Abstract: There has been an increase in the application of different biomaterials to repair hard tissues. Within these biomaterials, calcium phosphate (CaP) bioceramics are suitable candidates, since they can be biocompatible, biodegradable, osteoinductive, and osteoconductive. Moreover, during sintering, bioceramic materials are prone to form micropores and undergo changes in their surface topographical features, which influence cellular physiology and bone ingrowth. In this study, five geometrical properties from the surface of CaP bioceramic particles and their micropores were analyzed by data mining techniques, driven by the research question: what are the geometrical properties of individual micropores in a CaP bioceramic, and how do they relate to each other? The analysis not only shows that it is feasible to determine the existence of micropore clusters, but also to quantify their geometrical properties. As a result, these CaP bioceramic particles present three groups of micropore clusters distinctive by their geometrical properties. Consequently, this new methodological clustering assessment can be applied to advance the knowledge about CaP bioceramics and their role in bone tissue engineering.

Abstract: In the United States, approximately eight million osseous fractures are reported annually, of which 5-10% fail to create a bony union. Osteoblast-specific deletion of the gene Pten in mice has been found to stimulate bone growth and accelerate fracture healing. Healing rates at four weeks increased in femurs from Pten osteoblast conditional knock-out mice (Pten-CKO) compared to wild-type mice (WT) of the same genetic strain as measured by an increase in mechanical stiffness and failure load in four-point bending tests. Preceding mechanical testing, each femur was imaged using a Skyscan 1172 micro-computed tomography (mu CT) scanner (Skyscan, Kontich, Belgium). The present study used μCT image-based analysis to test the hypothesis that the increased femoral fracture force and stiffness in Pten-CKO were due to greater section properties with the same effective material properties as that of the WT. The second moment of area and section modulus were computed in ImageJ 1.46 (National Institutes of Health) and used to predict the effective flexural modulus and the stress at failure for fourteen pairs of intact and callus WT and twelve pairs of intact and callus Pten-CKO femurs. For callus and intact femurs, the failure stress and tissue mineral density of the Pten-CKO and WT were not different; however, the section properties of the Pten-CKO were more than twice as large 28 days post-fracture. It was therefore concluded, when the gene Pten was conditionally knocked-out in osteoblasts, the resulting increased bending stiffness and force to fracture were due to increased section properties. (C) 2014 Elsevier Ltd. All rights reserved.

Abstract: Laternula elliptica is a key bivalve species and widely distributed around the Antarctic continent. This bivalve has been the study subject in several studies centered on ecological, physiological, biochemical, and behavioral patterns. However, little is known about the chemistry and the biomechanical properties of the shells of this mollusk. Here, we present the first report of the intra-population variability in the organic composition and mechanical properties of L. elliptica shells. Further, we analyze different morphological traits and their association with the metabolism of a population of L. elliptica from King George Island, Western Antarctic Peninsula. The summer metabolic rates and the hepatosomatic index values indicate good health conditions of this clam's population. Shell periostracum chemistry is quite similar to bivalves from temperate regions, but the relative amount of protein increased ca. five-fold in shells of L. elliptica. The microhardness is approximately 32% lower than in bivalves from temperate regions. Our characterization of the L. elliptica shells suggests that periostracum chemistry could be specially fitted to avoid shell carbon exposure to dissolution (e.g., in corrosive acidified seawater). In contrast, the reduction in shell hardness may result from prioritizing behavioral (burial) and shell repairing strategies to confront biological (predators) and physical disturbances (e.g., ice scouring). Similar studies in other Antarctic mollusks will help understand the role of shell structure and function in confronting projected climate changes in the Antarctic ocean.

Abstract: The exposure to environmental variations in pH and temperature has proven impacts on benthic ectotherms calcifiers, as evidenced by tradeoffs between physiological processes. However, how these stressors affect structure and functionality of mollusk shells has received less attention. Episodic events of upwelling of deep cold and low pH waters are well documented in eastern boundary systems and may be stressful to mollusks, impairing both physiological and biomechanical performance. These events are projected to become more intense, and extensive in time with ongoing global warming. In this study, we evaluate the independent and interactive effects of temperature and pH on the biomineral and biomechanical properties of Argopecten purpuratus scallop shells. Total organic matter in the shell mineral increased under reduced pH (similar to 7.7) and control conditions (pH similar to 8.0). The periostracum layer coating the outer shell surface showed increased protein content under low pH conditions but decreasing sulfate and polysaccharides content. Reduced pH negatively impacts shell density and increases the disorder in the orientation of calcite crystals. At elevated temperatures (18 degrees C), shell microhardness increased. Other biomechanical properties were not affected by pH/temperature treatments. Thus, under a reduction of 0.3 pH units and low temperature, the response of A. purpuratus was a tradeoff among organic compounds (biopolymer plasticity), density, and crystal organization (mineral plasticity) to maintain shell biomechanical performance, while increased temperature ameliorated the impacts on shell hardness. Biopolymer plasticity was associated with ecophysiological performance, indicating that, under the influence of natural fluctuations in pH and temperature, energetic constraints might be critical in modulating the long-term sustainability of this compensatory mechanism.

