Similar damage initiation but different failure behavior in trabecular and cortical bone tissue

The mechanical properties of bone tissue are reflected in its micro- and nanostructure as well as in its composition. Numerous studies have compared the elastic mechanical properties of cortical and trabecular bone tissue and concluded that cortical bone tissue is stiffer than trabecular bone tissue. This study compared the progression of microdamage leading to fracture and the related local strains during this process in trabecular and cortical bone tissue. Unmachined single bovine trabeculae and similarly-sized cortical bovine bone samples were mechanically tested in three-point bending and concomitantly imaged to assess local strains using a digital image correlation technique. The bone whitening effect was used to detect microdamage formation and propagation. This study found that cortical bone tissue exhibits significantly lower maximum strains (trabecular 36.6%±14% vs. cortical 22.9%±7.4%) and less accumulated damage (trabecular 16100±8800 pix/mm2 vs. cortical 8000±3400 pix/mm2) at failure. However, no difference was detected for the maximum local strain at whitening onset (trabecular 5.8%±2.6% vs. cortical 7.2%±3.1%). The differences in elastic modulus and mineral distribution in the two tissues were investigated, using nanoindentation and micro-Raman imaging, to explain the different mechanical properties found. While cortical bone was found to be overall stiffer and more highly mineralized, no apparent differences were noted in the distribution of modulus values or mineral density along the specimen diameter. Therefore, differences in the mechanical behavior of trabecular and cortical bone tissue are likely to be in large part due to microstructural (i.e. orientation and distribution of cement lines) and collagen related compositional differences.     

Bone composition measured by x-ray scattering.

Ten composite samples consisting of cortical bone and adipose tissue, in known proportions, were made. The intensity of monochromatic x-rays (energy 8 keV) scattered by these samples was determined as a function of the modulus of the scattering vector, K. The ratio of the heights of peaks at K values of around 134 and 22 nm-1 provided a measure of the ratio of adipose tissue to bone mineral in these samples. This method was then used to determine the ratio of adipose tissue to mineral in samples of trabecular bone from 16 vertebral bodies. The results were correlated with measurements of the bone composition determined by ashing (r = 0.66) and histomorphometry (r = 0.66). Furthermore, the ashing and histomorphometry results were correlated with each other (r = 0.68). The feasibility of using higher energy x-rays (35-80 keV) for obtaining the same information from bone within the body is briefly discussed.     

The electrical and dielectric properties of human bone tissue and their relationship with density and bone mineral content

In this study, we examined the electrical properties of wet human cortical and cancellous bone tissue from distal tibia and their relationship to the wet, dry, and ash tissue densities. The resistivity and specific capacitance of both cortical and cancellous bone were determined for different frequencies and directions (orientation). The wet, dry, and ash tissue densities of the bone samples were measured, and the ash content was determined. Correlation and regression analysis was used to examine the possible relationships among the electrical properties and the tissue densities for cancellous and cortical bone specimens separately as well as for all of the bone specimens combined. Highly significant positive correlations (p<0.001) were found between the wet density of bone and the dry and ash densities. The specific capacitance of the cancellous bone specimens in all three orthogonal directions showed significant (p<0.01) positive correlations with the wet, dry, and ash densities. In general, the specific capacitance depended more on density for all bone specimens, and only a weak relationship was found between the resistivity of human cortical bone and density. Presented in part at the 13th Annual IEEE EMBS Conference held in Orlando, Florida, Oct. 31–Nov. 3, 1991, and at the 14th Annual Meeting of the Society for Physical Regulation in Biology and Medicine held in Arlington, Virginia, Oct. 13–16, 1994.     

