Biomedical subjects

V A Gibson

Publications and source records attributed to V A Gibson.

13 recordsLinked to original sources

PubMed2005

Osteonal effects on elastic modulus and fatigue life in equine bone.

We hypothesized that recently formed, incompletely mineralized, and thus, relatively deformable osteons in the equine third metacarpus enhance in vitro load-controlled fatigue life in two ways. Macroscopically, there is a compliance effect, because reduced tissue elastic modulus diminishes the stress required to reach a given strain. Microscopically, there is a cement line effect, in which new osteons and their cement lines more effectively serve as barriers to crack propagation. We studied 18 4 x 10 x 100 mm beams from the medial, lateral, and dorsal cortices of metacarpal bones from 6 thoroughbred racehorses. Following load-controlled fatigue testing to fracture in 4 point bending, a transverse, 100 microm thick, basic fuchsin-stained cross-section was taken from the load-bearing region. The number and diameter of all intact (and thus recently formed/compliant) secondary osteons in a 3.8 x 3.8 mm region in the center of the section were determined. The associated area fraction and cement line length of intact osteons were calculated, and the relationships between these variables, elastic modulus (E), and the logarithm of fatigue life (logN(F)) were analyzed. As expected, logN(F) was negatively correlated with E, which was in turn negatively correlated with intact osteon area fraction and density. (LogN(F))/E increased in proportion to intact osteon density and nonlinearly with cement line density (mm/mm(2)). These results support the hypothesis that remodeling extends load-controlled fatigue life both through the creation of osteonal barriers to microdamage propagation and modulus reduction.

Animals↗

PubMed2003

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 10(5) 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.

Animals↗

PubMed1999

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.

Bone and Bones↗

PubMed1997

Model of flexural fatigue damage accumulation for cortical bone.

Analytical models that predict modulus degradation in cortical bone subjected to uniaxial fatigue loading in tension and compression are presented. On the basis of experimental observations, damage was modeled as self-limiting for tension but not for compression. These mechanistic uniaxial damage models were then developed into a model for flexural fatigue of cortical bone based on laminated beam theory. The unknown coefficients in the uniaxial damage models were obtained by successfully fitting the resulting equations to uniaxial fatigue data from the literature on human cortical bone in tension and compression. Then, the predictions of the flexural model for the behavior of human cortical bone were compared with experimental results from a small but independent set of specimens tested at three different ranges of load in our laboratory. The behavior of the modulus degradation curves and the flexural fatigue lives of the specimens were in excellent agreement with the predictions of the model.

Bone and Bones↗

PubMed1997

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.

Analysis of Variance↗

PubMed1997

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.

Animals↗

PubMed1996

In vitro fatigue behavior of the equine third metacarpus: remodeling and microcrack damage analysis.

We studied remodeling and microcrack damage in specimens of Thoroughbred racehorse third metacarpal bone that had been subjected to monotonic or fatigue failure. We asked three questions. What effects does mechanical loading have on histologically observable microcrack damage? Are there regional variations in remodeling of the equine cannon bone, and do these variations correlate with mechanical properties? To what extent are remodeling and microcrack damage age-dependent? Machined beams from the medial, lateral, and dorsal cortices were loaded to fracture in four-point bending monotonically, or cyclically at a load initially producing 10,000 microstrain. Specimens were then bulk-stained in basic fuchsin, and cross sections were prepared from loaded and load-free regions of each beam. Current and past remodeling, porosity, and microcrack density and length were determined histomorphometrically. Strained and unstained microcracks were observed. Unstained cracks were associated with regions of woven bone and appeared to be damaged Sharpey's fibers. Their density (approximately 30/mm2) did not increase after failure, but their length (approximately 25 microns) did, especially near the surfaces of the beam. Stained cracks were wider and longer than unstained cracks and were located primarily near the fracture surface and on the compressed side of the beam. Stained cracks after failure were more numerous in those beams having a higher elastic modulus, a shorter fatigue life, or greater deformation at failure. The extent of past remodeling increased with age, especially in the medial region; the rate of current remodeling generally declined with age, but not in the dorsal region, which has the best fatigue resistance. In summary, while remodeling varied with age and region, its effects on bone structure did not appear to influence microdamage. Basic fuchsin staining of damage in fractured equine bone was independent of age and region and confined to near the fracture surfaces. Distributed microdamage consisted only of what appeared to be subtle disruptions of Sharpey's fibers.

Age Distribution↗

PubMed1996

Osteonal structure in the equine third metacarpus.

