New Methods and Applications in the Study of Bone Histomorphometry and Biomechanics

A special issue of Biology (ISSN 2079-7737). This special issue belongs to the section "Biotechnology".

Deadline for manuscript submissions: closed (15 February 2023) | Viewed by 23978

Special Issue Editors

Forensic Medicine Unit, Department of Forensic Sciences, Medical School, University of Crete, 71003 Heraklion, Greece
Interests: forensic medicine; forensic pathology; forensic anthropology
Special Issues, Collections and Topics in MDPI journals
Center for Anatomy and Human Identification (CAHID), School of Science and Engineering, University of Dundee, Dundee DD1 4HN, Scotland, UK
Interests: forensic anthropology; biological anthropology; bone histomorphometry; skeletal variation; aging; bone pathology
Research Associate in Forensic Taphonomy, School of Natural Sciences, University of Central Lancashire, Preston PR1 2HE, UK
Interests: forensic anthropology; forensic science; histomorphometry; biomechanics; molecular biology; aging

Special Issue Information

Dear Colleagues,

Bone is a resistant and dynamic tissue that undergoes constant changes throughout life, and is subject to various factors, such as growth, development, mechanical loading, diet and pathology. Bone is a valuable diagnostic tool in forensic medicine and forensic anthropology, due to its unique properties, and it can be used to extract crucial information when investigating unknown, heavily decomposed remains. Bone is analysed by observing its microscopic structure, as well as quantifying its structural elements and material properties, which provides an alternative methodological perspective that allows bone to be fully understood at the tissue level. Research has demonstrated that bone histomorphometry can assist in the identification process, with methods being developed for the purpose of species differentiation, age estimation and histo-taphonomy assessment. Research pertaining to bone biomechanics explores the material and mechanical properties of bone, providing insights into bone behavior in relation to fracture, trauma assessment and interpretation, as well as age-related changes. Moreover, both methodological approaches offer the possibility of evaluating the impact of pathological conditions on bone, intra and inter-individual skeletal variation, as well as other relevant information, such as physical activity. In addition to the recent developments in X-ray-based imaging, a number of techniques, based on metric, histomorphometric and simulated mechanical analyses, have attracted great interest for forensic purposes, due to their noninvasive nature. High-resolution imaging techniques can provide three-dimensional information, in addition to the classical two-dimensional techniques on bone remodeling and microstructure, and can also accommodate biomechanical studies of relevant nature. This Special Issue aims to collect important contributions in the field of histomorphometry and biomechanics, highlighting the potential of the latest state-of-the-art methodologies, in order to meet the challenges in forensic and medical sciences.

Dr. Elena F. Kranioti
Dr. Julieta Gómez García-Donas
Dr. Andrea Bonicelli
Guest Editors

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Keywords

  • age estimation
  • sex estimation
  • intra- and inter-population variation
  • trauma analysis
  • bone modelling and remodeling
  • bone histopathology
  • validation and testing
  • CT- and μCT-based histomorphometry
  • automated methods for histomorphometric analysis
  • bone biomechanics
  • bone diagenesis
  • simulation and modelling

Published Papers (8 papers)