Diffusion of ceftriaxone in human bone tissue

Ceftriaxone diffusion into spongious and cortical bone after single 2 g dose by intravenous injection was studied in 40 patients undergoing total hip replacement. Serum and tissue assays were performed by HPLC and microbiological method 2, 4, 12 and 24 hours after administration. Both methods produced similar results. Antibiotic levels (HPLC) were 19.3 +/- 7.3; 16.9 +/- 9.2; 11.2 +/- 4.7 and 5.6 +/- 2.7 micrograms/g in spongious bone at hours 2, 4, 12 and 24, and 6.5 +/- 1.6; 3.1 +/- 0.7; 3.3 +/- 1.1 and 2.2 +/- 0.4 micrograms/g in cortical bone at the same times. The tissue/serum ratios were 14% for spongious bone and 5% for cortical bone after 2 hours. At hour 24 these ratios were 21% and 8% in spongious and cortical bone respectively. These tissue levels remain higher than ceftriaxone MIC for most Enterobacteriaceae strains until hour 24.     

Long-term alcohol consumption and brown adipose tissue in man

Summary The purpose of the present work was to study whether long-term alcohol consumption in man affects the develeopment of brown adipose tissue. The adipose tissue around the thoracic aorta and common carotid arteries was collected at medicolegal autopsies on adults with a positive record of heavy alcohol consumption. Adults without any evident history of alcohol consumption served as controls. Histochemical reactions of the oxidative mitochondrial enzymes, cytochrome oxidase and succinate dehydrogenase were studied in samples of this adipose tissue and the activities of the enzymes were measured biochemically.There was histological evidence of some multilocular adipose tissue around the thoracic aorta and common carotid arteries of the alcohol consumers, whereas the adipose tissue from the non-drinkers was mostly unilocular resembling white adipose tissue. Histochemical evidence of brown adipose tissue was found in all alcohol consumers, but also in some of the controls. Biochemical cytochrome oxidase (CYO) and succinate dehydrogenase measurements in isolated mitochondria showed activity in 70% of the cases of drinkers and in one of the eight controls. Activity of CYO was measurable in the mitochondria from two other controls. The protein content of the samples from the alcoholics was twice that of the controls. The results suggest that chronic alcohol intake may induce a change in the white adipose tissue around the thoracic aorta and common carotid arteries of human adults into brown fat.     

A novel multiphase anodic spark deposition coating for the improvement of orthopedic implant osseointegration: an experimental study in cortical bone of sheep

The effect of a new three-step anodic spark deposition process, labeled TiSpark, including two consecutive treatments performed first in a P solution and second in Ca solution, followed by an additional alkali etching step, was investigated for the improvement of osseointegration of commercial grade 2 titanium, machined (Ti) or Al(2)O(3) sandblasted (Ti-SA), cylindrical implants (12 mm in length and 4 mm in diameter) in cortical bone of 12 adult sheep. Histomorphometric and microhardness measurements were carried out at each experimental time (4, 8, and 12 weeks) to quantify the bone-to-implant contact around the implants as well as the newly bone hardness and bone maturation index. TiSpark treated surfaces were covered by a thick layer of crystalline anatase TiO(2) and by a further Ca/P layer. Bone tissue extends and grows on the surface of the TiSpark treated implants without any fibrous tissue, enhancing the short-term osseointegration properties of implant. Bone mineralization rate was also influenced by the chemical composition of implants and sandblasted materials presented the lowest bone maturation rate at the interface. Data suggests that the TiSpark treatment produces a modification of the Ti surface, which presents good bioactivity and may be suitable for achieving a stable implant osseointegration.     

Mechanical properties of OI type III bone tissue measured by nanoindentation

Nanoindentation was used to characterize the intrinsic mechanical properties of bone tissue from eight (8) children with type III Osteogenesis Imperfecta (OI). The bone samples were harvested from the cortex portion at the site of bowing (the mid 2/3 of the shaft of the tibia/femur). Unlike normal bone tissue, OI type III cortical bone exhibited more isotropic material properties. Young's modulus and hardness values measured in the longitudinal direction did not show significant differences from the transverse measurements. No differences were observed in modulus or hardness in an analysis of the cortical and trabecular samples. The deformation patterns of the OI type III bone during nanoindentation were found to be similar to those of normal adult bone in an analysis of the ratio of modulus to hardness. No correlation was found between nanoindentation measurement and age in an analysis of regression.     

