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Dynamic stochastic simulation of cancellous bone resorption.

A stochastic simulation of cancellous bone resorption was developed and applied to a simple two-dimensional lattice structure representing the vertebral body. The simulation is based upon the concept of a basic multicellular unit (BMU) where net resorption (-deltaB.BMU) is considered at bone/marrow surfaces. The cancellous bone structure is defined as a binary matrix with the size of the pixels corresponding to a square element of approximately 20 microm dimension. The simulation considers both the probability that any surface pixel will be activated into a BMU and, if activated, the length of the resorption cavity. The relationship between relative stiffness and density for the simulation was predicted by finite element analysis. The stochastic simulation was iterated eight times with the mechanical properties assessed after each stage. Perforation of a single trabeculae was first observed at step 2, the structure completely lacking connectivity and mechanical integrity by step 8. The slope of the stiffness-porosity graph was greater than unity for the first five steps, but thereafter approached zero because the structure had lost connectivity and effectively collapsed. The eight-step simulation was repeated five times and demonstrated that, although the stiffness/density relationships were similar at the extremes of density, the dependence of stiffness upon density varied. This clearly demonstrates the stochastic nature of the simulation upon cancellous bone structure, and is probably indicative of a significant dependence of mechanical integrity upon perforation effects.

Biomechanical Phenomena↗

ADAMTS10 mutations in autosomal recessive Weill-Marchesani syndrome.

Weill-Marchesani syndrome (WMS) is characterized by the association of short stature; brachydactyly; joint stiffness; eye anomalies, including microspherophakia and ectopia of the lenses; and, occasionally, heart defects. We have recently mapped a gene for the autosomal recessive form of WMS to chromosome 19p13.3-p13.2, in a 12.4-cM interval. Here, we report null mutations in a member of the extracellular matrix protease family, the gene encoding ADAMTS10, a disintegrin and metalloprotease with thrombospondin motifs. A total of three distinct mutations were identified in two consanguineous families and in one sporadic WMS case, including one nonsense mutation (R237X) and two splice mutations (1190+1G-->A and 810+1G-->A). ADAMTS10 expression studies using reverse-transcriptase polymerase chain reaction, northern blot, and dot-blot analyses showed that ADAMTS10 is expressed in skin, fetal chondrocytes, and fetal and adult heart. Moreover, electron microscopy and immunological studies of the skin fibroblasts from the patients confirmed impairment of the extracellular matrix. We conclude, therefore, that ADAMTS10 plays a major role in growth and in skin, lens, and heart development in humans.

ADAM Proteins↗

[Value of pulp recession in inveterate hook fingers. Techniques/indications].

The aim of this operation is to preserve the sensitivity whenever when a finger has to be amputated. We have reviewed 9 patients operated according to this technique, out of 14 patients for whom an amputation or a shortening arthrodesis was discussed. The results of pulp recession were most encouraging. The incision were located on both sides of the finger and distant from the ungueal matrix. The dorsal skin was elevated in the sub-periosteal plane. Resection of the necessary amount of 2nd phalanx to obtain an extended finger was performed. An osteosynthesis with pins is preferred for a duration of 6 weeks. This operation can be performed for very stiff finger, hooked fingers or dystrophic fingers with a good pulp. In extreme cases of shortening, the nail with its bed ar sacrificed, and the entire pulp is used to cover the distal stump. This principal of pulp recession has been used in two other cases in emergency surgery. The postoperative was marked by finger swelling and some pain of the distal sutured skin. Overall the results were satisfactory and patients were very satisfied to keep their finger, which would otherwise have been doomed.

Adolescent↗

Short-term infliximab therapy improves symptoms of psoriatic arthritis and decreases concentrations of cartilage oligomeric matrix protein.

