Clodronate exerts an anabolic effect on articular chondrocytes mediated through the purinergic receptor pathway.

OBJECTIVE: Bisphosphonates are commonly used anti-osteoporotic drugs which have controversial effects on joint diseases including osteoarthritis. Certain bisphosphonates have been shown to have anabolic effects on cartilage which could have important ramifications for their proposed effects in vivo; however, the underlying mechanisms are poorly understood. Thus, the purpose of this study was to characterize the effects of clodronate on primary articular chondrocyte metabolism and to determine the underlying signaling pathways responsible. DESIGN: The effects of clodronate and pamidronate on extracellular matrix (ECM) biosynthesis, accumulation and MMP-13 activity were observed in high density, 3D cultures of bovine articular chondrocytes for up to 4 weeks were evaluated. Mechanisms were delineated by measuring intracellular Ca(2+) signaling and the effects of pharmacologic inhibition of the purinergic receptor pathway. RESULTS: Clodronate (100 µM) induced an anabolic effect (increased biosynthesis by 13-14%) which resulted in an 89-90% increase in ECM accumulation after 4 weeks of culture and without an associated effect on matrix turn-over. Stimulation by clodronate resulted in a 3.3-fold increase in Ca(2+) signaling and pharmacological inhibitor experiments suggested that the anabolic effects exerted by clodronate are transduced through the purinergic receptor pathway. CONCLUSIONS: These findings support the previous notion that certain bisphosphonates may be useful as adjunctive therapies to potentially ameliorate progression of cartilage degeneration and improve arthritis management.

The effect of moving point of contact stimulation on chondrocyte gene expression and localization in tissue engineered constructs.

Tissue engineering is a promising approach for articular cartilage repair. However, using current technologies, the developed engineered constructs generally do not possess an organized superficial layer, which contributes to the tissue's durability and unique mechanical properties. In this study, we investigated the efficacy of applying a moving point of contract-type stimulation (MPS) to stimulate the production of a superficial-like layer in the engineered constructs. MPS was applied to chondrocyte-agarose hydrogels at a frequency of 0.5, 1 or 2 Hz, under a constant compressive load of 10 mN for durations between 5 and 60 min over 3 consecutive days. Expression and localization of superficial zone constituents was conducted by qRT-PCR and in situ hybridization. Finite element modeling was also constructed to gain insight into the relationship between the applied stimulus and superficial zone constituent expression. Gene expression of superficial zone markers were affected in a frequency dependent manner with a physiologic frequency of 1 Hz producing maximal expression of PRG4, biglycan, decorin and collagen II. In situ hybridization revealed that localization of these markers predominantly occurred at 500-1000 µm below the construct surface which correlated to sub-surface strains between 10 and 25% as determined by finite element modeling. These results indicate that while mechanical stimuli can be used to enhance the expression of superficial zone constituents in engineered cartilage constructs, the resultant subsurface loading is a critical factor for localizing expression. Future studies will investigate altering the applied stimulus to further localize superficial zone constituent expression at the construct surface.

Chondrocyte repopulation of the zone of death induced by osteochondral harvest.

OBJECTIVE: Harvesting osteochondral grafts results in a zone of chondrocyte death (ZCD) in and around the periphery of the graft, creating a barrier for chondrocytes to migrate to the graft periphery, thus limiting cartilage-to-cartilage healing. The purpose of this study was to repopulate the induced ZCD through the combined effects of collagenase treatment and delivery of a chemotactic agent. DESIGN: In bovine cartilage, the ZCD induced by the OATS™ osteochondral harvesting system was determined, followed by a corresponding collagenase treatment to penetrate the developed ZCD. The chemotactic potential of platelet derived growth factor (PDGF-bb), insulin-like growth factor I (IGF-I), and basic fibroblast growth factor (bFGF) (2.5-100 ng/mL) was then assessed using a modified Boyden chamber assay to select an appropriate agent to induce chondrocyte migration. Afterwards, the combined effects of collagenase treatment and chondrocyte chemotaxis on the repopulation of an induced ZCD were examined in cartilage explants over a 4-week-period. RESULTS: The OATS™ osteochondral harvesting system induced a significant ZCD (173 µm, 95% CI: [72-274 µm]) in the grafts. Chondrocyte chemotaxis was induced by all agents investigated at concentrations greater than 25 ng/mL. After 4 weeks in culture, collagenase treatment alone reduced the ZCD by approximately 40% relative to untreated explants. Coupling the collagenase treatment with 25 ng/mL IGF-I reduced the ZCD by approximately 80% relative to untreated explants, and 65% relative to explants treated only with collagenase. CONCLUSION: Treating cartilage explants with collagenase and 25 ng/mL IGF-I resulted in a decreased ZCD after a 4-week-period, and increased chondrocyte density within the induced ZCD.

