Mechanics of bone/PMMA composite structures: an in vitro study of human vertebrae

The goal of this study was to provide material property data for the cement/bone composite resulting from the introduction of PMMA bone cement into human vertebral bodies. A series of quasistatic tensile and compressive mechanical tests were conducted using cement/bone composite structures machined from cement-infiltrated vertebral bodies. Experiments were performed both at room temperature and at body temperature. We found that the modulus of the composite structures was lower than bulk cement (p<0.0001). For compression at 37( composite function)C: composite =2.3+/-0.5GPa, cement =3.1+/-0.2GPa; at 23( composite function)C: composite =3.0+/-0.3GPa, cement =3.4+/-0.2GPa. Specimens tested at room temperature were stiffer than those tested at body temperature (p=0.0004). Yield and ultimate strength factors for the composite were all diminished (55-87%) when compared to cement properties. In general, computational models have assumed that cement/bone composite had the same modulus as cement. The results of this study suggest that computational models of cement infiltrated vertebrae and cemented arthroplasties could be improved by specifying different material properties for cement and cement/bone composite.     

Theoretical temperature calculation in the cutting area in drilling cortical bones

To date, analysis of temperature in the drill area of cortical bone have been limited to measurements with thermocouple systems at a certain distance from the drill hole. Many authors equate this temperature measurement with the drill--cortical bone interface temperature. In order show that there is a temperature difference, a drill hole was simulated with the aid of the "Finite Element Method". The interface temperature was calculated by the energy distribution. It was shown, that for "dry" and "watercooled" drilling, the drill hole temperature was 13 degrees C higher than the temperature measured with the thermocouple systems at a distance 0.5 mm of from the drill hole. In particular when using "watercooled" drills for bone and dental surgery, the temperature may be higher than the bone damage limit of 44 degrees C for lengthy and 50 degrees C for brief drilling.     

Thermal analysis of bone cement polymerisation at the cement-bone interface

The two major problems that have been reported with the use of polymethylmethacrylate (PMMA) cement are thermal necrosis of surrounding bone due to the high heat generation during polymerisation and chemical necrosis due to unreacted monomer release. Computer models have been used to study the temperature and monomer distribution after cementation. In most of these models, however, polymerisation is modelled as temperature independent and cancellous bone is modelled as a continuum. Such models thus cannot account for the expected important role of the trabecular bone micro-structure. The aim of this study is to investigate the distribution of temperature and monomer leftover at the cancellous bone–cement interface during polymerisation for a realistic trabecular bone—cement micro-structure and realistic temperature-dependent polymerisation kinetics behaviour.

A 3-D computer model of a piece of bovine cancellous bone that underwent pressurization with bone–cement was generated using a micro-computed tomography scanner. This geometry was used as the basis for a finite element model and a temperature-dependent problem for bone cement polymerisation kinetics was solved to simulate the bone cement polymerisation process in the vicinity of the interface. The transient temperature field throughout the interface was calculated, along with the polymerisation fraction distribution in the cement domain.

The calculations revealed that the tips of the bone trabeculae that are embedded in the cement attain temperatures much higher than the average temperature of the bone volume. A small fraction of the bone (10%) is exposed to temperatures exceeding 70°C, but the exposure time to these high temperatures is limited to 50 s. In the region near the bone, the cement polymerisation fraction (about 84%) is less than that in the centre (where it is reaching values of over 96%). An important finding of this study thus is the fact that the bone tissue that is subjected to the highest temperatures is also subjected to high leftover monomer concentration. Furthermore the maximum bone temperature is reached relatively early, when monomer content in the neighbouring cement is still quite high.     

Bone temperature estimation during orthopaedic round bur milling operations

Heat generation during orthopaedic bone cutting operations may cause thermal bone damage. During the bone cutting, the maximum temperature occurs at the point of contact between the bone and the cutting tool. However, this temperature is difficult to measure. Many researchers have attempted to measure this temperature using a thermocouple; however, limitations of the thermocouple makes it difficult to determine the maximum temperature at the point of contact. In order to solve this problem, in this study, two infrared thermometers are used to measure the fresh-milled surface temperature, and the maximum temperature was extrapolated by a moving plane heat source solution. The estimated maximum temperature increment varied from 49 to 115 degrees C under various cutting conditions. These results showed that the thermal damage may reach up to 1.9 mm in depth during round bur milling. A larger feed rate and a smaller cutting depth decreased the maximum temperature.     

