Skeletal development and bone functional adaptation.

The role of in vivo mechanical loading histories in normal skeletogenesis is related to the process of adaptive, stress-regulated bone remodeling in the adult. The results of many previous computer models for endochondral ossification and bone modeling and remodeling are reviewed. These studies support the view that simple stress-related mathematical algorithms or "construction rules" can be used to emulate normal skeletal development and architectural construction. Such mathematical rules presumably represent the net result of biophysical phenomena influencing cell metabolism and biosynthetic activity. These rules are also successful in describing the adaptation of adult bone to changes in tissue stresses. The findings suggest that stress-related functional adaptation in mature bones may be merely the adult manifestation of the same mechanical construction rules that guide and constrain normal development.

Hyperresponsiveness of the rat neuroendocrine system due to repeated exposure to stress.

Sequential exposure to stressors may elicit a period of endocrine hyperresponsiveness during which plasma hormone concentrations reach higher levels after repeated exposure to a stressor compared to levels after initial exposure. The present study was designed to further characterize hyperresponsiveness to repeated stress and determine if hyperresponsiveness is dependent upon repeated exposure to the same stressful stimuli. In Experiment 1, rats were stressed by inescapable tailshock, immobilization or exposure to shock chamber without shock for one, two, three, four or five consecutive days (15 min/day). In rats exposed to tailshock, corticosterone (CS) levels in plasma collected on days 2, 3, 4 and 5 were higher than CS levels following acute tailshock on day 1, demonstrating hyperresponsiveness to repeated tailshock. Hyperresponsiveness of CS secretion also occurred in groups of rats restrained for four or five days. No changes occurred in the CS response of animals repeatedly exposed to immobilization. Prolactin (PRL) levels were not affected by repeated exposure to the stressors. However, PRL values were different between the stress conditions and indicated that the order of stressor severity was tailshock greater than immobilization greater than exposure to shock chamber without shock. In Experiment 2, rats were exposed to either one or two consecutive days of tailshock or immobilization. Other rats were exposed to either tailshock or immobilization on the first day, then switched to the other stressor on the next day. Hyperresponsiveness to repeated tailshock, but not immobilization, was reflected in plasma levels of CS and adrenocorticotropic hormone (ACTH), but not PRL. Hyperresponsiveness of CS and ACTH secretion also was found in rats first stressed by immobilization then switched to tailshock, demonstrating that hyperresponsiveness is not dependent upon reexposure to familiar stressful stimuli. However, hyperresponsiveness did not occur in rats first exposed to tailshock then switched to immobilization. The data suggest that both immobilization and tailshock primed the organism to hyperrespond, but only the more severe stressor (tailshock) elicited hyperresponsiveness of the neuroendocrine system.

Compressive properties of cancellous bone defects in a rabbit model treated with particles of natural bone mineral and synthetic hydroxyapatite.

A rabbit model was developed to evaluate the compressive mechanical properties of cancellous bone defects treated with particles of selected bone graft substitute materials. A novel feature of the model was the precise retrieval of the site of implantation. A notable finding was a 9-fold increase in the modulus of elasticity of the defect implanted with a synthetic hydroxyapatite material after 26 weeks when compared to the modulus of the trabecular bone normally at the site. The compressive modulus of lesions treated with particles of a natural bovine bone mineral (anorganic bovine bone) was closer to the normal modulus of the cancellous bone at the site. While the compressive strength of the anorganic bone particles was less than that of normal bone, the site implanted with the bone mineral particles achieved compressive strength greater than normal after 6 weeks. Moreover, the anorganic bone particles accelerated the increase in strength of the lesion, at 6 weeks exceeding the strength achieved by the untreated defect after 26 weeks. The potential problem associated with the disparity in the compressive modulus between sites implanted with the synthetic HA particles and surrounding bone is discussed.

Stress analyses of joint arthroplasty in the proximal humerus.

Finite element stress analyses were performed on the proximal humerus before and after the simulated implantation of stemmed, metallic prosthetic components with porous sintered surfaces for direct bony attachment. Design geometries with surfaces at the prosthetic head/bone interface that were (a) convex, (b) flat, and (c) concave were studied. Analyses for each of the three geometries were conducted to reflect (a) bone ingrowth on all the prosthesis/bone surfaces and (b) bone ingrowth only along the underside of the prosthetic humeral head (assuming the stem was not coated with a porous material). Three loading conditions were used to model various degrees of abduction of the arm. Results indicated that in the normal humerus the compressive joint forces are transmitted from the articular surface through cancellous bone to the inferior cortical shell. Contraction of the rotator cuff muscles created tensile stresses in the superolateral cancellous bone and the superior cortical shell of the humerus. Results of the implanted humeral component models indicated that the use of a prosthesis with bone ingrowth along the stem would cause marked stress shielding proximally whereas the use of implants with porous ingrowth only on the underside of the humeral head replacement produced stress fields more similar to the normal humerus. The convex, flat, and concave surfaces provided similar load transfer from the component to the underlying bone in all loading cases. Other prosthetic head designs that may offer better initial stability produced stress fields similar to those of existing prostheses.