Effects of dehydration-induced structural and material changes on the apparent modulus of cancellous bone.

Dehydration is known to cause an increase in the elastic modulus of bone tissue. However, it also causes structural changes (i.e. shrinkage) which can themselves significantly alter the mechanical properties, particularly in cancellous bone. The current study attempts to estimate the contribution of these two competing factors to the net change of dehydration on the apparent modulus of bovine cancellous bone. Cylindrical cores from the lumbar vertebrae were tested in tension, while hydrated and again after dehydration. The bone volume fractions (BV/TV) were measured in both conditions. The results indicate that the average overall increase in the apparent modulus after dehydration is 14±14% (mean±SD), which represents the net effect of a 27% increase in modulus due to increased tissue modulus offset by a modulus decrease of 13% due to reductions in bone volume fraction. These observations underscore the need to consider both structural and material changes when comparing hydrated and dehydrated mechanical behaviour.

Specimen diameter and "side artifacts" in cancellous bone evaluated using end-constrained elastic tension.

In cancellous bone testing of cored samples, side artifacts are the underestimation of the true (i.e. in situ) mechanical properties due to the severing of the trabecular network during specimen preparation. Although other researchers have suggested correction factors derived from finite element method (FEM) models, it is proposed that side effects can be minimized by increasing the specimen diameter. Six different diameter specimens (3.1-10.6 mm), from two different anatomic sites (bovine femoral condyle and bovine lumbar vertebrae), were mechanically tested in elastic tension using an epoxy endcap protocol to eliminate end artifacts. Elastic modulus was found to be significantly affected by diameter in both sites. For example, the 5.1 mm samples underestimated the elastic modulus of the 10.6 mm samples by an average of roughly 20%. Yet no statistical difference was detected between the 8.3 and 10.6 mm samples in either anatomic site, suggesting that 8.3 mm diameter specimens were sufficiently large to avoid side artifacts. FEM models created from micro-CT images reveal that modulus approaches an asymptotic value with increasing diameter, and demonstrate an architecture-dependent drop in modulus at decreasing diameters. These results confirm, both experimentally and numerically, that side effects can be ignored given a suitably large specimen diameter and that this minimum diameter will be dependent on the cancellous architecture. An important implication of the latter result is that specimen diameters must be chosen appropriately when comparing test groups with different architectures (e.g. normal versus osteoporotic) to ensure that the magnitude of side artifacts does not confound the true differences between the groups.

Harnessing the purinergic receptor pathway to develop functional engineered cartilage constructs.

