giori et al., 1995)
404 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414
2.2. Bone functional adaptation
Bone adaptation to mechanical stimuli has been
studied extensively using in vivo models. This subject
is a subset of the broader examination of cell and tissue
differentiation and may be a more straightforward
problem as a starting point for our understanding,
because only a single tissue is under consideration, bone.
Bone adaptation has been studied using a wide variety
of experimental models. Approaches used to increase
loading include exercise, osteotomy and devices to apply
controlled loading (Biewener and Bertram, 1993;
Churches and Howlett, 1982). Decreased loading was
achieved by casting, neurectomy, hindlimb suspension
and space flight (Morey-Holten et al., 1996; Uhthoff and
Jaworksi, 1978; Vico et al., 2001). Several common
concepts emerge from the past century of in vivo
experimentation. We know that bone adaptation occurs
in response to cyclic, not static, loading. The outcome is
mainly changes in cortical bone quantity, not bone
quality. Cancellous adaptation is less well characterized
but has similar responses that are present as changes in
the tissue apparent density. In general, demonstrating a
definitive increase in bone mass or strength is more
difficult than demonstrating a loss. Increased properties
are also harder to achieve with physiological than
nonphysiological loading conditions. Finally, we know
that the adaptive response is generally greater in
growing than mature animals, but growth may con-
found the results.
While these qualitative observations generally hold
true, precise quantitative predictions of the skeletal
response to mechanical stimuli are often desirable and
currently impossible. Much of our inability to predict
adaptation arises from experimental logistics: until
recently, most skeletal functional adaptation experi-
ments could not identify the driving feature of the
mechanical stimulus nor the contributing elements of
the response pathway (Duncan and Turner, 1995). For
example, when the loading environment is altered by
exercise and a skeletal effect is observed, does the
response result from the increased applied forces, the
increased numbers of loading cycles, the increased
loading rates, another exercise-induced parameter, or a
combination of these and other factors? Can one design
an exercise study to distinguish these features of the
stimulus? Increasing bone mass and strength with in
vivo physiological loading such as exercise is notoriously
difficult. Exercise studies have produced contradictory
results: running chickens at one speed was shown to
increase cortical area and moments of inertia (Biewener
et al., 1986), while running for a longer duration at a
similar intensity suppressed bone growth and reduced
strength over a similar experimental period (Matsuda
et al., 1986). Differences from one skeletal site to the
next within the same animal were also found in these
studies (Li et al., 1991). Whether these differences were
caused by differences in the local mechanical environ-
ment or by fundamentally different adaptive responses is
unknown. Answering these questions experimentally is
difficult because the in vivo loads cannot be measured
directly. In adult animals, physiological exercise gen-
erally produced bone hypertrophy or no response. The
absence of a response may indicate that the exercise
protocol did not sufficiently elevate the loads above
habitual levels. These unknowns all point towards the
need for a better understanding of the applied mechan-
ical loads.
To better address the mechanical stimulus underlying
bone adaptation, noninvasive experimental models have
been developed recently, which control the stimulus
applied to the cortical diaphysis (Gross et al., 2000;
Mosley et al., 1997; Torrance et al., 1994; Turner et al.,
1991). The two most common, currently, are the ulnar
compression model of Lanyon and coworkers (Fig. 2)
and the tibial four-point bending approach developed at
Creighton by Turner and colleagues. The ulnar com-
pression model likely creates more physiological stresses
and strains within the bone and has load application
points that are farther from the point of interest than the
tibial four-point bending model. Both models were
initially developed for the rat and have been modified
for the mouse (Akhter et al., 1998; Fritton et al., 2001).
Neither model requires surgery as previous controlled
loading models often did (Churches and Howlett, 1982;
Hert et al., 1969; Rubin and Lanyon, 1984). In both
systems, dynamic loads are applied and the load
magnitude, rate, number of cycles and duration are
well controlled. These noninvasive models, combined
with increased computing power, higher resolution
Fig. 2. The ulnar compression model (from Mosley and Lanyon,
1998).