OBJECTIVE: The aim of the current study was to evaluate the short-term effects of anti-tumour necrosis factor alpha (infliximab) therapy on serum cartilage oligomeric matrix protein (COMP) levels, a possible biomarker of cartilage destruction. METHODS: Nine consecutive patients with active psoriatic arthritis (PsA) were treated with infliximab for 6 weeks. Serum COMP levels were measured and correlated to pre-established disease activity outcome variables: pain as assessed by the patient, using the 100 mm visual analogue scale (VAS), duration of morning stiffness (MGST), swollen joint count (SJC), tender joint count (TJC), erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP). RESULTS: Significant improvements in MGST, VAS, SJC and TJC values were observed after 6 weeks of therapy. Similar significant improvements were demonstrated in the ACR response rate and in eight (89%) patients the ACR20 was achieved. ESR and CRP decreased significantly over 6 weeks. Serum COMP levels also decreased significantly after 6 weeks of treatment (12.99 +/- 1.71 baseline, 10.22 +/- 1.1 after 6 weeks, P < 0.008). CONCLUSION: The results of our study suggest that short-term therapy with infliximab leads to decreased COMP levels in patients with PsA. COMP seems to be a good candidate for a biomarker reflecting cartilage response to this treatment in PsA patients.

Antibodies, Monoclonal↗

A fibril-network-reinforced biphasic model of cartilage in unconfined compression.

Cartilage mechanical function relies on a composite structure of a collagen fibrillar network entrapping a proteoglycan matrix. Previous biphasic or poroelastic models of this tissue, which have approximated its composite structure using a homogeneous solid phase, have experienced difficulties in describing measured material responses. Progress to date in resolving these difficulties has demonstrated that a constitutive low that is successful for one test geometry (confined compression) is not necessarily successful for another (unconfined compression). In this study, we hypothesize that an alternative fibril-reinforced composite biphasic representation of cartilage can predict measured material responses and explore this hypothesis by developing and solving analytically a fibril-reinforced biphasic model for the case of uniaxial unconfined compression with frictionless compressing platens. The fibrils were considered to provide stiffness in tension only. The lateral stiffening provided by the fibril network dramatically increased the frequency dependence of disk rigidity in dynamic sinusoidal compression and the magnitude of the stress relaxation transient, in qualitative agreement with previously published data. Fitting newly obtained experimental stress relaxation data to the composite model allowed extraction of mechanical parameters from these tests, such as the rigidity of the fibril network, in addition to the elastic constants and the hydraulic permeability of the remaining matrix. Model calculations further highlight a potentially important difference between homogeneous and fibril-reinforced composite models. In the latter type of model, the stresses carried by different constituents can be dissimilar, even in sign (compression versus tension) even though strains can be identical. Such behavior, resulting only from a structurally physiological description, could have consequences in the efforts to understand the mechanical signals that determine cellular and extracellular biological responses to mechanical loads in cartilage.

Animals↗

Defects in articular cartilage metabolism and early arthritis in fibroblast growth factor receptor 3 deficient mice.

Fibroblast growth factor (FGF) receptor 3 has been identified as a key regulator of endochondral bone development and of post-natal bone metabolism through its action on growth plate chondrocytes and osteoblasts, respectively. It has also been shown to promote chondrogenesis and cartilage production by cultured pre-chondrogenic cells in response to FGF18. In the current studies, we show that the absence of signaling through Fgfr3 in the joints of Fgfr3(-/-) mice leads to premature cartilage degeneration and early arthritis. Degenerative changes in cartilage matrix included excessive proteolysis of aggrecan core protein and type II collagen, as measured by neo-epitope immunoreactivity. These changes were accompanied by increased expression of metalloproteinase MMP13, type X collagen, cellular hypertrophy and loss of proteoglycan at the articular surface. Using a novel micro-mechanical indentation protocol, it was shown that articular cartilage in the humeral head of 4-month-old Fgfr3(-/-) mice was less resistant to compressive force and less stiff than that of littermate controls. These results identify Fgfr3 signaling as a potential target for intervention in degenerative disorders of cartilage metabolism.

Aggrecans↗

[Associations between viscoelastic properties of large arteries and their extracellular matrix composition in abdominal aortic aneurysms in humans].

