Results of the phase IIa RADICAL trial of the FGFR inhibitor AZD4547 in endocrine resistant breast cancer.

We conducted a phase IIa, multi-centre, open label, single arm study (RADICAL; NCT01791985) of AZD4547 (a potent and selective inhibitor of Fibroblast Growth Factor Receptor (FGFR)-1, 2 and 3 receptor tyrosine kinases) administered with anastrozole or letrozole in estrogen receptor positive metastatic breast cancer patients who had become resistant to aromatase inhibitors. After a safety run-in study to assess safety and tolerability, we recruited 52 patients. The primary endpoint was change in tumour size at 12 weeks, and secondary endpoints were to assess response at 6 weeks, 20 weeks and every 8 weeks thereafter and tolerability of the combined treatment. Two partial responses (PR) and 19 stable disease (SD) patients were observed at the 12-week time point. At 28 weeks, according to centrally reviewed Response Evaluation Criteria in Solid Tumours (RECIST) criteria, five PR and 8 SD patients were observed in 50 assessable cases. Overall, objective response rate (5 PR) was of 10%, meeting the pre-specified endpoint. Fourteen patients discontinued due to adverse events. Eleven patients had retinal pigment epithelial detachments which was asymptomatic and reversible in all but one patient. Exploratory ribonucleic acid sequencing (RNA-Seq) analysis was done on patients' samples: 6 differentially-expressed-genes could distinguish those who benefited from the addition of AZD4547.

Human articular cartilage is orthotropic where microstructure, micromechanics, and chemistry vary with depth and split-line orientation.

OBJECTIVE: Quantitative, micrometer length scale assessment of human articular cartilage is essential to enable progress toward new functional tissue engineering approaches, including utilization of emerging 3D bioprinting technologies, and for improved computational modeling of the osteochondral unit. Thus the objective of this study was to characterize the structural organization, material properties, and chemical composition of human skeletally mature articular cartilage with respect to depth and defined morphological features: normal to the articulating surface, parallel to the split-line, and transverse to the split-line. METHOD: Three samples from the lateral femoral condyles of 4 healthy adult donors (55-61 years old) were evaluated via histology, second harmonic generation, microindentation, and Raman spectroscopy. All metrics were evaluated as a function of depth and direction relative to the split-line. RESULTS: All donors presented with intact and healthy tissue. Collagen fiber orientation varied significantly between testing directions and with increasing depth from the articular surface. Both compressive and tensile modulus increased significantly with depth and differed across the middle and deep zones and depended on orthogonal direction relative to the split-line. Similarly, matrix components varied with both depth and direction, where chondroitin sulfate steadily increased with depth while collagen prevalence was highest in the surface layer. CONCLUSIONS: Microscale measurements of human articular cartilage demonstrate that properties are both depth-dependent and orthotropic and depend on the underlying tissue structure and composition. These findings improve upon existing knowledge establishing more accurate measurements, with greater degree of depth and spatial specificity, as inputs for tissue engineering and computational modeling.

A Small Deformation Thermoporomechanics Finite Element Model and Its Application to Arterial Tissue Fusion.

Understanding the impact of thermally and mechanically loading biological tissue to supraphysiological levels is becoming of increasing importance as complex multiphysical tissue-device interactions increase. The ability to conduct accurate, patient specific computer simulations would provide surgeons with valuable insight into the physical processes occurring within the tissue as it is heated or cooled. Several studies have modeled tissue as porous media, yet fully coupled thermoporomechanics (TPM) models are limited. Therefore, this study introduces a small deformation theory of modeling the TPM occurring within biological tissue. Next, the model is used to simulate the mass, momentum, and energy balance occurring within an artery wall when heated by a tissue fusion device and compared to experimental values. Though limited by its small strain assumption, the model predicted final tissue temperature and water content within one standard deviation of experimental data for seven of seven simulations. Additionally, the model showed the ability to predict the final displacement of the tissue to within 15% of experimental results. These results promote potential design of novel medical devices and more accurate simulations allowing for scientists and surgeons to quickly, yet accurately, assess the effects of surgical procedures as well as provide a first step toward a fully coupled large deformation TPM finite element (FE) model.