OBJECTIVE: Mechanical stimulation is a widely used method to enhance the formation and properties of tissue-engineered cartilage. While this approach can be highly successful, it may be more efficient and effective to harness the known underlying mechanotransduction pathways responsible. With this aim, the purpose of this study was to assess the effect of directly stimulating the purinergic receptor pathway through exogenous adenosine 5'-triphosphate (ATP) in absence of externally applied forces. METHODS: Isolated bovine articular chondrocytes were seeded in high density, 3D culture and supplemented with varying doses of ATP for up to 4 weeks. The effects on biosynthesis, extracellular matrix accumulation and mechanical properties were then evaluated. Experiments were also conducted to assess whether exogenous ATP elicited any undesirable effects, such as: inflammatory mediator release, matrix turn-over and mineralization. RESULTS: Supplementation with ATP had a profound effect on the growth and maturation of the developed tissue. Exogenous ATP (62.5-250 microM) increased biosynthesis by 80-120%, and when stimulated for a period of 4 weeks resulted in increased matrix accumulation (80% increase in collagen and 60% increase in proteoglycans) and improved mechanical properties (6.5-fold increase in indentation modulus). While exogenous ATP did not stimulate the release of inflammatory mediators or induce mineralization, high doses of ATP (250 microM) elicited a 2-fold increase in matrix metalloproteinase-13 expression suggesting the emergence of a catabolic response. CONCLUSIONS: Harnessing the ATP-purinergic receptor pathway is a highly effective approach to improve tissue formation and impart functional mechanical properties. However, the dose of ATP needs to be controlled as not to elicit a catabolic response.

Minimizing specimen length in elastic testing of end-constrained cancellous bone.

End-constraint is needed when performing tensile and compressive testing of cancellous bone so that frictional and structural end-effects are minimized and the mechanical properties are not underestimated. Since many end-constrained testing techniques are limited to certain sites and species by the large specimen sizes required, reductions in overall specimen length would be advantageous. The following study examines the effect of specimen gauge length and aspect ratio on the elastic tensile testing of cancellous bone from the bovine femoral condyle. Three different nominal gauge lengths (10, 16, 22 mm) were examined at two different diameters (5.1, 10.7 mm) which bound the range typically used for cancellous bone testing. No effect of gauge length or aspect ratio was observed, but evidence of a diameter effect was noted. Three-dimensional finite element method (FEM) models confirmed the experimental observations and demonstrated that an overestimation of modulus begins to occur for specimens shorter than 10 mm.

Mechanical vibrations increase the proliferation of articular chondrocytes in high-density culture.

Tissue engineering is a promising approach for articular cartilage repair; however, it still has proven a challenge to produce tissue from the limited number of cells that can be extracted from a single individual. Relatively few cell expansion methods exist without the problems of dedifferentiation and/or loss of potency. Previously, it has been shown that mechanical vibrations can enhance chondrocyte proliferation in monolayer culture. Thus, it was hypothesized that chondrocytes grown in high-density culture would respond in a similar fashion while maintaining phenotypic stability. Isolated bovine articular chondrocytes were seeded in high-density culture on Millicell filters and subjected to mechanical vibrations 48 h after seeding. Mechanical vibrations enhanced chondrocyte proliferation at frequencies above 350 Hz, with the peak response occurring at a 1g amplitude for a duration of 30 min. Under these conditions, the gene expression of cartilage-specific and dedifferentiation markers (collagen II, collagen I, and aggrecan) were unchanged by the imposed stimulus. To determine the effect of accumulated extracellular matrix (ECM) on this proliferative response, selected cultures were stimulated under the same conditions after varying lengths of preculture. The amount of accumulated ECM (collagen and proteoglycans) decreased this proliferative response, with the cultures becoming insensitive to the stimulus after 1 week of preculture. Thus, mechanical vibration can serve as an effective means preferentially to stimulate the proliferation of chondrocytes during culture, but its effects appear to be limited to the early stages where ECM accumulation is at a minimum.

Are micropatterned substrates for directed cell organization an effective method to create ordered 3D tissue constructs?

