Load applied on a bone-anchored transfemoral prosthesis: characterisation of a prosthesis : a pilot study

The objectives of this study were (A) to record the inner prosthesis loading during activities of daily living (ADL), (B) to present a set of variables comparing loading data, and (C) to provide an example of characterisation of two prostheses. The load was measured at 200 Hz using a multi-axial transducer mounted between the residuum and the knee of an individual with unilateral transfemoral amputation fitted with a bone-anchored prosthesis. The load was measured while using two different prostheses including a mechanically (PRO1) and a microprocessor controlled (PRO2) knee during six ADL. The characterisation of prosthesis was achieved using a set of variables split into four categories, including temporal characteristics, maximum loading, loading slopes and impulse. Approximately 360 gait cycles were analysed for each prosthesis. PRO1 showed a cadence improved by 19% and 7%, a maximum force on the long axis reduced by 11% and 19%, as well as an impulse reduced by 32% and 15% during descent of incline and stairs compared to PRO2, respectively. This work confirmed that the proposed apparatus and characterisation can reveal how changes of prosthetic components are translated into inner loading.

One step closer into the design of fall protective device for individuals fitted with a bone-anchored prosthesis

The consequences of falls are often dreadful for individuals with lower limb amputation using bone-anchored prosthesis.[1-5] Typically, the impact on the fixation is responsible for bending the intercutaneous piece that could lead to a complete breakage over time. .[3, 5-8] The surgical replacement of this piece is possible but complex and expensive. Clearly, there is a need for solid data enabling an evidence-based design of protective devices limiting impact forces and torsion applied during a fall. The impact on the fixation during an actual fall is obviously difficult to record during a scientific experiment.[6, 8-13] Consequently, Schwartze and colleagues opted for one of the next best options science has to offer: simulation with an able-bodied participant. They recorded body movements and knee impacts on the floor while mimicking several plausible falling scenarios. Then, they calculated the forces and moments that would be applied at four levels along the femur corresponding to amputation heights.[6, 8-11, 14-25] The overall forces applied during the falls were similar regardless of the amputation height indicating that the impact forces were simply translated along the femur. As expected, they showed that overall moments generally increased with amputation height due to changes in lever arm. This work demonstrates that devices preventing only against force overload do not require considering amputation height while those protecting against bending moments should. Another significant contribution is to provide, for the time, the magnitude of the impact load during different falls. This loading range is crucial to the overall design and, more precisely, the triggering threshold of protective devices. Unfortunately, the analysis of only a single able-bodied participant replicating falls limits greatly the generalisation of the findings. Nonetheless, this case study is an important milestone contributing to a better understanding of load impact during a fall. This new knowledge will improve the treatment, the safe ambulation and, ultimately, the quality of life of individuals fitted with bone-anchored prosthesis.

Bone-anchored prosthesis: Evidence that “gains” overcome the “pain”

Most of socket related discomforts leading to a significant decrease in quality of life of individuals with limb amputation can be overcome by surgical techniques enabling bone-anchored prostheses. To date, the OPRA two-stage procedure (i.e., S1, S2) is the most acknowledged treatment. However, surgical implantations of osseointegrated fixations are developing at an unprecedented pace worldwide.[1-18] Clearly, this option is becoming accessible to a wide range of individuals with limb amputations. The team led by Dr Rickard Branemark has published a number of landmark articles each focusing on a particular aspect (e.g., health related quality of life, functional outcomes, bone remodelling, infection rate). [1-3, 19-32] However, evidences presented in this prospective study are remarkable. Functional outcome, health-related quality of life and complications were considered concurrently for a large population (i.e., 51 participants) over an extended period of time (i.e., up to year follow up). Therefore, the “gain” and “pain” of the whole procedure were truly contrasted for the first time. The results confirmed that OPRA surgical and rehabilitation procedures improved significantly prosthetic use, mobility, global situation and fewer problems. Furthermore, the authors reported 47 episodes of infections for 63% (32) participants between post-op S1 and two years follow up. A total of 87% (41) were superficial infections recorded for 28 participants between post-op S2 and two years follow up, while 13% (6) were deep infections occurring for 4 participants during post-op S1 and S2. As expected, post-op S2 phase was the most prone to both infections. More importantly, the vast majority of infections were effectively treated with oral antibiotics. Clearly, this study provided definitive evidence that the benefits of OPRA fixation overcome complications. This article is also establishing reporting standards and benchmark data for future studies focusing on bone-anchored prostheses.