Effect of gamma irradiation on mechanical properties of human cortical bone: influence of different processing methods

The secondary sterilisation by irradiation reduces the risk of infectious disease transmission with tissue allografts. Achieving sterility of bone tissue grafts compromises its biomechanical properties. There are several factors, including dose and temperature of irradiation, as well as processing conditions, that may influence mechanical properties of a bone graft. The purpose of this study was to evaluate the effect of gamma irradiation with doses of 25 or 35?kGy, performed on dry ice or at ambient temperature, on mechanical properties of non-defatted or defatted compact bone grafts. Left and right femurs from six male cadaveric donors aged from 46 to 54?years, were transversely cut into slices of 10?mm height, parallel to the longitudinal axis of the bone. Compact bone rings were assigned to the eight experimental groups according to the different processing method (defatted or non-defatted), as well as gamma irradiation dose (25 or 35?kGy) and temperature conditions of irradiation (ambient temperature or dry ice). Axial compression testing was performed with a material testing machine. Results obtained for elastic and plastic regions of stress-strain curves examined by univariate analysis are described. Based on multivariate analysis it was found that defatting of bone rings had no significant effect on any mechanical parameter studied, whereas irradiation with both doses decreased significantly the ultimate strain and its derivative toughness. The elastic limit and resilience were significantly increased by irradiation with the dose 25?kGy, but not 35?kGy, when the time of irradiation was longer. Additionally, irradiation at ambient temperature decreased maximum load, elastic limit, resilience, and ultimate stress. As strain in the elastic region was not affected, decreased elastic limit resulted in lower resilience. The opposite phenomenon was observed in the plastic region, where in spite of the lower ultimate stress, the toughness was increased due to the increase in the ultimate strain. The results of our study suggest that there may be an association between mechanical properties of bone tissue grafts and the damage process of collagen structure during gamma irradiation. This collagen damage in cortical bone allografts containing water does not depends on the temperature of irradiation or defatting during processing if dose of gamma irradiation does not exceed 35?kGy.     

Heat resistance of allograft tissue

Lyophilized allograft musculoskeletal tissue is generally intended to be stored at “ambient” or “room” temperature, and usually is kept in climate controlled indoor storage areas. However, there is a question of what temperature extremes tissue may see, especially during transportation, in that these extremes may exceed even the limits of “ambient” conditions. Temperatures may become quite hot, but only for a few hours and only during daytime. Damage from high temperatures, if it occurs, is expected to be evident by damage to the collagen component of bone, soft tissue, and demineralized bone, as well as to the growth factors contained in demineralized bone. If damage is significant, then temperature monitoring requirements for lyophilized allograft tissue might be necessary. To answer this question, a literature review was carried out to look at the short-term temperature resistance of collagen and demineralized bone. Both collagen and the growth factors in demineralized bone show remarkable short term tolerance to elevated temperatures in the dry state, and it was concluded that temperature excursions of 50°C or less, lasting for a few days or less, would not cause any significant deterioration. This means that temperature monitoring also should not be required.     

Effect of temperature on the fracture toughness of compact bone

Bone is a composite composed mainly of organics, minerals, and water. Many researchers have studied effects such as crack velocity, density, orientation, storage media, porosity, and age on the fracture toughness (K(C), also called critical stress intensity factor) of compact bone. Most of these studies were conducted at room temperature. Considering that the body temperature of animals is greater than room temperature, and that bone has a large volumetric percentage of organics and water (generally, 55-65%), it is hypothesized that temperature has a significant effect on the fracture toughness of compact bone. Single-edge V-notched (SEVN) specimens were prepared to measure the fracture toughness of bovine femur and manatee rib in water at 0, 10, 23, 37, and 50 degrees C in four-point flexure. The fracture toughness values of bovine femur and manatee rib were found to decrease from 7.0 to 4.3MPam(1/2) and from 5.5 to 4.0MPam(1/2), respectively, as temperature increased over a temperature range of 50 degrees C. The results support the hypothesis that temperature has a significant effect on the fracture toughness of compact bone. Therefore, we suggest that study on fracture toughness of bone should be done at physiologically relevant temperatures.     

Stability of RNA isolated from human trabecular bone at post-mortem and surgery

To determine the reliability of gene expression studies in human post-mortem bone, it is important to evaluate the stability of RNA isolated from such tissues as a function of the post-mortem interval. The stability of total RNA and bone-specific mRNA species was examined in bone samples obtained from routine autopsies and at surgery. The optimal temperature for any storage and transport of the bone before RNA isolation was shown to be 4 degrees C, and RT-PCR analysis is the preferred technique for the analysis of gene expression in post-mortem bone as it tolerates partial RNA degradation. For gene expression studies in bone, post-mortem cases, with a post-mortem interval of less than 48 h, should be selected, and the time that bone is stored after retrieval at autopsy or surgery should be kept to a minimum. Overall, our findings indicate that with appropriate storage and handling, RNA can be reliably isolated from human bone obtained at post-mortem and surgery to study ex vivo the pattern of gene expression in healthy individuals and in patients with musculoskeletal diseases such as osteoporosis and osteoarthritis.     

