Bone 42 (2008) 775 – 787

www.elsevier.com/locate/bone

Correlations between grey-level variations in 2D projection images (TBS)

and 3D microarchitecture: Applications in the study of human
trabecular bone microarchitecture
Laurent Pothuaud a,⁎, Pascal Carceller a , Didier Hans b
a
Med-Imaps Plateforme Technologique d'Innovation Biomédicale-Xavier Arnozan Hospital, University Hospital of Bordeaux, Pessac, France
b
Department of Radiology, Geneva University Hospital, Geneva, Switzerland
Received 5 May 2007; revised 20 November 2007; accepted 28 November 2007
Available online 29 January 2008

Abstract

X-ray imaging remains a very cost-effective technique, with many applications in both medical and material science. However, the physical
process of X-ray imaging transforms (e.g. projects) the 3-dimensional (3D) microarchitecture of the object or tissue being studied into a complex 2D
grey-level texture. The 3D/2D projection process continues to be a difficult mathematical problem, and neither demonstrations nor well-established
correlations have positioned 2D texture analysis-based measurement as a valid indirect evaluation of 3D microarchitecture. The trabecular bone score
(TBS) is a new grey-level texture measurement which utilizes experimental variograms of 2D projection images. The aim of the present study was to
determine the level of correlation between the 3D characteristics of trabecular bone microarchitecture, as evaluated using µCT reconstruction, and
TBS, as evaluated using 2D projection images derived directly from 3D µCT reconstruction. Analyses were performed using sets of human cadaver
bone samples from different anatomical sites (lumbar spine, femoral neck, and distal radius). Significant correlations were established via standard
multiple regression analysis, and via the use of a generic mathematical 3D/2D relationship. In both instances, the correlations established a significant
relationship between TBS and two 3D characteristics of bone microarchitecture: bone volume fraction and mean bone thickness. In particular, it
appears that TBS permits to accurately differentiate between two 3D microarchitectures that exhibit the same amount of bone, but different trabecular
characteristics. These results demonstrate the existence of a robust and generic relationship, taking into consideration a simplified model of a 2D
projection image. Ultimately, this may lead to using TBS measurements directly on DXA images obtained in routine clinical practice.
© 2007 Elsevier Inc. All rights reserved.

Keywords: Trabecular bone microarchitecture; 2D projection-based evaluation; 3D/2D correlation; TBS (trabecular bone score); Mathematical modeling

Introduction 2D grey-level plain-projection image. Presently, only macro-

X-ray imaging remains a very cost-effective technique, with
many applications in both the medical and material science fields.
The digital (or digitized) radiograph saves the image that is
formed by X-rays which are more or less absorbed when passing
through various tissues or materials, depending upon the sub-
stance's composition, thickness, and density. This process trans-
forms (e.g. projects) 3 dimensional (3D) objects or tissues into a
⁎ Corresponding author. Med-Imaps Plateforme Technologique d'Innovation
Biomédicale, Hôpital Xavier Arnozan, Centre Hospitalier Universitaire de
Bordeaux, Avenue du Haut-Lévêque, 33600 Pessac, France. Fax: +33 5 57 10 28 56.
E-mail address: lpothuaud@medimaps.fr (L. Pothuaud).
8756-3282/$ - see front matter © 2007 Elsevier Inc. All rights reserved.
doi:10.1016/j.bone.2007.11.018
scopic geometry or defects in the 3D tissues or materials are
evaluated using plain-projection 2D images.
In medical X-ray imaging, bone tissue constitutes the major
natural contrast, due to its high content of calcium and other X-ray
absorbing minerals. The morphology of bone can be evaluated
from X-ray images directly. Examples of this are the cortical bone
index [1], vertebral morphology [2], and hip geometry [3]. Fur-
thermore, several X-ray-based techniques have been developed to
evaluate bone mineral density, based upon the relationship bet-
ween the grey level on 2D projection images and the attenuation
of an X-ray beam at a single point. The World Health Orga-
nization (WHO) Working Group defines osteoporosis based upon
measurements of Bone Mineral Density (BMD) obtained using
776 L. Pothuaud et al. / Bone 42 (2008) 775–787
Dual X-ray Absorptiometry (DXA). To date, however, the diag-
nosis of osteoporosis suffers from the lack of any evaluation of the
3D microarchitecture of bone tissue, which constitutes an im-
portant additional component of bone strength [4], as noted in the
definition of osteoporosis itself [5].
Two-dimensional projection-based images do not directly
portray a material's 3D microarchitecture. However, several kinds
of texture analysis, such as fractal analysis, have been proposed as
an indirect measurement of 3D trabecular bone microarchitecture
[6–12]. In the case of materials with surface fractal properties, a
mathematical relationship can be established between the fractal
dimension of 3D microarchitecture and the fractal dimension
evaluated on a 2D projection image. This 3D/2D relationship is
not adequate for trabecular bone microarchitecture, however,
because it does not satisfy surface fractal properties. Other limi-
tations of fractal analysis concern evaluation of the fractal di-
mension itself. An accurate estimate of the fractal dimension of a
2D projection image must take into account a large surface of
projection, and it depends upon the estimator using a certain size
of projection image in the 2-power scale. These constraints are not
compatible with small and/or irregular surfaces of analysis, such
as the standard regions of measurement contained in DXA images.
Based upon the physical process of X-ray absorption, varia-
tions in the grey levels measured in a 2D projection-based image
in a given region of interest reflect the global variations in X-ray
absorption properties that exist in the corresponding 3D tissue
microarchitecture. Porous materials constitute a class of materials
wherein such variations are maximal, since X-rays are alterna-
tively absorbed by the solid (mineralized) and void (porous)
phases. Hence, spatial variations in grey-level texture in 2D
projection-based images of porous materials should constitute an
indirect measurement of the boundary between the solid and
porous phases within the 3D microarchitecture. Trabecular bone
tissue is not exactly a porous material; nonetheless, it can be
considered such, in the sense that it is composed of solid (mine-
ralized bone) and minimally mineralized (bone marrow) phases.
In order to take advantage of 2D projection-based evalua-
tions of porous materials, it first is necessary to establish an
empirical relationship between 3D microarchitecture and 2D
image texture; that is, to understand the ‘nature and causes’ of
spatial variations in 2D images, in terms of how they relate to
3D microarchitecture. Two major approaches are available
when trying to elucidate the nature and causes of spatial varia-
tions in 2D projection-based images: empirical and theore-
tical. The empirical approach is based upon the definition of
parameters that measure 2D texture, followed by the study of
correlations between these 2D parameters and various 3D
microarchitectural characteristics. This approach is character-
ized by a need for large sets of images, and these images should
represent a wide variety of microarchitectural characteristics, so
as to identify robust and useful relationships. Ideally, images
wherein only the parameter of interest varies should be utilized
for this purpose. In this context, the theoretical approach, which
is based upon numerical simulations and artificial images, re-
presents a unique alternative, since it allows for the study of the
sensitivity of 2D textural information to detect a number of 3D
microarchitectural characteristics.
In the literature, almost all the studies exploring 3D/2D cor-
relations in trabecular bone tissue have been empirically-based
[7–9]; only a very few studies have explored the theoretical
approach [13,14]. To date, the 3D/2D projection remains a dif-
ficult mathematical problem [6], and neither demonstrations nor
well-established correlations have validated 2D texture analysis-
based measurement as a true indirect evaluation of 3D trabecular
bone microarchitecture. The key stumbling block is constructing
an empirical model that correlates 3D microarchitecture and 2D
projection texture.
The imaging techniques considered here are standard X-rays
and DXA. Both techniques are based upon the absorption of X-rays
by the object being examined, whether that object is an inanimate
object or live tissue, and the transformation of 3D microarchitecture
into complex 2D grey-level texture—a 2D projection image. Based
upon the Beer–Lambert law, the physical process is complex,
taking into account such parameters as the absorption and diffusion
of X-rays through the object, the geometry of acquisition (source/
detector), and the characteristics of the detection unit [15]. In order
to comprehend the effects of projection and the correlations bet-
ween 3D microarchitecture and 2D projection-based grey-level
texture, a simplified model of study consists of summing all binary
slices of the 3D microarchitecture into a parallel, plain-projection,
2D image [13,14]. This simplified projection process can be con-
sidered the first step in the real projection process [14]; if no
correlation is established in this first stage, no correlation will be
established in the final stage that is comprised of the real bone
radiographic 2D image.
The trabecular bone score (TBS) is a new grey-level texture
measurement that utilizes an experimental variogram of 2D
projection images [16]. TBS is not an estimator of fractal dimen-
sion. Rather, it measures the rate of local variation in grey levels
within 2D projection images. The aim of the present study was to
determine the levels of correlation between certain 3D character-
istics of trabecular bone microarchitecture, as evaluated using µCT
reconstruction, and TBS, as evaluated using 2D projection images
derived directly from 3D µCT reconstruction. This approach cons-
titutes a first step in our development and validation of this new
TBS measurement tool, the final objective being to validate TBS
for direct use on DXA images obtained in routine clinical practice.
Materials and methods
Bone samples
Fifty-seven human cadaver bone pieces were obtained from the Anatomy
Laboratory of the University Hospital of Bordeaux (France). These dried bone pieces,
deprived of marrow, had been dedicated for practice sessions with students in
Medicine and Orthopaedics. Bone pieces from three different anatomical sites were
selected for study: (1) spine samples, consisting of 13 lumbar and 7 low thoracic
vertebrae (n=20); (2) femoral neck samples (n=17); and (3) radius samples (n=20).
Three-dimensional reconstructions
Three-dimensional reconstruction image of trabecular bone volume was
obtained for each piece using high-resolution micro-Computed Tomography
(eXplore Locus, GE HealthCare), with cubic voxels comprised of 93 µm sides.
A specific region of measurement, excluding cortical bone, was defined for each
piece type (Fig. 1): (1) for spine samples, the vertebral body (Vbody); (2) for hip
 L. Pothuaud et al. / Bone 42 (2008) 775–787 777