Tissue-engineered constructs grown in vitro tend to have random arrangements of cells and extracellular matrix (ECM) and much research effort is aimed at developing long-range organization in tissue-engineered constructs. Contact guidance, which utilizes substrates with topographical patterns of the scale of single cells (0.1-100 microm) to limit cell adhesion to specific locations and to influence cell shape and orientation, is one popular method which has been used to generate order in cell cultures. The use of contact guidance to generate three-dimensional (3D) order relies on the assumption that a newly forming cell or tissue layer will be guided by the organization of the previous layer, which has been organized by the patterned substrate. However, the ability for cellular patterns to be coupled through organized cell layers from a patterned substrate has not been effectively demonstrated. The results of this study demonstrate that, although the patterned substrate induces initial organization and polarization, this organization is not sustained in the successive cell/tissue layers that form above the initial cell layer. This finding suggests that cells must be in direct contact with the patterned substrate to maintain their polarization, orientation and positional organization. Therefore, contact guidance does not appear to be a promising technique to create ordered 3D tissue-engineered constructs. Alternative techniques, in particular those involving the application of mechanical, electrical or flow fields, may be more useful in sustaining organization in multilayered constructs as the organizational influence extends as a field into 3D space.

A single application of cyclic loading can accelerate matrix deposition and enhance the properties of tissue-engineered cartilage.

OBJECTIVE: Mechanical stimulation is a widely used method to enhance the formation and properties of tissue-engineered cartilage. While studies have evaluated the responsiveness of chondrocytes to mechanical stimuli, little is known about how much stimulation is actually required. Thus, the purpose of this study was to investigate the effect of a single application of cyclic loading to chondrocytes on the formation and properties of in vitro-formed tissue. DESIGN: Isolated bovine articular chondrocytes were seeded on ceramic substrates in 3D culture and subjected to a single application of compressive cyclic loading at 1, 8 or 15 days after seeding. Once the time at which the chondrocytes were most sensitive to mechanical loading was determined, the effect of a single application on the synthesis and accumulation of matrix molecules as well as the mechanical properties of the in vitro-formed cartilage tissue was evaluated. RESULTS: Chondrocytes were more responsive to cyclic loading applied early in culture. Cyclic forces applied 24 h after the cultures were established increased collagen and proteoglycan syntheses (48 +/- 11% and 49 +/- 11%, respectively). This single application of cyclic loading also increased the accumulation of collagen (stimulated: 207 +/- 20 microg, control: 173 +/- 9 microg) and proteoglycans (stimulated: 302 +/- 24 microg, control: 270 +/- 14 microg) as well as improved the mechanical properties of the in vitro-formed tissue (twofold increase in equilibrium stress and modulus) determined 4 weeks after the applied stimulus. CONCLUSIONS: A single application of cyclic loading to chondrocytes early in culture increased matrix accumulation and enhanced the mechanical properties of the in vitro-formed tissue. This suggests that mechanical forces do not have to be applied intermittently over long periods of time to accelerate in vitro tissue formation.

Effect of zoledronate on bone quality in the treatment of aseptic loosening of hip arthroplasty in the dog.

Periprosthetic bone loss, which is a direct cause of aseptic loosening in total hip arthroplasty (THA), can be suppressed by bisphosphonates. It is unknown how the quality of this bone is affected in the presence of both wear debris (from implant) and bisphosphonates. The objective of this study was to evaluate the effect of zoledronate (ZLN) on bone quality in the presence of wear debris [polyethylene (PE) particles] in a canine model of uncemented THA. Thirty dogs underwent THA, and aseptic loosening was induced via implantation of PE particles packed into the femoral component. For 26 weeks until sacrifice, two groups (each n = 10) received weekly injections of ZLN (low dose 2 mug/kg, high dose 10 mug/kg) and the third group (control) received saline. Histological and radiographic examinations were performed to evaluate the degree of implant reaction. Histomorphometry (static/dynamic) was performed to evaluate bone turnover. Back-scattered electron imaging was used to quantify the newly formed bone and to evaluate the mineralization distribution. Density fractionation and X-ray diffraction were used to evaluate mineral properties, while four-point bending was used to determine mechanical properties. A dose-dependent presence of newly formed subperiosteal bone was found, which appeared to be less mineralized than the adjacent cortical bone. The high-dose ZLN group showed decreased cortical porosity and turnover and increased mineralization profile, failure strength, and modulus. We conclude that ZLN affects some of the material properties of cortical bone and allows the newly formed subperiosteal bone to remain and therefore affect the overall quality of the bone.

