Bone histomorphometry as a diagnostic tool. A review article

INTRODUCTION:Various diagnostic modalities are used in the study of bone structure and metabolism. These include radiological examinations, laboratory and biochemical testing, histological and histomorphometric assessments, immunohistochemistry, and some non-invasive techniques. Bone histomorphometry is regarded as the gold standard in the diagnostics of bone-related conditions. It is a reliable method for detailed quantitative assessment of bone microarchitecture and physiology. It allows observation of cell types and their activity and provides essential information on bone healing, modeling, and remodeling.    AIM:The present review aims to summarize the applications and limitations of bone histomorphometry and define its role in the diagnostics, monitoring, and treatment of various bone-related conditions. MATERIALS AND METHODS:An electronic search using Google Scholar, PubMed, Scopus, and ScienceDirect was conducted up to July 2022. The article is based on the existing scientific database and includes 198 studies. It summarizes the current knowledge on bone histomorphometry, highlights its advantages and limitations, and gives some recommendations for further research.RESULTS: Bone histomorphometry is a key diagnostic tool in the field of bone research. It is used for the detection and monitoring of metabolic bone diseases, for establishing the safety of the pharmaceutical agents that affect bone metabolism, and for the effects of different biomaterials, used for guided bone regeneration and implant treatment.CONCLUSION:Bone histomorphometry is applied in various scientific fields. Although some innovative non-invasive techniques have been suggested, the method remains a significant component in the study of bone structure and physiology

RELATIONSHIPS OF LONG-TERM BISPHOSPHONATE TREATMENT WITH MEASURES OF BONE MICROARCHITECTURE AND MECHANICAL COMPETENCE

Oral bisphosphonate drug therapy is a common and effective treatment for osteoporosis. Little is known about the long-term effects of bisphosphonates on bone quality. This study examined the structural and mechanical properties of trabecular bone following 0-16 years of bisphosphonate treatment. Fifty-three iliac crest bone samples of Caucasian women diagnosed with low turnover osteoporosis were identified from the Kentucky Bone Registry. Forty-five were treated with oral bisphosphonates for 1 to 16 years while eight were treatment naive. A section of trabecular bone was chosen from a micro-computed tomography (Scanco µCT 40) scan of each sample for a uniaxial linearly elastic compression simulation using finite element analysis (ANSYS 14.0). Morphometric parameters (BV/TV, SMI, Tb.Sp., etc.) were computed using µCT. Apparent modulus, effective modulus and estimated failure stress were calculated. Biomechanical and morphometric parameters improved with treatment duration, peaked around 7 years, and then declined independently of age. The findings suggest a limit to the benefits associated with bisphosphonate treatment and that extended continuous bisphosphonate treatment does not continue to improve bone quality. Bone quality, and subsequently bone strength, may eventually regress to a state poorer than at the onset of treatment. Treatment duration limited to less than 7 years is recommended

An Optimized Approach to Perform Bone Histomorphometry

Bone histomorphometry allows quantitative evaluation of bone micro-architecture, bone formation, and bone remodeling by providing an insight to cellular changes. Histomorphometry plays an important role in monitoring changes in bone properties because of systemic skeletal diseases like osteoporosis and osteomalacia. Besides, quantitative evaluation plays an important role in fracture healing studies to explore the effect of biomaterial or drug treatment. However, until today, to our knowledge, bone histomorphometry remain time-consuming and expensive. This incited us to set up an open-source freely available semi-automated solution to measure parameters like trabecular area, osteoid area, trabecular thickness, and osteoclast activity. Here in this study, the authors present the adaptation of Trainable Weka Segmentation plugin of ImageJ to allow fast evaluation of bone parameters (trabecular area, osteoid area) to diagnose bone related diseases. Also, ImageJ toolbox and plugins (BoneJ) were adapted to measure osteoclast activity, trabecular thickness, and trabecular separation. The optimized two different scripts are based on ImageJ, by providing simple user-interface and easy accessibility for biologists and clinicians. The scripts developed for bone histomorphometry can be optimized globally for other histological samples. The showed scripts will benefit the scientific community in histological evaluation

Bone marrow Fat - A Novel Quantification Method and Potential Clinical Applications

Ageing bone is characterised by increased marrow fat infiltration altering its composition and microstructure, thus predisposing the person to osteoporosis. Yet to date, non-invasive quantifications of marrow fat are limited to special MRI techniques, and clinical studies examining marrow fat in the ageing skeleton are scarce. Thus, the key aims of this thesis are to: · Validate a new non-invasive technique of marrow fat quantification using CT technology · Determine the effects of dietary fatty acids on marrow fat · Measure marrow fat content in different skeletal regions in healthy older men · Determine the effect of exercise and calcium on marrow fat. The imaging techniques employed in our animal and human studies were micro CT (µCT) and quantitative CT (QCT) respectively. All images were analysed with the imaging software Slice O Matic version 4.1 (Tomovision). Regions of interest [ROIs] were Volumes of interests (VOIs) of bone, fat and blood measured in µm3 or mm3. Individual tissue volumes, expressed as percentages of the total marrow volume, and ratios of tissue volumes were also used in the analysis. Global and local thresholds for individual tissue volumes were determined separately for µCT and QCT. Thresholds for µCT were those derived from the initial validation study, whereas those for QCT were based on previous published data. To account for partial volume averaging effects, further manual refinement of threshold ranges were undertaken by inspection of individual pixels and their neighbours. This manual process was carried out for both µCT and QCT to derive local thresholds for use in manual segmentation and computation of volumes. Our validation study showed that quantification of marrow fat using µCT was reliable and accurate compared to the gold standard technique- histology- when reliably defined thresholds were used. Good agreement between tissue volumes measured by histology and those computed by the imaging software was demonstrated. We applied this technique to quantify marrow fat in an animal model of senile osteoporosis, and showed that fatty acids (ω- 3 and ω-6) had dual effects on bone. With QCT studies, we confirmed the age related increase in marrow adiposity, and more significantly, different ratios between fat and bone in common fracture regions. Similarly, exercise affects marrow fat differently in different regions, and there was a trend to statistically significant changes to marrow fat with exercise. In conclusion, this body of work showed that quantification of marrow fat using CT is promising, and has future clinical implications. However, significantly more clinical studies are needed to confirm these findings and refine shortfalls in quantification capabilities