The objective of the present study was to determine the viscoelastic properties of the common carotid artery in 35 patients with aortic aneurysm before surgery (AAA) (age 71 years, range 61-84), in comparison with 48 patients with essential hypertension (HT: 50 years, range 24-88) and 44 normotensive subjects (NT: 44 years, range 23-85). The second objective was to establish the relations between common carotid artery (CCA) viscoelastic properties and histologic lesions observed on AAA segments, obtained after surgery. CCA diameter was larger and distensibility smaller in AAA patients than in HT and NT. Distensibility of the aortic aneurysm was smaller than that of upstream 'normal' aorta, itself being smaller than control aortas. AAA wall lesions were extensive, associating adventitial and medial fibrosis, elastolysis, smooth muscle rarefaction, neovascularization, inflammation and plaques. The grade of these lesions was not correlated with the mechanical properties of the aorta and CCA; however, they could explain their qualitative alterations. AAA is characterized by severe stiffening and dilatation of large arteries distant from the aneurysm location. Whether this pattern of arterial phenotype is explained by the increase in stiff material (collagen) and the rarefaction of distensible material (smooth muscle and elastin) remains to be determined.

Adult↗

Conjugate addition reactions combined with free-radical cross-linking for the design of materials for tissue engineering.

PEG-diacrylamide was synthesized and utilized to make materials for tissue engineering. Acrylamide groups readily react with thiol groups, and peptides containing a single thiol group were coupled to the PEG-diacrylamide in aqueous solution at room temperature in about 2 h. If only a portion of the acrylamide groups were targeted for reaction with peptide, sufficient amounts of PEG-diacrylamide remained to be polymerized by free-radical mechanisms via photoinitiation. The photopolymerization step can be performed in contact with cells, providing a means to produce bioactive scaffolds for tissue engineering. The photopolymerization conditions and precursor composition greatly affect the stiffness of the materials, which subsequently affected cell spreading. The kinetics and extent of cell spreading on the bioactive materials were measured and compared to cell spreading on tissue culture polystyrene. Although the PEG materials resist protein adsorption, the experiments suggest that the cells can secrete extracellular matrix that can adhere to the gels.

Acrylamides↗

Cytoskeletal mechanics in confluent epithelial cells probed through integrins and E-cadherins.

Mechanical forces associated with the cytoskeleton (CSK) and transmitted to adjacent cells or to the extracellular matrix (ECM) influence cellular functions. We investigated the force transfer across cell-to-ECM and cell-to-cell connections using magnetic twisting cytometry. We probed the CSK through integrins and E-cadherins in confluent epithelial cell lines (MCF7). At high applied stress (> 10 dyn/cm2), stiffness (stress/strain) of the CSK coupled through integrins was greater than stiffness coupled through E-cadherins. The stiffness reduction after microfilament or microtubule disruption with cytochalasin D or colchicine was greater for integrins. At low applied stress, disruption of microfilaments had very little effect on stiffness probed through either receptor type, indicating a correspondingly small contribution of microfilaments to the CSK mechanics in these confluent cells. This differs from results in nonconfluent MCF7 cells and from predictions that are based on prestressed models in which tensile stresses presumably associated with the microfilaments are the origin of prestress and, in consequence, cell stiffness. In addition, there was substantial cell spreading on collagen I-coated dishes, in contrast to little spreading on dishes coated with E-cadherin antibody. This result, together with observations of a relatively high cell stiffness probed through integrins compared with the small stiffness probed through E-cadherins, suggests that mechanical force transmission might also be important in regulating cell spreading. We conclude that the degree of confluency may be associated with different mechanics and functions of the CSK network.

Actin Cytoskeleton↗

Replacement of the knee meniscus by a porous polymer implant: a study in dogs.