Abstract: Birnbaum-Saunders (BS) models are receiving considerable attention in the literature. Multivariate regression models are a useful tool of the multivariate analysis, which takes into account the correlation between variables. Diagnostic analysis is an important aspect to be considered in the statistical modeling. In this paper, we formulate multivariate generalized BS regression models and carry out a diagnostic analysis for these models. We consider the Mahalanobis distance as a global influence measure to detect multivariate outliers and use it for evaluating the adequacy of the distributional assumption. We also consider the local influence approach and study how a perturbation may impact on the estimation of model parameters. We implement the obtained results in the R software, which are illustrated with real-world multivariate data to show their potential applications.

Abstract: Increased bone formation resulting from mechanical loading is well documented; however, the interactions of the mechanotransduction pathways are less well understood. Endothelin-1, a ubiquitous autocrine/paracrine signaling molecule promotes osteogenesis in metastatic disease. In the present study, it was hypothesized that exposure to big endothelin-1 (big ET1) and/or mechanical loading would promote osteogenesis in ex vivo trabecular bone cores. In a 2 x 2 factorial trial of daily mechanical loading (-2000 μepsilon,120 cycles daily, “jump” waveform) and big ET1 (25 ng/mL), 48 bovine sternal trabecular bone cores were maintained in bioreactor chambers for 23 days. The bone cores' response to the treatment stimuli was assessed with percent change in core apparent elastic modulus (Delta E-app), static and dynamic histomorphometry, and prostaglandin E2 (PGE2) secretion. Two-way ANOVA with a post hoc Fisher's LSD test found no significant treatment effects on Delta E-app (p = 0.25 and 0.51 for load and big ET1, respectively). The Delta E-app in the “no load + big ET1” (CE, 13 +/- 12.2%, p = 0.56), “load + no big ET1” (LC, 17 +/- 3.9%, p = 0.14) and “load + big ETI” (LE, 19 +/- 4.2%, p = 0.13) treatment groups were not statistically different than the control group (CC, 3.3% +/- 8.6%). Mineralizing surface (MS/BS), mineral apposition (MAR) and bone formation rates (BFR/BS) were significantly greater in LE than CC (p = 0.037, 0.0040 and 0.019, respectively). While the histological bone formation markers in LC trended to be greater than CC (p = 0.055, 0.11 and 0.074, respectively) there was no difference between CE and CC (p = 0.61, 0.50 and 0.72, respectively). Cores in LE and LC had more than 50% greater MS/BS (p = 0.037, p = 0.055 respectively) and MAR (p = 0.0040, p = 0.11 respectively) than CC. The BFR/BS was more than two times greater in LE (p = 0.019) and LC (p = 0.074) than CC. The PGE2 levels were elevated at 8 days post-osteotomy in all groups and the treatment groups remained elevated compared to the CC group on days 15,19 and 23. The data suggest that combined exposure to big ET1 and mechanical loading results in increased osteogenesis as measured in biomechanical, histomorphometric and biochemical responses. (C) 2016 Elsevier Inc. All rights reserved.

Abstract: Asphalt self-healing by encapsulated rejuvenating agents is considered a revolutionary technology for the autonomic crack-healing of aged asphalt pavements. This paper aims to explore the use of Bio-Oil (BO) obtained from liquefied agricultural biomass waste as a bio-based encapsulated rejuvenating agent for self-healing of bituminous materials. Novel BO capsules were synthesized using two simple dripping methods through dropping funnel and syringe pump devices, where the BO agent was microencapsulated by external ionic gelation in a biopolymer matrix of sodium alginate. Size, surface aspect, and elemental composition of the BO capsules were characterized by optical and scanning electron microscopy and energy-dispersive X-ray spectroscopy. Thermal stability and chemical properties of BO capsules and their components were assessed through thermogravimetric analysis (TGA-DTG) and Fourier-Transform Infrared spectroscopy (FTIR-ATR). The mechanical behavior of the capsules was evaluated by compressive and low-load micro-indentation tests. The self-healing efficiency over time of BO as a rejuvenating agent in cracked bitumen samples was quantified by fluorescence microscopy. Main results showed that the BO capsules presented an adequate morphology for the asphalt self-healing application, with good thermal stability and physical-chemical properties. It was also proven that the BO can diffuse in the bitumen reducing the viscosity and consequently self-healing the open microcracks.