Chronic alcohol exposure stimulates adipose tissue lipolysis in mice: role of reverse triglyceride transport in the pathogenesis of alcoholic steatosis

Alcohol consumption induces liver steatosis; therefore, this study investigated the possible role of adipose tissue dysfunction in the pathogenesis of alcoholic steatosis. Mice were pair-fed an alcohol or control liquid diet for 8 weeks to evaluate the alcohol effects on lipid metabolism at the adipose tissue-liver axis. Chronic alcohol exposure reduced adipose tissue mass and adipocyte size. Fatty acid release from adipose tissue explants was significantly increased in alcohol-fed mice in association with the activation of adipose triglyceride lipase and hormone-sensitive lipase. Alcohol exposure induced insulin intolerance and inactivated adipose protein phosphatase 1 in association with the up-regulation of phosphatase and tensin homolog (PTEN) and suppressor of cytokine signaling 3 (SOCS3). Alcohol exposure up-regulated fatty acid transport proteins and caused lipid accumulation in the liver. To define the mechanistic link between adipose triglyceride loss and hepatic triglyceride gain, mice were first administered heavy water for 5 weeks to label adipose triglycerides with deuterium, and then pair-fed alcohol or control diet for 2 weeks. Deposition of deuterium-labeled adipose triglycerides in the liver was analyzed using Fourier transform ion cyclotron mass spectrometry. Alcohol exposure increased more than a dozen deuterium-labeled triglyceride molecules in the liver by up to 6.3-fold. These data demonstrate for the first time that adipose triglycerides due to alcohol-induced hyperlipolysis are reverse transported and deposited in the liver.     

Poly(vinyl alcohol) gels for use as tissue phantoms in photoacoustic mammography

Materials for solid photoacoustic breast phantoms, based on poly(vinyl alcohol) hydrogels, are presented. Phantoms intended for use in photoacoustics must possess both optical and acoustic properties of tissue. To realize the optical properties of tissue, one approach was to optimize the number of freezing and thawing cycles of aqueous poly(vinyl alcohol) solutions, a procedure which increases the turbidity of the gel while rigidifying it. The second approach concentrated on forming a clear matrix of the rigid poly(vinyl alcohol) gel without any scattering, so that appropriate amounts of optical scatterers could be added at the time of formation, to tune the optical properties as per requirement. The relevant optical and acoustic properties of such samples were measured to be close to the average properties of human breast tissue. Tumour simulating gel samples of suitable absorption coefficient were created by adding appropriate quantities of dye at the time of formation; the samples were then cut into spheres. A breast phantom embedded with such 'tumours' was developed for studying the applicability of photoacoustics in mammography.     

Innovative magnetic scaffolds for orthopedic tissue engineering

The use of magnetism in tissue engineering is a very promising approach, in fact magnetic scaffolds are able not only to support tissue regeneration, but they can be activated and work like a magnet attracting functionalized magnetic nanoparticles (MNPs) injected close to the scaffold enhancing tissue regeneration. This study aimed to assess the in vivo biocompatibility and osteointegrative properties of novel magnetic scaffolds. Two hydroxyapatite/collagen (70/30 wt %) magnetic scaffolds were magnetized with two different techniques: direct nucleation of biomimetic phase and superparamagnetic nanoparticles (MNPs) on self-assembling collagen fibers (MAG-A) and scaffold impregnation in ferro-fluid solution (MAG-B). Magnetic scaffolds were implanted in rabbit distal femoral epiphysis and tibial mid-diaphysis. Histopathological screening showed no inflammatory reaction due to MNPs. Significantly higher bone healing rate (ΔBHR) results were observed in MAG-A in comparison to MAG-B. Significant differences were also found between experimental times with an increase in ΔBHR from 2 to 4 weeks for both scaffolds in trabecular bone, while only for MAG-B (23%, p < 0.05) in cortical bone. The proposed magnetic scaffolds seem to be promising for magnetic guiding in orthopedic tissue engineering applications and they will be suitable to treat also several pathologies in regenerative medicine area.     

