Quantification of the bisphosphonate alendronate using capillary electrophoresis mass spectrometry with dynamic pH barrage junction focusing.

A quantitative method was developed for the direct identity confirmation and quantification of alendronate using CE-MS combined with a pH-assisted focusing technique, dynamic pH barrage junction focusing. A pH-induced variation in electrophoretic mobility led to online focusing of alendronate at the sample/pH barrage boundary, significantly improving the detection sensitivity. In addition, the use of a flow-through microvial CE electrospray interface and the multiple reaction monitoring mode of MS further improved the specificity and quantification capability of this technology. This quantitative method presented a wide linear dynamic range over 8-2000 ng/mL and an LOD of 2 ng/mL. A 460-fold improvement in sensitivity was obtained when pH barrage junction focusing was applied during the CE process, in comparison to when normal CE was conducted without online sample stacking. The superior detection sensitivity over previously reported methods enables direct analysis of bisphosphonate compounds, eliminating tedious pre-column sample enrichment and derivatization. Validation of alendronate content in a commercial drug tablet further proved the reliability and power of this method. This simple method with no sample derivatization, superior sensitivity, and short run time (<8 min) is a promising alternative for accurate quantification of alendronate and other types of bisphosphonate compounds in both drug formulations and plasma samples.

Globular structure of the hypermineralized tissue in human femoral neck.

Bone becomes more fragile with ageing. Among many structural changes, a thin layer of highly mineralized and brittle tissue covers part of the external surface of the thin femoral neck cortex in older people and has been proposed to increase hip fragility. However, there have been very limited reports on this hypermineralized tissue in the femoral neck, especially on its ultrastructure. Such information is critical to understanding both the mineralization process and its contributions to hip fracture. Here, we use multiple advanced techniques to characterize the ultrastructure of the hypermineralized tissue in the neck across various length scales. Synchrotron radiation micro-CT found larger but less densely distributed cellular lacunae in hypermineralized tissue than in lamellar bone. When examined under FIB-SEM, the hypermineralized tissue was mainly composed of mineral globules with sizes varying from submicron to a few microns. Nano-sized channels were present within the mineral globules and oriented with the surrounding organic matrix. Transmission electron microscopy showed the apatite inside globules were poorly crystalline, while those at the boundaries between the globules had well-defined lattice structure with crystallinity similar to the apatite mineral in lamellar bone. No preferred mineral orientation was observed both inside each globule and at the boundaries. Collectively, we conclude based on these new observations that the hypermineralized tissue is non-lamellar and has less organized mineral, which may contribute to the high brittleness of the tissue.

Damage-tolerance strategies for nacre tablets.

Nacre, a natural armor, exhibits prominent penetration resistance against predatory attacks. Unraveling its hierarchical toughening mechanisms and damage-tolerance design strategies may provide significant inspiration for the pursuit of high-performance artificial armors. In this work, relationships between the structure and mechanical performance of nacre were investigated. The results show that other than their brick-and-mortar structure, individual nacre tablets significantly contribute to the damage localization of nacre. Affected by intracrystalline organics, the tablets exhibit a unique fracture behavior. The synergistic action of the nanoscale deformation mechanisms increases the energy dissipation efficiency of the tablets and contributes to the preservation of the structural and functional integrity of the shell.

Quantifying non-covalent binding affinity using mass spectrometry: a systematic study on complexes of cyclodextrins with alkali metal cations.