Stresses in plated long-bones: the role of screw tightness and interface slipping.

Using a three-dimensional finite element model of a plated long bone, we studied the influence of screw tightness, sliding frictional interfaces, and loading magnitude on the stresses within the plated bone. The model incorporated frictional interface elements that allowed stress-free separation under tensile loading to occur between the plate and bone and between the screw heads and the plate. The applied loading stimulated both static preloads created by tightening the screws that secure the plate to the bone and physiologic loads created by activity. Initial screw tightening with plate application created regions of bone hydrostatic compressive stress that may be partly responsible for ischemia under the plate. The inclusion of frictional interfaces resulted in a nonlinear relationship between physiologic loan and bone strain that was dependent on screw tightness. This nonlinear response correlated well with the results of previous in vitro studies showing that slippage between the plate and the bone can occur at physiologic load levels. The results showed that the effect of such slippage can be at least as important as plate material, rigidity, and placement in determining the degree of stress shielding. The results also indicated that previous plated bone models that assumed tight interfaces may have overestimated the extent of mechanical stress shielding.

An approach for time-dependent bone modeling and remodeling--theoretical development.

A time-dependent approach for emulating bone modeling and remodeling in response to the daily loading history is presented. We postulate that genotype, systemic metabolic conditions, and local tissue interactions establish the level of local tissue mechanical stimulation (attractor state) appropriate for the maintenance of bone tissue. The net daily rate of apposition or resorption on a bone surface is determined by the difference between the actual stimulus and the tissue attractor state and can be modulated by other biologic factors. In calculating the net change in local bone apparent density, the technique takes into account the bone surface area available for osteoblastic and osteoclastic activity. Endosteal, periosteal, haversian, and cancellous bone modeling and remodeling are thereby treated in a consistent, unified fashion.

An approach for time-dependent bone modeling and remodeling-application: a preliminary remodeling simulation.

In a companion paper, we presented a time-dependent theory for bone modeling and remodeling in response to a daily loading history. This paper represents a preliminary attempt to use the theory to determine the distribution of bone density within the adult proximal femur under an assumed normal loading history. Subsequent functional adaptation of the internal structure due to changes in the loading history are then determined. Throughout this preliminary study, the external geometry of the proximal femur is considered to be fixed, i.e., changes in the external shape are neither stimulated nor allowed. Linear and trilinear (dead-zone nonlinearity) rate remodeling laws were compared. Computer emulations using two-dimensional finite element models were successful in creating a normal-appearing distribution of bone tissue when remodeling was initiated from a solid structure of homogeneous bone density. Subsequent reduction in the loading history caused regional bone atrophy. Reinstatement of the normal loading history caused a generalized increase in bone mass but resulted in a slightly different bone distribution than was calculated for a constant loading history. These results demonstrate the utility of the remodeling theory and are consistent with the hypothesis that similar stress-related phenomena are responsible for both normal morphogenesis and functional adaptation in response to changes in the bone loading.

Acute immobilization stress disrupts testicular steroidogenesis in adult male rats by inhibiting the activities of 17 alpha-hydroxylase and 17,20-lyase without affecting the binding of LH/hCG receptors.

We have investigated the effect of acute immobilization (3 hours) stress on testicular steroidogenesis in the adult rat. Immobilization did not alter plasma luteizing hormone (LH) levels, but plasma testosterone (T) levels were reduced by 82%. Plasma levels of corticosterone in stressed rats were elevated more than ninefold over control levels. After 3 hours of stress, testicular levels of progesterone were elevated 33%, and levels of 17 alpha-hydroxyprogesterone and T were reduced 47% and 37%, respectively, compared to controls. Immobilization for 3 hours had no effect on the association or dissociation rate constants of LH/human chorionic gonadotropin (hCG) receptors of testicular interstitial cells and did not alter specific hCG binding. The effect of 3 hours of immobilization on testicular 17 alpha-hydroxylase and 17,20-lyase was assessed by incubating testicular microsomes from stressed and control animals in the presence of 21[14C]progesterone and [3H]17 alpha-hydroxyprogesterone. Immobilization of rats reduced the Vmax values of 17 alpha-hydroxylase and 17,20-lyase by 47% and 48%, respectively, but had no effect on the Km values. These results support the hypothesis that stress for 3 hours disrupts rat testicular steroidogenesis via a mechanism that is independent of changes in circulating levels of LH and the binding characteristics of LH/hCG receptors. The effects of immobilization on the content of testicular steroids and on the activities of 17 alpha-hydroxylase and 17,20-lyase suggest that stress inhibits the activities of both 17 alpha-hydroxylase and 17,20-lyase.