samples, 3/4 of the neck region (neck34); (3) for radius samples, 15 mm in the
most distal segment, termed the ultradistal segment (UD-15 mm). The grey-level
µCT reconstructions were binarized by means of global thresholding, with the
threshold value automatically determined inside the region of measurement
(MicroView 2.1.2, GE HealthCare: Bone Analysis/Auto Threshold plugin). The
volume of analysis (V [mm3]) was determined from the total number of voxels
inside the region of measurement.
Evaluation of three-dimensional parameters
The 3D microarchitecture of trabecular bone was characterized in terms of
bone volume fraction and two classes of parameter: (1) stereologic parameters,
based upon the parallel plate model; and (2) model-independent parameters,
based upon porous material characterization algorithms. The same region of
measurement was utilized for the evaluation of both parameter classes.
Stereologic parameters (parallel plate model)
Parfitt's stereologic parameters were calculated, using the parallel plate
model [17], following recommended nomenclature [18], and using specific
software: Advanced Bone Analysis plugin in MicroView software (MicroView
calculated as the mean length of many chords (n = 5.106), randomly distributed
throughout the 3D region of measurement. Mean pore thickness (Sp, expressed
in mm) was evaluated following the same algorithm, taking into consideration
the porous instead of the solid phase.
The density of connectivity ( χV , expressed in mm− 3) was evaluated based
upon the Euler–Poincare relationship [21]: χV =β0 +β2 − EPC, where EPC is the
Euler–Poincare characteristic [22]; β0 and β2 are the 0-order and 2nd-order Betti's
numbers, respectively; and V is the volume of the entire region of measurement.
Two-dimensional projection image
To each 3D reconstruction of trabecular bone volume (Ω) was associated a
2D projection image (P) (Fig. 1).
The coordinate system (x, y, z) in the 3D space was oriented so that the third
axis (Z) corresponded to the projection axis. To each point (pixel) of coordinates
(x, y) on the projection image, P was attributed a grey level P(x, y), which was
calculated as follows:
X
NZ
Pð x; yÞ ¼ Xð x; y; zÞ ð1Þ
z¼1

2.1.2, GE HealthCare). These parameters were trabecular thickness (TbTh, where

expressed in mm), trabecular spacing (TbSp, expressed in mm), and trabecular
number (TbN, expressed in mm− 1).
Bone volume fraction
Bone volume fraction was calculated as the ratio of bone volume (BV) to
total volume (TV) in the entire region of measurement. BV/TV was evaluated
using MicroView software, and expressed as a percentage.
Model-independent parameters
A chord length distribution algorithm [19,20] was used to evaluate model-
independent mean thicknesses. Mean solid thickness (Th, expressed in mm) was
evaluated as the mean chord length of the solid phase. A chord of solid was
determined as a linear segment with endpoints at the solid/pore interface. Th was
(x, y, z) coordinates in the X, Y, and Z axes, respectively
P the 2D projection image
P(x, y) the grey-level value corresponding to the pixel of coordinates (x, y)
in the 2D projection image
Ω the 3D binary image
Ω(x, y, z) the binary value (porous = 0, solid = 1) corresponding to the voxel of
coordinates (x, y, z) in Ω
NZ the projection thickness (size of Ω following the Z axis)
The projection axis (Z) was determined following the anterior–posterior axis
for all anatomical sites (Fig. 1). The 2D region of measurement for TBS evaluation
was defined as the entire surface of the projected 3D region of measurement. Under