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Research

10 pages, 1656 KiB  
Communication
Automatic Segmentation of Osteonal Microstructure in Human Cortical Bone Using Deep Learning: A Proof of Concept
Biology 2023, 12(4), 619; https://doi.org/10.3390/biology12040619 - 19 Apr 2023
Viewed by 1671
Abstract
Cortical bone microstructure assessment in biological and forensic anthropology can assist with the estimation of age-at-death and animal-human differentiation, for example. Osteonal structures within cortical bone are the key feature under analysis, with osteon frequency and metric parameters providing crucial information for the [...] Read more.
Cortical bone microstructure assessment in biological and forensic anthropology can assist with the estimation of age-at-death and animal-human differentiation, for example. Osteonal structures within cortical bone are the key feature under analysis, with osteon frequency and metric parameters providing crucial information for the assessment. Currently, the histomorphological assessment consists of a time-consuming manual process for which specific training is required. Our work investigates the feasibility of automatic analysis of human bone microstructure images through the application of deep learning. In this paper, we use a U-Net architecture to address the semantic segmentation of such images into three classes: intact osteons, fragmentary osteons, and background. Data augmentation was used to avoid overfitting. We evaluated our fully automatic approach using a sample of 99 microphotographs. The contours of intact and fragmentary osteons were traced manually to provide ground truth. The Dice coefficients were 0.73 for intact osteons, 0.38 for fragmented osteons, and 0.81 for background, giving an average of 0.64. The Dice coefficient of the binary classification osteon-background was 0.82. Although further refinement of the initial model and tests with larger datasets are needed, this study provides, to the best of our knowledge, the first proof of concept for the use of computer vision and deep learning for differentiating both intact and fragmentary osteons in human cortical bone. This approach has the potential to widen and facilitate the use of histomorphological assessment in the biological and forensic anthropology communities. Full article
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13 pages, 4438 KiB  
Article
Investigating the Timing and Extent of Juvenile and Fetal Bone Diagenesis in a Temperate Environment
Biology 2023, 12(3), 403; https://doi.org/10.3390/biology12030403 - 03 Mar 2023
Cited by 1 | Viewed by 7613
Abstract
It is well understood that intrinsic factors of bone contribute to bone diagenesis, including bone porosity, crystallinity, and the ratio of organic to mineral components. However, histological analyses have largely been limited to adult bones, although with some exceptions. Considering that many of [...] Read more.
It is well understood that intrinsic factors of bone contribute to bone diagenesis, including bone porosity, crystallinity, and the ratio of organic to mineral components. However, histological analyses have largely been limited to adult bones, although with some exceptions. Considering that many of these properties are different between juvenile and adult bone, the purpose of this study is to investigate if these differences may result in increased degradation observed histologically in fetal and juvenile bone. Thirty-two fetal (n = 16) and juvenile (n = 16) Sus scrofa domesticus femora subject to different depositions over a period of two years were sectioned for histological observation. Degradation was scored using an adapted tunneling index. Results showed degradation related to microbial activity in both fetal and juvenile remains across depositions as early as three months. Buried juvenile remains consistently showed the greatest degradation over time, while the blanket fetal remains showed more minimal degradation. This is likely related to the buried remains’ greater contact with surrounding soil and groundwater during deposition. Further, most of the degradation was seen in the subendosteal region, followed by the subperiosteal region, which may suggest the initial microbial attack is from endogenous sources. Full article
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11 pages, 14086 KiB  
Article
Osteonal Microcracking Pattern: A Potential Vitality Marker in Human Bone Trauma
Biology 2023, 12(3), 399; https://doi.org/10.3390/biology12030399 - 03 Mar 2023
Cited by 2 | Viewed by 1226
Abstract
In forensic anthropology, the differential diagnosis between peri- and postmortem bone fractures is mainly based on macroscopic criteria. In contrast, studies focusing on bone histology are very scarce. In a recent publication, we showed that (perimortem) fractures in fresh human bones exhibit a [...] Read more.
