Assessment of age-dependent sexual dimorphism in paediatric vertebral size and density using a statistical shape and statistical appearance modelling approach.

This work focuses on the growth patterns of the human fourth lumbar vertebra (L4) in a paediatric population, with specific attention to sexual dimorphism. The study aims to understand morphological and density changes in the vertebrae through age-dependent statistical shape and statistical appearance models, which can describe full three-dimensional anatomy. Results show that the main growth patterns are associated with isotropic volumetric vertebral growth, a decrease in the relative size of the vertebral foramen, and an increase in the length of the transverse processes. Moreover, significant sexual dimorphism was demonstrated during puberty. We observe significant age and sex interaction in the anterior vertebral body height (P = 0.005), where females exhibited an earlier increase in rates of vertebral height evolution. Moreover, we also observe an increase in cross-sectional area (CSA) with age (P = 0.020), where the CSA is smaller in females than in males (significant sex effect P = 0.042). Finally, although no significant increase in trabecular bone density with age is observed (P = 0.363), a trend in the statistical appearance model suggests an increase in density with age.

Anatomical variation is the norm: A novel curriculum framework.

Anatomical variation is an inherent part of every health curriculum, due in large to the negative clinical consequences that can ensue if anatomical variation is not thoroughly understood. However, current literature fails to describe any structured whole-of-course pedagogy for the teaching of anatomical variation in higher education. This study therefore aimed to (i) propose a whole-of-course curriculum framework to guide academic development and implementation of anatomical variation resources and assessment; and (ii) assess the depth of anatomical variation knowledge in a multiyear undergraduate health-science cohort (n = 152) at the Queensland University of Technology. Current anatomical variation pedagogy, and subsequently the need for the curriculum framework, were explored using a scoping review protocol. As part of this study, anatomical variation was novelly defined as macroscopic differences in morphology (shape and size), topography (location), developmental timing, or frequency (number) of an anatomical structure between individuals that form during embryological or subadult development and result in no substantive, observable interruption to physiological function. The framework incorporated three themes of anatomical variation learning outcomes: description of anatomical variation, theories of the professional implications of variation, and investigation of variant formation. These themes were strongly aligned with the concepts recommended for teaching identified through the scoping review. Significant deficits in anatomical variation student knowledge were identified, with the third-year cohort recording a mean total score of only 54.6%. A strong recommendation to implement the anatomical variation curriculum framework in all medical and health-science curricula is made to intentionally develop student understanding of anatomical variation and improve future clinical practice.

Identifying Calcaneal Anatomical Regions of Interest (ROI) for Quantitative Ultrasound Application in Subadults.

Whilst quantitative ultrasound can be reliably used to assess bone health in adults, the fixed location of the transducers in current devices may result in inaccurate and unreliable measurements in bone assessment in children due to the variation in foot size during growth. To improve positioning for paediatric assessment, Jaworski et al. (1995) created an anatomical method to identify the region of interest (ROI), however, there have been no medical imaging studies to confirm that the Jaworski method results in consistent placement of the transducer on the centre of the calcaneal body to avoid edge artefacts. In this study, computed tomography scans of the tarsus were collected from 498 individuals (258 females; 240 males) aged 2 to 20 years and used to create three novel anatomical methods to identify ROI on the calcaneus using palpable landmarks. In addition, the established Jaworski method was applied to the same scans and compared to our novel methods. The maximum ROI significantly increased with age with males having significantly greater diameters, supporting the recommendation that ½ inch diameter transducers should be used on individuals younger than 7 years of age. We identified that 79% of the 'Jaworski points' lied anterosuperior to the ROI centre point identified in this study, with 10% of the points lying outside the ROI. Of the three novel methods, only the calcaneal insertion method demonstrated small measurement variance between individuals of the same age in each sex and is therefore the preferred method for ultrasound clinical application.

Quantifying the ossification and fusion of the calcaneal apophysis using computed tomography.