Fig. 1. (a) Three-dimensional reconstruction of the trabecular bone microarchitecture in a human cadaver spine sample. The 3D region of measurement corresponded to
the entire vertebral body, excluding cortical bone. The 2D projection image was obtained by projecting the 3D volume following the anterior–posterior axis (b). TBS
was evaluated across the entire projected area. This sample was characterized in 3D by BV/TV = 31.8% and Th = 0.259 mm, and in 2D by TBS = 7.831 mm− 1.
778 L. Pothuaud et al. / Bone 42 (2008) 775–787

these conditions, the 2D regions of measurement were representative of the regions Non-linear data modeling
of standard clinical use for both the spine and radius (entire vertebral body, 15-mm
ultradistal area). With hip samples, the 2D region of measurement was defined as a An empirical, non-linear regression model linking TBS and 3D parameters was
15 mm-thick section of the femoral neck. This section was reduced in width (3/4) in determined, based upon numerical simulations and models of 3D porous materials
order to exclude lateral cortical bone projection. (see Appendix). In the present study, this mathematically-defined 3D/2D correlation
model was evaluated as a tool to analyze trabecular bone microarchitecture. Fit-
TBS (trabecular bone score) evaluation coefficients, as well as the predictive power and significance of this non-linear
regression model were determined using SigmaPlot software (version 9.0.1).
Each 2D projection image, P(x, y), was used for TBS evaluation. A point M
Y
of P was located from the corresponding vector M ð x; yÞ, where (x, y) were the
Results
spatial coordinates for P. The experimental variogram V(k) vs. k was calculated
by averaging the difference squared of P values over several pairs of points
located at a specified distance, k: Descriptive statistics at each anatomical site (Table 1)
h    i2
h Y
V ðk Þ ¼ P M 0 þ k T Y uh  P M 0
Y
i ð2Þ Mean value, standard deviation, and range of values were
where evaluated independently for each parameter at each anatomical
site (Table 1). TBS values, as expressed in mm− 1, varied
k the unit distance (varying from 0 to 10, in steps of 1 pixel) globally from 7.430 to 11.373 mm− 1, with different ranges of
V(k) the experimental variogram function, representing the grey level values across the three anatomical sites.
variations in function of the distance k;
Y Depending upon the anatomical site, projection thickness
M0 the initial point in the 2D projection image P
θ the angle defining a direction from the horizontal line passing through
(evaluated from the number of projected planes of the 3D
Y volume) varied from 14.1 to 21.2 mm for spine samples
M0
Y
uh the unit vector in the θ direction (anterior–posterior projection axis), from 27.0 to 38.5 mm for
femoral neck samples (anterior–posterior projection axis), and
6 from 18.8 to 26.5 mm for ultradistal radius samples (axial
 was
Averaging  performed across a large number of calculations (n = 5.10 ) with
Y projection axis). Hence, although globally the projection
different M 0 ; h initialization points. In order to perform mean evaluations of
grey-level differences between two points in the 2D projection 
image, independent thickness varied from 14.1 to 38.5 mm, the range of projection
Y
of the orientation of this pair of points, each initialization M 0 ; h was determined thickness at each anatomical site was about 10 mm. There was
randomly with iso-probability for each angle over the 2π rotation. no statistically-significant correlation between TBS and projec-
TBS was defined as the slope at the origin of the log–log representation of V(k),
tion thickness at any site: r = 0.26 ( p-value = 0.26) for spine
evaluated from least square regression line calculation. TBS characterized the rate
of variation of the grey levels of the 2D projection image, and was expressed in samples; r = − 0.06 ( p-value = 0.82) for femoral neck samples;
mm− 1 units, taking into consideration the size of the pixel (93 µm) (Fig. 2). and r = 0.27 ( p-value = 0.26) for distal radius samples.

Statistical analyses Correlation between parameters at each anatomical site

MedCalc software (version 9.0.1.1) was used to provide descriptive values
(mean, standard deviation, range) for each investigated parameter, and Pearson's High correlations were evaluated between parallel plate
correlation coefficients between parameters, and for multiple regression ana- model-based parameters and model-independent parameters
lyses (linear regression). (Table 2). For the spine samples, r = 0.94 ( p-value b 0.0001)

Fig. 2. Illustrating calculation of TBS as the slope at the origin of the experimental variogram, in the case of two samples with similar values of BV/TV: sample1 with
BV/TV = 36.1% and sample2 with BV/TV = 36.6%. The sample with the lower trabecular number (TbN1 = 1.373 mm− 1 vs. TbN2 = 1.575 mm− 1), which also means
more widely-spaced trabeculae, was characterized in the 2D projection image as having a higher TBS value (TBS1 N TBS2). In other words, the more widely-spaced the
trabeculae were in the 3D volume, the greater the degree of grey-level variation in the 2D projection image.
 L. Pothuaud et al. / Bone 42 (2008) 775–787 779

Table 1
Descriptive statistics at each anatomical site
Samples TBS BV/TV TbTh TbSp TbN Th Sp Cv
(mm− 1) (%) (mm) (mm) (mm− 1) (mm) (mm) (mm− 3)
Lumbar spine (n = 20)
Mean 8.376 30.6 0.233 0.551 1.309 0.268 0.623 3.325
SD 0.495 5.5 0.014 0.139 0.206 0.015 0.147 1.272
Range 7.430–9.118 18.8–38.7 0.210–0.263 0.389–0.909 0.893–1.616 0.238–0.295 0.456–1.012 1.396–5.494

Femoral neck (n = 17)

Mean 10.124 26.2 0.239 0.750 1.095 0.338 0.950 1.811
SD 0.462 6.7 0.023 0.378 0.251 0.035 0.168 0.608
Range 9.407–11.373 11.2–38.3 0.200–0.286 0.461–2.058 0.432–1.374 0.295–0.420 0.725–1.407 0.343–2.533

Ultradistal radius (n = 20)