Influence of early qualitative feed restriction and environmental temperature on long bone development of broiler chickens

This investigation was carried out to study the influence of early qualitative feed restriction and environmental rearing temperature on long bone development in broiler. Energy and protein restriction reduced femur width and humerus weight, but did not affect tibia parameters. Broilers kept at cold environmental temperature showed reduced femur, tibia and humerus length and tibia weight, but the calculated density was not affected by rearing temperature. These findings suggest that qualitative feed restriction and environmental temperature influenced the normal long bone growth; however, bone weight/bone length index (calculated density) was not affected by rearing temperature.     

Effect of accelerated electron beam on mechanical properties of human cortical bone: influence of different processing methods

Accelerated electron beam (EB) irradiation has been a sufficient method used for sterilisation of human tissue grafts for many years in a number of tissue banks. Accelerated EB, in contrast to more often used gamma photons, is a form of ionizing radiation that is characterized by lower penetration, however it is more effective in producing ionisation and to reach the same level of sterility, the exposition time of irradiated product is shorter. There are several factors, including dose and temperature of irradiation, processing conditions, as well as source of irradiation that may influence mechanical properties of a bone graft. The purpose of this study was to evaluate the effect e-beam irradiation with doses of 25 or 35?kGy, performed on dry ice or at ambient temperature, on mechanical properties of non-defatted or defatted compact bone grafts. Left and right femurs from six male cadaveric donors, aged from 46 to 54?years, were transversely cut into slices of 10?mm height, parallel to the longitudinal axis of the bone. Compact bone rings were assigned to the eight experimental groups according to the different processing method (defatted or non-defatted), as well as e-beam irradiation dose (25 or 35?kGy) and temperature conditions of irradiation (ambient temperature or dry ice). Axial compression testing was performed with a material testing machine. Results obtained for elastic and plastic regions of stress-strain curves examined by univariate analysis are described. Based on multivariate analysis, including all groups, it was found that temperature of e-beam irradiation and defatting had no consistent significant effect on evaluated mechanical parameters of compact bone rings. In contrast, irradiation with both doses significantly decreased the ultimate strain and its derivative toughness, while not affecting the ultimate stress (bone strength). As no deterioration of mechanical properties was observed in the elastic region, the reduction of the energy absorption capacity of irradiated bone rings apparently resulted from changes generated by irradiation within the plastic strain region.     

Measurement of transient and residual stresses during polymerization of bone cement for cemented hip implants

The initial fixation of a cemented hip implant relies on the strength of the interface between the stem, bone cement and adjacent bone. Bone cement is used as grouting material to fix the prosthesis to the bone. The curing process of bone cement is an exothermic reaction where bone cement undergoes volumetric changes that will generate transient stresses resulting in residual stresses once polymerization is completed. However, the precise magnitude of these stresses is still not well documented in the literature. The objective of this study is to develop an experiment for the direct measurement of the transient and residual radial stresses at the stem-cement interface generated during cement polymerization. The idealized femoral-cemented implant consists of a stem placed inside a hollow cylindrical bone filled with bone cement. A sub-miniature load cell is inserted inside the stem to make a direct measurement of the radial compressive forces at the stem-cement interface, which are then converted to radial stresses. A thermocouple measures the temperature evolution during the polymerization process. The results show the evolution of stress generation corresponding to volumetric changes in the cement. The effect of initial temperature of the stem and bone as well as the cement-bone interface condition (adhesion or no adhesion) on residual radial stresses is investigated. A maximum peak temperature of 70 degrees C corresponds to a peak in transient stress during cement curing. Maximum radial residual stresses of 0.6MPa in compression are measured for the preheated stem.     

Effects of freezing on bone histological morphology

Allograft bone has been widely used for reconstruction of different portions of the skeleton. The fragment of bone harvested must be kept under low temperatures. The cryopreservation also contributes to decrease the antigenic potential of the tissue. Although this technique is considered safe, there is little information about the morphological modifications that the medullary and cortical portions of bone suffer after freezing. Hence, the aim of this study was to investigate the morphology of bone tissue after freezing under different temperatures and periods. Twelve rabbits were used to analyze the effects of two temperatures, −20°C and −70°C, during four periods of time: 30, 60, 90, 120 days. Tissues were analyzed by HE, picro-sirius stains and also by Feulgen’s reaction, through qualitative and morphometric ways, considering the area occupied by cells and nuclei, medullary and cortical portions, as well as by collagen expression at cortical. The differences among the treatments were analyzed by Tukey′s test, at 5% significance level. Bone freezing increased cellular and nuclear areas at cancellous bone and diminished nuclear area at the cortical bone. Cortical bone collagen suffered denaturation proportionally to temperature decrease and to freezing duration. These alterations compromised the morphology of tissues after 90 or 120 days of freezing at the temperature of −70°C. Cells necrosed during freezing, contributing to reduce bone antigenicity.     

