Pub Date : 2025-10-29DOI: 10.1016/j.bonr.2025.101884
Mathieu Simon , Sasidhar Uppuganti , Jeffry S. Nyman , Philippe Zysset
Type 2 diabetes (T2D) is a chronic disease leading to an elevated glucose level in the blood and increased fracture risk. Because T2D individuals tend to have normal to higher areal bone mineral density (aBMD) than healthy individuals, their fracture risk is often underestimated. As an alternative, high-resolution peripheral quantitative computed tomography (HR-pQCT) is an attractive tool to investigate bone morphology in vivo and estimate fracture risk. Based on HR-pQCT scans, bone strength can be estimated using micro finite element (FE) analysis or homogenized finite element (hFE) analysis. While FE is computationally expensive, hFE provides an accurate estimation of bone mechanical properties within reasonable efforts. However, the hFE scheme is based on relationships between the local fabric (anisotropy) and elasticity. These relationships have been shown to hold for healthy controls as well as in the case of osteogenesis imperfecta. Nevertheless, whether these relationships are also valid for T2D-diagnosed patients remains unclear. Therefore, the present work aims to compare fabric–elasticity relationships between T2D and non-diabetic controls.
The present study collected 56 trabecular bone cores from the femoral head of 28 T2D and 28 control donors. These cadaveric samples were scanned in a micro-CT system at an isotropic 14.8 m voxel size. Three cubic regions of interest (ROIs) were selected in each scan. The resolution of these ROIs was coarsened by a factor of 4, mimicking clinical HR-pQCT resolution, and the ROIs were subsequently segmented. Standard morphometric parameters were computed from the segmented ROIs using medtool (v4.8; Dr. Pahr Ingenieurs e.U., Pfaffstätten, Austria). Additionally, their fabric tensor and their apparent stiffness tensors were computed. The ROIs were compared between T2D and control regarding their morphometric and mechanical properties. Finally, ROIs were matched between T2D and control for bone volume fraction () and degree of anisotropy (DA). The matched dataset allowed the comparison of fabric–elasticity relationships between T2D and control samples.
No significant difference was observed between T2D and control samples, both regarding their morphology and their mechanical properties. Specifically, fabric–elasticity relationships were shown to hold for both the control and the T2D groups. A comparison of the resulting exponents related to and DA has highlighted different trends but no important difference between T2D and control samples.
In conclusion, trabecular bone architecture was similar between T2D and non-T2D donors. Additionally, fabric–elasticity relationships, i.e. morphology-mechanical relationships, are also similar between donors with and without diabetes. Accordingly, HR-p
2型糖尿病（T2D）是一种导致血液中葡萄糖水平升高和骨折风险增加的慢性疾病。由于T2D患者的面骨矿物质密度（aBMD）往往高于健康人群，因此他们的骨折风险往往被低估。作为替代方案，高分辨率周边定量计算机断层扫描（HR-pQCT）是一种有吸引力的工具，用于研究体内骨形态和估计骨折风险。基于HR-pQCT扫描结果，可采用微有限元（μFE）分析或均质有限元（hFE）分析估算骨强度。虽然μFE在计算上是昂贵的，但hFE在合理的努力下提供了骨力学性能的准确估计。然而，hFE方案是基于局部结构（各向异性）和弹性之间的关系。这些关系已被证明适用于健康对照以及成骨不全症。然而，这些关系是否也适用于t2d诊断患者仍不清楚。因此，本研究旨在比较T2D和非糖尿病对照之间的织物弹性关系。本研究从28例T2D供体和28例对照供体的股骨头中收集了56个骨小梁核心。这些尸体样品在微ct系统中以各向同性14.8 μm体素尺寸进行扫描。在每次扫描中选择三个立方感兴趣区域（roi）。这些roi的分辨率被粗化4倍，模仿临床HR-pQCT分辨率，随后对roi进行分割。使用medtool （v4.8; Dr. Pahr Ingenieurs e.U, Pfaffstätten，奥地利）从分割的roi中计算标准形态计量参数。此外，还计算了它们的织物张量和表观刚度张量。比较了T2D和对照组在形态计量学和力学性能方面的roi。最后，在骨体积分数（ρ）和各向异性程度（DA）方面，将T2D与对照组的roi进行匹配。匹配的数据集允许比较T2D和对照样本之间的织物弹性关系。在T2D和对照样品之间，无论是形貌还是力学性能都没有观察到显著差异。具体来说，织物弹性关系被证明对对照组和T2D组都有效。与ρ和DA相关的结果指数的比较突出了不同的趋势，但在T2D和控制样本之间没有重要差异。总之，T2D和非T2D供体的骨小梁结构相似。此外，织物-弹性关系，即形态-力学关系，在有和没有糖尿病的供体之间也相似。因此，基于hr - pqct的hFE分析也可用于估计T2D患者的骨力学特性和评估其骨折风险。
{"title":"Fabric–elasticity relationships of femoral head trabecular bone are similar in Type 2 diabetes and non-diabetic individuals","authors":"Mathieu Simon , Sasidhar Uppuganti , Jeffry S. Nyman , Philippe Zysset","doi":"10.1016/j.bonr.2025.101884","DOIUrl":"10.1016/j.bonr.2025.101884","url":null,"abstract":"<div><div>Type 2 diabetes (T2D) is a chronic disease leading to an elevated glucose level in the blood and increased fracture risk. Because T2D individuals tend to have normal to higher areal bone mineral density (aBMD) than healthy individuals, their fracture risk is often underestimated. As an alternative, high-resolution peripheral quantitative computed tomography (HR-pQCT) is an attractive tool to investigate