RATIONALE: To date, the quantification of binding affinities for non-covalent complexes between cyclodextrin (CD) and alkali cations including Li(+) , Na(+) , K(+) , Rb(+) , and Cs(+) has not been investigated in detail by electrospray ionization mass spectrometry (ESI-MS) due to the unknown ionization efficiencies of the different species. In this study, the binding constants of CD-Cs(+) complexes were determined by an improved mass spectrometric titration methodology, which was based only on the peak intensities of equilibrium CD. Hence, the discrepancy of ionization efficiencies of CD, alkaline cation and their complex would not affect the measurement. Then the obtained lgKa values were provided as references for competitive ESI-MS. The binding constants for complexes of α-, ß- or γ-CD with Li(+) , Na(+) , K(+) and Rb(+) could be derived directly and quickly. METHODS: The lgKa values between α-, ß- or γ-CD and Cs(+) data were processed by curve fitting. These lgKa values were provided as references for competitive ESI-MS. In addition, linear fit equations for complexes of α-, ß- or γ-CD with Cs(+) were derived. Through the linear fit equations of competitive ESI-MS, the binding constants for complexes of Li(+) , Na(+) , K(+) and Rb(+) with α-, ß- or γ-CD were acquired. RESULTS: Results showed that the binding constant (lgKa ) values for the complexes of Cs(+) with α-, ß- and γ-cyclodextrins were 3.94, 3.88 and 3.80, respectively, revealing that the binding strength decreased with the increase in diameter of cyclodextrins. The competitive ESI-MS results showed a clear trend of decreasing affinity for complexes of cyclodextrins in the order of Li(+) , Na(+) , K(+) , Rb(+) . CONCLUSIONS: The binding constants of non-covalent cyclodextrin-alkali cation complexes have been systematically studied by an improved mass spectrometric titration and competitive ESI-MS. Also, the structural features of the complexes were discussed. Our results are valuable for better understanding of mechanisms driving inclusion chemistry under well-defined conditions.

Inelastic deformation and microcracking process in human dentin.

Dentin is a mineralized collagen tissue with robust mechanical performance. Understanding the mechanical behavior of dentin and its relations to the dentinal structure can provides insight into the design strategies to achieve tooth functions. This study focuses on the inelastic deformation of human dentin and its underlying mechanisms. By combining four-point bending tests with fluorescent staining and laser scanning confocal microscopy, it was found that human dentin, especially root dentin, exhibited significant inelastic deformation and developed extensive microdamage in the form of microcracks prior to fracture. Dense and wavy microcracks spread uniformly across the tensile surface of root dentin, while compressive microcracks formed cross-hatched patterns. The presence of peritubular dentin in coronal dentin dramatically decreased the extent of microcracking, reducing inelasticity. Dentinal tubules were found to be initiation sites of both tensile and compressive microcracks. A unique crack propagation process was observed in root dentin under tension: numerous ring-shaped cracks formed at each dentinal tubule ahead of a growing crack tip. The advance of the tensile microcracks occurred by the merging of those ring-shaped cracks. The current findings on the microcracking process associated with inelastic deformation helps to understand the nature of strength and toughness in dentin, as well as the mechanical significance for structural variations across the whole tooth.

A microstructural study of individual nacre tablet of Pinctada maxima.

Nacre tablets from the shell of Pinctada maxima were studied with SEM, TEM and STEM. The systematic nanolath morphology on the (001) surface of nacre tablets was observed after acidic etching and mechanical polishing. The nanolaths were along the [100] crystallographic orientation of aragonite crystal. The (010) and (100) cross section surfaces of the nacre tablets showed nanolath and nanograin morphologies, respectively, which was consistent with [100] crystallographic orientation of nanolath on the (001) surface. Sheet-like defects with low mass density were observed on the (001) plane inside nacre tablets and were considered to be the cause of nanolath morphology revealed on the surfaces by acidic etching and mechanical polishing. On the other hand, large block [110] twins that divide the nacre tablets into two sectors were identified. The implication of these twins on the understanding to the crystallization mechanism of nacre tablets was discussed.

Robust Nanoparticle-Derived Lubricious Antibiofilm Coating for Difficult-to-Coat Medical Devices with Intricate Geometry.