Mean 9.852 28.4 0.226 0.584 1.255 0.350 0.765 2.395
SD 0.411 4.3 0.018 0.105 0.173 0.034 0.100 0.695
Range 8.953–10.577 23.0–36.5 0.201–0.269 0.401–0.772 0.998–1.589 0.298–0.414 0.557–0.936 1.588–3.824

between TbTh and Th; r = 0.99 (p-value b 0.0001) between appeared to be a significant variable in all of the linear regression
TbSp and Sp; and r = 0.97 ( p-value b 0.0001) between TbN and models (Table 3).
Cv. For the femoral neck samples, r = 0.87 ( p-value b 0.0001)
between TbTh and Th; r = 0.91 ( p-value b 0.0001) between Generic 3D/2D correlations, with TBS as the dependent variable
TbSp and Sp; and r = 0.96 ( p-value b 0.0001) between TbN and
Cv. At the distal radius, r = 0.92 ( p-value b 0.0001) between The empirical non-linear regression model linking TBS with
TbTh and Th; r = 0.85 ( p-value b 0.0001) between TbSp and BV/TV and mean bone thickness, extrapolated from the Eq. 5,
Sp; and r = 0.95 ( p-value b 0.0001) between TbN and Cv. was evaluated at each anatomical site, with both TbTh and Th
Only moderate to low-level correlations were identified
between TbTh and BV/TV (0.38 ≤ r ≤ 0.59, depending upon the
anatomical site); correlations between Th and BV/TV were Table 2
moderate to high (0.53 ≤ r ≤ 0.73, again depending upon the Correlations between parameters at each anatomical site
anatomical site). Samples TBS BV/TV TbTh TbSp TbN Th Sp Cv
The correlation between TBS and 3D parameters was dependent Lumbar spine
upon the anatomical site. Among spine samples, more significant TBS 1.00 − 0.63 0.23 0.73 − 0.84 0.02 0.76 − 0.85
correlation was observed between TBS and TbN (resp. Cv): r= BV/TV 1.00 0.59 − 0.97 0.94 0.73 − 0.94 0.90
− 0.84, p-value b 0.0001 (resp. r = − 0.85, p-value b 0.0001). TbTh 1.00 − 0.44 0.28 0.94 − 0.36 0.22
TbSp 1.00 − 0.98 − 0.60 0.99 − 0.91
Among femoral neck samples, more significant correlation was
TbN 1.00 0.48 0.97 0.97
obtained between TBS and TbSp (resp. Sp): r = 0.62, p-value= Th 1.00 − 0.54 0.40
0.0079 (resp. r = 0.56, p-value = 0.0194). In the set of distal radius Sp 1.00 − 0.89
samples, higher correlation was obtained between TBS and TbTh Cv 1.00
(resp. Th): r =0.83, p-valueb 0.0001 (resp. r =0.69, p-value=
Femoral neck
0.0008).
TBS 1.00 − 0.25 0.57 0.62 − 0.52 0.64 0.56 − 0.53
BV/TV 1.00 0.42 − 0.86 0.92 0.53 − 0.91 0.89
Multiple regression analysis, with TBS as the dependent variable TbTh 1.00 − 0.00 0.04 0.87 − 0.07 0.07
TbSp 1.00 − 0.93 − 0.10 0.91 − 0.88
Stepwise multiple regression analysis was performed entering TbN 1.00 0.21 − 0.97 0.96
Th 1.00 − 0.18 0.18
two sets of independent variables: (1) {BV/TV, TbTh, TbSp,
Sp 1.00 − 0.96
TbN} in relation to parallel plate model-based 3D parameters; and Cv 1.00
(2) {BV/TV, Th, Sp, Cv} in relation to model-independent 3D
parameters. All analyses examined the significance of the model Ultradistal radius
and the relative contributions of the various independent vari- TBS 1.00 0.00 0.83 0.34 − 0.46 0.69 0.22 − 0.60
BV/TV 1.00 0.38 − 0.91 0.86 0.66 − 0.89 0.74
ables, while expressing TBS as a linear function of two 3D
TbTh 1.00 0.02 − 0.14 0.92 − 0.20 − 0.28
parameters. In the case of spine samples, TBS was assessed as a TbSp 1.00 − 0.98 − 0.32 0.85 − 0.91
function of TbTh and TbN (resp. Th and Cv); with femoral neck TbN 1.00 0.20 − 0.85 0.95
samples, as a function of TbTh and TbSp (resp. Th and Sp); and, Th 1.00 − 0.47 0.06
with the radius sample, as a function of TbTh and TbN (resp. Th Sp 1.00 − 0.82
Cv 1.00
and Cv). Mean bone thickness, as evaluated by TbTh or Th,
780 L. Pothuaud et al. / Bone 42 (2008) 775–787

Table 3
Multiple regression analysis, with TBS as the dependent variable
Samples TBS = F(BV/TV, TbTh, TbSp, TbN) TBS = F(BV/TV, Th, Sp, Cv)
Lumbar spine (n = 20)
Coefficient Std. error t p-value Coefficient Std. error t p-value
Constant 7.4294 – – – Constant 5.9054 – – –
TbTh 17.2771 2.1847 7.908 b0.0001 Th 14.1213 2.9997 4.708 0.0002
TbN − 2.3508 0.1523 − 15.438 b0.0001 Cv − 0.3966 0.0360 − 11.029 b0.0001

Adj-R2 RMSSD F-ratio p-value Adj-R2 RMSSD F-ratio p-value

Model 0.93 0.0854 126.0296 b0.001 Model 0.88 0.1190 60.8616 b0.001

Femoral neck (n = 17)

Coefficient Std. error t p-value Coefficient Std. error t p-value
Constant 6.8452 – – – Constant 4.8797 – – –
TbTh 11.3384 2.8252 4.013 0.0013 Th 10.1602 1.2949 7.846 b0.0001
TbSp 0.7610 0.1747 4.356 0.0007 Sp 1.9049 0.2655 7.175 b0.0001

Adj-R2 RMSSD F-ratio p-value Adj-R2 RMSSD F-ratio p-value

Model 0.67 0.1693 17.4715 b0.001 Model 0.85 0.1129 48.0614 b0.001

Ultradistal radius (n = 20)

Coefficient Std. error t p-value Coefficient Std. error t p-value
Constant 6.9660 – – – Constant 7.6622 – – –
TbTh 17.3649 2.4076 7.213 b0.0001 Th 8.8671 1.0233 8.665 b0.0001
TbN − 0.8335 0.2564 − 3.251 0.0047 Cv − 0.3797 0.0495 − 7.678 b0.0001

Adj-R2 RMSSD F-ratio p-value Adj-R2 RMSSD F-ratio p-value

Model 0.78 0.1247 35.3778 b0.001 Model 0.87 0.0974 63.3411 b0.001

Table 4
Generic 3D/2D correlation, with TBS as the dependent variable
Samples TBS = F1(BV/TV, TbTh) TBS = F1(BV/TV, Th)
Lumbar spine (n = 20)
Coefficient Std. error t p-value Coefficient Std. error t p-value
a00 11.3005 0.2169 52.0939 b0.0001 a00 11.8278 0.3208 36.8696 b0.0001
a10 0.2207 0.0262 8.4268 b0.0001 a10 0.2466 0.0382 6.4495 b0.0001
a11 0.2178 0.0201 10.8591 b0.0001 a11 0.2745 0.0340 8.0825 b0.0001

Adj-R2 RMSSD F-ratio p-value Adj-R2 RMSSD F-ratio p-value

Model 0.91 0.1441 102.7130 b0.0001 Model 0.86 0.1845 59.3618 b0.0001

Femoral neck (n = 17)