Prospective ten-month exercise intervention in premenarcheal girls: Positive effects on bone and lean mass

Enhancement of bone mineral acquisition during growth may be a useful preventive strategy against osteoporosis. The aim of this study was to explore the lean mass, strength, and bone mineral response to a 10-month, high-impact, strength-building exercise program in 71 premenarcheal girls, aged 9–10 years. Lean body mass, total body (TB), lumbar spine (LS), proximal femur (PF), and femoral neck (FN) bone mineral were measured using the Hologic QDR 2000+ bone densitometer. Strength was assessed using a grip dynamometer and the Cybex isokinetic dynamometer (Cybex II). At baseline, no significant difference in body composition, pubertal development, calcium intake, physical activity, strength, or bone mineral existed between groups. At completion, there were again no differences in height, total body mass, pubertal development, calcium intake, or external physical activity. In contrast, the exercise group gained significantly more lean mass, less body fat content, greater shoulder, knee and grip strength, and greater TB, LS, PF, and FN BMD (exercise: TB 3.5%, LS 4.8%, PF 4.5%, and FN 12.0%) compared with the controls (controls: TB 1.2%, LS 1.2%, PF 1.3%, and FN 1.7%). TB bone mineral content (BMC), LS BMC, PF BMC, FN BMC, LS bone mineral apparent density (BMAD), and FN bone area also increased at a significantly greater rate in the exercise group compared with the controls. In multiple regression analysis, change in lean mass was the primary determinant of TB, FN, PF, and LS BMD accrual. Although a large proportion of bone mineral accrual in the premenarcheal skeleton was related to growth, an osteogenic effect was associated with exercise. These results suggest that high-impact, strength building exercise is beneficial for premenarcheal strength, lean mass gains, and bone mineral acquisition.

The impact of intense training on endogenous estrogen and progesterone concentrations and bone mineral acquisition in adolescent rowers

The effect of 18 months of training on the ovarian hormone concentrations and bone mineral density (BMD) accrual was assessed longitudinally in 14 adolescent rowers and 10 matched controls, aged 14–15 years. Ovarian hormone levels were assessed by urinary estrone glucuronide (E1G) and pregnanediol glucuronide (PdG) excretion rates, classifying the menstrual cycles as ovulatory or anovulatory. Total body (TB), total proximal femur (PF), femoral neck (FN) and lumbar spine (LS) (L2–4) bone mass were measured at baseline and 18 months using dual-energy X-ray densitometry. Results were expressed as bone mineral content (BMC), BMD and bone mineral apparent density (BMAD). Five rowers had anovulatory menstrual cycles compared with zero prevalence for the control subjects. Baseline TB BMD was significantly higher in the ovulatory rowers, with PF BMD, FN BMD and LS BMD similar for all groups. At completion, the LS bone accrual of the ovulatory rowers was significantly greater (BMC 8.1%, BMD 6.2%, BMAD 6.2%) than that of the anovulatory rowers (BMC 1.1%, BMD 3.9%, BMAD 1.6%) and ovulatory controls (BMC 0.5%, BMD 1.1%, BMAD 1.1%). No difference in TB, PF or FN bone accrual was observed among groups. This study demonstrated an osteogenic response to mechanical loading, with the rowers accruing greater bone mass than the controls at the lumbar spine. However, the exercise-induced osteogenic benefits were less when rowing training was associated with low estrogen and progesterone metabolite excretion.