In studying the flexural fatigue properties of the equine third metacarpal (cannon) bone, we previously found that the dorsal region was weaker monotonically, but more fatigue resistant, than the lateral region. Fatigue resistance was associated with fracture surfaces which demonstrated that secondary osteons had "pulled out" of the surrounding matrix; this never happened in lateral specimens. We therefore became interested in the osteonal structure of this bone, and began to study its birefringence patterns in circularly polarized light. We found that the predominant type of secondary osteon was one in which only the outermost few lamellae were circumferential, with the inner lamellae being longitudinally oriented. This "hoop" pattern had not been described in Ascenzi's classic papers. Using basic fuchsin-stained, undecalcified cross-sections from the dorsal, medial, and lateral midshaft regions of 12 pairs of cannon bones, we classified 360 secondary osteons according to their birefringence patterns, and measured their inner and outer diameters. We found that variants of the hoop category comprised 60% of all osteons, but were significantly less common in the dorsal region, where the predominant types were Ascenzi's "longitudinal" or "alternating" patterns. The dorsal region also had smaller osteons (OD = 156 +/- 19 microns) than the medial (179 +/- 13 microns, p = 0.0004) and lateral (182 +/- 13 microns, p = 0.0001) regions. We postulate that these regional variations in osteonal size and structure, which are obviously produced by regional variations in remodeling, have important mechanical implications.

Analysis of Variance↗

PubMed1996

Collagen fiber organization is related to mechanical properties and remodeling in equine bone. A comparison 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.

Animals↗

PubMed1995

Fatigue behavior of the equine third metacarpus: mechanical property analysis.

This is the first in a series of experiments to study the fatigue properties of equine cannon (third metacarpal) bone specimens from Thoroughbred racehorses. Monotonic and fatigue tests to failure were performed in four-point bending on diaphyseal specimens in a 37 degrees C saline bath to answer three initial questions. (a) Will a linear variable differential transducer yield the same elastic modulus as strain gauges? (b) Will fatigue results depend on whether the periosteal or endosteal side of the beam is in tension? (c) Are there regional variations in the monotonic and fatigue properties of the cannon bone midshaft? Eighteen left-right pairs of specimens from six horses were used. One beam of each pair was fitted with strain gauges. Fatigue tests were conducted on 24 specimens under load control at 2 Hz; an initial range of 0-10,000 microstrain was used so as to produce failure in a reasonable period of time. There were no left-right differences in the fatigue or monotonic properties, and the presence of a gauge had no effect on modulus measured by a linear variable differential transducer. However, gauge-measured moduli were about 1 GPa less than transducer-measured values. Fatigue life was independent of which side of the beam was in tension, and there were significant variations in mechanical properties around the cortex. The lateral region was stiffer than the dorsal region but the latter had a longer fatigue life. The fixed cylindrical supports used in this experiment eventually produced slight wear grooves, causing artifactual stiffening at the end of the load cycle in some specimens. A second experiment using roller supports confirmed the reason for this stiffening. It also showed that fatigue life was shorter when roller supports were used but regional differences were similar.

Analysis of Variance↗

PubMed1992

Binding and functional selectivity of himbacine for cloned and neuronal muscarinic receptors.

The binding potencies for the putative M2-selective antagonist himbacine were determined in radioligand binding and in functional response assays in neuronal tissue and Chinese hamster ovary cells containing transfected muscarinic receptors. Himbacine was shown to bind to all five cloned muscarinic receptor subtypes in the order of potencies: hM2 = hM4 > hM3 > hM1 > hM5 (Kd values were 4, 7, 59, 83 and 296 nM, respectively). Himbacine was shown to bind to M2 receptors in rat heart and brain stem with Kd values of 6.9 and 4.6 nM, respectively. In rat brain tissues with complex mixtures of muscarinic receptors, and using the radioligand [3H] +/- -5,11-dihydro-11-([(2-(2-[(dipropylamino)methyl]-1- peperidinyl)ethyl)amino]carbonyl)-6H-pyrido(2,3-b)(1,4)benzodiazep ine-6-one to demarcate M2 and M4 receptors, himbacine was shown to bind to 80% of cortical or striatal receptors with Kd values of 4.5 and 3.8 nM, respectively, consistent with the involvement of M2 and/or M4 receptors in both these brain regions. Himbacine was a potent blocker of oxotremorine-M-mediated cyclic AMP inhibition in rat striatum (4.4 nM) and in N1E-115 neuroblastoma cells (10.6 nM), responses mediated by M4 receptors. Himbacine also reversed oxotremorine-M-mediated inhibition of evoked acetylcholine release from hippocampal tissue with a Kd value of 8.6 nM, a value consistent with the involvement of M2 or M4 receptors. At the cortical postsynaptic muscarinic receptors involved with phosphoinositide turnover (putative M1 and M3 receptors), himbacine was 21-fold less potent. Himbacine appears to be a potent muscarinic antagonist that displays selectivity for M2 or M4 receptors, as compared to M1 or M3 receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

PubMed1991

Interactions of agonists with M2 and M4 muscarinic receptor subtypes mediating cyclic AMP inhibition.