Knowledge of the anatomical development of the calcaneal apophysis is essential in clinical assessment and management of both paediatric and sub-adult patients presenting with heel pain. Despite this, the current understanding of calcaneal apophyseal development is constrained by the limitations of the imaging modalities used to examine the apophysis, with no current literature reporting the development of the medial and lateral processes. This study aimed to overcome these limitations by investigating the ossification and fusion of the calcaneal apophysis using three-dimensional computed tomography analysis, and statistically predicting the apophyseal developmental stage in contemporary Australian children. The development and fusion status of the apophysis was scored using a novel 11-stage scoring system on 568 multi-slice computed tomography scans (295 females; 274 males) and 266 lateral radiographic scans (119 females; 147 males) from the Queensland Children's Hospital. Multinomial logistic regression along with classification tables and predictive probabilities were then utilised to assess developmental stage likelihood from known age and sex. The apophysis commenced ossification at a mean age of 5.2 years for females and 7.2 years for males, and then elongated to form the apophyseal cap around 10 years for females and 12.4 years for males. Fusion of the apophysis commenced at a mean age of 11.18 years for females and 13.3 years for males, with the earliest age of complete fusion observed at 10 years for females and 14 years for males. The results demonstrate significant sexual dimorphism in ossification and fusion with females developing and fusing significantly earlier. Furthermore, the use of computed tomography in this study allowed for the first time evaluation of the ossification and fusion of the medial and lateral processes of the calcaneus. The medial process formed at a mean age of 9.5 years for females and 10.9 years for males while the lateral process formed at around 9.8 years for females and 11.7 years for males. The medial process demonstrated slower rates of fusion compared to the lateral process. The present study provides Queensland specific standards for assessing the calcaneal apophyseal developmental stage as well as novel predictive regression models for apophyseal stage estimation using known age and sex to aid in the diagnosis of heel pain conditions such as apophysitis or screen for developmental delays in children and subadults.

Temporal mapping of the closure of the anterior fontanelle and contiguous sutures using computed tomography, in silico models of modern infants.

The aim of this study is to quantify and statistically model the age-related decline in the fibrous connective tissue interface of the anterior fontanelle in modern Australian infants, using three-dimensional, semi-automated computed-assisted design protocols. Non-linear regression with variance models, using power functions, combined with quantile regression of the 5th and 95th population percentiles, were utilised to assess absolute anterior fontanelle surface area (AFSA) as a function of age, using multi-slice cranial computed tomography scans obtained from 256 infants aged < 30 months (males: n = 126, females: n = 109) from Brisbane children's hospitals. Normalised AFSA (NFSA), standardised for variation in cephalic size, followed a progressive decline from birth, the greatest velocity change occurring between the 3-6 and 6-9 month cohorts. Growth of the neurocranium is the most significant within the first 8 months postpartum, with a mean increase of 19.03 mm in maximum cranial length and 10.04 mm in breadth. Directionality of fontanelle closure, quantified using spline curves refutes fundamental assumptions that the anterior fontanelle is consistent with a quadrilateral, and contiguous sutures exhibit constant velocity of closure. The present study provides normative values for fontanelle size and diameters as well as new predictive non-linear models for age substantiation, screening of developmental abnormalities and indicators of suspected child maltreatment in modern infants aged birth to 30 months.

Partnering for Professional Development in the Galápagos Islands.

Nurses living in the Galápagos Islands face barriers to accessing continuing professional development (CPD), particularly in their native language and appropriate for their unique island culture due to their remote location, 1,000 km off the coast of Ecuador. This article reviews the evidence on providing professional development from high-resource countries to low-resource countries; describes a case example of the process of developing, implementing, and evaluating the first professional development program targeting nurses in the Galápagos; and presents recommendations to sustain a culturally relevant international professional nursing development partnership that can be a model for nurses in other remote locations with limited educational access. [J Contin Educ Nurs. 2019;50(9):417-422.].

Differential growth patterns of the abdominal aorta and vertebrae during childhood.

The adult vertebral level of the splanchnic branches of the abdominal aorta relies on a complex series of fusion and regression steps during embryological development, such that variation is common. Little is known however regarding the anatomy of the abdominal aorta in children. This study aimed to investigate the spatial relationship between the abdominal aorta and the vertebral column during childhood development to inform clinical management of pediatric patients. Retrospective multislice computed tomography abdominopelvic angiograms of children aged neonate to 19 years (n = 232) were used to examine vertebral levels of the celiac trunk (CoT), superior mesenteric artery (SMA), inferior mesenteric artery (IMA), and aortic bifurcation (AB) using multiplanar formatting views in OsiriX. The abdominal aorta length, AB angle, and displacement of the aorta from the midline were quantified with the effect of age and sex analyzed using multinomial logistic regression and general linear models. The most frequent origins of CoT, SMA, IMA, and AB were T12, L1, L3, and L4, respectively, with significant variation in vertebral level for each vessel. SMA level was significantly more proximal with age, and CoT and AB demonstrated marked sex differences in vertebral level. As the age of the child increased, AB angle decreased, aortic displacement increased, and the length of the abdominal aorta increased at a slower velocity to the vertebral column (P < 0.001). Our study highlights the variation of the location and geometry of the abdominal aorta in children; this knowledge will positively impact pediatric surgical approaches and endovascular procedures. Clin. Anat. 32:783-793, 2019. © 2019 Wiley Periodicals, Inc.