In forensic anthropology, the differential diagnosis between peri- and postmortem bone fractures is mainly based on macroscopic criteria. In contrast, studies focusing on bone histology are very scarce. In a recent publication, we showed that (perimortem) fractures in fresh human bones exhibit a different osteonal microcracking pattern than (postmortem) damage in dry bones. In the current work, we explored whether this osteonal microcracking pattern is distinctive of the vitality of (perimortem) fresh bone fractures. To this end, we compared the number, length and structural distribution of microcracks in vital humeral fractures from forensic autopsy cases with experimentally reproduced, three point-bending fractures in fresh and dry human humeri. Half of the fresh experimental bones were fractured whilst applying axial compression, i.e., attempting to simulate intra vitam conditions more accurately. The results showed a similar osteonal microcracking pattern between vital fractures and experimental fractures of fresh humeri subjected to axial compression. Interestingly, this pattern was significantly different from the one observed in the experimental fractures of fresh humeri without axial compression and dry humeri. This supports our hypothesis that the osteonal microcracking pattern can potentially be used as a marker for vital perimortem trauma, providing a histomorphometric tool for fracture timing. Full article
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14 pages, 8638 KiB  
Article
The Diagenetic Alterations of Historic Skeletons from the Crown Mines Cemetery, South Africa
Biology 2023, 12(3), 378; https://doi.org/10.3390/biology12030378 - 27 Feb 2023
Viewed by 2093
Abstract
Human skeletons associated with early gold mining in Johannesburg, South Africa are investigated. An unmarked cemetery was buried beneath a mine dump which resulted in macroscopically stained and poorly preserved bones. Histological assessments were conducted to understand the postmortem treatment of the remains, [...] Read more.
Human skeletons associated with early gold mining in Johannesburg, South Africa are investigated. An unmarked cemetery was buried beneath a mine dump which resulted in macroscopically stained and poorly preserved bones. Histological assessments were conducted to understand the postmortem treatment of the remains, determine the extent of bone degradation, and understand how this environment affected the bone’s microstructure. Various diagenetic alterations and the general histological index were assessed using normal and polarized light microscopy of thin anterior midshaft femur sections (n = 50). Degradation was identified in the periosteal and endosteal regions, while the intra-cortical region remained well-preserved. Bacterial bioerosion, microcracks, infiltrations, inclusions, and staining were found throughout the sample. Numerous non-Wedl micro-foci of destruction were observed, filled with exogenous material. The degradation suggested that the remains were buried in neutral soil that was subsequently covered by acidic mine dumps which resulted in a corrosive environment. Although the skeletons were poorly preserved, their histological integrity was more promising, especially the intra-cortical area. This is important for future investigations of archaeological bone, as this area can lead to more accurate descriptions of skeletal assemblages. Targeted sampling of this region could produce promising estimates of age, descriptions of pathology, and biomolecular results, which require further study. Full article
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21 pages, 8795 KiB  
Article
Quantitative Load Dependency Analysis of Local Trabecular Bone Microstructure to Understand the Spatial Characteristics in the Synthetic Proximal Femur
Biology 2023, 12(2), 170; https://doi.org/10.3390/biology12020170 - 20 Jan 2023
Cited by 1 | Viewed by 3115
Abstract
Analysis of the dependency of the trabecular structure on loading conditions is essential for understanding and predicting bone structure formation. Although previous studies have investigated the relationship between loads and structural adaptations, there is a need for an in-depth analysis of this relationship [...] Read more.
Analysis of the dependency of the trabecular structure on loading conditions is essential for understanding and predicting bone structure formation. Although previous studies have investigated the relationship between loads and structural adaptations, there is a need for an in-depth analysis of this relationship based on the bone region and load specifics. In this study, the load dependency of the trabecular bone microstructure for twelve regions of interest (ROIs) in the synthetic proximal femur was quantitatively analyzed to understand the spatial characteristics under seven different loading conditions. To investigate the load dependency, a quantitative measure, called the load dependency score (LDS), was established based on the statistics of the strain energy density (SED) distribution. The results showed that for the global model and epiphysis ROIs, bone microstructures relied on the multiple-loading condition, whereas the structures in the metaphysis depended on single or double loads. These results demonstrate that a given ROI is predominantly dependent on a particular loading condition. The results confirm that the dependency analysis of the load effects for ROIs should be performed both qualitatively and quantitatively. Full article
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22 pages, 3378 KiB  
Article
Estimation of Age-at-Death Using Cortical Bone Histomorphometry of the Rib and Femur: A Validation Study on a British Population
Biology 2022, 11(11), 1615; https://doi.org/10.3390/biology11111615 - 04 Nov 2022
Cited by 3 | Viewed by 1758
Abstract