The steroidal aromatase inhibitor exemestane prevents bone loss in ovariectomized rats.

The irreversible steroidal aromatase inhibitor exemestane (EXE) is one of three third generation aromatase inhibitors currently prescribed for advanced breast cancer in postmenopausal women. Its principal mechanism of action is to reduce estrogen by inhibiting its synthesis. In addition to its efficacy against breast cancer, its effects on other organs are important, especially when given to women with good-prognosis breast cancer or potentially to healthy women at increased risk of developing breast cancer. The purpose of this study was to evaluate the effects of EXE on bone and lipid metabolism in ovariectomized (OVX) rats. Ten-month-old Sprague-Dawley female rats were sorted into intact controls, intact + EXE, OVX controls, and OVX + EXE groups, and treated by weekly intramuscular injection with vehicle or 100 mg/kg EXE for 16 weeks. The bone mineral density (BMD), mechanical testing, histomorphometry, bone resorption marker-serum pyridinoline (PYD), and bone formation marker-serum osteocalcin (OC) were used to determine the effects of treatment on bone. In addition, total serum cholesterol, triglyceride, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were determined. BMD of the lumbar spine and femur were 11% and 7%, respectively, higher in OVX animals given EXE than in OVX controls (all Ps<0.001). Significant increases in the bending strength and toughness of the femora as well as the compressive strength and elastic modulus of the vertebrae were observed in OVX rats given EXE (all Ps<0.02 vs. OVX controls). Trabecular bone volume (BV) was significantly higher in OVX rats treated with EXE than in OVX controls (P<0.0001). In OVX animals, EXE reduced the OVX-induced increase of serum PYD by 96% (P<0.0001), and the OVX-induced increase of serum OC was completely prevented by treatment with EXE. In OVX animals, EXE resulted in a 28% reduction of serum cholesterol (P<0.0001) and reduced LDL by 64% compared with OVX controls (P<0.0001). The positive results of EXE on bone and lipid metabolism in the OVX rat model merit further investigation of the effects of EXE in postmenopausal women.

Polycyclic aromatic hydrocarbons present in cigarette smoke cause bone loss in an ovariectomized rat model.

A number of epidemiological studies have suggested that cigarette smoking is a risk factor for osteoporosis. Benzo(a)pyrene (BaP) and 7,12-dimethylbenz(a)anthracene (DMBA) are polycyclic aromatic hydrocarbons (PAHs) found in the tar fraction of cigarette smoke, as well as in car exhaust and furnace gases. We hypothesized that BaP and DMBA are responsible, through interaction with the aryl hydrocarbon receptor (AhR), for the bone loss and fragility seen in smoking-related osteoporosis. In this study four groups of 9-month-old Sprague-Dawley rats were examined. An intact group served as controls. A second control was the ovariectomized (ovx) group. The third group (ovx + E(2)) were ovariectomized and also given a continuous basal dose of estrogen by implanted estrogen pellet (0.085 mg of 17beta-estradiol per rat). The fourth group (ovx + E(2) + BaP/DMBA) was ovariectomized with an estradiol pellet, and received subcutaneous injections of 250 microg/kg of BaP/DMBA weekly for 15 weeks. The loss of ovarian function allowed the study of a direct effect of BaP/DMBA on bone while the concomitant estrogen repletion prevented ovx-related bone loss. Dual-energy X-ray absorptiometry (DEXA), histomorphometry, image analysis, and mechanical testing were used to determine the effect of the treatments on bone. The DEXA results showed a significant (p < 0.05) decrease in bone mineral density compared with intact controls with both ovx alone and with ovx + E(2) + BaP/DMBA treatment. The ovx + E(2) rats were similar to the intact controls. The osteoid parameters showed a significant increase (p < 0.05) with BaP/DMBA addition vs. intact controls, mimicking the ovx rats. The ovx + E(2) rats had osteoid parameters comparable to those of intact rats. Bone connectivity was decreased in the ovx and ovx + E(2) + BaP/DMBA animals. Connectivity of the ovx + E(2) rats was comparable to that of intact animals. A decrease in failure force was seen in three-point bending for the ovx + E(2) + BaP/DMBA group and in vertebral compression in both the ovx and ovx + E(2) + BaP/DMBA groups vs. intact controls. The mechanical properties of the ovx + E(2) rats were similar to those of intact rats. These results demonstrate that BaP/DMBA causes a loss of bone mass and bone strength, possibly through an increase in bone turnover. This is the first in vivo study linking environmental toxicants, found in the tar fraction of cigarette smoke and in urban air pollution, to loss of bone mass and strength in estrogen-replete ovx rats.