Fatty acid ethyl esters in adipose tissue. A laboratory marker for alcohol-related death

Fatty acid ethyl esters, a family of ethanol metabolites, are formed by esterification of ethanol with fatty acids and have been detected in human organs commonly damaged by ethanol abuse. Because alcohol-related deaths may occur up to six times as often as reported on death certificates, we undertook quantitation of these potentially longer-lived alcohol metabolites in postmortem human adipose tissue to assess their usefulness as a measure of recent ethanol exposure. After isolation and identification using sequential thin-layer and gas chromatography, fatty acid ethyl esters were present in adipose tissue of chronic alcoholics (mean +/- SEM equals 300 +/- 46 nmol/g), even though blood ethanol concentration at the time of death was undetectable. Unintoxicated nonalcoholic subjects who had no history of alcohol abuse had concentrations seven times lower (mean +/- SEM equals 43 +/- 13 nmol/g). In vitro studies demonstrate that fatty acid ethyl esters are synthesized by human adipose tissue in proportion to the ethanol concentration present and their half-life in adipose tissue of laboratory animals is 16 +/- 1.6 hours, ie, fourfold greater than that of alcohol. These results indicate that fatty acid ethyl esters are long-lived ethanol metabolites whose persistence and accumulation in adipose tissue may allow an accurate diagnosis of significant alcohol consumption even when ethanol has been completely eliminated from the body.     

Site-matched assessment of structural and tissue properties of cortical bone using scanning acoustic microscopy and synchrotron radiation muCT

200 MHz scanning acoustic microscopy (SAM) and synchrotron radiation muCT (SR-muCT) were used to assess microstructural parameters and tissue properties in site-matched regions of interest in cortical bone. Anterior and postero-lateral regions of ten cross sections from human cortical radius were explored. Structural parameters, including diameter and number of Haversian canals per cortical area (Ca.Dm, N.Ca/Ar) and porosity Po were assessed with both methods using a custom-developed image fusion and analysis software. Acoustic impedance Z and degree of mineralization of bone DMB were extracted separately for osteonal and interstitial tissues from the fused images. Structural parameter estimations obtained from radiographic and acoustic images were almost identical. DMB and impedance values were in the range between 0.77 and 1.28 g cm(-3) and 5.13 and 12.1 Mrayl, respectively. Interindividual and regional variations were observed, whereas the strongest difference was found between osteonal and interstitial tissues (Z: 7.2 +/- 1.1 Mrayl versus 9.3 +/- 1.0 Mrayl, DMB: 1.06 +/- 0.07 g cm(-3) versus 1.16 +/- 0.05 g cm(-3), paired t-test, p < 0.05). Weak, but significant correlations between DMB and Z were obtained for the osteonal (R(2) = 0.174, p < 10(-4)) and for the pooled (osteonal and interstitial) data. The regression of the pooled osteonal and interstitial tissue data follows a second-order polynomial (R(2) = 0.39, p < 10(-4)). Both modalities fulfil the requirement for a simultaneous evaluation of cortical bone microstructure and material properties at the tissue level. While SAM inspection is limited to the evaluation of carefully prepared sample surfaces, SR-muCT provides volumetric information on the tissue without substantial preparation requirements. However, SAM provides a quantitative estimate of elastic properties at the tissue level that cannot be captured by SR-muCT.     