Enhanced mechanical properties of photo-clickable thiol-ene PEG hydrogels through repeated photopolymerization of in-swollen macromer.

Current hydrogels used for tissue engineering are limited to a single range of mechanical properties within the replicated tissue construct. We show that repeated in-swelling by a single hydrogel pre-cursor solution into an existing polymerized hydrogel followed by photo-exposure increases hydrogel mechanical properties. The process is demonstrated with a photo-clickable thiol-ene hydrogel using a biocompatible precursor solution of poly(ethylene glycol) dithiol and 8-arm poly(ethylene glycol) functionalized with norbornene. The polymer fraction in the precursor solution was varied by 5, 10, and 20 percent by weight and an off-stoichiometric ratio of thiol : ene was used, leaving free enes available for subsequent reaction. Multiple swelling and exposure cycles for the same precursor solution were performed. The compressive modulus increased by a factor between three and ten (formulation dependent), while volume swelling ratio decreased by a factor of two, consistent with increased crosslink density. The modified hydrogels also demonstrate increased toughness by fracturing at compressive forces five times greater than the initial hydrogel. We attribute the increased toughness to subsequent increases in crosslink density created by the repeated photopolymerization of in-swollen macromer. This technique demonstrates the ability to significantly modify hydrogel network properties by exploiting swelling and polymerization processes that can be applied to traditional three-dimensional printing systems to spatially control local mechanical properties.

Reinforcement of Mono- and Bi-layer Poly(Ethylene Glycol) Hydrogels with a Fibrous Collagen Scaffold.

Biomaterial-based tissue engineering strategies hold great promise for osteochondral tissue repair. Yet significant challenges remain in joining highly dissimilar materials to achieve a biomimetic, mechanically robust design for repairing interfaces between soft tissue and bone. This study sought to improve interfacial properties and function in a bi-layer hydrogel interpenetrated with a fibrous collagen scaffold. 'Soft' 10% (w/w) and 'stiff' 30% (w/w) PEGDM was formed into mono- or bi-layer hydrogels possessing a sharp diffusional interface. Hydrogels were evaluated as single-(hydrogel only) or multi-phase (hydrogel + fibrous scaffold penetrating throughout the stiff layer and extending >500 µm into the soft layer). Including a fibrous scaffold into both soft and stiff mono-layer hydrogels significantly increased tangent modulus and toughness and decreased lateral expansion under compressive loading. Finite element simulations predicted substantially reduced stress and strain gradients across the soft-stiff hydrogel interface in multi-phase, bilayer hydrogels. When combining two low moduli constituent materials, composites theory poorly predicts the observed, large modulus increases. These results suggest material structure associated with the fibrous scaffold penetrating within the PEG hydrogel as the major contributor to improved properties and function-the hydrogel bore compressive loads and the 3D fibrous scaffold was loaded in tension thus resisting lateral expansion.

Biological and metabolic response in STS-135 space-flown mouse skin.

There is evidence that space flight condition-induced biological damage is associated with increased oxidative stress and extracellular matrix (ECM) remodeling. To explore possible mechanisms, changes in gene expression profiles implicated in oxidative stress and in ECM remodeling in mouse skin were examined after space flight. The metabolic effects of space flight in skin tissues were also characterized. Space Shuttle Atlantis (STS-135) was launched at the Kennedy Space Center on a 13-day mission. Female C57BL/6 mice were flown in the STS-135 using animal enclosure modules (AEMs). Within 3-5 h after landing, the mice were euthanized and skin samples were harvested for gene array analysis and metabolic biochemical assays. Many genes responsible for regulating production and metabolism of reactive oxygen species (ROS) were significantly (p < 0.05) altered in the flight group, with fold changes >1.5 compared to AEM control. For ECM profile, several genes encoding matrix and metalloproteinases involved in ECM remodeling were significantly up-/down-regulated following space flight. To characterize the metabolic effects of space flight, global biochemical profiles were evaluated. Of 332 named biochemicals, 19 differed significantly (p < 0.05) between space flight skin samples and AEM ground controls, with 12 up-regulated and 7 down-regulated including altered amino acid, carbohydrate metabolism, cell signaling, and transmethylation pathways. Collectively, the data demonstrated that space flight condition leads to a shift in biological and metabolic homeostasis as the consequence of increased regulation in cellular antioxidants, ROS production, and tissue remodeling. This indicates that astronauts may be at increased risk for pathophysiologic damage or carcinogenesis in cutaneous tissue.

