Quantitative backscattered electron imaging of bone in proximal femur fragility fracture and medical illness

Bone quality consists of a number of factors including the amount of bone, bone architecture and the degree of bone mineralization. Quantitative backscattered electron imaging is a technique that allows the degree of mineralization of trabeculae to be assessed and in this study is applied to inter‐trochanteric bone biopsies of the proximal femur. Biopsy cores from 22 controls, nine individuals with acute and chronic medical conditions and 22 fragility fracture individuals undergoing total hip replacement were processed into methyl methacrylate, polished and analysed in a Philips XL20 scanning electron microscope. A mean and distribution of weight percent calcium were determined for each individual, and for the control, medically ill and fragility fracture groups. All individuals and groups of individuals showed normal distributions of percent calcium with both the ill and fragility fracture groups being under mineralized relative to the control group. The shape and position of the mineralization distributions suggest that the fragility group resulted from increased bone turnover with a slow progression to under mineralization. In contrast, the ill group appears to have had a more rapid change in the mineralization dynamic. Clear distinctions between the control, fragility fracture and medically ill groups could be seen when the mineralization data were plotted as a scatter graph against age. Graphing the data in this way showed an age‐related increase in the degree of mineralization in control individuals with the under‐mineralized, fragility fracture group scattered below this normal trend. The medically ill group was similarly less mineralized which highlighted the degree to which medical conditions and treatments can alter bone matrix mineralization.     

Acoustic and ultrasonic tissue characterization--assessment of osteoporosis.

Osteoporosis, often termed the 'silent epidemic', has been defined as 'a decrease in bone mass and architectural deterioration of bone tissue, leading to enhanced bone fragility and consequent increase in fracture risk'. In the United Kingdom alone, the annual health costs are in excess of 750 million Pounds, with 60,000 patients suffering a hip fracture each year. A quarter of these will die within 12 months of their fracture, half of the remainder will never regain independent living. The established procedure for assessing the risk of osteoporotic fracture is via bone mineral density (BMD) assessment using dual-energy X-ray absorptiometry (DXA). However, DXA is an expensive technique and is not widely available. Within the past 15 years, ultrasound assessment of bone has rapidly advanced in scientific understanding, technical development and clinical utility. Measurements of cancellous bone (particularly at the calcaneus) are generally performed in preference to those of cortical bone (tibial cortex). There are currently 15 commercial systems available and over 3500 systems are in use world-wide. The low cost and portability offered by ultrasound systems should enable an integrated community-based screening programme to be established in the near future. Ultrasound measurements of bone are generally obtained using transmission rather than pulse-echo techniques owing to its highly attenuating nature. Ultrasound velocity and attenuation measurements are utilized. For velocity, there are well-defined fundamental relationships describing the dependence upon the elasticity and density of bone.     

Label‐free detection of fibrillar collagen deposition associated with vascular elements in glioblastoma multiforme by using multiphoton microscopy

Glioblastoma multiforme (GBM‐WHO grade IV) is the most common and the most aggressive form of brain tumors in adults with the median survival of 10–12 months. The diagnostic detection of extracellular matrix (ECM) component in the tumour microenvironment is of prognostic value. In this paper, the fibrillar collagen deposition associated with vascular elements in GBM were investigated in the fresh specimens and unstained histological slices by using multiphoton microscopy (MPM) based on two‐photon excited fluorescence (TPEF) and second harmonic generation (SHG). Our study revealed the existence of fibrillar collagen deposition in the adventitia of remodelled large blood vessels and in glomeruloid vascular structures in GBM. The degree of fibrillar collagen deposition can be quantitatively evaluated by measuring the adventitial thickness of blood vessels or calculating the ratio of SHG pixel to the whole pixel of glomeruloid vascular structure in MPM images. These results indicated that MPM can not only be employed to perform a retrospective study in unstained histological slices but also has the potential to apply for in vivo brain imaging to understand correlations between malignancy of gliomas and fibrillar collagen deposition.     