Effect of restraint stress on gonadal proopiomelanocortin peptides and the pituitary-testicular axis in rats.

We examined the effect of restraint on testicular interstitial fluid (TIF) concentrations of ACTH, beta-endorphin-lipotropin (beta-E-LI) and testosterone and correlated those changes with plasma concentrations of ACTH, beta-E-LI, corticosterone, LH and testosterone in adult rats. Animals were subjected to 1, 2, or 3 h of restraint and were killed immediately following the stress period. Plasma values of ACTH and beta-E-LI were elevated above control values after 1 and 2 h, but not after 3 h of restraint. Plasma corticosterone showed a similar response to restraint except that concentrations were also elevated after 3 h. Plasma testosterone concentrations were elevated after 1 h of restraint, but after 3 h of restraint had fallen below control values. Restraint reduced plasma testosterone concentrations without altering plasma LH concentrations. The decline in plasma testosterone during restraint was associated with a parallel decrease in testosterone in the TIF. Concentrations of ACTH and beta-E-LI were 6- and 3-fold greater in TIF than in the plasma. While 1 or 2 h of restraint did not affect ACTH and beta-E-LI in TIF, values of these hormones were elevated in rats exposed to 3 h of restraint. These data, coupled with recent reports that testicular proopiomelanocortin (POMC)-derived peptides may modulate testicular steroidogenesis, suggest that these factors may play an autocrine or paracrine role in mediating stressor-induced changes in testicular function.

Brain levels of amines and amino acids in taste aversion-prone and -resistant rats.

Possible biological contributions to taste aversion (TA) conditionability were explored by comparing whole-brain levels of five neurotransmitter amines and 14 common amino acids within TA-prone (TAP) and TA-resistant (TAR) rats. The selectively bred strains had been developed via 22 generations of bidirectional nonsibling matings based on susceptibility to cyclophosphamide-induced conditioned TAs. The target substances were separated by HPLC and were measured by electrochemical or fluorometric procedures. The TAP brains had higher levels of serotonin (5-HT) and lower levels of norepinephrine (NE) than TAR brains. No strain differences were found with respect to dihydroxyphenylalanine (DOPAC), dopamine (DA), or 5-hydroxyindoleacetic acid (5-HIAA). Among amino acids, TAP rats had lower levels of lysine than TARs: no other differences were detected. Therefore, higher levels of 5-HT and lower levels of NE and lysine were associated with enhanced TA conditionability. The 5-HT and NE results extend prior indications of their central neurotransmitter TA involvements. The functional role of lysine in TA or other brain functions remains obscure.

Musculoskeletal ontogeny, phylogeny, and functional adaptation.

Physical forces applied to connective tissues may cause significant changes in cell metabolism and gene expression. Theoretical investigations indicate that mechanical loading histories beginning very early in skeletal development may guide endochondral ossification patterns and the initial architectural construction of bones. Developmental patterns and structures of bones can be emulated using mathematical algorithms or "rules of construction" which relate developmental processes to tissue stress (or strain) histories. Skeletal forms and tissues are well-designed for their mechanical function primarily because their histomorphological construction has been guided by mechanical loading during growth and development. Construction rules of developmental mechanics can also be used to describe many of the histological and morphological adaptations of mature skeletal tissues to changes in customary physical activity. Over many generations, changes in the heritable genetic information occurs by mutation and genetic variability. The range of skeletal forms that are possible in evolution due to such variations, however, is constrained by the developmental rules of construction that reflect biophysical processes associated with the tissue mechanical loading.

Relationships between loading history and femoral cancellous bone architecture.