bone morphology in vivo and estimate fracture risk. Based on HR-pQCT scans, bone strength can be estimated using micro finite element (<span><math><mi>μ</mi></math></span>FE) analysis or homogenized finite element (hFE) analysis. While <span><math><mi>μ</mi></math></span>FE is computationally expensive, hFE provides an accurate estimation of bone mechanical properties within reasonable efforts. However, the hFE scheme is based on relationships between the local fabric (anisotropy) and elasticity. These relationships have been shown to hold for healthy controls as well as in the case of osteogenesis imperfecta. Nevertheless, whether these relationships are also valid for T2D-diagnosed patients remains unclear. Therefore, the present work aims to compare fabric–elasticity relationships between T2D and non-diabetic controls.</div><div>The present study collected 56 trabecular bone cores from the femoral head of 28 T2D and 28 control donors. These cadaveric samples were scanned in a micro-CT system at an isotropic 14.8 <span><math><mi>μ</mi></math></span>m voxel size. Three cubic regions of interest (ROIs) were selected in each scan. The resolution of these ROIs was coarsened by a factor of 4, mimicking clinical HR-pQCT resolution, and the ROIs were subsequently segmented. Standard morphometric parameters were computed from the segmented ROIs using medtool (v4.8; Dr. Pahr Ingenieurs e.U., Pfaffstätten, Austria). Additionally, their fabric tensor and their apparent stiffness tensors were computed. The ROIs were compared between T2D and control regarding their morphometric and mechanical properties. Finally, ROIs were matched between T2D and control for bone volume fraction (<span><math><mi>ρ</mi></math></span>) and degree of anisotropy (DA). The matched dataset allowed the comparison of fabric–elasticity relationships between T2D and control samples.</div><div>No significant difference was observed between T2D and control samples, both regarding their morphology and their mechanical properties. Specifically, fabric–elasticity relationships were shown to hold for both the control and the T2D groups. A comparison of the resulting exponents related to <span><math><mi>ρ</mi></math></span> and DA has highlighted different trends but no important difference between T2D and control samples.</div><div>In conclusion, trabecular bone architecture was similar between T2D and non-T2D donors. Additionally, fabric–elasticity relationships, i.e. morphology-mechanical relationships, are also similar between donors with and without diabetes. Accordingly, HR-p","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101884"},"PeriodicalIF":2.6,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145424383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hyperglycemia in diabetes leads to the formation of methylglyoxal (MG) and accumulation of advanced glycation end-products (AGEs). We previously reported that exogenous MG exposure deteriorated osteoblastic differentiation in vitro and diabetic mice showed delayed bone defect healing along with elevated MG-derived AGE levels. However, whether endogenously formed MG is involved in impaired bone repair and remodeling in diabetes remains unclear. In this study, we investigated the effects of hyperglycemia-induced MG formation on bone quality and defect healing in mice. Using a synthetic MG probe [Ir(ppy)2(DA-phen)], we found that endogenous MG formation deteriorated osteoblastic and osteoclastic differentiation under hyperglycemic conditions in cultured cells. In the bone defect site of streptozotocin (STZ)-induced diabetic mice, along with impaired defect healing, we observed elevated endogenous MG levels and downregulation of alkaline phosphatase (ALP) compared with that in non-diabetic control mice; however, these alterations were alleviated by managing blood glucose levels through insulin supplementation. Furthermore, treatment with pyridoxamine (PM), an MG scavenger, ameliorated these impairments by suppressing MG elevation and upregulating the expression of osteocalcin, osteoprotegerin, and osteoclast-associated receptor genes without improving the diabetic status. These findings suggest that endogenously formed MG is detrimental to hyperglycemia-related delayed bone defect healing in type 1 diabetes mellitus (T1DM). Collectively, this study suggests that MG scavenging by PM and suppression of MG formation by glycemic control are potential therapeutic strategies for T1DM-associated bone disorders.