Mass spectrometry for the study of protein glycation in disease

The structural elucidation of advanced glycation end-product (AGE)-modified proteins and quantitative analysis of free AGEs have been successfully performed, by use of mass spectrometry (MS) in plasma and tissues of patients with AGE-related diseases, such as diabetes mellitus, uremia, cataract, and liver cirrhosis. Matrix-assisted laser desorption/ionization (MALDI)-MS made it possible to directly analyze the AGE-modified proteins such as albumin and IgG. However, because the direct structural analysis of intact AGE-modified proteins is often not easy due to the formation of broad and poorly resolved peaks, peptide mapping after enzymatic hydrolysis was introduced into the analysis of AGE-modified proteins and the site-specific analysis of defined AGEs by MALDI-MS. Liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) has been employed not only for the structural elucidation of enzymatically hydrolyzed AGEs-modified peptides but also for simultaneous quantification of free AGEs in plasma and tissues of patients. Based on many studies that use MS for the analysis of AGEs, there is no doubt as to the important role of protein-linked AGEs in several diseases.     

Experimental investigation on cutting mechanisms in fixed diamond wire sawing of bone

Bone sawing has been widely used in performing bone surgery. However, thermal necrosis, loss of cutting precision and surface damage may occur in cutting process. The primary objective of this research is to improve cutting performance of bone by advantages of diamond wire sawing. Mechanism of material removal, cutting force, temperature and surface quality are analyzed based on experimental results. It is indicated that wire sawing provides small depth of cut, which is effective to obtain ductile material removal mode. Due to small material removal rate per abrasive, thermal energy is low and most of the heat can be taken away by the cyclic wire and bone chips. Consequently, cutting force and temperature in cutting zone are lower than that of traditional sawing. Due to the high efficiency of chip ejection, burrs and fracture are reduced and a significant improvement in surface quality is achieved. Based on cutting experiments with various values of cutting parameters, it is observed that better performance is achievable at higher wire speeds. These results provide a valuable basis for application of wire sawing and understanding of bone cutting mechanisms.     

Dynamic three-dimensional visualization of collagen matrix remodeling and cytoskeletal organization in living corneal fibroblasts

The remodeling of extracellular matrices by cells plays a defining role in developmental morphogenesis and wound healing, as well as in tissue engineering. Three-dimensional (3-D) type I collagen matrices have been used extensively as an in vitro model for studying cell-induced matrix reorganization at the macroscopic level. However, few studies have directly assessed the dynamic process of 3-D matrix remodeling at the cellular and subcellular level. We recently developed an experimental model for investigating cell-matrix mechanical interactions by plating green fluorescen protein (GFP)-zyxin transfected cells inside fibrillar collagen matrices and performing high-magnification time-lapse differential interference microscopy (DIC) and wide-field fluorescent imaging. In this study, we extend this experimental model by performing four-dimensional (4-D) reflected light and fluorescent confocal imaging (using either visible light or multiphoton excitation) of living corneal fibroblasts transfected to express GFP-zyxin or GFP-alpha-actinin, 18 h after plating inside 3-D collagen matrices. Reflected light confocal imaging allowed detailed visualization of the cells and the fibrillar collagen surrounding them. By overlaying maximum intensity projections of reflected light and GFP-zyxin or GFP-alpha-actinin images and generating stereo pair reconstructions, 3-D interactions between focal adhesions and collagen fibrils in living cells could be visualized directly. Focal adhesions were generally oriented parallel to the direction of collagen fibril alignment in front of the cell. Killing the cells induced relaxation of transient cell-induced tension on the matrix; however, significant permanent remodeling always remained. Time-lapse 3-D imaging demonstrated an active response to the Rho-kinase inhibitor Y-27632, as indicated by cell elongation, extracellular matrix relaxation, and extension of pseudopodial processes. It is interesting that, at higher cell densities, groups of collagen fibrils were compacted and aligned into straps between neighboring cells. Overall, the continued development and application of this new approach should provide important insights into the basic underlying biochemical and biomechanical regulatory mechanisms controlling matrix remodeling by corneal fibroblasts.     

The bone diagnostic instrument II: indentation distance increase

The bone diagnostic instrument (BDI) is being developed with the long-term goal of providing a way for researchers and clinicians to measure bone material properties of human bone in vivo. Such measurements could contribute to the overall assessment of bone fragility in the future. Here, we describe an improved BDI, the Osteoprobe IItrade mark. In the Osteoprobe IItrade mark, the probe assembly, which is designed to penetrate soft tissue, consists of a reference probe (a 22 gauge hypodermic needle) and a test probe (a small diameter, sharpened rod) which slides through the inside of the reference probe. The probe assembly is inserted through the skin to rest on the bone. The distance that the test probe is indented into the bone can be measured relative to the position of the reference probe. At this stage of development, the indentation distance increase (IDI) with repeated cycling to a fixed force appears to best distinguish bone that is more easily fractured from bone that is less easily fractured. Specifically, in three model systems, in which previous mechanical testing and/or tests reported here found degraded mechanical properties such as toughness and postyield strain, the BDI found increased IDI. However, it must be emphasized that, at this time, neither the IDI nor any other mechanical measurement by any technique has been shown clinically to correlate with fracture risk. Further, we do not yet understand the mechanism responsible for determining IDI beyond noting that it is a measure of the continuing damage that results from repeated loading. As such, it is more a measure of plasticity than elasticity in the bone.     