A theory relating bone maintenance to mechanical loading history has been applied to successfully predict the distribution of bone density and trabecular orientation in the adult proximal femur. The loading history was simulated by determining the stress fields in a two-dimensional finite element model exposed to various discrete loading cases and making assumptions about the relative number of loading cycles associated with each load case. The total stimulus to bone maintenance was then calculated by a linear superposition of the stimulus of each loading case. Based on the calculated total stimulus, the apparent density and material properties of each element were changed and the stress solutions were again determined. Using this iterative technique, the bone apparent density and orientation characteristics were predicted. The results indicate that the trabecular morphology of the femur can only be explained by considering the joint loadings from multiple directions. Contrary to the 'trajectorial theory' promoted by Wolff (The Law of Bone Remodelling, 1892), trabecular orientations predicted from our multiple-load analyses are not necessarily perpendicular and do not correspond to the principal stress directions of any one loading condition. Our predicted orientations correspond better to the drawing of bone trabecular morphology by von Meyer (Archs Anat. Physiol. wiss. Med. 34, 615-628, 1867) than to the classic drawing by Wolff and suggest that further study of the trajectorial theory is warranted.

Animation of in vitro biomechanical tests.

Interdisciplinary communication of three-dimensional kinematic data arising from in vitro biomechanical tests is challenging. Complex kinematic representations such as the helical axes of motion (HAM) add to the challenge. The difficulty increases further when other quantities (i.e. load or tissue strain data) are combined with the kinematic data. The objectives of this study were to develop a method to graphically replay and animate in vitro biomechanical tests including HAM data. This will allow intuitive interpretation of kinematic and other data independent of the viewer's area of expertise. The value of this method was verified with a biomechanical test investigating load-sharing of the cervical spine. Three 3.0 mm aluminium spheres were glued to each of the two vertebrae from a C2-3 segment of a human cervical spine. Before the biomechanical tests, CT scans were made of the specimen (slice thickness=1.0 mm and slice spacing=1.5 mm). The specimens were subjected to right axial torsion moments (2.0 Nm). Strain rosettes mounted to the anterior surface of the C3 vertebral body and bilaterally beneath the facet joints on C3 were used to estimate the force flow through the specimen. The locations of the aluminium spheres were digitised using a space pointer and the motion analysis system. Kinematics were measured using an optoelectronic motion analysis system. HAMs were calculated to describe the specimen kinematics. The digitised aluminium sphere locations were used to match the CT and biomechanical test data (RMS errors between the CT and experimental points were less than 1.0 mm). The biomechanical tests were "replayed" by animating reconstructed CT models in accordance with the recorded experimental kinematics, using custom software. The animated test replays allowed intuitive analysis of the kinematic data in relation to the strain data. This technique improves the ability of experts from disparate backgrounds to interpret and discuss this type of biomechanical data.

In vitro axial preload application during spine flexibility testing: towards reduced apparatus-related artefacts.

Presently, there is little consensus about how, or even if, axial preload should be incorporated in spine flexibility tests in order to simulate the compressive loads naturally present in vivo. Some preload application methods are suspected of producing unwanted "artefact" forces as the specimen rotates and, in doing so, influencing the resulting kinematics. The objective of this study was to quantitatively compare four distinct types of preload which have roots in contemporary experimental practice. The specific quantities compared were the reaction moments and forces resulting at the intervertebral disc and specimen kinematics. The preload types incorporated increasing amounts of caudal constraint on the preload application vector ranging from an unconstrained dead-load arrangement to an apparatus that allowed the vector to follow rotations of the specimen. Six human cadaveric spine segments were tested (1-L1/L2, 3-L2/L3, 1-L3/L4 and 1-L4/L5). Pure moments were applied to the specimens with each of the four different types of compressive preload. Kinematic response was measured using an opto-electronic motion analysis system. A six-axis load cell was used to measure reaction forces and moments. Artefact reaction moments and shear forces were significantly affected by preload application method and magnitude. Unconstrained preload methods produced high artefact moments and low artefact shear forces while more constrained methods did the opposite. A mechanical trade-off is suggested by our results, whereby unwanted moment can only be prevented at the cost of shear force production. When comparing spine flexibility studies, caution should be exercised to ensure preload was applied in a similar manner for all studies. Unwanted moments or forces induced as a result of preload application method may render the comparison of two seemingly similar studies inappropriate.

Serotonin reuptake is less efficient in taste aversion resistant than in taste aversion-prone rats.

We have previously reported the development of rat lines bred selectively for differences in taste aversion conditionability. Earlier studies demonstrated that the taste aversion resistant (TAR) animals exhibited lower concentrations of brain serotonin and consumed greater amounts of ethanol than their taste aversion prone (TAP) counterparts. In the present study, TAR rats demonstrated significantly less efficient brain serotonin transport compared to TAP rats, but the rat lines demonstrated similar levels of serotonin transporter or V(max) and similar whole brain paroxetine (a specific serotonin reuptake inhibitor) binding (B(max)). These results suggest that the rat lines differ in the mechanisms that transport serotonin into nerve endings, but do not differ in the binding of serotonin to the transporter or in the number of serotonin transport sites. The data support the hypothesis that genetically determined differences in the serotonin system contribute to individual differences in taste aversion conditionability. The findings further suggest that differences in serotonin transport may influence the propensity to self-administer ethanol.