Assessing the fusion of the ischiopubic synchondrosis using predictive modeling.

Flaring of the ischiopubic synchondrosis at the time of fusion is a common clinical observation in pediatrics and represents a normal physiological process in skeletal maturation. When presenting unilaterally, this flaring can mimic a range of serious pathological conditions such as osteomyelitis, osteal tumors, and traumatic injury. An improved understanding of ischiopubic synchondrosis fusion is therefore critical to avoid potential misdiagnosis. Retrospective multi-slice computed tomography pelvic scans of Australian individuals aged neonate to 24 years (n = 184) were assessed using a novel five stage morphological classification system of the maturation and fusion of the ischiopubic synchondrosis. Maturation scoring was conducted using both multiplanar formatting views and volume-rendered reconstructions in OsiriX™. Maturational stage was strongly related to age (P < 0.001) with fusion of the ischiopubic synchondrosis observed between the ages of 4 and 9 years in females and 7 and 13 years for males. The highest probability of fusion in our Queensland Australian population based on multinomial regression predictive modeling was between 7 and 10 years of age. We documented three variants of fusion: pubic and ischial outgrowths, appearance of a secondary ossification center, and a fusiform-shaped enlargement. This study provides the first predictive modeling of the timing of fusion of the ischiopubic synchondrosis using a reliable morphological classification system. The significant variation in timing and progression of fusion of the ischiopubic synchondrosis reported in this study, will aid in minimizing misdiagnosis and unnecessary treatment in children presenting with asymmetrical or delayed ischiopubic synchondrosis anomalies. Clin. Anat. 32:851-859, 2019. © 2019 Wiley Periodicals, Inc.

Limited short-term effects on human prostate cancer xenograft growth and epidermal growth factor receptor gene expression by the ghrelin receptor antagonist [D-Lys3]-GHRP-6.

PURPOSE: The ghrelin axis regulates many physiological functions (including appetite, metabolism, and energy balance) and plays a role in disease processes. As ghrelin stimulates prostate cancer proliferation, the ghrelin receptor antagonist [D-Lys3]-GHRP-6 is a potential treatment for castrate-resistant prostate cancer and for preventing the metabolic consequences of androgen-targeted therapies. We therefore explored the effect of [D-Lys3]-GHRP-6 on PC3 prostate cancer xenograft growth. METHODS: NOD/SCID mice with PC3 prostate cancer xenografts were administered 20 nmoles/mouse [D-Lys3]-GHRP-6 daily by intraperitoneal injection for 14 days and tumour volume and weight were measured. RNA sequencing of tumours was conducted to investigate expression changes following [D-Lys3]-GHRP-6 treatment. A second experiment, extending treatment time to 18 days and including a higher dose of [D-Lys3]-GHRP-6 (200 nmoles/mouse/day), was undertaken to ensure repeatability. RESULTS: We demonstrate here that daily intraperitoneal injection of 20 nmoles/mouse [D-Lys3]-GHRP-6 reduces PC3 prostate cancer xenograft tumour volume and weight in NOD/SCID mice at two weeks post treatment initiation. RNA-sequencing revealed reduced expression of epidermal growth factor receptor (EGFR) in these tumours. Further experiments demonstrated that the effects of [D-Lys3]-GHRP-6 are transitory and lost after 18 days of treatment. CONCLUSIONS: We show that [D-Lys3]-GHRP-6 has transitory effects on prostate xenograft tumours in mice, which rapidly develop an apparent resistance to the antagonist. Although further studies on [D-Lys3]-GHRP-6 are warranted, we suggest that daily treatment with the antagonist is not a suitable treatment for advanced prostate cancer.