Monte Carlo based water/medium stopping-power ratios for various ICRP and ICRU tissues

Water/medium stopping-power ratios, s(w,m), have been calculated for several ICRP and ICRU tissues, namely adipose tissue, brain, cortical bone, liver, lung (deflated and inflated) and spongiosa. The considered clinical beams were 6 and 18 MV x-rays and the field size was 10 x 10 cm(2). Fluence distributions were scored at a depth of 10 cm using the Monte Carlo code PENELOPE. The collision stopping powers for the studied tissues were evaluated employing the formalism of ICRU Report 37 (1984 Stopping Powers for Electrons and Positrons (Bethesda, MD: ICRU)). The Bragg-Gray values of s(w,m) calculated with these ingredients range from about 0.98 (adipose tissue) to nearly 1.14 (cortical bone), displaying a rather small variation with beam quality. Excellent agreement, to within 0.1%, is found with stopping-power ratios reported by Siebers et al (2000a Phys. Med. Biol. 45 983-95) for cortical bone, inflated lung and spongiosa. In the case of cortical bone, s(w,m) changes approximately 2% when either ICRP or ICRU compositions are adopted, whereas the stopping-power ratios of lung, brain and adipose tissue are less sensitive to the selected composition. The mass density of lung also influences the calculated values of s(w,m), reducing them by around 1% (6 MV) and 2% (18 MV) when going from deflated to inflated lung.     

Fat metabolism in brown adipose tissue in vivo

1. Noradrenaline infusions (I.V. for 30 min) in new-born and 1-week-old unanaesthetized rabbits caused a rapid and sustained rise in oxygen consumption and an increase in blood glucose, free fatty acid and glycerol concentrations. Similar changes occurred in anaesthetized 1-week-old rabbits.2. A simple direct method was used to measure the net exchange of metabolites across brown adipose tissue in anaesthetized young rabbits. It was found that the brown adipose tissue of rabbits reared from birth in a thermoneutral environment and studied when their next feed was due took glucose from and released fatty acids and glycerol into the circulation.3. Similar rabbits kept unfed for a further 48 hr in a warm environment released fatty acids and glycerol from their brown adipose tissue at a far greater rate, but the rate of glucose uptake was reduced.4. On the other hand the brown adipose tissue of rabbits kept unfed in a cold environment (20 degrees C) took up circulating free fatty acids as well as glucose and did not release glycerol. The brown adipose tissue of these rabbits was depleted of fat.5. The rate of blood flow through brown adipose tissue and the exchange of all three metabolites increased fourfold during noradrenaline infusion in the three groups of rabbits.6. It is concluded (i) that brown adipose tissue releases significant amounts of fatty acids and glycerol into the circulation and that this contribution is greatly increased with noradrenaline infusion and presumably cold exposure, and (ii) that brown adipose tissue depleted of fat produces heat by drawing free fatty acids as well as glucose from the circulation.     

猪源性骨支架材料的制备及性能研究

目的:制备猪源性骨支架材料,并与人源性骨支架材料对比检测其理化性能和组织相容性。方法:经低温深冻、超声清洗、H2O2、酒精浸泡、冻干、辐照制备猪源性骨支架材料和人源性骨支架材料。扫描电镜观察,测定材料孔隙率、蛋白质含量、钙磷含量及弹性模量。2种材料的浸提液与脂肪源间充质干细胞复合培养,观察细胞一般形态,流式细胞仪PI法检测细胞生命周期。皮下植入2种材料,在植入4、8、12、16周取材做病理切片观察和扫描电镜观察。结果:2种材料均具有骨本身的天然网状三维支架系统。猪源性骨支架材料的孔隙率高于人源性骨支架材料,蛋白含量低于人源性骨支架材料,弹性模量分别为无显著差异。材料浸提液组及空白对照组的细胞生长状态良好。流式细胞仪PI法检测细胞周期见G1期、G2期细胞百分率接近。皮下植入试验表明,随着植入时间的延长,炎症反应逐步减轻,材料降解增加,新生软骨样结构逐渐增多。结论:猪源性异种骨支架材料在理化性能和材料毒性等方面与同种异体骨支架材料接近,具有良好的应用前景。     