 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414
imaging and new molecular and genetic techniques
enable systematic evaluation of loading parameters to
understand the nature of the osteogenic stimuli and
pathways. For the cortical diaphysis, stimulus charac-
teristics that induce bone modeling are becoming
evident. The applied strain rates and magnitudes need
to be high (Mosley et al., 1997; Mosley and Lanyon,
1998), the strain rate contributes to the morphology of
the bone formed (Turner et al., 1994), distinct,
temporally separated loading ‘episodes’ are required,
not just increased numbers of loading cycles (Robling
et al., 2000), and gender and genetic background
influence the nature of the response (Akhter et al.,
1998; Pedersen et al., 1999).
Similar questions need to be examined in cancellous
bone, a site of more clinical relevance than the cortical
diaphyses. Applying controlled loading to cancellous
bone is more difficult than to the cortical diaphyses.
Trabecular bone adaptation models include direct
stimulation of the site of interest as well as in
encapsulating bone chambers (Chambers et al., 1993;
Goldstein et al., 1991, 1997b; Guldberg et al., 1997b;
Hollister et al., 1996; Lamerigts et al., 2000; Morgan
et al., 2001). Nearly all models require surgical inter-
vention at the trabecular site; the rat-tail vertebra model
is an exception, since only the adjoining vertebral bodies
are operated on. Systematic application of dynamic
loading needs to be performed to address the same
questions posed above for cortical bone. For example,
the rodent ulna model can presumably be extended also
to examine cancellous bone, and perhaps also the
response of the cartilage in the growth plate to
mechanical loading.
Compared to questions about the nature of the
stimulus, even less is known regarding the transduction
of osteogenic signals to stimulate bone formation or
resorption. This pathway is hard to delineate, particu-
larly when using in vivo models. However, recent
molecular advances allow examination of whether
individual genes and molecules contribute to the adapta-
tion pathways. The mouse provides a unique opportunity
to examine genetic factors through manipulation of
murine embryonic stem cells and because significant
portions of the genome have been characterized (Clark
and Rowe, 1996). Transgenic technology has led to the
creation of both knockout and transgenic animals, either
missing or over-expressing genes, and previously re-
corded natural mutations can now be identified. One can
now directly determine whether a given protein is
important to skeletal adaptation by removing it.
Transgenic technology allows the role of potential
regulatory factors (Erlebacher and Derynck, 1996), cell
surface receptors (Zimmerman et al., 2000) and matrix
constituents (Khillan et al., 1991) to be examined.
Increasingly, mouse genetic studies are demonstrating
extreme redundancy within the genome under normal
405
conditions; simply characterizing the normal phenotype
may be unrevealing or insufficient. However, manipulat-
ing the system by changing the applied loads can require
cellular/tissue adaptation above and beyond normal
development and can demonstrate significant phenoty-
pic effects. Examples include the osteopontin-knockout
mouse and the bone morphogenetic protein-5 deficient
short ear mouse. The former has no obvious phenotype,
but does not lose bone when subjected to hindlimb
suspension (Ishijima et al., 2001). The phenotype of the
latter is an overall reduced body size and these mice
respond to increased loads more slowly than hetero-
zygous controls (Mikic et al., 1996; van der Meulen,
1999). Unfortunately, developing transgenic or knock-
out mice and examining all the potential regulatory
factors will consume many research careers. However,
further strengths of these genetic models will become
evident when we start to examine genetic control of
skeletal responses by not only expression but also
linkage analyses (Beamer et al., 2001; Klein et al.,
2001; Yershov et al., 2001).
In summary, many questions remain regarding the
role of mechanics in the formation and adaptation of
skeletal tissues. For functional adaptation, we need to
be able to globally describe the adaptive response for
different skeletal sites and ages, for example. To do so
will require understanding the nature of an osteogenic
mechanical stimulus and characterizing the molecular
pathway that this stimulus activates. More generally, in
skeletal tissue formation these same questions apply,
focused on the mechanical stimuli that induce cell and
tissue differentiation. Answering these questions will
require carefully controlled experiments with well-
characterized mechanical environments. When design-
ing and interpreting experiments, consideration must be
given to contributing factors, including the type of
model (in vitro or in vivo), species, type of loading
(physiological or invasive), the nature of loading
(tension/compression, cyclic/static, controlled/natural,
etc.) and level of analysis (cell, organ or tissue level).
The discussion here has focused on in vivo models with
an emphasis on controlled applied loading. A similar
discussion could be presented for in vitro experiments,
which are used to answer similar questions, and subject
to different limitations.