Spaceflight and hind limb unloading induce similar changes in electrical impedance characteristics of mouse gastrocnemius muscle.

OBJECTIVE: To assess the potential of electrical impedance myography (EIM) to serve as a marker of muscle fiber atrophy and secondarily as an indicator of bone deterioration by assessing the effects of spaceflight or hind limb unloading. METHODS: In the first experiment, 6 mice were flown aboard the space shuttle (STS-135) for 13 days and 8 earthbound mice served as controls. In the second experiment, 14 mice underwent hind limb unloading (HLU) for 13 days; 13 additional mice served as controls. EIM measurements were made on ex vivo gastrocnemius muscle. Quantitative microscopy and areal bone mineral density (aBMD) measurements of the hindlimb were also performed. RESULTS: Reductions in the multifrequency phase-slope parameter were observed for both the space flight and HLU cohorts compared to their respective controls. For ground control and spaceflight groups, the values were 24.7±1.3°/MHz and 14.1±1.6°/MHz, respectively (p=0.0013); for control and HLU groups, the values were 23.9±1.6°/MHz and 19.0±1.0°/MHz, respectively (p=0.014). This parameter also correlated with muscle fiber size (ρ=0.65, p=0.011) for spaceflight and hind limb aBMD (ρ=0.65, p=0.0063) for both groups. CONCLUSIONS: These data support the concept that EIM may serve as a useful tool for assessment of muscle disuse secondary to immobilization or microgravity.

The impact of off the visual axis retinoscopy on objective central refractive measurement in adult clinical practice: a prospective, randomized clinical study.

PURPOSE: The objective of this study was to examine the effect of the off axis retinoscopy on objective central refractive measurement in adult clinical practice. METHODS: In all, 40 subjects underwent undilated retinoscopy in a randomly selected eye both on and off the visual axis by a single masked examiner. Off axis retinoscopy was defined as retinoscopy performed with the testing eye of the examiner aligned with the contralateral (non-test) eye of the subject resulting in an off axis deviation in the nasal horizontal visual field. Retinoscopy was performed in negative cylinder only and spherocylindrical measurements were converted to power vectors for analysis. Paired t-test was used to assess differences in M, J(0) and J(45) power vectors including differences between mean aided and unaided LogMar acuities. RESULTS: In all, 14 subjects were myopic (SE≤-0.5 D), 13 subjects were emmetropic (SE between -0.49 and 1.0 D) and 13 subjects were hyperopic (SE >1.0 D). Mean angle of deviation was 5.58° in the nasal horizontal visual field. J(0) showed a significant negative shift in those with myopia (P<0.001) and emmetropia (P=0.049) following off axis retinoscopy. No significant differences in M, J(0) and J(45) were found in the hyperopes. Mean aided LogMar acuities after on and off axis retinoscopy were both significantly better than mean unaided LogMar VA (P<0.001 in both cases). CONCLUSION: Small degrees of off axis retinoscopy encountered in everyday clinical practice can induce errors in objective central refractive measurement.

Deformation partitioning provides insight into elastic, plastic, and viscous contributions to bone material behavior.