A major medical device-associated complication is the biofilm-related infection post-implantation. One promising approach to prevent this is to coat already commercialized medical devices with effective antibiofilm materials. However, developing a robust high-performance antibiofilm coating on devices with a nonflat geometry remains unmet. Here, we report the development of a facile scalable nanoparticle-based antibiofilm silver composite coating with long-term activity applicable to virtually any objects including difficult-to-coat commercially available medical devices utilizing a catecholic organic-aqueous mixture. Using a screening approach, we have identified a combination of the organic-aqueous buffer mixture which alters polycatecholamine synthesis, nanoparticle formation, and stabilization, resulting in controlled deposition of in situ formed composite silver nanoparticles in the presence of an ultra-high-molecular-weight hydrophilic polymer on diverse objects irrespective of its geometry and chemistry. Methanol-mediated synthesis of polymer-silver composite nanoparticles resulted in a biocompatible lubricious coating with high mechanical durability, long-term silver release (∼90 days), complete inhibition of bacterial adhesion, and excellent killing activity against a diverse range of bacteria over the long term. Coated catheters retained their excellent activity even after exposure to harsh mechanical challenges (rubbing, twisting, and stretching) and storage conditions (>3 months stirring in water). We confirmed its excellent bacteria-killing efficacy (>99.999%) against difficult-to-kill bacteria (Proteus mirabilis) and high biocompatibility using percutaneous catheter infection mice and subcutaneous implant rat models, respectively, in vivo. The developed coating approach opens a new avenue to transform clinically used medical devices (e.g., urinary catheters) to highly infection-resistant devices to prevent and treat implant/device-associated infections.

An Ammonia-Induced Calcium Phosphate Nanostructure: A Potential Assay for Studying Osteoporosis and Bone Metastasis.

Osteoclastic resorption of bones plays a central role in both osteoporosis and bone metastasis. A reliable in vitro assay that simulates osteoclastic resorption in vivo would significantly speed up the process of developing effective therapeutic solutions for those diseases. Here, we reported the development of a novel and robust nanostructured calcium phosphate coating with unique functions on the track-etched porous membrane by using an ammonia-induced mineralization (AiM) technique. The calcium phosphate coating uniformly covers one side of the PET membrane, enabling testing for osteoclastic resorption. The track-etched pores in the PET membrane allow calcium phosphate mineral pins to grow inside, which, on the one hand, enhances coating integration with a membrane substrate and, on the other hand, provides diffusion channels for delivering drugs from the lower chamber of a double-chamber cell culture system. The applications of the processed calcium phosphate coating were first demonstrated as a drug screening device by using alendronate, a widely used drug for osteoporosis. It was confirmed that the delivery of alendronate significantly decreased both the number of monocyte-differentiated osteoclasts and coating resorption. To demonstrate the application in studying bone metastasis, we delivered a PC3 prostate cancer-conditioned medium and confirmed that both the differentiation of monocytes into osteoclasts and the osteoclastic resorption of the calcium phosphate coating were significantly enhanced. This novel assay thus provides a new platform for studying osteoclastic activities and assessing drug efficacy in vitro.

Improving fretting corrosion resistance of CoCrMo alloy with TiSiN and ZrN coatings for orthopedic applications.

Total hip replacement is the most effective treatment for late stage osteoarthritis. However, adverse local tissue reactions (ALTRs) have been observed in patients with modular total hip implants. Although the detailed mechanisms of ALTRs are still unknown, fretting corrosion and the associated metal ion release from the CoCrMo femoral head at the modular junction has been reported to be a major factor. The purpose of this study is to increase the fretting corrosion resistance of the CoCrMo alloy and the associated metal ion release by applying hard coatings to the surface. Cathodic arc evaporation technique (arc-PVD) was used to deposit TiSiN and ZrN hard coatings on CoCrMo substrates. The morphology, chemical composition, crystal structures and residual stress of the coatings were characterized by scanning electron microscopy, energy dispersive X-ray spectroscopy, and X-ray diffractometry. Hardness, elastic modulus, and adhesion of the coatings were measured by nano-indentation, nano-scratch test, and the Rockwell C test. Fretting corrosion resistance tests of coated and uncoated CoCrMo discs against Ti6Al4V spheres were conducted on a four-station fretting testing machine in simulated body fluid at 1Hz for 1 million cycles. Post-fretting samples were analyzed for morphological changes, volume loss and metal ion release. Our analyses showed better surface finish and lower residual stress for ZrN coating, but higher hardness and better scratch resistance for TiSiN coating. Fretting results demonstrated substantial improvement in fretting corrosion resistance of CoCrMo with both coatings. ZrN and TiSiN decreased fretting volume loss by more than 10 times and 1000 times, respectively. Both coatings showed close to 90% decrease of Co ion release during fretting corrosion tests. Our results suggest that hard coating deposition on CoCrMo alloy can significantly improve its fretting corrosion resistance and could thus potentially alleviate ALTRs in metal hip implants.