Coefficient Std. error t p-value Coefficient Std. error t p-value
a00 11.2594 0.3463 32.5178 b0.0001 a00 11.5879 0.2427 47.7412 b0.0001
a10 0.1625 0.0424 3.8348 0.0018 a10 0.1201 0.0191 6.2925 b0.0001
a11 0.1443 0.0327 4.4086 0.0006 a11 0.1634 0.0208 7.8563 b0.0001

Adj-R2 RMSSD F-ratio p-value Adj-R2 RMSSD F-ratio p-value

Model 0.55 0.3089 10.8562 0.0014 Model 0.80 0.2053 33.4388 b0.0001

Ultradistal radius (n = 20)

Coefficient Std. error t p-value Coefficient Std. error t p-value
a00 10.9221 0.3064 35.6446 b0.0001 a00 12.3057 0.3477 35.3904 b0.0001
a10 0.2354 0.0282 8.3359 b0.0001 a10 0.1224 0.0148 8.2451 b0.0001
a11 0.1841 0.0207 8.8887 b0.0001 a11 0.1996 0.0196 10.1996 b0.0001

Adj-R2 RMSSD F-ratio p-value Adj-R2 RMSSD F-ratio p-value

Model 0.80 0.1828 39.5042 b0.0001 Model 0.84 0.1628 52.0157 b0.0001
 L. Pothuaud et al. / Bone 42 (2008) 775–787 781

Table 5
Generic 3D/2D correlation, with TbTh (or, Th) as the dependent variable
Samples TbTh = F2(BV/TV,TBS) Th = F2(BV/TV,TBS)
Lumbar spine (n = 20)
Coefficient Std. error t p-value Coefficient Std. error t p-value
a00 11.4778 0.2752 41.7145 b0.0001 a00 12.1963 0.3825 31.886 b0.0001
a10 0.2922 0.0413 7.0764 b0.0001 a10 0.3421 0.0538 6.3571 b0.0001
a11 0.2710 0.0320 8.4789 b0.0001 a11 0.3566 0.0480 7.4313 b0.0001

R2 RMSSD F-ratio p-value R2 RMSSD F-ratio p-value

Model 0.86 0.0053 60.5882 b0.0001 Model 0.88 0.0053 68.9240 b0.0001

Femoral neck (n = 17)

Coefficient Std. error t p-value Coefficient Std. error t p-value
a00 11.8862 0.2490 47.7490 b0.0001 a00 11.9713 0.1659 72.1713 b0.0001
a10 0.3183 0.0702 4.5361 0.0005 a10 0.1819 0.0297 6.1219 b0.0001
a11 0.2707 0.0502 5.3888 b0.0001 a11 0.2347 0.0286 8.1998 b0.0001

R2 RMSSD F-ratio p-value R2 RMSSD F-ratio p-value

Model 0.65 0.0139 15.5465 0.0003 Model 0.81 0.0148 36.223 b0.0001

Ultradistal radius (n = 20)

Coefficient Std. error t p-value Coefficient Std. error t p-value
a00 11.1456 0.3822 29.1609 b0.0001 a00 12.6177 0.4176 30.2183 b0.0001
a10 0.2804 0.0345 8.1234 b0.0001 a10 0.1388 0.0175 7.9233 b0.0001
a11 0.2197 0.0258 8.5251 b0.0001 a11 0.2258 0.0232 9.7182 b0.0001

R2 RMSSD F-ratio p-value R2 RMSSD F-ratio p-value

Model 0.82 0.0078 44.4551 b0.001 Model 0.91 0.0102 93.3428 b0.0001

used to estimate mean solid thickness. Results were obtained
under the generic condition a01 = 0, which did not change the
nature of the generic 3D/2D correlation, but meant that all
regression lines in the (TBS, BV/TV) plot intersected each other
at the same point. All analyses identified significance of the
model and co-factors (Table 4).
Generic 3D/2D correlation, with TbTh (or Th) as the dependent
variable
The empirical non-linear regression model, which linked
mean bone thickness with BV/TV and TBS extrapolated from
Eq. 6, was evaluated at each anatomical site, with both TbTh

Fig. 3. Relationship between TBS and BV/TV. The linear regression model expressed TBS as a function of BV/TV, but only explained a minority of the variance in
TBS (R2 = 0.39). The use of Th as a co-factor in the explanation of TBS allowed for determination of a unique value of TBS, just by knowing BV/TV and Th,
significantly enhancing the predictive power of the model (R2 = 0.86): The network of dashed lines (calculated from Eq. 5) helped to explain the dispersion of the
experimental points across the linear regression line.
782 L. Pothuaud et al. / Bone 42 (2008) 775–787
Fig. 4. Relationship between Th and BV/TV. The linear regression model expressed Th as a function of BV/TV, but only explained about half of the variance in Th
(R2 = 0.53). The use of TBS as a co-factor to explain Th allowed for determination of a unique value of Th, from the knowledge of both BV/TV and TBS, significantly
enhancing the predictive power of the model (R2 = 0.88): The network of dashed lines (calculated from Eq. 6) helped to explain the dispersion of the experimental
points across the linear regression line.
and Th to estimate mean solid thickness. Results were obtained
under the generic condition a01 = 0, which did not change the
nature of the generic 3D/2D correlation (see above). All
analyses identified significance of the model and co-factors
(Table 5).
Interpretation of the generic 3D/2D correlation model
As previously reported, BV/TV did not appear to be a
significant co-factor in multiple regression analysis (Table 3).
However, it was found to be a significant co-factor, in com-
plement with mean solid thickness, when imposed as a co-factor
via a generic 3D/2D correlation model. For example, in the set
of spine samples, the linear regression model expressing TBS as
a function of BV/TV (Fig. 3) was determined to have relatively
poor predictive power (R2 = 0.39). The use of Th as a co-factor
for TBS allowed for the determination of a unique value of
TBS, derived from knowledge of both BV/TV and Th, with
significantly enhanced predictive power (R2 = 0.86): TBS was
defined as ‘a score of microarchitecture that can be evaluated as
a function of bone volume fraction and mean bone thickness’.
The network of dashed lines, calculated from the mathematical
3D/2D model (Eq. 5), helped to explain the dispersion of the
experimental points across the linear regression line. In parti-
cular, two samples with similar BV/TV (for example, sample1
with BV/TV = 36.1% and sample2 with BV/TV = 36.6%) and
different TBS values were characterized by different mean bone
thicknesses (Th). In such an instance, the higher the value of
TBS was, the higher the mean bone thickness was.
Similarly, the linear regression model expressing Th as a
function of BV/TV (Fig. 4) only explained about half the variance
(R2 =0.53), whereas the additional insertion of TBS into the model
raised R2 to 0.88. Hence, TBS was defined as “a differentiating
factor in the relationship between mean bone thickness and bone
volume fraction.” The network of dashed lines, calculated from
the mathematical 3D/2D model (Eq. 6), helped to explain the
dispersion of experimental points across the linear regression line.
Again, two samples with close BV/TV (for example, sample1
with BV/TV = 36.1% and sample2 with BV/TV = 36.6%) and
different mean bone thicknesses (Th) were distinguished by
means of different TBS values. In such cases, the higher the mean
bone thickness was, the higher the value of TBS was.
TBS was calculated as the slope at the origin of the expe-
rimental variogram. In that same case of two samples with close
values of BV/TV (sample1 with BV/TV = 36.1% and sample2
with BV/TV = 36.6%), the difference in grey-level variation
in the 2D projection images depended upon the number of
trabeculae or, similarly, the space between trabeculae (Fig. 2).
In other words, the more space between trabeculae in the 3D
image, the larger the grey-level variations were in the 2D
projection image.
Discussion
In this study, we evaluated the correlation between grey-level
variations in 2D projection images (TBS) and certain para-
meters of 3D microarchitecture. Significant correlations were
identified by means of two distinct approaches: (1) standard
multiple regression analysis; and (2) a generic mathematical 3D/
2D relationship. In both instances, the levels of correlation were
high enough to establish a significant relationship between TBS
and two 3D characteristics of bone microarchitecture: bone
volume fraction and mean bone thickness. The mean bone
thickness did not correlate well with bone volume fraction;
consequently, the traditional belief that more bone means
thicker bone is inaccurate. On the other hand, using TBS in
addition to bone volume fraction allowed for highly accurate
predictions of variance in mean solid thickness. In particular, it
appears that using TBS accurately differentiates between two
3D samples in which the amount of bone is identical, but
trabecular characteristics differ, as in the number, thickness or
spacing of trabeculae.
 L. Pothuaud et al. / Bone 42 (2008) 775–787 783