3. Computational mechanobiology
The purpose of computational mechanobiology is to
determine the quantitative rules that govern the effects
of mechanical loading on tissue differentiation, growth,
adaptation and maintenance by trial-and-error. From a
mechanics point of view, the task is considered a
‘boundary value problem’, whereby the boundary loads
of a domain are translated into local mechanical
406 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414
variables within the domain, depending on the geometry
and mechanical properties of its materials. Such
problems are usually solved with finite element analysis
(FEA). The biological side of the computation is based
on the premise that local mechanical variables stimulate
cell expression to regulate matrix composition, density
or structure. These two parts of the problem are
combined in a computer simulation model. Modeling
considerations include force application at the bound-
ary, force transmission through the tissue matrix,
mechanosensation by cells, mechanotransduction by
the cells, cell gene expression, and transformation of
extracellular matrix characteristics. These processes
must be represented in variables, parameters and
mathematical relationships. Some of these are known,
or can be measured (e.g. morphology, mechanical tissue
properties, external loading characteristics), but others
cannot. So how are these unknowns dealt with in the
computations?
There is a difference between the output of a
computation and the output of a problem to be solved.
For example, the local mechanical variables that might
stimulate the cells are usually not known and neither are
the mathematical relationships between a given stimulus
value and its effect on local cell expression and matrix
adaptation. Potential stimuli include strain, fluid flow,
strain rate and strain energy. Although these unknowns
must be entered into the computer simulation, they also
represent precisely the quantitative rules that we are
trying to determine. Computational mechanobiologists
hypothesize a potential rule and determine if the
outcome of this hypothesis produces realistic tissue
structures and morphologies, hence ‘trial-and-error’. If
the results correspond well, they might be an explana-
tion for the mechanism being modeled. This method of
research is common practiceFand productiveFin
physics, less common in biology (Huiskes, 1995);
although ‘theoretical biology’ is based on this type of
approach. Computer simulation has recently been cited
as the ‘third method of science’, after logic and
experimentation (Kelly, 1998). Below we give some
examples from skeletal development, repair, mainte-
nance and adaptation, particularly to demonstrate the
general approach, its limitations and its progress in
recent years.
3.1. Tissue differentiation
Tissue differentiation occurs in bone development and
regeneration, fracture healing and adaptation to skeletal
implants. In 1941, Pauwels proposed a theoretical
framework in which the effects of mechanical forces
on tissue differentiation pathways occur through me-
chanical deformation of the tissues. The local deforma-
tions in a tissue can be described by a strain tensor,
which has first and second invariants representing the
dilatational and distortional strain components. The
first invariant represents a change in volume (as caused
by dilatational or hydrostatic stress), the second a
change in shape (as caused by distortional or shear
stress). Pauwels proposed that the combination of these
variables determines the differentiation pathway from
the mesenchymal origin to fibrous, fibro-cartilaginous or
cartilaginous tissue. Endochondral bone formation
would be stimulated in areas of low distortional and
high dilatational strain (Prendergast and van der
Meulen, 2001).1 Pauwels’ theory was based on clinical
observation and logic, but he did not have the capacity
to measure or calculate the tissue strains or stresses
accurately. In the following discussion we will show how
the application and evolution of FEA has affected our
thoughts about Pauwels’ ideas, and how computational
mechanobiology is gradually maturing.
Carter and colleagues tested Pauwels’ theory relative
to endochondral bone formation using FEA. Endo-
chondral ossification was defined by the peak cyclic
hydrostatic stress (D) and the peak cyclic octahedral
shear stress (S), combined in an ‘osteogenic index’ I, as
I ¼ S þ kD;
summed over a number of loading cycles; k is an
empirical constant. High values of the index were
hypothesized to stimulate ossification; no specific
threshold value was specified. Note that S is always
positive, but D can both be positive (tension) or negative
(compression). This tissue differentiation rule was tested
with an FEA model of a developing joint to determine
whether this approach could predict the emergence of
secondary ossification centers (Carter and Wong, 1988).
High values of the osteogenic index were found in the
cartilage precisely where osteogenesis occurs during in
vivo skeletal development. Comparing the results with
biological examples, the most predictive values for the
constant k were determined at 0:3pkp1:0 (Fig. 3).