Perivascular lymphocytic aggregates in hip prosthesis-associated adverse local tissue reactions demonstrate Th1 and Th2 activity and exhausted CD8+ cell responses.

Hip implants are a successful solution for osteoarthritis; however, some individuals with metal-on-metal (MoM) and metal-on-polyethylene (MoP) prosthetics develop adverse local tissue reactions (ALTRs). While MoM and MoP ALTRs are presumed to be delayed hypersensitivity reactions to corrosion products, MoM- and MoP-associated ALTRs present with different histological characteristics. We compared MoM- and MoP-associated ALTRs histopathology with cobalt and chromium levels in serum and synovial fluid. We analyzed the gene expression levels of leukocyte aggregates and synovial fluid chemokines/cytokines to resolve potential pathophysiologic differences. In addition, we classified ALTRs from 79 patients according to their leukocyte infiltrates as macrophage-dominant, mixed, and lymphocyte-dominant. Immune-related transcript profiles from lymphocyte-dominant MoM- and MoP-associated ALTR patients with perivascular lymphocytic aggregates were similar. Cell signatures indicated predominantly macrophage, Th1 and Th2 lymphocytic infiltrate, with strong exhausted CD8+ signature, and low Th17 and B cell, relative to healthy lymph nodes. Lymphocyte-dominant ALTR-associated synovial fluid contained higher levels of induced protein 10 (IP-10), interleukin-1 receptor antagonist (IL-1RN), IL-8, IL-6, IL-16, macrophage inflammatory protein 1 (MIP-1α), IL-18, MCP-2, and lower cell-attracting chemokine levels, when compared with prosthetic revisions lacking ALTRs. In addition, the higher levels of IP-10, IL-8, IL-6, MIP-1α, and MCP-2 were observed within the synovial fluid of the lymphocyte-dominant ALTRs relative to the macrophage-dominant ALTRs. Not all cytokines/chemokines were detected in the perivascular aggregate transcripts, suggesting the existence of other sources in the affected synovia. Our results support the hypothesis of common hypersensitivity pathogenesis in lymphocyte-dominant MoM and MoP ALTRs. The exhausted lymphocyte signature indicates chronic processes and an impaired immune response, although the cause of the persistent T-cell activation remains unclear. The cytokine/chemokine signature of lymphocyte-dominant-associated ATLRs may be of utility for diagnosing this more aggressive pathogenesis.

Cobalt ions induce metabolic stress in synovial fibroblasts and secretion of cytokines/chemokines that may be diagnostic markers for adverse local tissue reactions to hip implants.

Adverse local tissue reactions (ALTRs) are a prominent cause of hip implant failure. ALTRs are characterized by aseptic necrosis and leukocyte infiltration of synovial tissue. The prevalence of ALTRs in hips with failing metal implants, with highest rates occurring in patients with metal-on-metal articulations, suggests a role for CoCrMo corrosion in ALTR formation. Although hypersensitivity reactions are the most accepted etiology, the precise cellular mechanism driving ALTR pathogenesis remains enigmatic. Here we show that cobalt ions released by failing hip implants induce mitochondrial stress and cytokine secretion by synovial fibroblasts: the presumptive initiators of ALTR pathogenesis. We found that in-vitro treatment of synovial fibroblasts with cobalt, but not chromium, generated gene expression changes indicative of hypoxia and mitophagy responses also observed in ALTRs biopsies. Inflammatory factors secreted by cobalt-exposed synovial fibroblasts were among those most concentrated in ALTR synovial fluid. Furthermore, both conditioned media from cobalt-exposed synovial fibroblasts, and synovial fluid from ALTRs patients, elicit endothelial activation and monocyte migration. Finally, we identify the IL16/CTACK ratio in synovial fluid as a possible diagnostic marker of ALTRs. Our results provide evidence suggesting that metal ions induce cell stress in synovial fibroblasts that promote an inflammatory response consistent with initiating ALTR formation. STATEMENT OF SIGNIFICANCE: We demonstrate that the cytotoxic effects of cobalt ions on the synovial cells (fibroblast) is sufficient to trigger inflammation on hip joints with metal implants. Cobalt ions affect mitochondrial function, leading to the auto phagocytosis of mitochondria and trigger a hypoxic response. The cell's hypoxic response includes secretion of cytokines that are capable of trigger inflammation by activating blood vessels and enhancing leukocyte migration. Among the secreted cytokines is IL-16, which is highly concentrated in the synovial fluid of the patients with adverse local tissue reactions and could be use as diagnostic marker. In conclusion we define the cells of the hip joint as key players in triggering the adverse reactions to hip implants and providing biomarkers for early diagnosis of adverse reactions to hip implants.