Technical advances in the sectioning of dental tissue and of on‐section cross‐linked collagen detection in mineralized teeth

Immunohistochemical detection of cross‐linked fibrillar collagens in mineralized tissues is much desired for exploring the mechanisms of biomineralization in health and disease. Mineralized teeth are impossible to section when embedded in conventional media, thus limiting on‐section characterization of matrix proteins by immunohistochemistry. We hypothesized that by using an especially formulated acrylic resin suitable for mineralized dental tissues, not only sectioning of teeth would be possible, but also our recently developed immunofluorescence labeling technique would be amenable to fully calcified tissues for characterization of dentinal fibrillar collagens, which remains elusive. The hypothesis was tested on fixed rodent teeth embedded in Technovit 9100 New®. It was possible to cut thin (1 μm) sections of mineralized teeth, and immunofluorescence characterization of cross‐linked type I fibrillar collagen was selected due to its abundance in dentine. Decalcified samples of teeth embedded in paraffin wax were also used to compare immunolabeling from either method using the same immunoreagents in equivalent concentrations. In the decalcified tissue sections, type I collagen labeling in the dentine along the tubules was “patchy” and the signal in the predentine was very weak. However, enhanced signal in mineralized samples with type I collagen was detected not only in the predentine but also at the limit between intertubular dentine, within the elements of the enamel organ and subgingival stroma. This report offers advances in sectioning mineralized dental tissues and allows the application of immunofluorescence not only for on‐section protein detection but importantly for detecting cross‐linked fibrous collagens in both soft and mineralized tissue sections. Microsc. Res. Tech. 73:741–745, 2010. © 2009 Wiley‐Liss, Inc.     

Accumulation of in-vivo fatigue microdamage and its relation to biomechanical properties in ageing human cortical bone

Bone matrix accumulates microdamage in the form of microcracks as a result of everyday cyclic loading activities. In two very recent studies, which used conventional histological stains and light microscopy techniques, the amount of this in-vivo microdamage in the cortices of long bones has been shown to increase with age. These articles have suggested that in-vivo microcracks may have an effect on the material properties of the tissue. However, a precise quantitative relationship between the number of microcracks and the mechanical properties of these same bones has not been produced before, and in particular the way the microcracks may affect the stiffness, the strength or possibly the toughness of the tissue. This article presents an examination of the in-vivo microdamage in human bones by the use of laser scanning confocal microscopy, which offers better discrimination and allows examination of the cracks in-situ . Quantification of in-vivo fatigue microcracks was performed by counting the microcrack numerical density and surface density in specimens for which we have previously derived a full set of mechanical properties as a function of age. It is shown that bone microdamage relates more to the toughness (measured by three different measures) of ageing bone tissue than to its stiffness and strength. The result allows us (i) to re-evaluate the fragility of ageing human bone and put more emphasis on its energy-related resistance to fracture than perhaps on its stiffness or strength and also (ii) to understand more fully the causal relationship and interactions between microcracks and tissue toughness.     

Osteoblasts in bone tissue engineering

Osteoblasts are integral to the development, growth, function, repair and maintenance of bone. The osteoblast forms organic, non-mineralized bone matrix and is involved in complex interactions with a variety of factors, mediators and cell types. Degeneration, pathology, and trauma cause disruption and destruction of the normal skeletal environment and may lead to bone loss. There is a rise in active populations involved in trauma, elderly patients with fragility fractures and an overall increase in primary, revision and reconstructive bone and joint surgery. Despite the rapid evolution of implant technologies and bone grafting techniques, there is still a great demand for novel bone replacement strategies. Bone tissue engineering is the state of the art science with the potential to regenerate bone with natural form and function. This review presents the biology of osteoblasts and their current applications in bone tissue engineering biotechnologies and role in stem cell, bioactive factor, recombinant signalling molecule and gene therapy research.     

