Synchrotron X-Ray Scattering reveals a pivotal role of water in the ultrastructural mechanics of

bone.
Jitin Samuel, Xiaodu Wang.
University of Texas at San Antonio

Introduction: Bone fragility fracture is a major cause of healthcare concern in the aging population. It is
generally accepted that the structural and compositional characteristics of bone tissue at the ultra-structure level
play an important role in governing bone fragility [1]. Different bone constituents and their interactions impart
bone its unique mechanical properties at the bulk level. Water, an important bone constituent is a known
contributor to the overall toughness in bone. A recent study revealed that the bone mechanical properties were
greatly influenced by the presence of water in small gap regions (penetrable by molecules of size < 0.4 nm).
Although it is clear that presence of water in small ultrastructure regions is an important factor in determining the
bulk mechanical behavior of bone, the underlying mechanisms are not clearly understood. This study aims at
determining the role of water in influencing the mechanical behavior of bone at ultrastructural levels.

Materials and Methods: In this study, human cadaveric bone specimens (Φ3.05.0mm compression, 2x2mm
tension), from middle-aged male donors (51.6±1.8 years old), were tested in tension and compression using
progressive loading schemes. The specimens were tested in two groups, dehydrated and control (n=6, each
group). Wide Angle X-ray Scattering and Small Angle X-ray Scattering patterns were recorded simultaneously
with mechanical loading to measure the in situ deformation in the mineral and collagen phases.

Results and Discussion: Dry specimens exhibited drastically different deformation behavior compared to the wet
specimens. In the absence of water, bone did not exhibit
COMPRESSION TENSION
any post yield deformation [Figs. 1a - 1f], indicating that -250 dry

Applied Stress(MPa)
120
wet
Applied Stress(MPa)

water acts as a plasticizer at the ultrastructural levels -200

-150
100
80
a b
allowing bone to deform to much higher strains than -100
60
40
possible in the absence of water. -50 20
0
0
Mineral crystals and collagen fibrils exhibited co- 0 -1 -2 -3 -4 -5 -6 0 0.5 1 1.5
Applied Strain (%) Applied Strain (%)
operative deformation in both wet and dry conditions. 0.6
-1.2

Fibril Strain %
Under dry condition, the co-operation was higher for
Fibril Strain %

-1 0.5
0.4
both tension and compression (increased εm/εf)
-0.8 c d
-0.6 0.3
-0.4 0.2
indicating that in the absence of water at the -0.2 0.1
0
ultrastructural spaces, mineral and collagen fibrils 0 -1 -2 -3 -4 -5 -6
0
0 0.5 1
deform in a more concerted manner. However, under Applied Strain (%) e. Applied Strain %
0.35
wet condition, the cooperative deformation exhibited
Mineral Strain %

-0.8
Mineral Strain %

0.3
0.25
opposite trends in tension and compression. Under -0.6 e
0.2
f
-0.4 0.15
compression, the mineral-fibril deformation became -0.2 0.1
0.05
more co-operative and saturated at higher applied strains 0 0
0 -1 -2 -3 -4 -5 -6
whereas under tension, the mineral-fibril deformation Applied Strain (%)
0 0.5
Applied Strain %
1

became less co-operative with increasing load until 1 1

saturation at higher applied strains [Fig. 1g, 1h]. This 0.8 g 0.8 h
0.6
εm/εf

0.6
important observation indicates two distinct underlying
εm/εf

0.4 0.4
deformation mechanisms at the ultrastructure level in 0.2 0.2
0 0
bone. 0 1 2 3 4 5 6 0 0.5 1 1.5
Applied Strain Applied Strain

Conclusions: This study revealed distinct deformation

mechanisms under tension and compression in bone. Fig. 1. Dry vs Wet specimens. a. Compression Stress-Strain
curve. b. Stress-Strain curve Tension. c. Fibril Strain in
Water played a pivotal role as a plasticizer in both compression. d. Fibril Strain in tension. e. Mineral Strain in
loading conditions. The underlying mechanisms need to compression. f. Mineral Strain in tension. g. Mineral-Fibril
be explored further. cooperative deformation under compression. h. Mineral-Fibril
cooperative deformation under tension.
Acknowledgements: NIH/NIAMS grant
(1R01AR055955)
References: 1. Samuel, J., Sinha, D., Zhao, J. C.-G., & Wang, X. (2014). Bone, 59, 199–206.