Changes in cervical spine bone mineral density in response to flight training

BACKGROUND High magnitude loads and unusual loading regimes are two important determinants for increasing bone mass. Past research demonstrated that positive Gz-induced loading, providing high loads in an unaccustomed manner, had an osteogenic effect on bone. Another determinant of bone mass is that the bone response to loading is site specific. This study sought to further investigate the site specific bone response to loading, examining the cervical spine response, the site suspected of experiencing the greatest loading, to high performance flight. METHODS Bone mineral density (BMD) and bone mineral content (BMC) was monitored in 9 RAAF trainee fighter pilots completing an 8-mo flight training course on a PC-9 and 10 age-height-weight-matched controls. RESULTS At completion of the course, the pilots had a significant increase in cervical spine BMD and total body BMC. No significant changes were found for the control group. CONCLUSIONS This study demonstrated that the physical environment associated with flight training may have contributed to a significant increase in cervical spine bone mass in the trainee PC-9 pilots. The increase in bone mass was possibly a response to the strain generated by the daily wearing of helmet and mask assembly under the influence of positive sustained accelerative forces.

A humanized tissue-engineered in vivo model to dissect interactions between human prostate cancer cells and human bone

Currently used xenograft models for prostate cancer bone metastasis lack the adequate tissue composition necessary to study the interactions between human prostate cancer cells and the human bone microenvironment. We introduce a tissue engineering approach to explore the interactions between human tumor cells and a humanized bone microenvironment. Scaffolds, seeded with human primary osteoblasts in conjunction with BMP7, were implanted into immunodeficient mice to form humanized tissue engineered bone constructs (hTEBCs) which consequently resulted in the generation of highly vascularized and viable humanized bone. At 12 weeks, PC3 and LNCaP cells were injected into the hTEBCs. Seven weeks later the mice were euthanized. Micro-CT, histology, TRAP, PTHrP and osteocalcin staining results reflected the different characteristics of the two cell lines regarding their phenotypic growth pattern within bone. Microvessel density, as assessed by vWF staining, showed that tumor vessel density was significantly higher in LNCaP injected hTEBC implants than in those injected with PC3 cells (p\0.001). Interestingly, PC3 cells showed morphological features of epithelial and mesenchymal phenotypes suggesting a cellular plasticity within this microenvironment. Taken together, a highly reproducible humanized model was established which is successful in generating LNCaP and PC3 tumors within a complex humanized bone microenvironment. This model simulates the conditions seen clinically more closely than any other model described in the literature to date and hence represents a powerful experimental platform that can be used in future work to investigate specific biological questions relevant to bone metastasis.

Bone-anchored prosthesis: Update in international developments

This symposium will provide hand-on update on the current development of the load sensors measuring the inner prosthetic loading that can strongly contribute the ever increasing demand for evidence-based clinical practice. Surgical implantations of osseointegrated fixations for bone-anchored prosthesis are developing at an unprecedented pace worldwide (e.g., Australia, UK, Sweden, US). This option is becoming accessible to a wide range of individuals with limb loss. With these new developments come new potential challenges and opportunities for all the stakeholders involved in the prosthetic care of these patients. Clearly, there is a need for those stakeholders, particularly those attending the ISPO, to be informed of the current and upcoming international developments in bone-anchored prostheses. The objectives of this symposium will be: • To present an overview of the current growth of the procedures worldwide (e.g., identification of key players, centers of activities, growth trend) with a strong focus on the introduction of the framework to evaluate the availability of the procedure at national level (e.g., number of patients treated, range of the levels of implantation, number of commercial fixations accessible), • To provide first-hand updates on the latest cutting-edge scientific and clinical developments of fixations and rehabilitations programs (e.g., Innovative design of implant, cost-effectiveness, long-terms rehabilitation outcomes for screw-type fixation, current developments in US, comparative analysis for press-fit type of implant, potential moves toward single-stage surgeries).

Tissue engineering of a morphologically and functionally intact humanized bone organ for bone metastasis research

The aim of this thesis was to establish an individualized, patient-specific diagnostic and therapeutic preclinical disease model for bone metastasis research. Tissue engineering of humanized bone within mice allowed the development of a humanized immune system in the host animal. This novel platform makes it possible to analyze the growth of human cancer cells in human bone in the presence of human immune cells.