Irregular Shaped,Assumably Semi‐Crystalline Calciumphosphate Platelet Deposition at the Mineralization Front of Rabbit Femur Osteotomy: A HR‐TEM Study

Although bone minerals have been widely studied by various techniques in previous studies, crystal structures, morphology of bone minerals and its building pathway remained still controversy. In this work, the ultrastructure of the mineralization front of rabbit femur has been studied by conventional and high‐resolution (HR) transmission electron microscopy (TEM). In order to induce a healing and demineralization process the animals were subjected to a standardized osteotomy stabilized with titan screws and sonic pins. After 84 days follow‐up time the newly build bone was investigated. The mineralization front of rabbit femur osteotomy contains partly mineralized collagen fibrils with a pronounced striped pattern together with a large number of agglomerated apatite platelets. The striation is caused by mineralization in the hole zones of the collagen fibrils, corresponding to the early stage of mineralization. In the TEM micrographs, the mineralization zone appears denser and compact when compared with fully mineralized bone, although most of the collagen fibrils are completely mineralized in the latter (higher concentration of interfibrillar apatite platelets within the mineralization zone). In bone some partly mineralized collagen fibrils are also observed, revealing the same arrangement, regular shape, and size of apatite platelets as collagen fibrils in the mineralization zone. Apatite platelets with irregular shapes are observed at the vortex‐shaped outer boundary of the mineralization zone, i.e. at the interfaces with nonmineralized collagen or osteoblasts. HR TEM micrographs reveal that the platelets are assumably semicrystalline and that within the platelet nanocrystalline domains of apatite are embedded in an amorphous calciumphosphate matrix. SCANNING 35: 169‐182, 2013. © 2012 Wiley Periodicals, Inc.     

Trabecular bone microarchitecture is related to the number of risk factors and etiology in osteoporotic men

Microarchitecture of trabecular bone is a very important component of bone quality in osteoporosis and a determinant of vertebral fracture in men with low bone mineral density (BMD). In contrast to women, male osteoporosis is, in most cases, secondary. The relationships between microarchitecture and different risk factors have never been evaluated in men. About 152 men with low BMD at the lumbar spine or hip (BMD, T-score < -2.5) were included in this study. Risk factors were: age, BMI, alcohol intake, corticosteroid therapy, hypogonadism, and chronic diseases. Transiliac bone biopsies were obtained and histomorphometry was done on an image analyzer; the following parameters were measured: cortical thickness (Ct.Th), trabecular bone volume (BV/TV), trabecular thickness (Tb.Th), separation (Tb.Sp) and number (Tb.N), interconnectivity Index (ICI), star volume of the bone marrow, and strut analysis with node and free-end count. The 50 men with two risk factors had a lower BMD, lower Ct.Th and a significant higher star volume than those with one factor or idiopathic osteoporosis. The 26 men with at least three risk factors, had a lower BMD, a reduction of BV/TV and Ct.Th and a marked disorganization of the trabecular network (increased Tb.Sp, ICI, star volume, and free-end to free-end struts). The prevalence of vertebral fractures was higher in these patients. When the main risk factor was considered, a marked decrease in trabecular bone connectivity was observed in hypogonadic men. In osteoporotic men, higher the number of risk factors, lower the connectivity of trabecular network and higher the vertebral fracture risk.     

焊缝金属中氢、氮含量与氢裂敏感性的关系

试验和分析结果表明．铁素体中氢致裂纹敏感性的大小取决于氢原子在裂纹尖端局部区域扩散聚集的速度。原始氢含量的提高对断裂应力的影响是由于其降低裂纹尖端局部区域材料的断裂韧性，含氢量越高，Ｋ下降的速度越快。含氮量较低时，随含氮量提高，铁素体的氢裂敏感性增加，含氮量超过一定值后，提高含氮量会降低铁素体的氢裂敏感性。     