脂肪与骨髓来源间充质干细胞生物学特性的比较

背景：近几年来脂肪来源的间充质干细胞因其取材容易也被广泛研究。 目的：比较脂肪来源和骨髓来源间充质干细胞的生物学特性。 方法：分离及体外培养人骨髓源间充质干细胞和脂肪源间充质干细胞,比较它们的表型、细胞倍增时间及分泌因子水平等。 结果与结论：脂肪来源和骨髓来源的间充质干细胞在细胞表型上类似,只有CD106的表达有差异。脂肪来源间充质干细胞增殖速率比骨髓来源的间充质干细胞快。在相同体积的脂肪组织中能够得到的干细胞前体细胞的数量是骨髓的10倍以上。提示脂肪来源和骨髓来源的间充质干细胞具有相同功能,但脂肪组织是一个更有应用前景的干细胞来源。     

Bone response to three different chemical compositions of fluorcanasite glass-ceramic

The aim of this study was to evaluate the bone response to three fluorcanasite glass-ceramic compositions with different solubilities (K3, K5, and K8) after implantation in a femur rabbit model. Fluorcanasite glass-ceramic rods were implanted bilaterally in the mid-shafts rabbit femurs. Implants were harvested at 8 and 12 weeks and prepared for histological and histomorphometric analyses at the light microscope level. Bioglass 45S5 rods were used as a control material. At 8 weeks, all fluorcanasite glass-ceramics were entirely surrounded by a nonmineralized connective tissue. At 12 weeks, reduced areas of bone tissue were observed in the cortical area in direct contact with the K3 and K5 fluorcanasite glass-ceramics compared to Bioglass 45S5, whereas no bone tissue was observed in direct contact with the K8 surface. Bone-to-implant contact in the cortical area was affected by the material chemical composition and ranked as follows: Bioglass 45S5>K3>K5>K8 (p=0.001). In the bone marrow, a layer of fibrous connective tissue formed in direct contact with the fluorcanasite glass-ceramics and Bioglass 45S5, and only rarely exhibited contact osteogenesis. All the fluorcanasite glass-ceramics appeared to degrade in the biological environment. The solubility ratio did not alter significantly the biological reply of the fluorcanasite glass-ceramics in vivo. Further modifications of the chemical composition of the fluorcanasite glass-ceramic are required to increase the stability of the material in vivo.     

Contact with existing adipose tissue is inductive for adipogenesis in matrigel

The effect of adipose tissue on inductive adipogenesis within Matrigel (BD Biosciences) was assessed by using a murine chamber model containing a vascular pedicle. Three-chamber configurations that varied in the access to an adipose tissue source were used, including sealed- and open-chamber groups that had no access and limited access, respectively, to the surrounding adipose tissue, and a sealed-chamber group in which adipose tissue was placed as an autograft. All groups showed neovascularization, but varied in the amount of adipogenesis seen in direct relation to their access to preexisting adipose tissue: open chambers showed strong adipogenesis, whereas the sealed chambers had little or no adipose tissue; adipogenesis was restored in the autograft chamber group that contained 2- to 5-mg fat autografts. These showed significantly more adipogenesis than the sealed chambers with no autograft ( p < 0.01). Autografts with 1mg of fat were capable of producing adipogenesis but did so less consistently than the larger autografts. These findings have important implications for adipose tissue engineering strategies and for understanding de novo production of adipose tissue.     

Varying the mechanical properties of bone tissue by changing the amount of its structurally effective bone mineral content

The effect of fluoride ions on the mechanical properties of bone tissue in tension was investigated with an in vitro model. Structurally effective Bone Mineral Content (BMC) of bovine bone tissue was changed by fluoride ion treatment. First, bovine cortical bone specimens were treated with a detergent solution in order to increase the diffusion rates of the treatment ions across the samples. After the initial treatment, different ion solutions were used to treat the tension samples (fluoride, sodium and chloride). Ionic strength and pH were varied. Experimental results showed that the sodium chloride solutions of different ionic strengths, at physiological and high pH, do not affect the mechanical properties of bone tissue in tension. However, uniform fluoride treatment across the samples reduced the mechanical strength of bone tissue by converting small amounts of bone mineral to mostly calcium fluoride. This action reduces the structurally effective BMC and also possibly effects the interface bonding between the bone mineral and the organic matrix of the bone tissue.