{"title":"Scavenging methylglyoxal improves bone quality and defect healing in diabetic mice","authors":"Toshifumi Hikichi , Kumi Kimura , Seiichi Munesue , Yu Oshima , Ai Harashima , Tomo Hamada , Kanu Shimokawa , Yuki Fushitani , Hisanori Goto , Hiroyuki Tsuchiya , Satoru Demura , Hidenori Matsubara , Yasuhiko Yamamoto","doi":"10.1016/j.bonr.2025.101883","DOIUrl":"10.1016/j.bonr.2025.101883","url":null,"abstract":"<div><div>Hyperglycemia in diabetes leads to the formation of methylglyoxal (MG) and accumulation of advanced glycation end-products (AGEs). We previously reported that exogenous MG exposure deteriorated osteoblastic differentiation in vitro and diabetic mice showed delayed bone defect healing along with elevated MG-derived AGE levels. However, whether endogenously formed MG is involved in impaired bone repair and remodeling in diabetes remains unclear. In this study, we investigated the effects of hyperglycemia-induced MG formation on bone quality and defect healing in mice. Using a synthetic MG probe [Ir(ppy)<sub>2</sub>(DA-phen)], we found that endogenous MG formation deteriorated osteoblastic and osteoclastic differentiation under hyperglycemic conditions in cultured cells. In the bone defect site of streptozotocin (STZ)-induced diabetic mice, along with impaired defect healing, we observed elevated endogenous MG levels and downregulation of alkaline phosphatase (ALP) compared with that in non-diabetic control mice; however, these alterations were alleviated by managing blood glucose levels through insulin supplementation. Furthermore, treatment with pyridoxamine (PM), an MG scavenger, ameliorated these impairments by suppressing MG elevation and upregulating the expression of osteocalcin, osteoprotegerin, and osteoclast-associated receptor genes without improving the diabetic status. These findings suggest that endogenously formed MG is detrimental to hyperglycemia-related delayed bone defect healing in type 1 diabetes mellitus (T1DM). Collectively, this study suggests that MG scavenging by PM and suppression of MG formation by glycemic control are potential therapeutic strategies for T1DM-associated bone disorders.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101883"},"PeriodicalIF":2.6,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145473900","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-20DOI: 10.1016/j.bonr.2025.101882
Ellie H. Northall , Liam M. Grover , Helen M. McGettrick , Matthew Newton Ede , Amy J. Naylor , Simon W. Jones
Adolescent Idiopathic Scoliosis (AIS) is a common paediatric spinal disorder of incompletely understood aetiology. Current interventions include bracing and invasive surgery. However, determining which patients will benefit from observation, bracing, or surgery remains challenging due to the difficulty in predicting disease progression. A number of factors have previously been purported to play a causative role including hormones and biomechanics. However, recently GWAS and in vitro studies have provided insight into the underlying signalling pathways and intrinsic factors that act as drivers of AIS pathology across different tissue types, including spinal bone tissue, paraspinal muscles and cartilage. This review will explore these recent findings and evaluate their links to systemic factors as possible intrinsic drivers underpinning AIS pathophysiology and development.
{"title":"Integrating intrinsic musculoskeletal pathology and genetics: Recent advances in unravelling the causative factors of adolescent idiopathic scoliosis","authors":"Ellie H. Northall , Liam M. Grover , Helen M. McGettrick , Matthew Newton Ede , Amy J. Naylor , Simon W. Jones","doi":"10.1016/j.bonr.2025.101882","DOIUrl":"10.1016/j.bonr.2025.101882","url":null,"abstract":"<div><div>Adolescent Idiopathic Scoliosis (AIS) is a common paediatric spinal disorder of incompletely understood aetiology. Current interventions include bracing and invasive surgery. However, determining which patients will benefit from observation, bracing, or surgery remains challenging due to the difficulty in predicting disease progression. A number of factors have previously been purported to play a causative role including hormones and biomechanics. However, recently GWAS and in vitro studies have provided insight into the underlying signalling pathways and intrinsic factors that act as drivers of AIS pathology across different tissue types, including spinal bone tissue, paraspinal muscles and cartilage. This review will explore these recent findings and evaluate their links to systemic factors as possible intrinsic drivers underpinning AIS pathophysiology and development.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101882"},"PeriodicalIF":2.6,"publicationDate":"2025-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145358577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-06DOI: 10.1016/j.bonr.2025.101879
Charu Jain , Neharika Bhadouria , Justin Tiao , Jonathan J. Huang , Yunsoo Lee , Saad Chaudhary , Andrew C. Hecht , Nilsson Holguin , James C. Iatridis
Background
Human bone quality varies with age and spinal level, with upper lumbar vertebrae more prone to fracture. Mouse studies of bone health often focus on a single lumbar vertebral level and limited regional analysis with no comprehensive study evaluating how sex, lumbar level, vertebral region, and reconstruction method contribute to patterns of age-related vertebral bone loss in mice.