高效液相色谱-串联质谱法测定小鼠肝、肺、肾中Ⅰ、Ⅲ型胶原蛋白含量

建立了高效液相色谱-串联质谱(HPLC-MS/MS)法测定小鼠肝、肺、肾中Ⅰ、Ⅲ型胶原蛋白含量。首先识别小鼠Ⅰ、Ⅲ型胶原蛋白特征多肽,分别为GSEGPQGVR、GPSGFR。采用子离子及保留时间定性,以GLAGMK为内标对小鼠肝、肺、肾中Ⅰ、Ⅲ型胶原蛋白含量进行分析。结果表明,2个特征多肽在1～500 mg/L浓度范围内的线性关系良好,相关系数均大于0.99,精密度相对标准偏差小于3.7%,加标回收率在86%～118%之间。小鼠生长过程中,肝、肺、肾中Ⅰ型胶原蛋白的比例呈增加趋势,Ⅰ、Ⅲ型胶原蛋白总量呈先增加后降低的趋势。该方法前处理简便高效、精密度高、重现性好,可用于小鼠肝、肺、肾中Ⅰ、Ⅲ型胶原蛋白的精确定量。     

Atomic force microscopy of collagen structure in bone and dentine revealed by osteoclastic resorption

Mineralised tissues such as bone consist of two material phases: collagen protein fibrils, secreted by osteoblasts, form model structures for subsequent deposition of mineral, calcium hydroxyapatite. Collagen and mineral are removed in a three-dimensional manner by osteoclasts during bone turnover in skeletal growth or repair. Bone active drugs have recently been developed for skeletal diseases, and there is revived interest in changes in the structure of mineralised tissues seen in disease and upon treatment. The resolution of atomic force microscopy and use of unmodified samples has enabled us to image bone and dentine collagen exposed by the natural process of cellular dissolution of mineralised matrix. The morphology of bone and dentine has been analysed when fully mineralised and after osteoclast-mediated bone resorption, and compared with results from other microscopy techniques. Banded type I collagen, with 66.5+/-1.4 nm axial D-periodicity and 62.2+/-7.0 nm diameter, has been identified within resorption lacunae in bone and 69.4+/-4.3 nm axial D-periodicity and 140.6+/-12.4 nm diameter in dentine substrates formed by human and rabbit osteoclasts, respectively. This observation suggests a route by which the material and morphological properties of bone collagen can be analysed in situ, compared with collagen from non-skeletal sites, and contrasted in diseases of medical importance, such as osteoporosis, where skeletal tissue is mechanically weakened.     

Advanced glycation end-products change placental barrier function and tight junction in rats with gestational diabetes mellitus via the receptor for advanced glycation end products/nuclear factor-κB pathway

The placenta plays an important role in nutrient transport to maintain the growth and development of the embryo. Gestational diabetes mellitus (GDM), the most common complication during pregnancy, highly affects placental function in late gestation. Advanced glycation end-products (AGEs), a complex and heterogeneous group of compounds engaged by the receptor for AGEs (RAGE), are closely associated with diabetes-related complications. In this study, AGEs induced a decrease in the expression of tight junction (TJ) proteins in BeWo cells and increased the paracellular permeability of trophoblast cells by regulating RAGE/NF-κB. Sprague-Dawley (SD) rats injected with 100 mg/kg AGEs-rat serum albumin (RSA) via the tail vein from embryo day 2 were set as the placental barrier dysfunction model group (n = 10). The effect of AGEs on placental permeability was determined using the EvansBlue dye extravasation method. The ultrastructure of the placenta samples was observed by transmission electron microscopy. The effects of AGEs on the placenta were confirmed by treating rats with RAGE antagonist FPS-ZM1 and soluble forms of RAGE (sRAGE). AGEs treatment increased placental permeability and disrupted the tight junctions in pregnant rat placenta, but has no effect on blood glucose. The expression of TJ-related proteins, including ZO-1, Occludin, and Claudin 5, were downregulated after AGEs treatment. Further, AGEs treatment increased the expression of RAGE and nuclear factor-κB in the placenta of rats and upregulated the levels of vascular endothelial growth factor. The effects of AGEs on the placenta were blocked by RAGE antagonist FPS-ZM1 and sRAGE. This study demonstrates the mechanism underlying AGEs-induced disturbance in placental function in pregnant rats and highlights the potential of AGEs in the treatment of GDM.     

A device for improved reduction of tibial fractures treated with external fixation

A widely used method of treatment for unstable tibial shaft fractures is unilateral external fixation. The majority of fixators act as three distinct devices: an intra-operative reduction device, a device to maintain fracture alignment during healing and an aid to healing by allowing movement at the fracture site. Conventional operative techniques require the surgeon to manipulate a number of degrees of freedom at once, making reduction of the fracture difficult, and results in the fixator being out of alignment with the long axis of the bone. An operative method has been developed that separates reduction and fixation. A dedicated device has been designed to improve the per-operative control of fracture fragments during fracture reduction. The device has been used in clinical trials for the reduction of 22 diaphyseal tibial fractures. Compared with previous operative techniques there has been a saving of 53 per cent in fracture reduction time and an overall saving of 10 per cent in operating time. Fracture alignment has been improved compared with reductions achieved with a fixator which potentially improves healing and lowers the rate of malunion. In each case the fixator has been applied in alignment with the bone, improving dynamization and reducing the likelihood of malunion due to fixator cam slippage.     