CoCrMo metal release in metal-on-highly crosslinked polyethylene hip implants.

Increasing cases of adverse local tissue reactions (ALTRs) associated with metal release have been observed in patients with metal-on-highly crosslinked polyethylene (MoP) hip implants, the most common design in total hip replacements. Studies have demonstrated the metal release from fretting corrosion at the head-neck junction, but rarely investigated tribocorrosion associated metal release at articulating surfaces in MoP hip implants. The objective of this study is to investigate both tribocorrosion at the articulating surfaces and fretting corrosion at the head-neck junction in CoCrMo femoral heads, as well as their association with metal species released in periprosthetic tissues and body fluids in MoP hip systems. Twenty-three patients with ALTRs associated with MoP implants were included. Systematic analyses were performed on the wear damage in articulation, corrosion at the head-neck junction and their correlation with degradation products observed in synovial fluid, periprosthetic tissues, and serum. Results showed that tribocorrosion at the articulating surfaces contributed to the elevated concentration of both Co and Cr ions in serum, while fretting corrosion at the head-neck junction mainly released Co ions to serum. Both tribocorrosion at the articulating surfaces and fretting corrosion at the head-neck junction released particles rich in chromium and phosphate, the dominant particles found in synovial fluids and tissues. This study provides strong evidence that tribocorrosion at the articulating surfaces in MoP hip implants could result in significant metal release. This information should be taken into account when studying the mechanisms of ALTRs and developing strategies of preventing metal release in total hip replacements.

Detecting human articular cartilage degeneration in its early stage with polarization-sensitive optical coherence tomography.

Detecting articular cartilage (AC) degeneration in its early stage plays a critical role in the diagnosis and treatment of osteoarthritis (OA). Polarization-sensitive optical coherence tomography (PS-OCT) is sensitive to the alteration and disruption of collagen organization that happens during OA progression. This study proposes an effective OA evaluating method based on PS-OCT imaging. A slope-based analysis is applied on the phase retardation images to segment articular cartilage into three zones along the depth direction. The boundaries and birefringence coefficients (BRCs) of each zone are quantified. Two parameters, namely phase homogeneity index (PHI) and zonal distinguishability (Dz), are further developed to quantify the fluctuation within each zone and the zone-to-zone variation of the tissue birefringence properties. The PS-OCT based evaluating method then combines PHI and Dz to provide a G PS score for the severity of OA. The proposed method is applied to human hip joint samples and the results are compared with the grading by histology images. The G PS score shows very strong statistical significance in differentiating different stages of OA. Compared to using the BRC of each zone or a single BRC for the entire depth, the G PS score shows great improvement in differentiating early-stage OA. The proposed method is shown to have great potential to be developed as a clinical tool for detecting OA.

The effect of adhesion promoter on the adhesion of PDMS to different substrate materials.

The effect of the adhesion promoter GE SS4120 on the adhesion strength of PDMS to different substrates was tested. The adhesion to silicon, glass and aluminium was significantly increased, while adhesion of PDMS to Teflon remained poor, and the adhesion strength of PDMS to PDMS decreased.

Mechanisms of Adverse Local Tissue Reactions to Hip Implants.

