V. A. Gibson

COLLAGEN FIBER ORGANIZATION IS RELATED TO MECHANICAL PROPERTIES AND REMODELING IN EQUINE BONE. A COMPARSION OF TWO METHODS

We studied birefringence as an indicator of collagen fiber orientation in the diaphysis of the equine third metacarpal bone. We had previously shown that tissue from the lateral cortex of this bone is stronger monotonically, but less fatigue resistant, than tissue from the medial and dorsal regions. To learn whether collagen fiber orientation might play a role in this regional specialization, we tested three hypotheses using the same specimens: (1) collagen fiber orientation is regionally dependent; (2) remodeling changes collagen fiber orientation; (3) longitudinal collagen fibers correlate positively with modulus and monotonic bending strength and negatively with flexural fatigue life. Beams (N = 36) cut parallel to the long axes of six pairs of bones had been tested to determine elastic modulus (N = 36), and fatigue life (N= 24) or monotonic strength (N = 12) in four-point bending. Subsequently, histologic cross-sections were prepared, and porosity, active remodeling and past remodeling were quantified. Birefringence was measured as an indicator of transverse collagen orientation using plane-polarized light (PPL), and again using circularly polarized light (CPL). The CPL measurement was less variable than the PPL measurement. Both birefringence measures indicated that collagen was more longitudinally oriented in the lateral cortex than in the other two cortices. Longitudinally disposed collagen correlated with greater modulus and monotonic strength, but did not correlate with fatigue life. Remodeling was associated with more transverse collagen. Neither measure of birefringence was significantly correlated with porosity. It was concluded that, in the equine cannon bone, longitudinal collage fiber orientation is regionally variable, contributes to increased modulus and strength but not fatigue life, and is reduced by osteonal remodeling.

CALCIUM BUFFERING IS REQUIRED TO MAINTAIN BONE STIFFNESS IN SALINE SOLUTION

This work determined whether mineral dissolution due to prolonged testing or storage of bone specimens in normal saline would alter their elastic modulus. In one experiment, small pieces of equine third metacarpal bone were soaked in normal saline supplemented with varying amounts of CaCl2. Changing Ca ion concentrations in the bath were monitored and the equilibrium concentration was determined. In a second experiment, the elastic moduli of twenty 4 x 10 x 100 mm equine third metacarpal beams were determined non-destructively in four-point bending. Half the beams were then soaked for 10 days in normal saline, and the other half in saline buffered to the bone mineral equilibrium point with Ca ions. Modulus measurements were repeated at 6 and 10 days. The equilibrium Ca ion concentration for bone specimens was found to be 57.5 mg l-1. The modulus of bone specimens soaked in normal saline significantly diminished 2.4%, whereas the modulus of those soaked in calcium-buffered saline did not change significantly.

Osteon pullout in the equine third metacarpal bone: effects of ex vivo fatigue

An important concept in bone mechanics is that osteons influence mechanical properties in several ways, including contributing to toughness and fatigue strength by debonding from the interstitial matrix so as to „bridge”︁ developing cracks. Observations of „pulled out„ osteons on fracture surfaces are thought to be indicative of such behavior. We tested the hypothesis that osteon pullout varies with mode of loading (fatigue vs. monotonic), cortical region, elastic modulus, and fatigue life. Mid‐diaphseal beams from the dorsal, medial, and lateral regions of the equine third metacarpal bone were fractured in four point bending by monotonic loading to failure under deflection control, with or without 105 cycles of previous fatigue loading producing 5000 microstrain (15–20% of the expected failure strain) on the first cycle; or sinusoidal fatigue loading to failure, under load or deflection control, with the initial cycle producing 10,000 microstrain (30–40% of the expected failure strain). Using scanning electron microscopy, percent fracture surface area exhibiting osteon pullout (%OP.Ar) was measured. Monotonically loaded specimens and the compression side of fatigue fracture surfaces exhibited no osteon pullout. In load‐controlled fatigue, pullout was present on the tension side of fracture surfaces, was regionally dependent (occurring to a greater amount dorsally), and was correlated negatively with elastic modulus and positively with fatigue life. Regional variation in %OP.Ar was also significant for the pooled (load and deflection controlled) fatigue specimens. %OP.Ar was nearly significantly greater in deflection controlled fatigue specimens than in load‐controlled specimens (p < 0.059). The data suggest that tensile fatigue loading of cortical bone eventually introduces damage that results in osteonal debonding and pullout, which is also associated with increased fatigue life via mechanisms that are not yet clear.

Residual strength of equine bone is not reduced by intense fatigue loading: Implications for stress fracture

Fatigue or stress fractures are an important clinical problem in humans as well as racehorses. An important question in this context is, when a bone experiences fatigue damage during extreme use, how much is it weakened compared to its original state? Since there are very limited data on this question and stress fractures are common in racehorses, we sought to determine the effect of fatigue loading on the monotonic strength of equine cortical bone. Beams were machined from the dorsal, medial and lateral cortices of the third metacarpal bones of six thoroughbred racehorses. Beams from left and right bones were assigned to control and fatigue groups, respectively (N = 18 each). The fatigue group was cyclically loaded in three-point bending at 2 Hz for 100,000 cycles at 0-5000 microstrain while submerged in saline at 37 degrees C. These beams, as well as those in the control group, were then monotonically loaded to failure in three-point bending. The monotonic load-deflection curves were analyzed for differences using three-factor (fatigue loading, anatomic region, and horse) analysis of variance. The mean failure load was 3% less in the fatigue group, but this reduction was only marginally significant. Neither elastic modulus nor yield strength was significantly affected by the fatigue loading. The principal effects of fatigue loading were on post-yield behavior (yield being based on a 0.02% offset criterion). The work done and the load increase between yield and failure were both significantly reduced. All the variables except post-yield deflection were significantly affected by anatomic region. In summary, loading equivalent to a lifetime of racing does not significantly weaken equine cortical bone ex vivo. The clinical implication of this may be that the biological repair of fatigue damage can actually contribute to stress fracture if pressed too far.

The effects of testing methods on the flexural fatigue life of human cortical bone.

A flexural model of four-point bending fatigue that has been experimentally validated for human cortical bone under load control was used to determine how load and displacement control testing affects the fatigue behavior of human cortical bone in three-point and symmetric four-point bending. Under load control, it was predicted that three-point bending produced no significant differences in fatigue life when compared to four-point bending. However, three-point bending produced less stiffness loss with increasing cycles than four-point bending. In four-point bending, displacement control was predicted to produce about one and a half orders of magnitude greater fatigue life when compared to load control. This prediction agrees with experimental observations of equine cannon bone tested in load and displacement control (Gibson et al., 1998). Displacement controlled three-point bending was found to produce approximately a 25% greater fatigue life when compared to load control. The prediction of longer fatigue life under displacement control may have clinical relevance for the repair of damaged bone. The model can also be adapted to other geometric configurations, including modeling of whole long bones, and with appropriate fatigue data, other cortical bone types.

Artifactual nonlinearity due to wear grooves and friction in four-point bending experiments of cortical bone.

Experiments and analyses were performed to determine the cause of a nonlinear force-deflection response observed in four-point flexural fatigue of beams of cortical bone machined from the mid-diaphysis of the equine third metacarpus. Observable grooves which formed on the beam surface at supports and load noses were found to be the primary cause of the nonlinearity. An additional geometric nonlinearity at large deflections revealed by finite element modeling may be minimized by using the smallest diameter supports and load noses recommended in ASTM 790. However, frictional constraint of the beams at the load noses and supports can occur at low load levels and should be avoided by using roller-bearing supports and load noses, or some equivalent method.