Optimization of Picrosirius red staining protocol to determine collagen fiber orientations in vaginal and uterine cervical tissues by Mueller polarized microscopy

Polarized microscopy provides unique information on anisotropic samples. In its most complete implementation, namely Mueller microscopy, this technique is well suited for the visualization of fibrillar proteins orientations, with collagen in the first place. However, the intrinsic optical anisotropy of unstained tissues has to be enhanced by Picrosirius Red (PR) staining to enable Mueller measurements. In this work, we compared the orientation mapping provided by Mueller and second harmonic generation (SHG) microscopies on PR stained samples of vaginal and uterine cervix tissues. SHG is a multiphoton technique that is highly specific to fibrillar collagen, and was taken as the “gold standard” for its visualization. We showed that Mueller microscopy can be safely used to determine collagen orientation in PR stained cervical tissue. In contrast, in vaginal samples, Mueller microscopy revealed orientations not only of collagen but also of other anisotropic structures. Thus PR is not fully specific to collagen, which necessitates comparison to SHG microscopy in every type of tissue. In addition to this study of PR specificity, we determined the optimal values of the staining parameters. We found that staining times of 5 min, and sample thicknesses of 5 µm were sufficient in cervical and vaginal tissues. Microsc. Res. Tech. 78:723–730, 2015. © 2015 Wiley Periodicals, Inc.     

A preliminary dual-energy X-ray absorptiometry-based finite element model for assessing osteoporotic hip fracture risk

To more accurately assess osteoporotic hip fracture risk in a specific patient, a dual-energy X-ray absorptiometry (DXA)-based finite element model was constructed from the patient's femur DXA image. The outermost contour of the femur bone segmented from the DXA image was used to generate a finite element mesh. Bone mechanical properties, such as Young's modulus, are correlated with areal bone mineral density (BMD) captured in the DXA image. A quasi-static loading condition representing a sideway fall was applied to the finite element model. Three fracture risk indices were introduced and expressed as ratios of internal forces caused by impact forces occurring in sideway fall to bone ultimate cross-section strength at the three critical locations, i.e. the femoral neck, the intertrochanteric region, and the subtrochanteric region. The proposed finite element modelling procedure was validated against six representative clinical cases extracted from the Manitoba BMD database, where initial and follow-up DXA images have been taken to monitor longitudinal variation of areal BMD in individual patients. It was found from the clinical validation that variations in the proposed fracture risk indices have the same trends as those indicated by the conventional areal BMD and T-score. In addition, by the three proposed fracture risk indices it is possible to further identify the specific fracture location. It was also found that for the same subject, the variations in the three fracture risk indices have quite different magnitudes, with intertrochanteric region the largest and subtrochanteric region the smallest, which is probably owing to the different content of trabecular and cortical bones in the three regions. With further development, it is promising that the proposed DXA-based finite element model will be a useful tool for accurate assessment of osteoporosis development and for treatment monitoring.     

Some failure modes of four clinical bone cements

The fracture or failure behaviours of four commercial acrylic-based bone cements have been examined in tensile, bending and compression modes, and their mechanical properties are reviewed. It was found that Palacos R-40 bone cement had high radiopaque agent concentration, with high surface hardness. It exhibited a much lower bending strength and bending modulus compared with the other three bone cements (CMW1, CMW2000 and Simplex P). The textures of tensile fracture surfaces produced were similar for the four bone cements studied. The fracture surface was fragmented by crevices, which developed through the matrix and around large undissolved polymethylmethacrylate (PMMA) beads. Three bands with different features existed on the bending fracture surfaces, with an abrupt transition between them. It appears that the agglomerates of zirconium dioxide particles are implicated in Palacos R-40 bone cement fracture surface. The examination of compressive failed specimens revealed that a 'yielded crack band' existed across the transverse section. Plastic deformation resulted in the PMMA beads being squashed in the longitudinal direction and dilated in the transverse direction.