Adverse Local Tissue Reactions (ALTRs) are one of the main causes of hip implant failures. Although the metal release from the implants is considered as a main etiology, the mechanisms, and the roles of the released products are topics of ongoing research. The alloys used in the hip implants are considered biocompatible and show negligible corrosion in the body environment under static conditions. However, modularity and its associated mechanically assisted corrosion have been shown to release metal species into the body fluids. ALTRs associated with metal release have been observed in hip implants with metal-on-metal articulation initially, and later with metal-on-polyethylene articulation, the most commonly used design in current hip replacement. The etiological factors in ALTRs have been the topics of many studies. One commonly accepted theory is that the interactions between the metal species and body proteins and cells generate a delayed type IV hypersensitivity reaction leading to ALTRs. However, lymphocyte reactions are not always observed in ALTRS, and the molecular mechanisms have not been clearly demonstrated. A more accepted mechanism is that cell damage generated by metal ions may trigger the secretion of cytokines leading to the inflammatory reactions observed in ALTRs. In this inflammatory environment, some patients would develop hypersensitivity that is associated with poor outcomes. Concerns over ALTRS have brought significant impact to both the clinical selection and development of hip implants. This review is focused on the mechanisms of ALTRs, specifically, the metal release process and the roles of the metal species released in the etiology and pathogenesis of the disease. Hopefully, our presentation and discussion of this biological process from a material perspective could improve our current understanding on the ALTRs and provide useful guidance in developing preventive solutions.

Hypermineralization in the femoral neck of the elderly.

Hip fragility depends on the decline in bone mass as well as changes in bone microstructure and the properties of bone mineral and organic matrix. Although it is well-established that low bone mass or osteoporosis is a key factor in hip fracture risk, it is striking to observe that 92% of 24 patients who have sustained an intracapsular hip fracture showed hypermineralization at the superior-anterior quadrant, a critical region associated with increased hip fracture risk. In-depth material studies on a total of 12 human cadaver femurs revealed increased degree of mineralization in the hypermineralized tissue: calcium weight percentage as measured by quantitative backscattered electron imaging increased by approximately 15% compared with lamellar bone; mineral-to-matrix ratio obtained by Raman microspectroscopy imaging also increased. Immunohistochemistry revealed localized type II collagen in the hypermineralized region, implying its cartilaginous nature. At the ultrastructural level, X-ray scattering revealed significantly smaller (on average 2.3 nm thick and 15.6 nm long) and less ordered bone minerals in the hypermineralized tissue. Finally, the hypermineralized tissue was more brittle than lamellar bone under hydrated state - cracks propagated easily in the hypermineralized region but stopped at the lamellar boundary. This study demonstrates that hypermineralization of femoral neck cortical bone is a source of bone fragility which is worth considering in future fracture risk assessment when the origin of hip fracture is unclear based on current evaluation standards. STATEMENT OF SIGNIFICANCE: Hypermineralization of femoral cortical bone in older adults might occur in many more hip fracture cases than presently known. Yet, this tissue remains largely unknown to the orthopedic community possibly due to coarse resolution of clinical imaging. The current study showed the hypermineralized tissue had reduced fracture resistance which could be attributed to the material changes in mineral content, organic matrix, and mineral platelets properties. It thus could be a source for fracture initiation. Consequently, we believe hypermineralization of femoral neck cortical bone should be considered in hip fragility assessment, especially when low bone mass cannot be identified as a primary contributor to hip fracture.

Porous scaffold of gelatin-starch with nanohydroxyapatite composite processed via novel microwave vacuum drying.