No effect of unacylated ghrelin administration on subcutaneous PC3 xenograft growth or metabolic parameters in a Rag1-/- mouse model of metabolic dysfunction.

Ghrelin is a peptide hormone which, when acylated, regulates appetite, energy balance and a range of other biological processes. Ghrelin predominately circulates in its unacylated form (unacylated ghrelin; UAG). UAG has a number of functions independent of acylated ghrelin, including modulation of metabolic parameters and cancer progression. UAG has also been postulated to antagonise some of the metabolic effects of acyl-ghrelin, including its effects on glucose and insulin regulation. In this study, Rag1-/- mice with high-fat diet-induced obesity and hyperinsulinaemia were subcutaneously implanted with PC3 prostate cancer xenografts to investigate the effect of UAG treatment on metabolic parameters and xenograft growth. Daily intraperitoneal injection of 100 µg/kg UAG had no effect on xenograft tumour growth in mice fed normal rodent chow or 23% high-fat diet. UAG significantly improved glucose tolerance in host Rag1-/- mice on a high-fat diet, but did not significantly improve other metabolic parameters. We propose that UAG is not likely to be an effective treatment for prostate cancer, with or without associated metabolic syndrome.

Bayesian modeling predicts age and sex are not required for accurate stature estimation from femoral length.

Despite the recognized flaws in applying traditional stature estimation equations such as those of Trotter and Gleser (1952) [5] to a contemporary population, there are currently no available alternatives for stature estimation in Australia that address these limitations. Post mortem computed tomography (PMCT) DICOM scans of the left and right femora were acquired from 76 Australian deceased individuals aged 17-76 years for metric analysis. Femoral bicondylar length, femoral epicondylar breadth and anterior-posterior (AP) diameter, medial-lateral (ML) diameter, circumference and cortical area at the femoral midshaft were measured on three-dimensional (3D) models to build statistical models for estimating stature. In addition, Australian individuals aged 16-63 years (n=111) were measured in standing and supine positions to aid in the adjustment of supine stature of deceased individuals utilized in this study to standing stature. The results of this preliminary evaluation strongly indicate that the optimal model for estimating stature includes bicondylar femoral length and epicondylar breadth, that the effect of sex as an independent variable is very low, and there is limited practical benefit in including age in the estimation of stature. Our study indicates that the Australian population sampled represents a small yet significant shift in stature from the original Trotter and Gleser sample. Additionally, in the case of fragmentary remains, it was found that epicondylar breadth and AP diameter had the highest probability of accurate stature estimation in the absence of bicondylar femoral length. As stature forms a significant component of a biological profile and therefore aids in the personal identification of human remains, it is important that forensic anthropologists utilize the most accurate methodologies available. Stature estimation of Australian individuals is therefore achieved with higher accuracy through utilizing the femoral equations proposed in this study.

Standardized anthropological measurement of postcranial bones using three-dimensional models in CAD software.

This study introduces a standardized protocol for conducting linear measurements of postcranial skeletal elements using three-dimensional (3D) models constructed from post-mortem computed tomography (PMCT) scans. Using femoral DICOM datasets, reference planes were generated and plane-to-plane measurements were conducted on 3D surface rendered models. Bicondylar length, epicondylar breadth, anterior-posterior (AP) diameter, medial-lateral (ML) diameter and cortical area at the midshaft were measured by four observers to test the measurement error variance and observer agreement of the protocol (n=6). Intra-observer error resulted in a mean relative technical error of measurement (%TEM) of 0.11 and an intraclass correlation coefficient (ICC) of 0.999 (CI=0.998-1.000); inter-observer error resulted in a mean %TEM of 0.54 and ICC of 0.996 (CI=0.979-1.000) for bicondylar length. Epicondylar breadth, AP diameter, ML diameter and cortical area also yielded minimal error. Precision testing demonstrated that the approach is highly repeatable and is recommended for implementation in anthropological investigation and research. This study exploits the benefits of virtual anthropology, introducing an innovative, standardized alternative to dry bone osteometric measurements.

Near-field fault slip of the 2016 Vettore Mw 6.6 earthquake (Central Italy) measured using low-cost GNSS.