BACKGROUND: Meniscectomy will lead to articular cartilage degeneration in the long term. Therefore, the authors developed an implant to replace the native meniscus. HYPOTHESIS: The porous polymer meniscus implant develops into a neomeniscus and protects the cartilage from degeneration. STUDY DESIGN: Controlled laboratory study. METHODS: In a dog model, a porous polymer scaffold with optimal properties for tissue infiltration and regeneration of a neomeniscus was implanted and compared with total meniscectomy. The tissue infiltration and redifferentiation in the scaffold, the stiffness of the scaffold, and the articular cartilage degeneration were evaluated. RESULTS: Three months after implantation, the implant was completely filled with fibrovascular tissue. After 6 months, the central areas of the implant contained cartilage-like tissue with abundant collagen type II and proteoglycans in their matrix. The foreign-body reaction remained limited to a few giant cells in the implant. The compression modulus of the implant-tissue construct still differed significantly from that of the native meniscus, even at 6 months. Cartilage degeneration was observed both in the meniscectomy group and in the implant group. CONCLUSION: The improved properties of these polymer implants resulted in a faster tissue infiltration and in phenotypical differentiation into tissue resembling that of the native meniscus. However, the material characteristics of the implant need to be improved to prevent degeneration of the articular cartilage. CLINICAL RELEVANCE: The porous polymer implant developed into a polymer-tissue construct that resembled the native meniscus, and with improved gliding characteristics, this prosthesis might be a promising implant for the replacement of the meniscus.

Animals↗

Functions of lumican and fibromodulin: lessons from knockout mice.

Lumican and fibromodulin are collagen-binding leucine-rich proteoglycans widely distributed in interstitial connective tissues. The phenotypes of lumican-null (Lum(-/-)), Fibromodulin-null (Fmod(-/-)) and compound double-null (Lum(-/-)Fmod(-/-)) mice identify a broad range of tissues where these two proteoglycans have overlapping and unique roles in modulating the extracellular matrix and cellular behavior. The lumican-deficient mice have reduced corneal transparency and skin fragility. The Lum(-/-)Fmod(-/-) mice are smaller than their wildtype littermates, display gait abnormality, joint laxity and age-dependent osteoarthritis. Misaligned knee patella, severe knee dysmorphogenesis and extreme tendon weakness are the likely cause for joint-laxity. Fibromodulin deficiency alone leads to significant reduction in tendon stiffness in the Lum(+/+)Fmod(-/-) mice, with further loss in stiffness in a lumican gene dose-dependent way. At the level of ultrastructure, the Lum(-/-) cornea, skin and tendon show irregular collagen fibril contours and increased fibril diameter. The Fmod(-/-) tendon contains irregular contoured collagen fibrils, with increased frequency of small diameter fibrils. The tendons of Lum(-/-)Fmod(-/-) have an abnormally high frequency of small and large diameter fibrils indicating a de-regulation of collagen fibril formation and maturation. In tissues like the tendon, where both proteoglycans are present, fibromodulin may be required early in collagen fibrillogenesis to stabilize small-diameter fibril-intermediates and lumican may be needed at a later stage, primarily to limit lateral growth of fibrils

Animals↗

Theoretical model for myocardial trabeculation.

During the morphogenetic process of myocardial trabeculation, most of the cardiac jelly of the initially smooth-walled heart is replaced by sponge-like muscle. The mechanisms that drive and regulate this important process are poorly understood. Using a theoretical model, we examined the possible role that cytoskeletal contraction plays during the initial stages of trabeculation. The myocardium is modeled as a thin viscoelastic membrane consisting of contractile (stress) fibers embedded in an isotropic incompressible matrix, with the interaction of myocardial cells and cardiac jelly fibers providing long-range mechanical effects. The stress fibers are assumed to behave like smooth muscle and to normally operate on the descending limb of their stress-stretch curve. Mechanical instability due to the effectively negative stiffness then leads to the creation of pattern. As a first approximation, computations were carried out for a flat rectangular membrane with stress fibers aligned along a single direction. The computed deformation patterns depend strongly on the magnitude and anisotropy of the long-range effects. Given plausible assumptions about the mechanical properties of the embryonic heart, the model predicts trabecular patterns similar to those observed in the embryo, including the development of circumferential ridges and relatively thin regions ("holes") in the trabecular sheets.

Animals↗

Early changes in material properties of rabbit articular cartilage after meniscectomy.