Hydroxyapatite (HA) is a fundamental mineral-based biomaterial, used for preparing composites for bone repair and regeneration. Gelatin blended with starch results in scaffold composites with enhanced mechanical properties. A gelatin-starch blend reinforced with HA nanocrystals (nHA) gave biocompatible composites with enhanced mechanical properties. In this study, a porous scaffold of gelatin-starch-nHA composites was fabricated through microwave vacuum drying and crosslinking using trisodium citrate. Three different composite scaffolds were prepared at three different percentages of nHA: 20%, 30% and 40%. The microstructures and compositions of the composites were analyzed. Within the porous structure, the nHA crystals were observed to precipitate. The interaction between the gelatin-starch network film and nHA crystalline material was studied using Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction analysis (XRD). XRD reflections showed that there are two different minerals present in the scaffold composite. There were strong reflection peaks close to the 26 degrees and 32 degrees 2theta angles of HA, and close to the 8 degrees and 49 degrees 2theta angles for sodium citrate minerals. The FTIR result suggested that carboxyl groups, C=O and amino groups play crucial roles in HA formation on the surface of a gelatin network.

Clinical hip fracture is accompanied by compression induced failure in the superior cortex of the femoral neck.

Hip fractures pose a major health problem throughout the world due to their devastating impact. Current theories for why these injuries are so prevalent in the elderly point to an increased propensity to fall and decreases in bone mass with ageing. However, the fracture mechanisms, particularly the stress and strain conditions leading to bone failure at the hip remain unclear. Here, we directly examined the cortical bone from clinical intra-capsular hip fractures at a microscopic level, and found strong evidence of compression induced failure in the superior cortex. A total of 143 sections obtained from 24 femoral neck samples that were retrieved from 24 fracturing patients at surgery were examined using laser scanning confocal microscopy (LSCM) after fluorescein staining. The stained microcracks showed significantly higher density in the superior cortex than in the inferior cortex, indicating a greater magnitude of strain in the superior femoral neck during the failure-associated deformation and fracture process. The predominant stress state for each section was reconstructed based on the unique correlation between the microcrack pattern and the stress state. Specifically, we found clear evidence of longitudinal compression and buckling as the primary failure mechanisms in the superior cortex. These findings demonstrate the importance of microcrack analysis in studying clinical hip fractures, and point to the central role of the superior cortex failure as an important aspect of the failure initiation in clinical intra-capsular hip fractures.

Strain redistribution and cracking behavior of human bone during bending.

Long bones often fail due to bending loads. Understanding the fracture process during bending is of great importance to the prevention and treatment of bone fractures. In this study, we investigated the origin of long bone's bending strength through the study of the dynamic strain redistribution happening during the post-yield stage of deformation and its relation to microdamage at the microstructural level. This was accomplished by comparing the behaviors of human long bones with standard cortical bone specimens in terms of strain redistribution, Poisson's ratios, microdamage morphologies, and macro-scale fracture patterns. It was found that human tibia failure in bending was very similar to that of standard beam cortical bone specimens with respect to the four previous aspects. Also, the examination of bone's Poisson's ratio indicated very different inelastic deformation mechanisms under tension and compression: bone volume expanded in tension but was nearly conserved in compression. Finally, as a result of strain redistribution, bone's bending strength mainly depended on its compressive strength, which was significantly influenced by the osteonal "porous" microstructure of human bone as compared to its tensile behavior. Thus, we concluded that bone microstructure at the Haversian system level plays an important role in bone deformation and fracture.

Enamel inspired nanocomposite fabrication through amelogenin supramolecular assembly.

Fabricating the structures similar to dental enamel through the in vitro preparation method is of great interest in the fields of dentistry and material sciences. Developing enamel is composed of calcium phosphate mineral, water, and enamel matrix proteins, mainly amelogenins. To prepare a material mimicking such composition a novel approach of simultaneously assembling amelogenin and calcium phosphate precipitates by electrolytic deposition (ELD) was established. It was found that recombinant full-length amelogenin (rP172) self-assembled into nanochain structures during ELD (following increase in solution pH), and had significant effect on the induction of the parallel bundles of calcium phosphate nanocrystals, grown on semiconductive silicon wafer surface. When a truncated amelogenin (rP148) was used; no nanochain assembly was observed, neither parallel bundles were formed. The coating obtained in the presence of rP172 had improved elastic modulus and hardness when compared to the coating incorporated with rP148. Our data suggest that the formation of organized bundles in amelogenin-apatite composites is mainly driven by amelogenin nanochain assembly and highlights the potential of such composite for future application as dental restorative materials.