Nasal morphology and shape parameters as predictors of nasal esthetics in individuals with complete unilateral cleft lip and palate.

OBJECTIVE: The purpose of this study was to assess the ability of shape parameters of nasal morphology to predict esthetics in individuals with complete unilateral cleft lip and palate (CUCLP). METHODS: This retrospective study involved 28 patients with repaired CUCLP. Nostril morphology was analyzed using nose casts and a video-imaging technique. Calculated shape parameters included area, perimeter, centroid, angle of the principal axis, major and minor moments of area, anisometry, bulkiness, lateral offset, and three-dimensional internostril angles. Esthetics was assessed using a panel of six orthodontists who rated nasal esthetics from frontal, lateral, basal, and three-quarters view slides and from nose casts. Correlations between esthetics and the shape parameters were completed using the entire group as well as using two statistically determined subsets: those with the best and those with the worst esthetics. RESULTS: Nasal esthetics was related to only the perimeter and bulkiness parameter ratios. Symmetry of the perimeters between the right and left nostrils positively correlated with better esthetics using the entire sample group while symmetry of bulkiness between the right and left nostrils positively correlated with better esthetics using both the entire sample group and the best and worst subsets. CONCLUSIONS: Only perimeter and bulkiness showed positive correlations with nasal esthetics. The group of parameters used to assess nostril morphology had neither significant correlation with-nor predictive power for-esthetics. Thus, an assessment of the entire nasal surface topography in three dimensions needs to be completed and assessed with respect to predictability of nasal esthetics.

Video-imaging assessment of nasal morphology in individuals with complete unilateral cleft lip and palate.

OBJECTIVE: The purpose of this study was to develop a video-imaging mathematical method to assess nostril morphology. DESIGN: This retrospective study involved two age-matched groups: 28 subjects with complete unilateral cleft lip and palate (CUCLP) and 19 noncleft controls. Nose casts were reproducibly oriented in a jig such that the casts could be rotated about the coronal axis. Video images of the nostrils were captured and then analyzed for area, perimeter, centroid, principal axis, moments about the major and minor axes (I11, I22), anisometry, bulkiness, lateral offset, internostril angle, and rotational angle. RESULTS: All parameters identified nostril asymmetry in both groups. The results of the analyses using anisometry, I11, and I22 showed that, in both groups, one nostril was rounder and one was more elliptical. This asymmetry, however, differed between the two groups, and the difference was primarily based on the degree of ellipticity of the nostrils. Maximum dimension, perimeter, lateral offset, I11, and I22 were more asymmetric in the cleft group. In the control group, the right nostril was more elliptical and had a greater perimeter, and the left-side nostril had a greater bulkiness (enfolding). CONCLUSIONS: The method developed was validated for assessment of nasal morphology in cleft and noncleft samples. Nostril morphology was asymmetric in both groups but more asymmetric in the cleft group than the control group. The dominant influence of the cleft resulted in more elliptical noncleft nostrils and greater nostril shape asymmetry in the cleft group. The validated video-imaging method can now be used to assess the efficacy of treatment on nasal morphology.

Diabetic neuropathy: diagnosis and treatment for the pain management specialist.