We have correlated early material and biochemical changes in articular cartilage in a surgical model for cartilage degeneration. Medial meniscectomy was performed on the left knee of 17 adult, female New Zealand white rabbits. The equilibrium Young's modulus of cartilage was assessed by an indentation test in situ at defined sites on the medial and lateral tibial plateaus of the operated and control knees; the cartilage was then excised and analyzed biochemically. Focal changes were consistently observed in the medial surface of the operated knee. The equilibrium modulus and the glycosaminoglycan content fell rapidly, reaching a minimum by 2 weeks after surgery; the lateral tibial surface was essentially unaffected. Six months after surgery, the glycosaminoglycan content had returned to normal and the modulus to near normal. Independent measurements on cored plugs from the medial surface 2 weeks after surgery revealed a significant decrease in both the dynamic stiffness and the streaming potential in the operated knee compared with the control. The findings suggest that normal ambulatory loads in vivo will deform the affected medial cartilage much more than normal. It remains to be seen if altered mechanical stresses are solely responsible for initiating and sustaining matrix remodeling by the chondrocytes.

Animals↗

Osteoprotegerin mitigates tail suspension-induced osteopenia.

Osteoprotegerin (OPG) is a recently discovered protein related to the tumor necrosis factor receptor family. It has been shown to inhibit ovariectomy (ovx)-induced resorption in rats and increase bone mineral density in young mice. Tail suspension is a procedure that inhibits bone formation in maturing rodents. This study was designed to quantify OPG's effect on cortical bone formation. Fifty-four mice were assigned to one of five groups (n = 10-11/group). A baseline control group was killed on day 0 of the 10 day study. The remaining groups were: vivarium housed (nonsuspended) control mice receiving 0.3 mg/kg per day OPG; vivarium control mice receiving daily placebo injections; tail-suspended mice receiving 0. 3 mg/kg per day OPG; and tail-suspended mice receiving placebo injections. Tetracycline was administered on days 0 and 8. OPG treatment of tail-suspended mice produced mechanical properties similar to those of placebo-treated, vivarium-housed mice: structural stiffness (8.5%, 20.7%) and elastic (13.9%, 10.1%) and maximum (4.7%, 8.1%) force were increased compared with placebo controls (vivarium, suspended groups). Percent mineral composition was highly significantly greater (p < 0.001 for all comparisons) for OPG-treated mice in the femur, tibia, and humerus, relative to placebo treatment. Matrix mass was also significantly increased in the femur, although not to the same degree as mineral mass. OPG decreased the amount of femoral endocortical resorption compared with the placebo-treated groups for both vivarium (27%) and suspended (24%) mice. Administration of OPG significantly decreased endocortical formation of the tibia. Periosteal bone formation rates were not altered by OPG. OPG-mitigated tail suspension induced osteopenia not by returning bone formation to normal levels, but by inhibiting resorption and increasing percent mineral composition.

Animals↗

Collected studies on interfaces and interphases as related to the behaviour of fibre-reinforced aluminium alloy composites

This paper is an essentially practical treatment of interphases and interfaces and of their influence on the properties of a number of metal matrix composites (MMCs). The illustrations are drawn from the authors' experiences and have been chosen to underline the importance of detailed microstructural analysis for elucidating the fabrication behaviour and the mechanical performance of this group of materials. The work involves a series of MMCs based upon different combinations of aluminium alloy and ceramic/carbon fibre (both continuous and short) and made using the method of low-pressure liquid metal infiltration (LMI). Detailed analyses of the composite microstructures are given, with particular attention being paid to the interface regions. The data are used to categorize an interface according to the type of bond, that is a mechanical bond resulting from thermal mismatch between the fibre and metal matrix, or a chemical bond, with or without second phase, caused by chemical reaction. The information is then employed to account for aspects of composite fabrication, such as the cast microstructure produced by the LMI method and the effect of heat treatment, and to elucidate composite properties such as stiffness, yield stress and failure strength.

Journal Article↗

Pharmacokinetics of an injectable sustained-release formulation of morphine for use in dogs.