High-throughput bone and cartilage micropellet manufacture, followed by assembly of micropellets into biphasic osteochondral tissue

Engineered biphasic osteochondral tissues may have utility in cartilage defect repair. As bone-marrow-derived mesenchymal stem/stromal cells (MSC) have the capacity to make both bone-like and cartilage-like tissues, they are an ideal cell population for use in the manufacture of osteochondral tissues. Effective differentiation of MSC to bone-like and cartilage-like tissues requires two unique medium formulations and this presents a challenge both in achieving initial MSC differentiation and in maintaining tissue stability when the unified osteochondral tissue is subsequently cultured in a single medium formulation. In this proof-of-principle study, we used an in-house fabricated microwell platform to manufacture thousands of micropellets formed from 166 MSC each. We then characterized the development of bone-like and cartilage-like tissue formation in the micropellets maintained for 8–14 days in sequential combinations of osteogenic or chondrogenic induction medium. When bone-like or cartilage-like micropellets were induced for only 8 days, they displayed significant phenotypic changes when the osteogenic or chondrogenic induction medium, respectively, was swapped. Based on these data, we developed an extended 14-day protocol for the pre-culture of bone-like and cartilage-like micropellets in their respective induction medium. Unified osteochondral tissues were formed by layering 12,000 osteogenic micropellets and 12,000 chondrogenic micropellets into a biphasic structure and then further culture in chondrogenic induction medium. The assembled tissue was cultured for a further 8 days and characterized via histology. The micropellets had amalgamated into a continuous structure with distinctive bone-like and cartilage-like regions. This proof-of-concept study demonstrates the feasibility of micropellet assembly for the formation of osteochondral-like tissues for possible use in osteochondral defect repair.

Increased intra-cortical porosity reduces bone stiffness and strength in pediatric patients with osteogenesis imperfecta

Osteogenesis imperfecta (OI) is a heritable disease occurring in one out of every 20,000 births. Although it is known that Type I collagen mutation in OI leads to increased bone fragility, the mechanism of this increased susceptibility to fracture is not clear. The aim of this study was to assess the microstructure of cortical bone fragments from patients with osteogenesis imperfecta (OI) using polarized light microscopy, and to correlate microstructural observations with the results of previously performed mechanical compression tests on bone from the same source. Specimens of cortical bone were harvested from the lower limbs of three (3) OI patients at the time of surgery, and were divided into two groups. Group 1 had been subjected to previous micro-mechanical compression testing, while Group 2 had not been subjected to any prior testing. Polarized light microscopy revealed disorganized bone collagen architecture as has been previously observed, as well as a large increase in the areal porosity of the bone compared to typical values for healthy cortical bone, with large (several hundred micron sized), asymmetrical pores. Importantly, the areal porosity of the OI bone samples in Group 1 appears to correlate strongly with their previously measured apparent Young's modulus and compressive strength. Taken together with prior nanoindentation studies on OI bone tissue, the results of this study suggest that increased intra-cortical porosity is responsible for the reduction in macroscopic mechanical properties of OI cortical bone, and therefore that in vivo imaging modalities with resolutions of ~ 100 μm or less could potentially be used to non-invasively assess bone strength in OI patients. Although the number of subjects in this study is small, these results highlight the importance of further studies in OI bone by groups with access to human OI tissue in order to clarify the relationship between increased porosity and reduced macroscopic mechanical integrity.

AICAR administration affects glucose metabolism by upregulating the novel glucose transporter, GLUT8, in equine skeletal muscle

Equine metabolic syndrome is characterized by obesity and insulin resistance (IR). Currently, there is no effective pharmacological treatment for this insidious disease. Glucose uptake is mediated by a family of glucose transporters (GLUT), and is regulated by insulin-dependent and -independent pathways, including 5-AMP-activated protein kinase (AMPK). Importantly, the activation of AMPK, by 5-aminoimidazole- 4-carboxamide-1-D-ribofuranoside (AICAR) stimulates glucose uptake in both healthy and diabetic humans. However, whether AICAR promotes glucose uptake in horses has not been established. It is hypothesized that AICAR administration would enhance glucose transport in equine skeletal muscle through AMPK activation. In this study, the effect of an intravenous AICAR infusion on blood glucose and insulin concentrations, as well as on GLUT expression and AMPK activation in equine skeletal muscle (quantified by Western blotting) was examined. Upon administration, plasma AICAR rapidly reached peak concentration. Treatment with AICAR resulted in a decrease (P < 0.05) in blood glucose and an increase (P < 0.05) in insulin concentration without a change in lactate concentration. The ratio of phosphorylated to total AMPK was increased (P < 0.05) in skeletal muscle. While GLUT4 and GLUT1 protein expression remained unchanged, GLUT8 was increased (P < 0.05) following AICAR treatment. Up-regulation of GLUT8 protein expression by AICAR suggests that this novel GLUT isoform plays an important role in equine muscle glucose transport. In addition, the data suggest that AMPK activation enhances pancreatic insulin secretion. Collectively, the findings suggest that AICAR acutely promotes muscle glucose uptake in healthy horses and thus its therapeutic potential for managing IR requires investigation.