The relative contributions of elastic, plastic, and viscous material behavior are poorly described by the separate extraction and analysis of the plane strain modulus, E('), the contact hardness, H(c) (a hybrid parameter encompassing both elastic and plastic behavior), and various viscoelastic material constants. A multiple element mechanical model enables the partitioning of a single indentation response into its fundamental elastic, plastic, and viscous deformation components. The objective of this study was to apply deformation partitioning to explore the role of hydration, tissue type, and degree of mineralization in bone and calcified cartilage. Wet, ethanol-dehydrated, and PMMA-embedded equine cortical bone samples and PMMA-embedded human femoral head tissues were analyzed for contributions of elastic, plastic and viscous deformation to the overall nanoindentation response at each site. While the alteration of hydration state had little effect on any measure of deformation, unembedded tissues demonstrated significantly greater measures of resistance to plastic deformation than PMMA-embedded tissues. The PMMA appeared to mechanically stabilize the tissues and prevent extensive permanent deformation within the bone material. Increasing mineral volume fraction correlated with positive changes in E('), H(c), and resistance to plastic deformation, H; however, the partitioned deformation components were generally unaffected by mineralization. The contribution of viscous deformation was minimal and may only play a significant role in poorly mineralized tissues. Deformation partitioning enables a detailed interpretation of the elastic, plastic, and viscous contributions to the nanomechanical behavior of mineralized tissues that is not possible when examining modulus and contact hardness alone. Varying experimental or biological factors, such as hydration or mineralization level, enables the understanding of potential mechanisms for specific mechanical behavior patterns that would otherwise be hidden within a more complex set of material property parameters.

Exercise does not affect stiffness and mineralisation of third metacarpal condylar subarticular calcified tissues in 2 year old thoroughbred racehorses.

Impact exercise has a profound effect in increasing volumetric density of epiphyseal bone, as clearly shown in 2 year old thoroughbred racehorses from which we derived the tissue studied in the present investigation. Here, we asked the question whether the fabric-level properties of the mineralised tissues immediately below hyaline articular cartilage which transmit the extra loads are themselves altered in consequence. We therefore studied the nanoindentation elastic modulus and its relationship to the concentration of mineral determined by quantitative backscattered electron imaging in the heavily loaded palmar medial and lateral condyles of the distal third metacarpal bone (Mc3) of 4 untrained and 4 trained 2-year old Thoroughbred racehorses. We found no difference between trained and untrained horses in either subchondral bone or calcified cartilage in the mean stiffness or mineral content or their correlation. Thus neither articular calcified cartilage nor the immediately adjacent subchondral bone were affected by exercise, even though they transmitted the higher load associated with athletic training through to the deeper bone, which itself responded floridly to exercise. Under the circumstances of this experiment and at least in the very small regions studied, therefore, the structure of these two tissues was apparently optimised to function.

Comparison of tail-suspension and sciatic nerve crush on the musculoskeletal system in young-adult mice.

Musculoskeletal unloading and disuse result in significant muscle and bone loss. These phenomena can be modeled using sciatic nerve crush or tail-suspension. Mature animals eliminate the complication of growth superimposed on bone and muscle loss. In the current study, young-adult (12-week old male) C57BL/6J mice were subjected to sciatic nerve crush (NC; n = 9) or tail-suspension (TS; n = 9) for 14 days, with a normal ambulatory control (n = 10). The soleus, gastrocnemius, and EDL muscles were collected and weighed at sacrifice. Femurs were analyzed in three-point bending for stiffness, elastic force and maximum force. Muscle masses in tail suspended mice were reduced by 41.9% (p < 0.001), 17.5% (p < 0.001), and 9.1% (N.S.) for the soleus, gastrocnemius, and EDL, respectively. In NC mice, muscle masses were reduced by 18.6% (p = 0.004), 37.2% (p < 0.001), and 22.5% (p = 0.003). Femur stiffness, elastic and maximum forces were reduced by 20.9% (p = 0.014), 14.7% (N.S.), and 11.6% (N.S.) in TS, compared to NC where masses were reduced by 15.5% (p = 0.022), 0.2% (N.S.) and 11.2% (N.S.) in the crushed leg compared to the contralateral control. NC resulted in a greater reduction of muscle mass in the gastrocnemius and EDL muscle; whereas tail-suspension had a greater effect on the soleus. Tail-suspension had the greatest effect on bone mechanical properties. When comparing these results to actual spaceflight data, it appears as though TS most closely models muscle loss, and NC most closely models changes in bone mechanical properties. These unloading models have tissue-specific effects that impact their applications for musculoskeletal research.