Effect of gamma radiation and accelerated electron beam on stable paramagnetic centers induction in bone mineral: influence of dose,irradiation temperature and bone defatting

Ionizing radiation has been found to induce stable defects in the crystalline lattice of bone mineral hydroxyapatite, defined as CO₂ ^? radical ions possessing spins. The purpose of our study was to evaluate CO₂ ^? radical ions induced in non-defatted or defatted human compact bone by gamma radiation (G) and accelerated electron beam (EB), applied with two doses at different temperatures. Moreover, the potential effect of free radical ion formation on mechanical parameters of compact bone, tested under compression in the previous studies, was evaluated. Bone rings from femoral shafts of six male donors (age 51 ± 3 years) were collected and assigned to sixteen experimental groups according to different processing methods (non-defatted or defatted), G and EB irradiation dose (25 or 35 kGy), and irradiation temperature [ambient temperature (AT) or dry ice (DI)]. Untreated group served as control. Following grinding under LN₂ and lyophilization, CO₂ ^? radical ions in bone powder were measured by electron paramagnetic resonance spectrometry. We have found that irradiation of bone with G and EB induces formation of enormous amounts of CO₂ ^? radical ions, absent from native tissue. Free radical ion formation was dose-dependent when irradiation was performed at AT, and significantly lower in EB as compared to G-irradiated groups. In contrast, no marked effect of dose was observed when deep-frozen (DI) bone samples were irradiated with G or EB, and free radical ion numbers seemed to be slightly higher in EB-irradiated groups. Irradiation at AT induced much higher quantities of CO₂ ^? radical ions then on DI. That effect was more pronounced in G-irradiated bone specimens, probably due to longer exposure time. Similarly, bone defatting protective effect on free radical ion formation was found only in groups irradiated for several hours with gamma radiation at ambient temperature. Ambient irradiation temperature together with exposure time seem to be key parameters promoting CO₂ ^? radical ion formation in bone mineral and may mask the opposite effect of defatting and the possible effect of irradiation type. Significant weak negative correlations between CO₂ ^? radical ion number and some mechanical properties of compact bone rings (Young’s modulus and ultimate stress) were found.     

Validation of Radiation Dose Received by Frozen Unprocessed and Processed Bone during Terminal Sterilisation

Fresh frozen femoral heads (FH) and frozen processed bone (FP) are widely used as a source of allograft bone. The FP bone and some of the FH are terminally sterilised by the National Blood Service Tissue Services (NSBTS), via application of a minimum 25 kGy gamma radiation dose. To comply with the Guidelines for the Blood Transfusion Services in the United Kingdom (2002), frozen musculoskeletal tissue must be maintained below −40 °C during storage and transit. In practice, NBSTS stores bone long-term in −80 °C freezers. During transport for irradiation, a temperature of circa −79 °C is maintained by packing the bone in dry ice. An evaluation of the radiation dose received by bone has previously been made via dosimeters located within the tissue and dry ice, however, some evidence suggests that low temperature can influence the accuracy of the dosimeter readings. The aim of this study was to determine the actual radiation dose received by FH and FP bone during the irradiation process. This was accomplished by comparing radiation dose readings from dosimeters placed in dry ice with dosimeters placed in a dry ice substitute of similar dimensions and density i.e., polytetrafluoroethylene (PTFE) at ambient temperature. New packing formats were developed for both FH and FP bone such that 15 FH or 3 kg of FP bone could be irradiated in one transport box at any given time in a standardised fashion. The data show that low temperature consistently increased dosimeter readings 10–27%, and that radiation dose always fell within the range of 25–40 kGy (FH = 25.1–35.7 kGy; FP bone = 25.2–32.4 kGy).     