The aim of the present study was to evaluate the level of
correlation between the 3D characteristics of trabecular bone
microarchitecture, as evaluated via µCT reconstruction, and
TBS, as evaluated via a 2D projection image directly derived
from 3D µCT reconstruction. Using a large sample of human
cadaver bone pieces from different anatomical sites, we iden-
tified a robust and generic relationship that takes into consi-
deration a simplified model of a 2D projection image. Our final
objective in this process is ultimately to use this new TBS
measurement tool directly on DXA images obtained within the
context of clinical practice. The correlation models established
in the present study will be useful to determine and evaluate
specific 3D/2D correlations adapted for the DXA imaging of
bone pieces.
The experimental variogram expresses variance in the grey
levels of the analyzed image as a function of distance. It is a
positively-correlated variable, in that variations in grey level are
statistically higher at higher distances. Beyond a certain
distance (called the ‘range’) the function reaches a constant
value (called the ‘sill’). The range determines the distance
beyond which there is no further correlation between grey
levels. The sill determines the global variance in the grey levels.
Depending upon the scale of the sample, the range (and
consequently, the sill) may or may not appear in the observation
window. The slope at the origin constitutes an estimate of the
roughness of the grey levels: the lower the slope, the higher the
regularity of the grey levels. Experimental variograms have
been used widely as a primary tool for measuring spatial
variations in images, particularly in geostatistics [23], and to
characterize forest parameters from remotely visualized ima-
gery [24,25]. A simplified representation of a forest scene has
been developed by Jupp et al. [26,27]. The model contains a
scene that is composed of discs randomly distributed over an
unbounded contrasting background. The discs can overlap, but
the brightness value of the discs does not change in the
overlapped area. Though simple, the model is very effective at
illustrating the relationship between scene structure and the
spatial characteristics of an image. Via a simulation study

Fig. 5. Examples of 3D volumes generated by numerical simulation, randomly placing spherical holes in an initially solid block. The 3D characteristics were dependent
upon the size of the holes and the fixed solid volume fraction (BV/TV): BV/TV = 35% and Th = 13.5 pixels for a 3D volume generated with holes of size = 12 pixels
(left); BV/TV = 35% and Th = 7.4 pixels for a 3D volume generated with holes of size = 7 pixels (right). Such 3D volumes with the same solid volume fraction and
different mean solid thicknesses were distinguishable by means of different grey-level variations in the 2D projection images; in other words, the greater the mean solid
thickness, the higher the value of TBS: (left) TBS = 0.775 pixels− 1 (Th = 13.5 pixels), (right) TBS = 0.328 pixels− 1 (Th = 7.4 pixels).
784 L. Pothuaud et al. / Bone 42 (2008) 775–787

conducted based upon the previous simplified model, it has
been determined that variogram parameters (range, sill, slope at
the origin) are dependent upon forest parameters, in the
following ways: (1) where it exists, the sill is related to the
proportion of the area covered by an object that is determined by
its density [28]; (2) range is strongly associated with the size of
objects in the scene [28–30]; in particular, range can be used as
an efficient estimator of the diameter of the objects in the scene,
and has been shown to be related to tree density and thinning
and damage by insects [30]; and (3) the slope at the origin
correlates to the extent of the boundary between the objects
within the scene and the background [27]. Woodcock et al. [25]
achieved a similar result analyzing simulated images: “the slope
at the origin depends upon the amount of boundary between
discs and background.”
A 2D projection-based evaluation of trabecular bone micro-
architecture, such as what might be obtained from a standard
radiograph or from DXA, would have several advantages clini-
cally, in that it is relatively inexpensive and convenient, and is
associated with low radiation exposure. Using TBS measure-
ments associated with the mathematical relationship Th′ = F2(BV/
TV, TBS) would constitute a solution if, for example, bone
mineral density could be used in place of BV/TV. In this case, both
BMD and TBS could be evaluated from the same 2D projection-
based DXA image, and Th – a characteristic of 3D micro-
architecture – could be estimated directly from BMD and TBS.
An evaluation of mean bone thickness would be complementary
to BMD, and potentially useful when one wishes to distinguish
patients with the same BMD but different bone status.
At this stage in the development of this technology, however,
several effects and parameters must be clarified before either
standard radiographs or DXA images can be used in conjunc-
tion with TBS measurement in routine clinical practice. The
effects of resolution, geometry of acquisition (fan-beam, pencil-
beam, etc.), and image quality (contrast, luminosity, noise) on
TBS measurement must be studied. For a particular class of
imaging system (with the same resolution and geometry of
acquisition), we must speculate regarding good settings for the
calculation of TBS, in order to optimize the accuracy of the 3D/
2D correlation mathematical equation. Cross-calibration proce-
dures will be necessary to extend TBS measurements to diffe-
rent categories of system. Based upon preliminary data, it seems
that evaluation of TBS is appropriate with classical DXA sys-
tems. Using a sample of 59 clinical DXA images (QDR4500A,
Hologic) of the hip, we assessed TBS measurements for five
standard regions (femoral neck, trochanter, inter-trochanter,
Ward's triangle, and total hip). Contrast and luminosity were set
between 40 and 60 on a 100-scale prior to TBS evaluation; and,
for each combination of contrast and luminosity settings, the
relative error was evaluated, using 50/50 settings as the refe-
rence. Mean relative errors across the 59 images varied from
0.42% (for inter-trochanteric measurements) to 0.75% (for
Ward's triangle). Hence, modest alterations in contrast and/or
luminosity do not seem to influence TBS measurements to any
great degree. Further studies are warranted to evaluate such
image quality effects, however, because they might be clinically
significant in certain patients (for example, in patients who are
morbidly obese). Furthermore, an ex-vivo exactitude study must
be conducted, evaluating the significance and accuracy of the
generic 3D/2D correlation equations F1 and F2 in lumbar spine
and femoral neck samples with DXA images obtained using
different DXA systems.
In summary, we have demonstrated here that grey-level
variations in a 2D projection-based image – as evaluated by TBS
measurements – correlate with certain characteristics of 3D
microarchitecture, such as solid volume fraction and mean solid
thickness. Applying TBS measurements to clinical DXA images
could be an effective and efficient solution to the routine clinical
evaluation of trabecular bone microarchitecture. Other applica-
tions – like the detection of microscopic decalcification, 3D