Similarly successful predictions of endochondral ossifi-
cation sites were performed for other skeletal sites, for
bone regeneration in fracture healing and for healing
around orthopaedic implants (Carter et al., 1998; Carter
and Beaupre! , 2001; Giori et al., 1995).
A vital issue for mechanobiological FEA models is to
what extent they can be simplified without loosing their
potential to obtain meaningful results. On the one hand,
simplicity is dictated by contemporary computational
technology, and on the other it depends on the
1
The inclusion of enchondral ossification in a theoretical framework
for skeletal differentiation pathways is not entirely justified, as
mineralization and bone modeling are very different kinds of processes,
which actually terminate differentiation. Mineralized tissue cannot de-
differentiate, only resorb; or rather be resorbed. However, mineraliza-
tion occurs when particular conditions in cartilage are met and the
creation of those conditions can be seen as a part of the differentiation
pathway.
 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414
Fig. 3. The distributions of the osteogenic index as computed in the
cartilaginous part of developing bone with a 2-D FEA model, using 3
different values for the empirical constant k (Carter and Wong, 1988).
Ossification is predicted where the index is high (from Carter, 1987).
hypothesis to be tested. In that sense a model takes for
granted the matter in dispute, or ‘begs the question’,
because its assumptions determine its answers. One can
only make conclusions about phenomena accounted for
in the model, not about what is omitted or assumed not
to contribute. This is equally true for experimental
models of course. The above FE analyses of the cartilage
in the models assumed homogeneity, isotropy and linear
elasticity; while in reality the tissue is multi-phasic,
nonlinear and highly structured at many levels of scale.
Hydrostatic pressure and octahedral stress in biphasic
tissues seemed to be well captured by single-phasic linear
elastic FEA, provided that the permeability is low, as is
the case in cartilage (Mow et al., 1980; Tanck et al.,
2000; Yuan et al., 2000). Hence, relative to the
hypothesis the authors wanted to test, the conclusions
about the predictive quality of the osteogenic index may
not have been any different had they described the
cartilage as biphasic. But, although the proposed
definition of the osteogenic index produces realistic
results, another choice for other mechanical variables,
or an alternative mathematical rule, might produce
equal or potentially even better results.
One important consideration is fluid flow in cartilage.
There are several reasons that interstitial fluid flow
could be a more realistic mechanical variable for
feedback information to the cells during tissue differ-
entiation than hydrostatic pressure. Flow is known to
stimulate anabolic cell expressions during in vitro testing
(Jacobs et al., 1998). The hydrostatic stress, computed in
linear-elastic FEA models, is believed to be indicative of
the interstitial fluid pressure in vivo. However, biphasic
407
analyses of bone ossification suggested that the true fluid
pressure in the tissue might be sensitive to the solid
matrix compressibility, another poorly understood
parameter (Tanck et al., 1999). Based on a comparative
analysis of experimental tissue mineralization studies
(Burger et al., 1991) with biphasic FEA, the hypothesis
that hydrostatic stress was a modulating variable was
refuted (Tanck et al., 2000). In a biphasic analysis of a
tissue differentiation experiment around a moving
piston in the femoral condyles of dogs (Sballe et al.,
1992), Prendergast and Huiskes (1996), Prendergast and
colleagues (1997) found that local tissue fluid pressure
does not change as the tissue differentiates. Yuan et al.
(2000) came to a similar conclusion from a biphasic
analysis of soft-tissue membranes under tibial knee
implants, comparing results to those of an earlier linear-
elastic analysis of the same problem (Giori et al., 1995).
Prendergast and Huiskes (1996), Prendergast et al.
(1997) also found that the stresses on cells are not only
generated by the tissue matrix, but also to a large extent
by the drag forces from interstitial fluid flow. This
conclusion indicates the need for dynamically loaded,
biphasic models, because these effects cannot be
examined with static or linear elastic ones. Interstitial
fluid flow and pressure need to be investigated as
potential signaling variables in the simulations.