Exercise prevention of unloading-induced bone and muscle loss in adult mice.

Skeletal unloading causes bone and muscle loss that may be reversed by post-unloading exercise. This study examines the effects of unloading and exercise, using tail-suspension for 14 days combined with a week of post-suspension cage wheel running in mice. Twenty-four adult, male, C57BL/6J mice were divided into four groups (n = 6 mice/group); unsuspended non-running, tail-suspended non-running, unsuspended running, and tail-suspended running. At sacrifice, the calf (soleus, gastrocnemeius and plantaris complex), heart, tibia and femur were collected and weighed. The femora and tibiae were cleaned of non-osseous tissue, subjected to 3-point bending (femurs only), and weighed for dry (105 degrees C; 24h) and ash mass (800 degrees C; 24h). The mean calf mass from the tail-suspended groups (157.13 +/- 2.83 mg) was significantly less than in the unsuspended groups (167.33 +/- 2.83 mg; p = 0.019), with no significant effect of cage wheel running. The mean heart mass in running groups (166.58 +/- 4.78 mg) was significantly greater than the non-running groups (148.17 +/- 4.78 mg; p = 0.013), with no effect of hindlimb suspension. The mean femur ash mass from tail-suspended groups (24.02 +/- 0.38 mg) were significantly less than the unsuspended groups (25.11 +/- 0.34 mg; p = 0.050), and the running groups (25.13 +/- 0.38 mg) were significantly greater than the non-running groups (24.00 +/- 0.34 mg; p = 0.043). No effect was observed for the femur dry mass or percent mineralization. Measurements of mechanical length tended to be lower in tail-suspension, with no significant affects do to cage wheel running. This study suggests that tail-suspension in adult mice significantly decreases skeletal muscle and bone mass, with no change in percent mineralization. Furthermore, one week of running does not reverse the effects on the skeletal muscle and bone mass.

A novel combination of methods to assess sarcopenia and muscle performance in mice.

A novel combination of assays was developed to assess sarcopenia and muscle performance. Three techniques were tested to assess muscle function both during and upon termination of treatments designed to induce sarcopenia. In unsuspended (US) and hindlimb suspended (HS) mice, a Hindlimb Exertion Force Test (HEFT), cage wheel running, and in vitro muscle electrophysiology were performed. Twelve-week old, mature male C57BL/6J mice were HS (n = 24) for two weeks, or served as US controls (n = 26). Both groups were subjected to a HEFT on day 13; that is, the maximum force exerted against a beam force transducer (2 lb. linear range, Transducer Techniques, Temecula CA) following applied tail shock stimulus (0.15 mA, 300 msec). This test primarily evaluated the hindlimb muscles used for an escape response (i.e., hamstrings, quadriceps and calf muscles). Mice (n = 10-11/group) were given voluntary access to running wheels for 7 days post treatment to evaluate muscle endurance. On day 13, HS mice showed a mean 18.9% (p = 0.002) decrease in the maximum force exerted compared to US mice. After 7 days of wheel running, HS running distance tended to decrease (13.2%, p = 0.084). HS mice ran an average of 2.0 km/day less than US control mice, with similar running patterns: distance declined on day 2 following completion of HS but increased steadily thereafter. With in vitro testing, the maximum soleus tetanus response decreased by 31.8% (p = 0.01) with HS, in agreement with the changes observed by the other assays. These three assays, combined, appear to provide effective and complementary ways to measure muscle performance and functional differences.