A qualitative and quantitative investigation of structural morphology in the neonatal ilium

Cortical and trabecular bone characteristics can be used to make predictions regarding previous loading regimes and developmental milestones which a bone has encountered. This has led to the suggestion that in the adult pelvis, bone patterning is related to the remodeling forces generated during bipedal locomotion. However, during the neonatal period the pelvic complex is non-load bearing, therefore, structural organisation of the ilium cannot reflect direct stance related forces. This study considers the cortical and trabecular bone structure in the ilium of the fetal and newborn infant, a structural configuration which until now has remained largely neglected in the literature. Only recently, with the advent of imaging modalities, has a greater insight and understanding of previously unexplored human bone structural composition and developing bone structure been made possible. In this study, multiple imaging techniques were applied to establish the optimal modality for application to the assessment of bone microstructure. Plain plate macroradiography and micro-computed tomography were identified as the gold standard imaging modalities for bone structural analysis for respective qualitative and quantitative assessment. These techniques were applied to gain a perspective of bone form from a sample of fetal and neonatal ilia selected from the Scheuer collection of juvenile remains. Initially, qualitative analysis highlighted consistent and well-defined patterns of cortical and trabecular bone organisation within the fetal and neonatal ilium, which corresponded with previously recognised regions in the adult that have been attributed directly to forces associated with bipedal locomotion. This was highly unexpected as the early developmental ilium is non-load bearing. Subsequently, quantification of the neonatal cortical and trabecular structure reinforced radiographic observations by identifying regions of significant architectural arrangement. Further investigation of this precocious patterning led to a revised proposal for the mode of growth in the human ilium during the neonatal developmental period. Analysis revealed statistically significant differences in regional trabecular characteristics and cortical thicknesses which have formed the basis of a proposed growth model for the ilium. The presence of ‘progressive growth regions’ and ‘restricted growth regions’ which appear to relate to metaphyseal and non-metaphyseal borders of the ilium have been demonstrated. Analysis of the early iliac bone pattern is important for understanding the relationship between trabecular bone patterning and cortical bone structure during the earliest stages of development in response to the specific functional forces acting during this period. It is suggested that the seemingly organised rudimentary scaffold observed in the early developmental ilium may be attributable to early ossification patterning, non-weight bearing anatomical interactions or even to a predetermined genetic blueprint. It must also be postulated that whilst the observed patterning may be indicative of a predetermined inherent template, early non-load bearing locomotive influences may subsequently be superimposed upon this scaffolding and perhaps reinforced and likely remodelled at a later age. Ultimately, the analysis of this fundamental primary pattern has core implications for understanding the earliest changes in iliac trabecular architecture and provides a baseline insight into future ontogenetic development and bipedal capabilities. Finally, the structural data and statistical analysis presented challenge the current concept of implied centrifugal ossification within the human ilium and present evidence of an alternative pattern of ossification that is largely dictated and controlled by basic anatomical principles.EThOS - Electronic Theses Online ServiceLeng TrustWenner-Gren FoundationBiotechnology and Biological Sciences Research CouncilGBUnited Kingdo

Automated 3D trabecular bone structure analysis of the proximal femur—prediction of biomechanical strength by CT and DXA

The standard diagnostic technique for assessing osteoporosis is dual X-ray absorptiometry (DXA) measuring bone mass parameters. In this study, a combination of DXA and trabecular structure parameters (acquired by computed tomography [CT]) most accurately predicted the biomechanical strength of the proximal femur and allowed for a better prediction than DXA alone. An automated 3D segmentation algorithm was applied to determine specific structure parameters of the trabecular bone in CT images of the proximal femur. This was done to evaluate the ability of these parameters for predicting biomechanical femoral bone strength in comparison with bone mineral content (BMC) and bone mineral density (BMD) acquired by DXA as standard diagnostic technique. One hundred eighty-seven proximal femur specimens were harvested from formalin-fixed human cadavers. BMC and BMD were determined by DXA. Structure parameters of the trabecular bone (i.e., morphometry, fuzzy logic, Minkowski functionals, and the scaling index method [SIM]) were computed from CT images. Absolute femoral bone strength was assessed with a biomechanical side-impact test measuring failure load (FL). Adjusted FL parameters for appraisal of relative bone strength were calculated by dividing FL by influencing variables such as body height, weight, or femoral head diameter. The best single parameter predicting FL and adjusted FL parameters was apparent trabecular separation (morphometry) or DXA-derived BMC or BMD with correlations up to r = 0.802. In combination with DXA, structure parameters (most notably the SIM and morphometry) added in linear regression models significant information in predicting FL and all adjusted FL parameters (up to R adj = 0.872) and allowed for a significant better prediction than DXA alone. A combination of bone mass (DXA) and structure parameters of the trabecular bone (linear and nonlinear, global and local) most accurately predicted absolute and relative femoral bone strength