Improvements in computer capacity now enable FEA
computer simulations in a dynamic sense, as opposed to
the earlier computer analyses. In the latter case the
mechanical variables are computed for a particular
initial configuration and predict a steady state, final
homeostatic configuration. The result is determined
from the initial conditions, but the process is not
simulated over time. The results of Carter and associates
(Carter and Beaupre! , 2001) relative to the prediction of
ossification areas are quite realistic. However, these
analyses cannot predict the progression of ossification
based on a single set of initial conditions. To simulate
the time-varying mineralization process, the evolution of
the tissue material properties must also be simulated
with time. The process of ossification itself affects the
load transfer in the cartilaginous parts while time
transpires (Claes and Heigele, 1999; Gardner et al.,
2000; Lerner et al., 1998; Prendergast et al., 1997;
Stevens et al., 1999). Another advantage of time-
dependent simulations is the ability to incorporate
dynamic loading, including both fluctuating loads at a
particular instant and overall variation in load over
time.
Based on the above considerations, and the studies of
Prendergast and Huiskes (1996), Prendergast et al.
(1997), an alternative rule was proposed for differentia-
tion from loose connective tissue to cartilage or bone.
This was tested in dynamic, biphasic computer simula-
tions as a mechano-regulation index, deciding which
tissue phenotype would prevail as an effect of mechan-
408 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414
ical stimulation. For that purpose a mechano-regulation
index (M) was formulated as (Huiskes et al., 1997)
g n
M¼ þ ;
a b
where g ðdimensionlessÞ is the maximal distortional (or
octahedral shear) strain, v (mm/s) the interstitial fluid-
flow velocity, a ¼ 0:0375 and b ¼ 3 (mm/s). Based on
experimental data (Sballe et al., 1992), it was proposed
that where M > 3; fibrous tissue would prevail (char-
acterized by a modulus of 2.0 MPa and a permeability of
1.0  1014 m4 N/ s), where 1pMp3; cartilage would
prevail (modulus 10.0, permeability 5.0  1015) and
where Mo1; bone would form (modulus 4590, perme-
ability 3.7  1013). This theory has two empirical
thresholds which determine when transitions in the
tissue type occur, and two empirical constants a and b
which are, however, interdependent. In this model the
value b=a plays a similar role as k in the osteogenic
index. The rule was tested in a mechano-regulatory
computer simulation, using the FEA model of the
Sballe experiment discussed above (Huiskes et al.,
1997). Iteratively, cyclic loading was applied to the
piston and the tissue modulus and permeability were
adjusted locally per iteration, in accordance with the
distribution of the mechano-regulation index in the gap
tissue, until differentiation converged to a final tissue
phenotype (Fig. 4). Using this approach, the differentia-
tion of gap-tissue phenotype from fibrous connective to
cartilage to bone could be predicted, simulating the
experiment (Sballe et al., 1992). In the earlier analysis
of this experiment the ingrowth process was successful
New Tissue Phenotype
Finite Element Model Mechanical Characteristics
Implant Loading Conditions
Mechano-Regulation Diagram
Fluid Velocity
FIBROUS
GAP
FIBRO
TON
CARTILAGE
PIS
BONE
BONE
Strain
Tissue Strain
Fluid-Solid Velocity
Fig. 4. The regulatory scheme used in the computer simulation. The
strain and fluid-velocity distributions are calculated in the FEA model.
Based on the transition criteria, illustrated in a phase diagram
representing the differentiation rule, tissue phenotype characteristics
are updated in every iteration. The simulation is continued until no
more change occurs and homeostasis is reached (from Huiskes et al.,
1997).
because the piston displacement was force controlled
(Prendergast et al., 1997). Force control reduces the
excursion of the piston when the tissue gets stiffer,
allowing the tissue to control its differentiation stimulus.
To test this conclusion, simulations of motion control
were performed using the maximal total excursion as in
the force-controlled experiment, regardless of the force
required. While the tissue ossified fully in the force-
controlled simulation, motion control produced a
largely fibro-cartilaginous tissue, with only a few islands
of bone (Fig. 5). This lack of ingrowth under large
displacements is also seen around loose orthopedic
implants.
These results demonstrate the importance of temporal
simulation for the questions posed. In both the force-
controlled and motion-controlled simulations of tissue
ingrowth the differentiation patterns predicted after the
first iterations were identical, because both the force and
excursion of the piston were equal (Fig. 5). Therefore, in
an analysis based on the first iteration only, the same
outcome would have been predicted for both cases. The
same regulatory model was used to simulate tissue
differentiation in fracture healing, using the same values
for the empirical constants and tissue characteristics
(Lacroix et al., 2000). In these simulations the differ-
entiation pathways were also not monotonic, again
demonstrating the importance of simulation over
analyses based on the initial conditions only.