Diabetic neuropathy is the name used by clinicians to describe a heterogeneous group of diseases that affect the autonomic and peripheral nervous systems of patients suffering from diabetes mellitus. This article provides the pain management specialist with an overview of the pathophysiology and current trends in the diagnosis and treatment of these diseases. Because of the significant morbidity and suffering associated with diabetic neuropathy, this article emphasizes practical steps to prevent, treat, and manage diabetic neuropathy to assist the pain management specialist in caring for patients suffering from this common malady.

Imposed state of deformation determines local collagen fibre orientation but not apparent mechanical properties.

In a previous study, we have shown that the observed biaxial material behaviour of planar connective tissues is influenced by the sample gripping method. Commonly used suture attachments produced an apparently more compliant and extensible material compared to the same sample with clamped edges. We hypothesized that these differences were due to the imposed collagen fibre constraint under each method. In this study, we have directly compared the collagen fibre orientations which result under both gripping schemes. Small angle light scattering (SALS) was used to determine collagen fibre orientations in square bovine pericardial samples, before and after a 10% equibiaxial stretch. Local fibre distributions were determined at the sample centre and at the grip-sample interface. Resulting scattering patterns were statistically compared using repeated measures ANOVA. After stretch, collagen fibre distributions were identical at the sample centre where deformation is measured--but not at the sample boundaries. Therefore, the central fibre orientation distribution appears to be exclusively determined by the imposed deformation state. However, it is important to note that the loads necessary to achieve a given deformation depended strongly on gripping method. The resulting apparent differences in mechanical properties must be due to the method of load transmission. Indeed, fibres were observed to arc around suture attachment points, suggesting a discontinuous load transfer to the specimen which produced an apparent increase in extensibility and compliance. By contrast, only smooth transitions were observed at the clamped edges. Direct transmission of load from grip-to-grip in clamped samples (away from the sample centre) increased apparent stiffness.

Thermomechanical analysis of collagen crosslinking in the developing lamb pericardium.

In a developing lamb model, we have used hydrothermal isometric tension (HIT) techniques to assess collagen crosslink stability and its contribution to the mechanical properties of the pericardium. Strip samples of tissue were either: (i) heated to a 90 degrees C isotherm or (ii) heated in 5 degrees C increments between 60-90 degrees C and then 93 and 97 degrees C isotherms. The half-life of stress relaxation associated with peptide bond hydrolysis (t1/2) was calculated at each isotherm. The activation energy, Eact, for the hydrolysis-associated relaxation process was also calculated using the data from the stepwise HIT tests--a technical improvement which significantly reduces the experimental time required to develop statistically valid measurements. Crosslinking in the pericardium increased during development and was demonstrated both by thermoelasticity and by resistance to enzymatic solubilization. We observed greater conversation to thermally stable crosslinks upon maturation, the ratio of the NaBH4-stabilized/unstabilized half-lives peaking at 21 days postnatal. Whereas tissue from lambs (119 day fetal, and 3 day and 21 day postpartum) showed an early maximum and rapid decay of force, NaBH4 stabilization significantly increased thermal stability and yielded profiles similar to those in adult tissue.

Dynamic contact stress and rolling resistance model for total knee arthroplasties.

Problems associated with premature failure of total knee replacements (TKR's) include: wear, creep, and oxidation of ultrahigh-molecular-weight polyethylene (UHMWPe) as well as adverse tissue reactions to polyethylene wear debris. These problems are associated in part with the mechanical behavior of UHMWPe. In TKR's, contact stress analyses have been performed on the UHMWPe tibial component; however, most have employed simplified material properties and not accounted for joint kinematics. A nonlinear viscoelastic rolling model was developed for TKR's to predict the contact stress and rolling friction for varying rolling speed, conformity, applied load, and tibial plateau thickness. Results indicated that the contact stress increased and rolling friction decreased with increasing rolling speed. Effects of conformity, applied load, and tibial plateau thickness were consistent with previous models. At large rolling speeds, predicted peak contact stresses were almost twice their static value, resulting in a compound fatigue problem in UHMWPe components due to normal cyclic loading, moving point of contact, and velocity-dependent stresses.