In this study the similarities and differences between the M2 and M4 subtypes in their recognition of agonists were explored. A CHO-K1 cell line transfected with the human m2 receptor was used as a homogeneous M2 tissue for comparison with two putative M4 systems (rat striatum and the N1E-115 mouse neuroblastoma cell line). The equilibrium binding dissociation constants and intrinsic efficacies for seven muscarinic agonists were determined for their stimulation of cyclic AMP inhibition via the M2 and M4 receptors. Partial receptor occlusion with propylbenzilylcholine mustard was used to determine binding constants for the more efficacious drugs and the reference agonist oxotremorine-M. The binding dissociation constants and relative efficacies for other agonists were then determined in reference to oxotremorine-M by a null method. For the M2 receptor the agonist binding dissociation constants ranged in potency from oxotremorine (1.5 microM) to bethanechol (171 microM), whereas relative efficacies varied from that of muscarine (relative efficacy = 0.9) to the value for McN-A343 (relative efficacy = 0.04). In general, most agonists bound with similar potencies to M2 and M4 receptors (Kd values within a factor of 2-3). However, oxotremorine bound to the N1E-115 and striatal M4 receptors about 3-fold and 10-fold less potently, respectively, than it did to the M2 receptor. Another exception was pilocarpine, which bound to the N1E-115 receptor (1.9 microM) with 8-fold and 12-fold higher potency than to the CHO-K1 M2 receptor and the striatal M4 receptor, respectively. Despite the low affinity of bethanechol for the M2 receptor, it was an efficacious agonist (maximal response equivalent to that of oxotremorine-M; relative efficacy = 0.6) at this subtype, whereas it was a partial agonist (60%) with lesser efficacy in the clonal M4 system. In contrast, McN-A343 and arecoline were significantly more efficacious at the two M4 receptors than they were at the M2 receptor. The M4 system in the rat striatum displayed some similarity to the N1E-115 M4 system, with regard to the efficacy ranking for certain agonists (arecoline greater than bethanechol greater than McN-A343 greater than or equal to pilocarpine). This rank order was different from the ranking of these four agonists in the M2 system, indicating that these two M4 receptors are more similar to each other in efficacy ranking than they are to the M2 receptor. However, the rat striatal and N1E-115 M4 receptor differed in their binding of oxotremorine and pilocarpine, indicating that these two M4 systems were not pharmacologically identical.(ABSTRACT TRUNCATED AT 400 WORDS)

(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethy↗

PubMed1991

Binding of [3H]AF-DX 384 to cloned and native muscarinic receptors.

The binding selectivity of [3H]AF-DX 384 [(+-)-5,11-dihydro-11- ([(2-(2-[(dipropylamino)methyl]-1- piperidinyl)ethyl)amino]carbonyl)-H-pyrido(2,3-b)(1,4)benzodiazepine-6-o ne] was evaluated with cloned human muscarinic receptors (M1-M4) in Chinese hamster ovary (CHO-K1) cell lines as well as in rat heart and brain. There were uniform classes of sites for the radioligand in the M2-rich tissues, heart (Kd = 2.3 nM) and brainstem (Kd = 2.4 nM). However, [3H]AF-DX 384 bound to all four cloned receptor subtypes. Using kinetic methods, the calculated Kd values were M2 (1 nm) greater than M4 (2.2 nM) greater than M3 (15 nM) greater than M1 (55 nM). Scatchard analysis with the CHO cells confirmed the high affinity of this radioligand for the M2 (1.8 nM) and M4 (2.5 nM) receptors. To evaluate the potential for selectively binding to M2 and M4 receptors in cortex and striatum, low concentrations (0.5-0.8 nM) of the radioligand were used and a two-site competition model was used to derive the binding constants for pirenzepine and AF-DX 116 [(+-)-11-2((-((diethylamino) methyl)-1-piperidinyl)acetyl)-5,11-dihydro-6H- pyrido(2,3-b)(1,4)-benzodiazepine-6-one] and to compare them with values obtained with cloned M2 and M4 receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