While this simulation model captures the dynamic
aspects of the system and accounts for the time-
dependent properties of the tissues, limitations are
present as well. The fluid pressure and flow velocity
can be evaluated, but osmotic effects and charged-
density flows in the tissue, for example, are not
accounted for (Mow et al., 1999). Hence, this model
also ‘begs the question’. Similar to the linear elastic,
homeostatic models, the relevant mechanical variables
are evaluated at a macroscopic (homogenized) con-
tinuum level and cannot describe what individual cells
experience. Cells exist at a much lower scale, surrounded
by a quite irregular micro-morphology. These models
are ‘mechanistic’ with respect to fluid flow, but very
crude and approximate with regard to the cellular
environment (Wang et al., 2001). But, as stated earlier,
from a scientific point of view, these models could still
produce realistic hypotheses, because the importance of
these details is unknown. The physiological cellular
mechanisms are not understood, so the appropriateness
of macroscopic or microscopic formulations is un-
known. Perhaps the extracellular matrix acts as a filter
for the local environment, making a continuum model
equally appropriate. To capture every detail of the
system from cells to whole organs, we may need to build
FEA models of whole bones with refinement to the
cellular level, including the cascade of biochemical
reactions in and around the cells, but current computer
 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414 409

3
10

fluid-solid velocity [um/s]

2
10

1
10
Element status after first iteration
0
10 of both force-controlled and motion-
-1
controlled piston actuation
10 -4 -3 -2 -1
10 10 10 10
strain [-]

Final after force-control Final after motion-control

3 3
10 10

fluid-solid velocity [um/s]

fluid-solid velocity [um/s]

2 2
10 Fibrous tissue 10

1 1
10 10
Fibro-cartilage
0 0
10 10
Bone
-1
-1 10 -4
10 -4 -3 -2 -1 10
-3
10 10
-2 -1
10
10 10 10 10
strain [-] strain [-]
Fig. 5. Graphs illustrating the phenotypical stages of the gap tissue during the simulation; every dot represents an element. Initially all elements are
fibrous. After the first iteration two elements were predicted to become bone, a large number to become fibro-cartilaginous and the majority to be still
fibrous, based on the strain and fluid velocity history computed. This first phase developed identical in both the force and motion controlled
simulations. In the final homeostatic stage of the force-controlled simulation all elements turned to bone (Huiskes et al., 1997). In the motion-
controlled simulation, however, the gap tissue was predicted as a final mixture of fibrous and fibro-cartilaginous, with just a few islands of bone.

capacity still defies that. There is nothing scientifically
incorrect about the current approaches, however. This
discussion relates to the extent a model is ‘mechanistic’,
i.e. to what detail is the biological cascade of processes,
and their biochemical actors, represented or necessary to
be represented? In computational mechanobiology
mechanistic detail is not a goal by itself; the research
question dictates the level of sophistication required.
A final complicating issue for computational models
in mechanobiology is the presence of empirical con-
stants, whose values must be determined by comparison
to a biological reality. The osteogenic ossification rule
(Carter and Beaupre! , 2001), for example, contains the
constant k, which weights the sensitivity of the tissue for
hydrostatic stress relative to shear stress in the index. In
the mechano-regulation rule (Huiskes et al., 1997;
Lacroix et al., 2000) the constant b/a weights the relative
sensitivities for flow velocity and strain. In addition,
both theories require transitional values for their
indexes, which determine when the tissue differentiates;
the ossification rule requires a single value for this
purpose and the differentiation rule requires two. The
values of these constants can only be determined by
comparing computational results to similar experiments.