The origin of electrokinetic potentials in bone tissue: the organic phase

The purpose of this study was to determine the relative contributions of the organic and mineral phases of cow cortical bone to its electrokinetic response at room temperature. The technique of particle electrophoresis permitted electrokinetic (zeta) potentials to be calculated and plotted as a function of pH. Control and demineralized bone particles exhibited similar isoelectric points at pH approximately 5.1 (pH at which the zeta potential is zero), well below the isoelectric point of the bone mineral (pH approximately 8.6). In addition, the use of phosphate-containing buffers resulted in a zeta potential sign reversal of the bone mineral but had no effect on both the control and demineralized bone. These key results form the basis from which we suggest that the bone mineral lies within the organic phase (e.g., the mineral is not exposed to the fluid phase) and that the electrokinetic behavior of bone tissue is dominated by its organic ultrastructure.     

In vitro and in vivo evaluation of xenogeneic bone putty with the carrier of hydrogel derived from demineralized bone matrix

The demineralized bone matrix (DBM) putty is a traditional bone graft utilized to facilitate the repair and reconstruction of bone. Recent studies indicated the DBM putties with the various carriers were different in bone repairing ability. In order to prepare a kind of DBM putty with a good biocompatibility and bioactivity, the DBM gel was processed from the DBM and the feasibility as a carrier for the DBM putty was evaluated. After the bovine DBM gel was prepared, the BMPs content as well as the ability to promote osteogenic differentiation of MC3T3-E1 cells in vitro were investigated. Then the DBM putty was prepared and filled into the rat calvarial defect model to evaluate the bone repairing ability by micro-CT and histology. The result showed there was 2.953?±?0.054 ng BMP contained in per gram of the DBM gel. And the ALP production of MC3T3-E1 cells in the DBM gels group increased with prolonged culturing, the mineralized nodules formed in MC3T3-E1 cells on 14th day after co-culture. The putty prepared by DBM gel was easy to handle without loss of DBM particles at room temperature. In the rat calvarial bone defect experiment, histological observation showed more mature bone formed in the DBM putty group than that in the type I collagen group at 12 weeks, which indicated the bone putty prepared by DBM gel exhibited a better bone repair capability.     

The effect of gamma-irradiation on the temperature dependence of D.C. electrical conductivity of dry bone.

The effect of gamma-irradiation with doses from 10 to 500 kGy on the electrical conductivity (g) of dry bone was studied. Temperature measurement of electrical conductivity were made from 393 to 533 K. The dependence obtained indicates the increase in g with temperature. An increase in irradiation dose resulted in a decreased g value for each dose up to temperature 462 K. Temperature 462 K was interpreted as the temperature of collagen melting point in dry bone. Above 462 K, g values were dose independent. A dose of 500 kGy shifted the melting point to lower temperature. In addition, the activation energy for the charge conduction process was calculated. Obtained values for electrical conductivity and activation energy were typical for dielectrics and indicated degradation of the organic component of bone.     

The temperature dependence of the deformation of bone

The deformation characteristics of compact bone have been measured as a function of temperature in the range from −58 to 90°C. At temperatures below 25°C elastic, anelastic and plastic contributions to the strain could be distinguished. The Young's modulus increased as the temperature was reduced, while the nonelastic component was approximately independent of temperature. An interpretation of these results is made in terms of an idealised hydroxyapatite-collagen composite. At temperatures higher than 50°C the deformation was compounded by non-equilibrium recovery and an irreversible change in the structure of bone.     

Exercise mitigates the stunting effect of cold temperature on limb elongation in mice by increasing solute delivery to the growth plate

Ambient temperature and physical activity modulate bone elongation in mammals, but mechanisms underlying this plasticity are a century-old enigma. Longitudinal bone growth occurs in cartilaginous plates, which receive nutritional support via delivery of solutes from the vasculature. We tested the hypothesis that chronic exercise and warm temperature promote bone lengthening by increasing solute delivery to the growth plate, measured in real time using in vivo multiphoton microscopy. We housed 68 weanling female mice at cold (16°C) or warm (25°C) temperatures and allowed some groups voluntary access to a running wheel. We show that exercise mitigates the stunting effect of cold temperature on limb elongation after 11 days of wheel running. All runners had significantly lengthened limbs, regardless of temperature, while nonrunning mice had shorter limbs that correlated with housing temperature. Tail length was impacted only by temperature, indicating that the exercise effect was localized to limb bones and was not a systemic endocrine reaction. In vivo multiphoton imaging of fluoresceinated tracers revealed enhanced solute delivery to tibial growth plates in wheel-running mice, measured under anesthesia at rest. There was a minimal effect of rearing temperature on solute delivery when measured at an intermediate room temperature (20°C), suggesting that a lasting increase in solute delivery is an important factor in exercise-mediated limb lengthening but may not play a role in temperature-mediated limb lengthening. These results are relevant to the study of skeletal evolution in mammals from varying environments and have the potential to fundamentally advance our understanding of bone elongation processes.