Effect of MPC-11 myeloma and MPC-11 + IL-1 receptor antagonist treatment on mouse bone properties.

This study examines the effects of an IL-6-producing murine multiple myeloma cell line on trabecular and cortical mouse bone, and evaluates the efficacy of interleukin-1 receptor antagonist (IL-1ra) in mitigating bone destruction. Six-week-old BALB/c mice were assigned to two groups: normal controls and myeloma animals (5 x 10(7) MPC-11 cells on day 0). Myeloma animals were further assigned to three unique groups: MPC-11 only; MPC-11 treated with hyaluronic acid (HA); and MPC-11 + IL-1ra/HA (100 mg/kg). Disease development was assessed at 14 and 21 days via spleen, liver, and proximal tibia histology; histomorphometry at the femoral middiaphysis; and long bone composition and mechanical testing. Histologic analysis revealed marked myeloma infiltration into organs and bone marrow and gross bone resorption of the proximal tibia. IL-1ra tended to decrease bone resorption at the proximal tibia; however, it had no effect on quantitatively measured bone parameters. Whole femur and tibia, and tibial epiphysis, percent mineralization was decreased (3.0%, 2.9%, and 6.3%, respectively) in all MPC-11 groups. The presence of myeloma did not affect long bone stiffness, strength, or length over the 3 week study. The percent of the femoral endosteal perimeter showing excessive resorption ( approximately 60%) in the MPC-11 groups increased significantly after 21 days. MPC-11 cell presence caused no change in bone formation or morphology. Normal growth mechanisms were not impacted, as the bones lengthened and increased in size and mass despite the presence of myeloma. IL-1 does not appear to be a primary factor in in vivo bone destruction caused by the MPC-11 cell line. These findings reveal the stochastic nature of bone lesions in multiple myeloma and suggest that IL-1 is not a cytokine critical to this disease pathology.

Osteoprotegerin ameliorates sciatic nerve crush induced bone loss.

This study examines the ability of osteoprotegerin (OPG) to prevent the local bone resorption caused by sciatic nerve damage. Sixty-five 18-week-old male mice were assigned to one of six groups (n = 10-11/group). A baseline control group was sacrificed on day zero of the 10-day study. The remaining groups were placebo sham operated, placebo nerve crush (Plac NC) operated, 0.1 mg/kg/day OPG + nerve crush (LOW), 0.3 mg/kg/day OPG + nerve crush (MED), and 1.0 mg/kg/day OPG + nerve crush (HI). Nerve crush or sham operations were performed on the right leg. The left leg served as a contralateral control to the nerve crushed (ipsilateral) leg. The difference in mass between the right and left femur and tibia was examined. Additionally, quantitative histomorphometry was performed on the right and left femur and tibia diaphyses. Nerve crush resulted in a significant loss of bone mass in the ipsilateral side compared to the contralateral side. Bone mass for the ipsilateral bones of the Plac NC group were significantly reduced by 3.8% in the femur and 3.5% in the tibia compared to the contralateral limb. The percent diminution was reduced for OPG treated mice compared to the Plac NC group for both the femur and tibia. In the femur, the percent reduction of ipsilateral bone mass was reduced to 1.0% (LOW), 1.3% (MED) and 1.6% (HI) compared to the contralateral limb. In the tibia, loss of bone mass in the ipsilateral limb was reduced to 1.4% (LOW), 1.4% (MED), and 2.4% (HI) compared to the contralateral. OPG also decreased the amount of tibial endocortical resorption compared to the Plac NC group. In summary, OPG mitigated bone loss caused by damage to the sciatic nerve.

Psoriasis, psoriatic arthropathy and relapsing orbital myositis.

Orbital myositis is an inflammatory disorder of the orbital muscles causing orbital pain and restriction of eye movements. Although rare in children, it is most frequently seen after orbital trauma or as a post-infectious process. We describe a child with chronic relapsing psoriasis, juvenile psoriatic arthritis and relapsing bilateral orbital myositis.