If one assumes that the cell mediated processes are
independent of cell location, then the empirical con-
stants should have unique values for all differentiation
processes. If the empirical constants must be adjusted
per configuration or problem definition, then a theory is
not falsifiable, and has little predictive power (Prender-
gast, 2001). Comparing results of several different FEA
models to evaluate the ossification index to bone-
formation patterns in ontogenesis, growth-plate orienta-
tion, fracture healing, and fibrous-tissue characteristics
around orthopaedic implants, the ossification rule
delivered best-fitting k-values between 0.0 and 2.0
(Carter and Beaupre! , 2001; Ribble et al., 2001). As can
be seen in Fig. 3, the value of k can have a large effect on
the predicted distribution of the osteogenic index. The
differentiation rule was only tested for ingrowth of
implants, as shown above, and fracture healing (Lacroix
et al., 2000), using the same values for the empirical
constants. Much more experimentation will be needed
to validate these values.
410 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414

Fig. 6. A model to simulate trabecular modeling, remodeling and adaptation governed by strain-energy density rate as produced by external loads,
assuming osteocyte mechanosensation and signal transduction to lining cells at the trabecular surface, which stimulates osteoblastic bone formation.
Osteoclasts are assumed to remove microcracks in a spatially random way; resorption cavities produce the local strain concentrations attracting
osteoblastic repair. An example is shown in which, after 2000 iterations with the FEA model, a homeostatic structure, adapted to magnitude and
orientation of the external load, was formed from an arbitrary initial structure (from Huiskes et al., 2000).

Tissue differentiation encompasses a wide area of
scientific efforts, including many computational studies
that were not discussed here; relevant ones can be found
in the articular cartilage area (Wang et al., 2001). The
testing of theories for tissue differentiation with simula-
tion models has spread to ligaments and tendons (Wren
et al., 1998) and development of articular joints
(Heegaard et al., 1999). Currently the theories proposed
can only be used to predict the past, allowing the
formulations to be validated against known experiments
or biological realities. Our ultimate objective should be
to predict the future.
3.2. Bone adaptation
Computer-simulation of bone adaptation is more
mature than in the tissue differentiation area, not in the
least because the problems are better defined and
concern only bone, as mentioned earlier. For this case
the computational studies also started by analysis, with
true simulation following more recently. One could say
that Wolff’s Law (1892) was already based on the
comparison between trabecular morphology and the
results of a ‘computational’ analysis (Huiskes, 2000).
The philosophy behind the computer simulation studies
in this area is similar to those discussed above for tissue
differentiation. Hart (2001) recently wrote an excellent
historical overview of theories and computational
models. Modern developments started with the ‘theory
of adaptive elasticity’ for cortical bone (Cowin and
Hegedus, 1976), which was later used for an FEA
simulation model by Hart et al. (1984). These studies
examined the adaptation of cortical form to external
forces. Alternative theories were proposed and used by
Huiskes et al. (1987), Mattheck et al. (1998), Prender-
gast and Taylor (1994) and van der Meulen et al. (1993).
Adaptation of (trabecular) bone density was simulated
by several groups, using different isotropic (Beaupre!
et al., 1990; Carter et al., 1987; Huiskes et al., 1987;
Weinans et al., 1992a, b) or anisotropic theories (Hart
and Fritton, 1997; Jacobs et al., 1997; Luo et al., 1995).
All these trabecular simulation models were phenomen-
ological in nature and simulated the outcome of
coordinated osteoclastic and osteoblastic activity as
either a net increase or decrease in density. A more
mechanistic approach is necessary to represent the true
physiology of the adaptation process, which occurs only
on trabecular surfaces (Huiskes, 2000). A model to
simulate local trabecular adaptation that does allow this
surface mechanism to be simulated, and also includes a
biological osteocyte mechanosensory and signaling
function, was developed by Mullender and Huiskes
 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414
(1995). A model that also includes separate descriptions
of osteoclastic resorption and osteoblastic formation,
enabling simulation of mechanobiologically modulated
growth, adaptation and maintenance (remodeling),
illustrated in Fig. 6, was published recently (Huiskes
et al., 2000; Ruimerman et al., 2001). Adachi et al.
(2001) proposed a phenomenological adaptation model
for the trabecular structure using nonuniformity of the
local surface stress distribution as the regulatory feed-
back signal.
The models incorporating trabecular morphology
have been used only to study adaptation in small cubes
of bone. Analyses of whole bone FEA models with
refinement down to the trabecular level are feasible now
(van Rietbergen et al., 1999). However, there is not
enough computer capacity yet to use those in iterative
simulations to study trabecular adaptation based on a
(mechanistic) cell modulation scheme. Future models
will need to include both natural boundary conditions at
external bone surfaces and refined trabecular architec-
ture.