Abstract: Acrylic bone cement is often used in total joint replacement procedures to anchor an orthopaedic implant to bone. Bone cement is a viscoelastic material that exhibits creep and stress relaxation properties, which have been previously characterized using a variety of techniques such as flexural testing. Nanoindentation has become a popular method to characterize polymer mechanical properties at the nanoscale due to the technique's high sensitivity and the small sample volume required for testing. The purpose of the present work therefore was to determine the mechanical properties of bone cement using traditional macroscale techniques and compare the results to those obtained from nanoindentation. To this end, the quasi-static and viscoelastic properties of two commercially available cements, Palacos and Simplex, were assessed using a combination of three-point bending and nanoindentation. Quasi-static properties obtained from nanoindentation tended to be higher relative to three-point bending. The general displacement and creep compliance trends were similar for the two methods. These findings suggest that nanoindentation is an attractive characterization technique for bone cement, due to the small sample volumes required for testing. This may prove particularly useful in testing failed/ retrieved cement samples from patients where material availability is typically limited. (C) 2016 Wiley Periodicals, Inc.

Abstract: Direct ink writing (DIW) is a promising extrusion-based 3D printing technology, which employs an ink-deposition nozzle to fabricate 3D scaffold structures with customizable ink formulations for tissue engineering applications. However, determining the optimal DIW process parameters such as temperature, pressure, and speed for the specific ink is essential to achieve high reproducibility of the designed geometry and subsequent mechano-biological performance for different applications, particularly for porous scaffolds of finite sizes (total volume > 1000 mm3) and controlled pore size and porosity. The goal of this study was to evaluate the feasibility of fabricating Polycaprolactone (PCL) and bio-active glass (BG) composite-based 3D scaffolds of finite size using DIW. 3D-scaffolds were fabricated either as cylinders (10 mm diameter; 15 mm height) or cubes (5 × 5 × 5 mm3) with height/width aspect ratios of 1.5 and 1, respectively. A rheological characterization of the PCL-BG inks was performed before printing to determine the optimal printing parameters such as pressure and speed for printing at 110 °C. Microstructural properties of the scaffolds were analyzed in terms of overall scaffold porosity, and in situ pore size assessments in each layer (36 pores/layer; 1764 pores per specimen) during their fabrication. Measured porosity of the fabricated specimens&#65533;PCL: =46.94%, SD = 1.61; PCL-10 wt%BG: = 48.29%, SD = 5.95; and PCL-20 wt% BG: =50.87%, SD = 2.45&#65533;matched well with the designed porosity of 50%. Mean pore sizes&#65533;PCL [ = 0.37 mm (SD = 0.03)], PCL-10%BG [ = 0.38 mm (SD = 0.07)] and PCL-20% BG [ = 0.37 mm (SD = 0.04)]&#65533;were slightly fairly close to the designed pore size of 0.4 mm. Nevertheless there was a small but consistent, statistically significant (p < 0.0001) decrease in pore size from the first printed layer (PCL: 0.39 mm; PCL-10%BG: 0.4 mm; PCL-20%BG: 0.41 mm) to the last. SEM and micro-CT imaging revealed consistent BG particle distribution across the layers and throughout the specimens. Cell adhesion experiments revealed similar cell adhesion of PCL-20 wt% BG to pure PCL, but significantly better cell proliferation &#65533; as inferred from metabolic activity &#65533; after 7 days, although a decrease after 14 days was noted. Quasi-static compression tests showed a decrease in compressive yield strength and apparent elastic modulus with increasing BG fraction, which could be attributed to a lack of adequate mechanical bonding between the BG particles and the PCL matrix. The results show that the inks were successfully generated, and the scaffolds were fabricated with high resolution and fidelity despite their relatively large size (>1000 mm3). However, further work is required to understand the mechano-biological interaction between the BG particle additives and the PCL matrix to improve the mechanical and biological properties of the printed structures.

Abstract: 3D printing has gained great popularity due to its main feature of manufacturing complex geometries. The building process by adding successive layers generates mechanical properties that depend on the printing parameters, where build orientation is one of the most relevant factors. Due to this, the characterization of the mechanical response of these pieces is a challenging task of practical importance to estimate their lifespan. The aim of this study is to characterize the mechanical behavior and define a 3D constitutive model of polymer materials commonly used in 3D printing manufacturing. Hence, ABS and PLA were used with a low-cost desktop printer with which specimens were manufactured in two orthogonal orientations: flat and upright. Tensile and compression tests were performed to this end, where the Young's modulus, yield, and maximum stresses were determined. In the tensile tests, the samples with vertical (upright) orientation showed lower values in the evaluated mechanical properties than the corresponding to the horizontal (flat) orientation. However, no significant difference caused by the printing orientations was observed in the compression tests. Different values of Young's modulus and maximum strength were found between tensile and compression tests for the same material and orientation. Moreover, in order to describe the observed material response, a linear isotropic bimodular model is proposed. This constitutive model, which is fed with the previously obtained tensile and compression data, is used in the simulation of a four-point bending test where it is found to adequately represent the experimentally measured elastic behavior in the load-deflection curve. Thus, the combination of experiments and a bimodular constitutive model contributes to making better predictions of the mechanical response of structures made with 3D printing.