Histomorphometry constitutes a valuable tool for age estimation. Histological interpopulation variability has been shown to affect the accuracy of age estimation techniques and therefore validation studies are required to test the accuracy of the pre-existing methodologies. The present research constitutes a validation study [...] Read more.
Histomorphometry constitutes a valuable tool for age estimation. Histological interpopulation variability has been shown to affect the accuracy of age estimation techniques and therefore validation studies are required to test the accuracy of the pre-existing methodologies. The present research constitutes a validation study of widely known histological methods on the sixth rib and the femoral midshaft of a 19th century British population originating from Blackburn, England. An evaluation of the histomorphometric features of eleven ribs and five femora was performed and used to test the accuracy of selected methods. Results indicated that osteon area and circularity were the only histomorphometric variables that presented significant interpopulation variability. Cho et al.’s method for the ribs and the average value produced using Kerley and Ubelaker’s method for intact osteon and percentage of lamellar bone equations for femur were considered the only reliable markers for estimating the age on the Blackburn sample. In the case of old individuals, Goliath et al.’s method provided more satisfactory results. Overall, the present study provides evidence on the applicability of the aging histomorphometric methods on a British sample and highlights the limitations of applying histomorphometric methods developed on different reference populations than the one under investigation. Full article
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21 pages, 3361 KiB  
Article
The Effects of Experimental Whole-Body Burning on Histological Age-at-Death Estimation from Human Cortical Bone and Dental Cementum
Biology 2022, 11(11), 1569; https://doi.org/10.3390/biology11111569 - 26 Oct 2022
Cited by 2 | Viewed by 2855
Abstract
Whole-body donations (n = 6) were placed in various experimental fire-death scenarios to understand the histological effects of thermal alteration on bones and teeth. Midshaft samples of the femur, 6th rib, and metatarsal were removed from each donor pre- and post-burning to [...] Read more.
Whole-body donations (n = 6) were placed in various experimental fire-death scenarios to understand the histological effects of thermal alteration on bones and teeth. Midshaft samples of the femur, 6th rib, and metatarsal were removed from each donor pre- and post-burning to examine histomorphometric differences and test established age-at-death estimation methods. Dental samples were taken post-burning to test the applicability of dental cementum analysis for age-at-death estimation. Significant differences in osteon area or Haversian canal area between some pre- and post-burn samples were found although no patterns related to temperature or element were observable. The femoral age estimates across pre- and post-burn samples were 91% accurate across all donors. The point age estimates from the ribs compared to known age were significantly different (t(10) = 6.88, p < 0.001) with an average difference of −18.53 years. Dental age estimates of post-burn samples were not significantly different from the known donor age (t(3) = −0.74, p = 0.512) with an average difference of −3.96 years. Overall, the results of this study show that thermally altered remains can be used for histologic age-at-death analysis of cortical bone and dental cementum, within certain burning parameters. Full article
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21 pages, 2761 KiB  
Article
Ontogenetic Patterning of Human Subchondral Bone Microarchitecture in the Proximal Tibia
Biology 2022, 11(7), 1002; https://doi.org/10.3390/biology11071002 - 01 Jul 2022
Cited by 5 | Viewed by 2025
Abstract
High-resolution computed tomography images were acquired for 31 proximal human tibiae, age 8 to 37.5 years, from Norris Farms #36 cemetery site (A.D. 1300). Morphometric analysis of subchondral cortical and trabecular bone architecture was performed between and within the tibial condyles. Kruskal–Wallis and [...] Read more.
High-resolution computed tomography images were acquired for 31 proximal human tibiae, age 8 to 37.5 years, from Norris Farms #36 cemetery site (A.D. 1300). Morphometric analysis of subchondral cortical and trabecular bone architecture was performed between and within the tibial condyles. Kruskal–Wallis and Wilcoxon signed-rank tests were used to examine the association between region, age, body mass, and each morphometric parameter. The findings indicate that age-related changes in mechanical loading have varied effects on subchondral bone morphology. With age, trabecular microstructure increased in bone volume fraction (p = 0.033) and degree of anisotropy (p = 0.012), and decreased in connectivity density (p = 0.001). In the subchondral cortical plate, there was an increase in thickness (p < 0.001). When comparing condylar regions, only degree of anisotropy differed (p = 0.004) between the medial and lateral condyles. Trabeculae in the medial condyle were more anisotropic than in the lateral region. This research represents an innovative approach to quantifying both cortical and trabecular subchondral bone microarchitecture in archaeological remains. Full article
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