Fig. 6. Repartition of experimental mean models in the (TBS, BV/TV) plot. Each experimental point was reported with mean values for TBS and BV/TV evaluated
over a set of 30 individual models, generated using different initialization points selected by the random generator. The range of variation among TBS values within the
same set of 30 individual models is depicted using a vertical error-bar. Regrouping of the mean model characteristics by class of Th value allowed for the determination
of a specific linear regression model for each class of Th. Hence, the 3D/2D mathematical model was defined as a linear relationship between TBS and BV/TV (Eq. 3),
the coefficients of this linear relationship dependent upon the mean value of Th over the considered Th class (Eq. 4, Fig. 7).
 L. Pothuaud et al. / Bone 42 (2008) 775–787 785

Fig. 7. The coefficients a0 and a1 in linear regression models evaluated within each Th class (i.e., TBS = a0 + a1 ⁎ BV/TV, Fig. 6) were reported as a function of the
mean value of Th within the considered Th class. Hence, logarithmic regression models were established between the coefficients (a0, a1) and the mean value of Th
(Eq. 4). These logarithmic regression models permit mathematical determination of the relationship between TBS and both 3D characteristics, BV/TV and Th (Eq. 5).

object quality control, and the global characterization of porous
material– might be appropriate for TBS measurements, as well.
Acknowledgments
Sincere thanks are due to all personnel in the Anatomy
Laboratory of the University Hospital of Bordeaux who assisted
with the bone samples: Prs Dominique Midy and Jean-Marc
Vital; Drs Benoît Lavignolle and Mathieu de Sèze; Mrs Jean-
Jacques Barbouteau, Etienne Delamarre and José Prata. Thanks
also go to Dr. Kevin White, a professional editor, who
thoroughly edited this document and provided helpful feedback
regarding statistical analysis.
Appendix A. 3D/2D mathematical relationship
A specific algorithm was developed to generate 3D nume-
rical volumes of porous materials with various characteristics of
3D microarchitecture. The porosity network of each model was
constructed by randomly inserting spherical holes in an initially
solid block (Fig. 5). A numerical matrix of 512 × 512 × 512
voxels was used to generate each model, with (a) a pre-
determined hole size; (b) a pre-determined solid volume fraction
(BV/TV); and (c) a specific initialization of the random gene-
rator (init). For each pair of characteristics (size, BV/TV), a set
of 30 models was generated with different initialization points
of the random generator (Fig. 6).
The mean 3D characteristics, BV/TV and Th, were evaluated
over the 30 models, and mean TBS was calculated over the
30 corresponding models of the 2D projection. The mean charac-
teristics (BV/TV, Th, TBS) then were used for data modeling. The
total set of mean models (BV/TV, Th, TBS) was categorized
according to mean Th values, and then partitioned into 15 dis-
jointed classes. Each class [Th]i (i = 1 to 15) of the mean model
was characterized by the number (N) of mean models and Th
values ranging from minimal (min) to maximal (max). The mean

Fig. 8. Mean variations of TBS values due to reduction of projection thickness reported as a function of the solid fraction (BV/TV). The extent of variation is reported
as an error-bar varying from min to max values of TBS, for projection thicknesses varying from 512 to 292 slices.
786 L. Pothuaud et al. / Bone 42 (2008) 775–787

value (mean) in each class was evaluated over the N mean
models. The 15 classes were regrouped into a total of 121 mean
models with their own mean characteristics (BV/TV, Th, TBS) —
to which there corresponded a total of 3630 (=121⁎30) individual
models (size, BV/TV, init). For each class of Th, the relationship
between mean characteristics TBS and BV/TV was fitted with a
linear regression model (Fig. 6):
TBS V¼ a0 þ a1 T BV=TV ð3Þ
models, 2D projection images were generated with different
projection thicknesses (i.e. numbers of slices) from 512 to 272
slices. TBS was evaluated in each of these 2D images, and the
extent [min–max] of variations of TBS values due to reduction of
the projection thickness was evaluated in function of the solid
volume fraction (BV/TV) (Fig. 8). For examples, TBS varied
from 0.766 to 0.779 [pixels− 1] for 3D models with BV/TV = 10%,
and from 0.606 to 0.612 [pixels− 1] for 3D models with BV/
TV = 40%.