This study investigated the pharmacokinetics of morphine sulphate in an injectable chitosan-based gel. Gels were made from a combination of N-O-carboxymethylchitosan (NOCC) and chitosan and were easily injectable via a 22 gauge needle and appeared stable during long-term storage. Groups of six beagles were injected subcutaneously (s.c.) with 1.2 mg/kg morphine sulphate, either in sterile saline or in sterilized gels, and serial blood samples were withdrawn via a jugular catheter and later analysed for morphine concentrations using radioimmunoassay. Data were analysed according to non-compartmental pharmacokinetics. NOCC-based gels resulted in significantly lower serum morphine concentrations at 10 and 30 min following injection but significantly higher concentrations at all points from 120 to 480 min post-injection. Dogs receiving morphine gel exhibited equivalent or lesser variability in serum morphine concentrations than dogs receiving conventional morphine sulphate. Pharmacokinetic analysis revealed that morphine release from the gel matrix was significantly prolonged but fully bioavailable. There were no significant differences in either distribution (Vd) or terminal elimination (t 1/2). Dogs experienced no adverse effects other than those normally associated with morphine administration at the time of injection but all dogs receiving the gel presented with an undefined stiffness the next day that resolved spontaneously within 48 h. We conclude that carboxymethylchitosan-based gels hold considerable promise for the development of injectable sustained-release formulations of opioid analgesics.

Animals↗

Rat airway smooth muscle cell during actin modulation: rheology and glassy dynamics.

Although changes of cytoskeleton (CSK) stiffness and friction can be induced by diverse interventions, all mechanical changes reported to date can be scaled onto master relationships that appear to be universal. To assess the limits of the applicability of those master relationships, we focused in the present study on actin and used a panel of actin-manipulating drugs that is much wider than any used previously. We focused on the cultured rat airway smooth muscle (ASM) cell as a model system. Cells were treated with agents that directly modulate the polymerization (jasplakinolide, cytochalasin D, and latrunculin A), branching (genistein), and cross linking (phallacidin and phalloidin oleate) of the actin lattice. Contractile (serotonin, 5-HT) and relaxing (dibutyryl adenosine 3',5'-cyclic monophosphate, DBcAMP) agonists and a myosin inhibitor (ML-7) were also tested for comparison, because these agents may change the structure of actin indirectly. Using optical magnetic twisting cytometry, we measured elastic and frictional moduli before and after treatment with each agent. Stiffness increased with frequency as a weak power law, and changes of friction paralleled those of stiffness until they approached a Newtonian viscous limit. Despite large differences in the mechanism of action among the interventions, all data collapsed onto master curves that depended on a single parameter. In the context of soft glassy systems, that parameter would correspond to an effective temperature of the cytoskeletal matrix and reflect the effects of molecular crowding and associated molecular trapping. These master relationships demonstrate that when the mechanical properties of the cell change, they are constrained to do so along a special trajectory. Because mechanical characteristics of the cell shadow underlying molecular events, these results imply special constraints on the protein-protein interactions that dominate CSK mechanical properties.

Actins↗

Sex differences in bone growth of broiler chickens.

Females are often supposed to have a lighter skeleton than males, even in avian domestic species. However, in broiler chickens, females are less susceptible to bone deformities than males. In order to better understand these conflicting facts, male and female broilers were compared for the growth of cortical bone. Morphology, histomorphometry, composition and biomechanical properties of the tibiotarsi were analysed in both sexes at 1, 12, 26 and 42 days of age. The quantity of bone tissue of the tibiotarsus (weight, volume, diameter of the diaphysis, area of the cortex) was smaller in females, although the occurrence of varus-valgus deformations of the intertarsal joint was largely reduced in female chickens (8.8% versus 19.9% in males at 42 days of age). The tibia became significantly lighter in females from 26 d of age. Differences in tibia length and volume became significant at 42 d of age only, while cross-sections of the diaphysis were smaller in females from the hatching, leading to thinner bones in females. The percentage of dry matter of tibiae was higher in females from hatching. From 12 days old, tibiotarsi of females tended to be less porous and were more mineralized (higher ash/dry matter ratio). In females, mineralization proceeded at a higher rate (MAR) until 26 d of age and MAR became higher in males afterwards. The stiffness of the tibia diaphysis was similar in males and females all along the growth. In conclusion, the growth of cortical bone is very different in male and female broilers. In female broilers, the thinness of bone diaphysis is counter-balanced by modifications in the composition of the matrix and in the porosity of the cortex, leading to equal biomechanical characteristics of tibiotarsi in both sexes.

Aging↗