Three-dimensional measurement of cemented femoral stem stability: an in vitro cadaver study.

OBJECTIVE: To compare the in vitro stability of two cemented hip stem designs: Stem I was a collarless, double-tapered, highly polished implant; Stem II had a collar and matt finish. BACKGROUND: Stability of the femoral component of a hip implant is important for its long-term clinical success. Excessive migration or cyclic motion can increase the risk of early implant failure. METHODS: The stems were implanted in paired human cadaver femurs, and custom-designed micromotion sensors were used to measure three-dimensional motions of the stems at proximal, middle and distal locations during simulated in vivo loading cycles. RESULTS: This study found that despite 'rigid' fixation, cemented stems exhibit detectable motions under a limited number of cycles of simulated physiologic loads. At four times the donor body weight, Stem I showed a subsidence of 90 microm, compared to 25 microm of Stem II (P<0.05). In contrast, the proximal end of Stem II exhibited greater cyclic motions in the medial-lateral direction (P<0.05). CONCLUSIONS: The different motion patterns could be due to the design differences, such as surface finish and geometry. RelevanceImplant design is an important factor related to the behavior of the cement/bone interface and the overall success of the implant. This study compares in vitro micromotion of two cemented femoral prostheses with differing proximal designs.

Compliance of the L5-S1 spinal unit: a comparative study between an unconstrained and a partially constrained system.

A comparison between an unconstrained and a partially constrained system for in vitro biomechanical testing of the L5-S1 spinal unit was conducted. The objective was to compare the compliance and the coupling of the L5-S1 unit measured with an unconstrained and a partially constrained test for the three major physiological motions of the human spine. Very few studies have compared unconstrained and partially constrained testing systems using the same cadaveric functional spinal units (FSUs). Seven human L5-S1 units were therefore tested on both a pneumatic, unconstrained, and a servohydraulic, partially constrained system. Each FSU was tested along three motions: flexion-extension (FE), lateral bending (LB) and axial rotation (AR). The obtained kinematics on both systems is not equivalent, except for the FE case, where both motions are similar. The directions of coupled motions were similar for both tests, but their magnitudes were smaller in the partially constrained configuration. The use of a partially constrained system to characterize LB and AR of the lumbosacral FSU decreased significantly the measured stiffness of the segment. The unconstrained system is today's "gold standard" for the characterization of FSUs. The selected partially constrained method seems also to be an appropriate way to characterize FSUs for specific applications. Care should be taken using the latter method when the coupled motions are important.

Effects of restraint stress on plasma LH and testosterone concentrations, Leydig cell LH/hCG receptors, and in vitro testicular steroidogenesis in adult rats.

We examined the effect of restraint stress (3 hr) on plasma LH and testosterone levels, on the Leydig cell LH/hCG receptor, and on the activity of enzymes in the testicular steroidogenic pathway of the adult rat. Restraint stress caused a 47% reduction in plasma testosterone concentrations, but had no effect on plasma LH levels. The binding capacity and affinity of Leydig cell LH/hCG receptors were not affected by restraint. Stress did not affect the testicular activity of 20,22 desmolase or 3 beta-hydroxysteroid dehydrogenase, but testicular interstitial cells of stressed rats incubated in vitro with progesterone as a substrate produced more 17 alpha-hydroxyprogesterone but less testosterone than control cells, and when incubated with 17 alpha-hydroxypregnenolone, produced 39% less androstenedione and 40% less testosterone than control cells. These results suggest that restraint stress inhibited 17,20 desmolase but not 17 alpha-hydroxylase activity. When the delta 4 pathway was blocked with cyanoketone (3 beta-HSD inhibitor), stress did not alter the production of pregnenolone or 17 alpha-hydroxypregnenolone, but the production of dehydroepiandrosterone by cells from stressed rats was subnormal, suggesting again a reduction of 17,20 desmolase activity. The data suggest that a major site of the inhibitory action of restraint stress on testicular steroidogenesis is the 17,20 desmolase step. The disruption of androgen production by restraint appears to be LH independent since stress did not affect plasma LH levels, the binding capacity or affinity of LH/hCG receptors, or the activity of 20,22 desmolase.