where References

TBS′ the estimated value of TBS [1] Nanyan P, Pothuaud L, Benhamou L, Courteix D. Semi-automated
evaluation of the cortico-medullar index on radius radiographs: a study in
BV/TV the solid volume fraction
prepubertal girls. Eur J Radiol 2003;47:47–53.
a0, a1 fit-coefficients of the linear regression model [2] Genant H, Wu CY, van Kuijk C, Newitt MC. Vertebral fracture assessment
using a semi-quantitative approach. J Bone Min Res 1993;8:1137–48.
The fit-coefficients (a0, a1) were dependent upon the class of [3] Faulkner KG, Cummings SR, Black D, Palermo L, Gluer CC, Genant HK.
Th considered. These coefficients were plotted as functions of Simple measurement of femoral geometry predicts hip fracture: the study
of osteoporotic fractures. J Bone Miner Res 1993;8:1211–7.
the mean solid thickness [Th]imean evaluated over the whole
[4] Majumdar S. A review of magnetic resonance (MR) imaging of trabecular
class [Th]i (Fig. 7). Logarithmic fit–regression relationships bone micro-architecture: contribution to the prediction of biomechanical
were determined, as follows: properties and fracture prevalence. Technol Health Care 1998;6:321–7.
  [5] Organization WH. Assessment of fracture risk and its application to
a0 ¼ a00 þ a01 Tln½Thmean screening for postmenopausal osteoporosis [Report of the WHO Study
mean 
i
a1 ¼ a10 þ a11 Tln ½Thi ð4Þ Group]. World Health Organ Tech Rep Ser 1994;843:785–95.
[6] Chappard D, Guggenbuhl P, Legrand E, Basle MF, Audran M. Texture
analysis of X-ray radiographs is correlated with bone histomorphometry.
where J Bone Miner Metab 23, 24–29.
[7] Caligiuri P, Giger ML, Favus MJ, Jia H, Doi K, Dixon LB. Computerized
[Th]imean the mean solid thickness evaluated over the whole radiographic analysis of osteoporosis: preliminary evaluation. Radiology
1993;186:471–4.
class [Th]i
[8] Gregory JS, Stewart A, Undrill PE, Reid DM, Aspden RM. Identification of
(a00, a01) fit-coefficients relative to a0 following the logarith- hip fracture patients from radiographs using Fourier analysis of
mic model the trabecular structure: a cross-sectional study. BMC Med Imaging
(a10, a11) fit-coefficients relative to a1 following the logarith- 2004;4:4.
mic model [9] Brunet-Imbault B, Lemineur G, Chappard C, Harba R, Benhamou CL. A
new anisotropy index on trabecular bone radiographic images using the
fast Fourier transform. BMC Med Imaging 2005;5:4.
Based upon Eqs. 3 and 4, TBS′ – the estimated value of TBS – [10] Ruttimann UE, Webber RL, Hazelrig JB. Fractal dimension from
was expressed as a function of the 3D characteristics, BV/TV radiographs of peridental alveolar bone. A possible diagnostic indicator
and Th: of osteoporosis. Oral Surg Oral Med Oral Pathol 1992;74:98–110.
[11] Samarabandu J, Acharya R, Hausmann E, Allen K. Analysis of bone X-rays
TBS V¼ F1 ðBV=TV; ThÞ ¼ ½a00 þ a01 TlnðThÞ using morphological fractals. Medical Imaging IEEE 1993;12: 466–70.
þ ½a10 þ a11 TlnðThÞTBV=TV [12] Prouteau S, Ducher G, Nanyan P, Lemineur G, Benhamou L, Courteix D.
Fractal analysis of bone texture: a screening tool for stress fracture risk?
ð5Þ Eur J Clin Invest 2004;34:137–42.
[13] Pothuaud L, Benhamou CL, Porion P, Lespessailles E, Harba R, Levitz P.
A second empirical 3D/2D mathematical relationship, linking Fractal dimension of trabecular bone projection texture is related to three-
the 3D characteristics BV/TV and Th and the 2D characteristic dimensional microarchitecture. J Bone Miner Res 2000;15:691–9.
TBS, was established to transform Eq. 5, such that Th′ – the [14] Luo G, Kinney JH, Kaufman JJ, Haupt D, Chiabrera A, Siffert RS.
Relationship between plain radiographic patterns and three-dimensional tra-
estimated value of Th – became a function of BV/TV and TBS: becular architecture in the human calcaneus. Osteoporos Int 1999;9: 339–45.
[15] Della Rocca AB, Ferriani S, La Porta L. Computer simulation of the
ThV¼ F2 ðBV=TV;
 TBSÞ  radiographic image forming process: implementation and applications.
TBS  a00  a10 TBV=TV NDT & E Intl 1995;28:163–70.
¼ exp ð6Þ
a01 þ a11 Tfs [16] Pothuaud L. Process of interpretation of two-dimensional densitometry
images for the prediction of bone mechanical strength. Lecture notes in
computer science. Springer-Verlag GmbH; 2004. p. 1079–80.
Appendix B. Effect of projection thickness on TBS value [17] Parfitt AM. The stereologic basis of bone histomorphometry. Theory of
quantitative microscopy and reconstruction of the 3rd dimension. In:
Recker, R. editor. Bone histomorphometry technique and interpretation.
In order to evaluate the effect of projection thickness on TBS
Boca Raton. CRC Press, p. 53–87.
value, we have exploited 110 of the previous 3D models [18] Parfitt AM, Drezner MK, Glorieux FG, Kanis JA, Malluche H, Meunier
generated in 3D matrix of 512 × 512 × 512 voxels with different PJ, Ott SM, Recker RR. Bone histomorphometry: standardization of
characteristics (size, BV/TV, random). From each of these 3D nomenclature. J Bone Miner Res 2:595–610.
 L. Pothuaud et al. / Bone 42 (2008) 775–787
[19] Levitz P, Tchoubar D. Disordered porous solids: from chord distributions
to small angle scattering. J Phys II 1992;2:771–90.
[20] Hildebrand T, Ruegsegger P. Quantification of bone microarchitecture
with the structure model index. Comput Meth Biomech Engng 1997;1:
15–23.
[21] Feldkamp LA, Goldstein SA, Parfitt AM, Jesion G, Kleerekoper M. The
direct examination of three-dimensional bone architecture in vitro by
computed tomography. J Bone Miner Res 1989;4:3–11.
[22] Pothuaud L, Porion P, Lespessailles E, Benhamou CL, Levitz P. A new method
for three-dimensional skeleton graph analysis of porous media: application
to trabecular bone microarchitecture. J Microsc 2000;199: 149–61.
[23] Atkinson PM, Dunn R, Harrison AR. Measurement error in reflectance
data and its implications for regularizing the variogram. Int J Remote
Sensing 1996;17:3735–50.
[24] Bruniquel-Pinel V, Gastellu-Etchegorry JP. Sensitivity of texture of high
resolution images of forest to biophysical and acquisition parameters.
Remote Sensing Env 1998;65:61–85.
787
[25] Woodcock CE, Strahler AH, Jupp DL. The use of variograms in remote
sensing. I. Scene models and simulated images. Remote Sensing Env
1988;25:323–48.
[26] Jupp DLB, Strahler AH, Woodcock CE. Autocorrelation and regulariza-
tion in digital images. I. Basic theory. IEEE Trans Geos Remote Sensing
1988;26:463–73.
[27] Jupp DLB, Strahler AH, Woodcock CE. Autocorrelation and regulariza-
tion in digital images. II. Simple image models. IEEE Trans Geos Remote
Sensing 1989;27:247–58.
[28] Woodcock CE, Strahler AH, Jupp DL. The use of variograms in remote
sensing. II. Real digital images. Remote Sensing Env 1988;25:349–79.
[29] Cohen WB, Spies TA, Bradshaw GA. Semivariograms of digital imagery
for analysis of conifer canopy structure. Remote Sensing Env 1990;34:
167–78.
[30] St-Onge BA, Cavayas F. Estimating forest stand structure from high
resolution imagery using the directional variogram. Int J Remote Sensing
1995;16:1999–2021.