Oldest pathology in a tetrapod bone illuminates the origin of terrestrial vertebrates

The origin of terrestrial tetrapods was a key event in vertebrate evolution, yet how and when it occurred remains obscure, due to scarce fossil evidence. Here, we show that the study of palaeopathologies, such as broken and healed bones, can help elucidate poorly understood behavioural transitions such as this. Using high-resolution finite element analysis, we demonstrate that the oldest known broken tetrapod bone, a radius of the primitive stem tetrapod Ossinodus pueri from the mid-Viséan (333 million years ago) of Australia, fractured under a high-force, impact-type loading scenario. The nature of the fracture suggests that it most plausibly occurred during a fall on land. Augmenting this are new osteological observations, including a preferred directionality to the trabecular architecture of cancellous bone. Together, these results suggest that Ossinodus, one of the first large (>2m length) tetrapods, spent a significant proportion of its life on land. Our findings have important implications for understanding the temporal, biogeographical and physiological contexts under which terrestriality in vertebrates evolved. They push the date for the origin of terrestrial tetrapods further back into the Carboniferous by at least two million years. Moreover, they raise the possibility that terrestriality in vertebrates first evolved in large tetrapods in Gondwana rather than in small European forms, warranting a re-evaluation of this important evolutionary event.

Light history-dependent respiration explains the hysteresis in the daily ecosystem metabolism of seagrass

Oxygen flux between aquatic ecosystems and the water column is a measure of ecosystem metabolism. However, the oxygen flux varies during the day in a “hysteretic” pattern: there is higher net oxygen production at a given irradiance in the morning than in the afternoon. In this study, we investigated the mechanism responsible for the hysteresis in oxygen flux by measuring the daily pattern of oxygen flux, light, and temperature in a seagrass ecosystem (Zostera muelleri in Swansea Shoals, Australia) at three depths. We hypothesised that the oxygen flux pattern could be due to diel variations in either gross primary production or respiration in response to light history or temperature. Hysteresis in oxygen flux was clearly observed at all three depths. We compared this data to mathematical models, and found that the modification of ecosystem respiration by light history is the best explanation for the hysteresis in oxygen flux. Light history-dependent respiration might be due to diel variations in seagrass respiration or the dependence of bacterial production on dissolved organic carbon exudates. Our results indicate that the daily variation in respiration rate may be as important as the daily changes of photosynthetic characteristics in determining the metabolic status of aquatic ecosystems.

Genome-wide association study using extreme truncate selection identifies novel genes affecting bone mineral density and fracture risk

Osteoporotic fracture is a major cause of morbidity and mortality worldwide. Low bone mineral density (BMD) is a major predisposing factor to fracture and is known to be highly heritable. Site-, gender-, and age-specific genetic effects on BMD are thought to be significant, but have largely not been considered in the design of genome-wide association studies (GWAS) of BMD to date. We report here a GWAS using a novel study design focusing on women of a specific age (postmenopausal women, age 55-85 years), with either extreme high or low hip BMD (age- and gender-adjusted BMD z-scores of +1.5 to +4.0, n = 1055, or -4.0 to -1.5, n = 900), with replication in cohorts of women drawn from the general population (n = 20,898). The study replicates 21 of 26 known BMD-associated genes. Additionally, we report suggestive association of a further six new genetic associations in or around the genes CLCN7, GALNT3, IBSP, LTBP3, RSPO3, and SOX4, with replication in two independent datasets. A novel mouse model with a loss-of-function mutation in GALNT3 is also reported, which has high bone mass, supporting the involvement of this gene in BMD determination. In addition to identifying further genes associated with BMD, this study confirms the efficiency of extreme-truncate selection designs for quantitative trait association studies. © 2011 Duncan et al.