The temporal evolution of slip on surface ruptures during an earthquake is important for assessing fault displacement, defining seismic hazard and for predicting ground motion. However, measurements of near-field surface displacement at high temporal resolution are elusive. We present a novel record of near-field co-seismic displacement, measured with 1-second temporal resolution during the 30th October 2016 Mw 6.6 Vettore earthquake (Central Italy), using low-cost Global Navigation Satellite System (GNSS) receivers located in the footwall and hangingwall of the Mt. Vettore - Mt. Bove fault system, close to new surface ruptures. We observe a clear temporal and spatial link between our near-field record and InSAR, far-field GPS data, regional measurements from the Italian Strong Motion and National Seismic networks, and field measurements of surface ruptures. Comparison of these datasets illustrates that the observed surface ruptures are the propagation of slip from depth on a surface rupturing (i.e. capable) fault array, as a direct and immediate response to the 30th October earthquake. Large near-field displacement ceased within 6-8 seconds of the origin time, implying that shaking induced gravitational processes were not the primary driving mechanism. We demonstrate that low-cost GNSS is an accurate monitoring tool when installed as custom-made, short-baseline networks.

Insights from engraftable immunodeficient mouse models of hyperinsulinaemia.

Hyperinsulinaemia, obesity and dyslipidaemia are independent and collective risk factors for many cancers. Here, the long-term effects of a 23% Western high-fat diet (HFD) in two immunodeficient mouse strains (NOD/SCID and Rag1 -/-) suitable for engraftment with human-derived tissue xenografts, and the effect of diet-induced hyperinsulinaemia on human prostate cancer cell line xenograft growth, were investigated. Rag1 -/-and NOD/SCID HFD-fed mice demonstrated diet-induced impairments in glucose tolerance at 16 and 23 weeks post weaning. Rag1 -/- mice developed significantly higher fasting insulin levels (2.16 ± 1.01 ng/ml, P = 0.01) and increased insulin resistance (6.70 ± 1.68 HOMA-IR, P = 0.01) compared to low-fat chow-fed mice (0.71 ± 0.12 ng/ml and 2.91 ± 0.42 HOMA-IR). This was not observed in the NOD/SCID strain. Hepatic steatosis was more extensive in Rag1 -/- HFD-fed mice compared to NOD/SCID mice. Intramyocellular lipid storage was increased in Rag1 -/- HFD-fed mice, but not in NOD/SCID mice. In Rag1 -/- HFD-fed mice, LNCaP xenograft tumours grew more rapidly compared to low-fat chow-fed mice. This is the first characterisation of the metabolic effects of long-term Western HFD in two mouse strains suitable for xenograft studies. We conclude that Rag1 -/- mice are an appropriate and novel xenograft model for studying the relationship between cancer and hyperinsulinaemia.

Apophyseal Ossification of the Iliac Crest in Forensic Age Estimation: Computed Tomography Standards for Modern Australian Subadults.

This study contrasts the ontogeny of the iliac crest apophysis using conventional radiography and multislice computed tomography (MSCT), providing probabilistic information for age estimation of modern Australian subadults. Retrospective abdominopelvic MSCT data acquired from 524 Australian individuals aged 7-25 and surveillance radiographs of adolescent idiopathic scoliosis patients included in the Paediatric Spine Research Group Progression Study (n = 531) were assessed. Ossification scoring of pseudo-radiographs and three-dimensional (3D) volume-rendered reconstructions using Risser (1958) quantitative descriptors indicate discrepancies in age estimates, stage allocation, and conflicting morphological progression. To mitigate visualization limitations associated with two-dimensional radiographs, we provide and validate a modified 3D-MSCT scoring tier of ossification, demonstrating complete fusion between 17.3-19.2 and 17.1-20.1 years in males and females. Legal demarcation for doli incapax presumption and age of majority (18 years) can be achieved using probability estimates from a fitted cumulative probit model for apophyseal fusion using the recalibrated standards.

Effect of Head Position on Facial Soft Tissue Depth Measurements Obtained Using Computed Tomography.

Facial soft tissue depth (FSTD) studies employing clinical computed tomography (CT) data frequently rely on depth measurements from raw 2D orthoslices. However, the position of each patient's head was not standardized in this method, potentially decreasing measurement reliability and accuracy. This study measured FSTDs along the original orthoslice plane and compared these measurements to those standardized by the Frankfurt horizontal (FH). Subadult cranial CT scans (n = 115) were used to measure FSTDs at 18 landmarks. Significant differences were observed between the methods at eight of these landmarks (p < 0.05), demonstrating that high-quality data are not generated simply by employing modern imaging modalities such as CT. Proper technique is crucial to useful results, and maintaining control over head position during FSTD data collection is important. This is easily and most readily achieved in CT techniques by rotating the head to the FH plane after constructing a 3D rendering of the data.