4. Discussion
So why mechanobiology? While the term is the inverse
of biomechanics, the definition of biomechanics clearly
encompasses the experimental and computational mod-
els described here: ‘The science that studies the effects of
forces on biological tissues, organs and organisms, in
relation to biological and medical problems’’ (ESB,
1978). As a word, however, mechanobiology moves the
emphasis from mechanics to biology and to determining
the mechanisms behind ‘form follows function’. But
then what does ‘function’ follow? In an evolutionary
sense, one could consider function to follow form, a
strategy that has enabled vertebrates to successfully
breed and survive. This then produces ‘yform
follows function follows formy’ Considering this
cycle of repetition, one can now distinguish between
the two terms: biomechanics focuses on the latter part,
whether function follows form, whereas mechanobiol-
ogy emphasizes the former, whether function determines
form.
Both experimental and computational studies are
critical to advancing our understanding of form and
function, and even more important is the integration of
models and experiments. Yet we have separated experi-
mental and computational studies in this article, a
distinction that reflects contemporary science and
presumably arises from the cultural roots of both
activities. Experimental studies, particularly in vivo
models, originated in biology, while computational
work is fundamentally based on physics and engineer-
ing. To make significant progress, the objective needs to
be the integration of these two types of studies: models
411
for interpreting experiments and experiments for in-
sights and observation to develop models; yet another
circular mechanism! The challenge is to educate
scientists who can understand and contribute to both
ends of the spectrum and help to eliminate the
distinction. Many other fields rely upon this integration.
In mechanobiology, however, the interdisciplinary
nature of these questions cuts across many additional
spectra besides experiments and simulations: in vivo to
in vitro, biology to engineering, laboratory to clinic, etc.
This not only produces great challenges but also
potentially significant rewards.
Beyond the integration within the field itself, we must
ask where mechanobiology fits in a broader scientific
and social context. Scientifically, mechanobiology is
central to unraveling ‘nature versus nurture’ distinc-
tions. With the sequencing of the human genome
complete (Venter et al., 2001) it is now apparent that
the genetic code is only the beginning and provides few
answers to how form is generated and maintained. The
limitations of genomics and proteomics emphasize the
importance of understanding the role of environmental
influences, particularly biophysical factors (Stewart,
1998). Scientific interest in form and function is not
new, but recent advances in cell and molecular biology
allow examination of phenomena that were previously
inaccessible.
On Earth, gravity is ubiquitous and is likely one of the
most pervasive factors in the development of tissues.
This particularly holds true for the musculoskeletal
system, which plays a central role in vertebrate’s ability
to function in Earth’s gravitational environment. The
potential reward for successful quantitative models and
insightful experiments is enormous. The challenge lies in
disentangling environmental modulation and genetic
predisposition in the skeleton. The conservation of
osseous microstructure and morphology speak, to a high
degree, of predisposition. Yet, the action of gravity and
the resulting mechanical stimuli are also highly con-
served, making a case for modulation. Successfully
distinguishing adaptation from genetic programming of
cell metabolism will be a great mechanobiological feat,
requiring intimate interaction between experimental and
computational efforts.
The potential for mechanobiology to contribute to
clinical progress is also promising. Mechanically based
diseases such as osteoporosis and osteoarthritis are
already areas of intense research activity. In addition,
the development of successful synthetic and engineered
organs and tissues, ‘tissue engineering’, will depend on
mechanobiological progress. This is especially true for
functional tissue engineering (Butler et al., 2000). Not
only is an intimate knowledge of the natural tissues
necessary, but controlling and modulating mechanical
stimuli will be essential to develop appropriately
engineered organs and to integrate and function with
412 M.C.H. van der Meulen, R. Huiskes / Journal of Biomechanics 35 (2002) 401–414
the host. The realization of this promise will be
particularly rewarding.
Acknowledgements
This article synthesizes two presentations from the
Mechanobiology Symposium of the 12th Conference of
the European Society of Biomechanics in Dublin,
Ireland, August 28–30, 2000. We thank the ESB and
the organizing committee for the opportunity to present
our work. Funding is acknowledged from the National
Institutes of Health (USA) and the Netherlands
Foundation for Research (NWO-MW).
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