Introducing Computed Tomography Standards for Age Estimation of Modern Australian Subadults Using Postnatal Ossification Timings of Select Cranial and Cervical Sites(.).

Contemporary, population-specific ossification timings of the cranium are lacking in current literature due to challenges in obtaining large repositories of documented subadult material, forcing Australian practitioners to rely on North American, arguably antiquated reference standards for age estimation. This study assessed the temporal pattern of ossification of the cranium and provides recalibrated probabilistic information for age estimation of modern Australian children. Fusion status of the occipital and frontal bones, atlas, and axis was scored using a modified two- to four-tier system from cranial/cervical DICOM datasets of 585 children aged birth to 10 years. Transition analysis was applied to elucidate maximum-likelihood estimates between consecutive fusion stages, in conjunction with Bayesian statistics to calculate credible intervals for age estimation. Results demonstrate significant sex differences in skeletal maturation (p < 0.05) and earlier timings in comparison with major literary sources, underscoring the requisite of updated standards for age estimation of modern individuals.

Ontogeny of the spheno-occipital synchondrosis in a modern Queensland, Australian population using computed tomography.

Due to disparity regarding the age at which skeletal maturation of the spheno-occipital synchondrosis occurs in forensic and biological literature, this study provides recalibrated multislice computed tomography (MSCT) age standards for the Australian (Queensland) population, using a Bayesian statistical approach. The sample comprises retrospective cranial/cervical MSCT scans obtained from 448 males and 416 females aged birth to 20 years from the Skeletal Biology and Forensic Anthropology Research Osteological Database. Fusion status of the synchondrosis was scored using a modified six-stage scoring tier on an MSCT platform, with negligible observer error (κ = 0.911 ± 0.04, intraclass correlation coefficient = 0.994). Bayesian transition analysis indicates that females are most likely to transition to complete fusion at 13.1 years and males at 15.6 years. Posterior densities were derived for each morphological stage, with complete fusion of the synchondrosis attained in all Queensland males over 16.3 years of age and females aged 13.8 years and older. The results demonstrate significant sexual dimorphism in synchondrosis fusion and are suggestive of intrapopulation variation between major geographic regions in Australia. This study contributes to the growing repository of contemporary anthropological standards calibrated for the Queensland milieu to improve the efficacy of the coronial process for medicolegal death investigation. As a stand-alone age indicator, the basicranial synchondrosis may be consulted as an exclusion criterion when determining the age of majority that constitutes 17 years in Queensland forensic practice.

Fracture healing via periosteal callus formation requires macrophages for both initiation and progression of early endochondral ossification.

The distribution, phenotype, and requirement of macrophages for fracture-associated inflammation and/or early anabolic progression during endochondral callus formation were investigated. A murine femoral fracture model [internally fixed using a flexible plate (MouseFix)] was used to facilitate reproducible fracture reduction. IHC demonstrated that inflammatory macrophages (F4/80(+)Mac-2(+)) were localized with initiating chondrification centers and persisted within granulation tissue at the expanding soft callus front. They were also associated with key events during soft-to-hard callus transition. Resident macrophages (F4/80(+)Mac-2(neg)), including osteal macrophages, predominated in the maturing hard callus. Macrophage Fas-induced apoptosis transgenic mice were used to induce macrophage depletion in vivo in the femoral fracture model. Callus formation was completely abolished when macrophage depletion was initiated at the time of surgery and was significantly reduced when depletion was delayed to coincide with initiation of early anabolic phase. Treatment initiating 5 days after fracture with the pro-macrophage cytokine colony stimulating factor-1 significantly enhanced soft callus formation. The data support that inflammatory macrophages were required for initiation of fracture repair, whereas both inflammatory and resident macrophages promoted anabolic mechanisms during endochondral callus formation. Overall, macrophages make substantive and prolonged contributions to fracture healing and can be targeted as a therapeutic approach for enhancing repair mechanisms. Thus, macrophages represent a viable target for the development of pro-anabolic fracture treatments with a potentially broad therapeutic window.

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.