Methods
This micro-computed tomography (μCT) study measured lumbar spinal level- and vertebral region of interest (ROI)-specific patterns of bone quantity, structure, and density in relation to age and sex in mice. Lumbar vertebrae (L1–L6; n = 4/sex/age) from young adult (4-month), middle-aged (12-month), and old (24-month) mice were analyzed for trabecular bone volume fraction (BV/TV), volumetric bone mineral density (vBMD), cortical thickness (Ct.Th), and tissue mineral density (TMD). Parameters were measured for full vertebrae, cranial, middle, and caudal vertebral regions, comparing both 1/3-vertebral and standardized 30-slice reconstruction approaches.
Results
Age was associated with a reduction in vBMD and BV/TV in both sexes, with bone volume, structure, and density most significantly reduced at L1 and cranial vertebral regions. Females exhibited greater declines than males, particularly in BV/TV and vBMD. Ct.Th was the greatest mid-spine but declined with age at L1 and L5. Regional differences were observed for both cortical parameters.
Conclusions
Trabecular bone is affected by age, sex, lumbar level, and vertebral region in mice. The greatest sensitivity to age-related bone loss was detected in the L1 lumbar level and cranial vertebral regions in female mice. These findings highlight the need for region- and method- specific analysis in bone research and exhibit age-related changes in vertebrate by level and region that exhibit similarities to humans.
人的骨骼质量随年龄和脊柱水平而变化，上腰椎更容易骨折。对小鼠骨骼健康的研究通常集中在单个腰椎水平和有限的区域分析上，没有全面的研究来评估性别、腰椎水平、椎体区域和重建方法如何影响小鼠年龄相关的椎体骨质流失模式。方法微计算机断层扫描（μCT）研究测量了小鼠腰椎水平和椎体感兴趣区（ROI）特异性骨数量、结构和密度与年龄和性别的关系。对年轻成年（4个月）、中年（12个月）和老年（24个月）小鼠腰椎（L1-L6, n = 4/性别/年龄）进行骨小梁体积分数（BV/TV）、体积骨矿物质密度（vBMD）、皮质厚度(Ct。Th)和组织矿物质密度（TMD）。测量全椎体、颅骨、中段和尾椎区域的参数，比较1/3椎体和标准化30层重建方法。结果年龄与男女vBMD和BV/TV的降低有关，L1和颅椎区的骨体积、结构和密度降低最为显著。女性表现出比男性更大的下降，尤其是BV/TV和vBMD。Ct。腰椎中部最大，但随着L1和L5年龄的增长而下降。两种皮质参数均存在区域差异。结论小鼠的骨梁受年龄、性别、腰椎水平和椎体区域的影响。在雌性小鼠中，L1腰椎水平和颅椎区域对年龄相关的骨质流失最敏感。这些发现强调了在骨骼研究中对区域和方法进行特定分析的必要性，并展示了脊椎动物在水平和区域上与人类相似的年龄相关变化。
{"title":"Age-related bone loss in mouse lumbar vertebrae is affected by region, sex, and level: Implications for spinal loading and analysis methods","authors":"Charu Jain , Neharika Bhadouria , Justin Tiao , Jonathan J. Huang , Yunsoo Lee , Saad Chaudhary , Andrew C. Hecht , Nilsson Holguin , James C. Iatridis","doi":"10.1016/j.bonr.2025.101879","DOIUrl":"10.1016/j.bonr.2025.101879","url":null,"abstract":"<div><h3>Background</h3><div>Human bone quality varies with age and spinal level, with upper lumbar vertebrae more prone to fracture. Mouse studies of bone health often focus on a single lumbar vertebral level and limited regional analysis with no comprehensive study evaluating how sex, lumbar level, vertebral region, and reconstruction method contribute to patterns of age-related vertebral bone loss in mice.</div></div><div><h3>Methods</h3><div>This micro-computed tomography (μCT) study measured lumbar spinal level- and vertebral region of interest (ROI)-specific patterns of bone quantity, structure, and density in relation to age and sex in mice. Lumbar vertebrae (L1–L6; <em>n</em> = 4/sex/age) from young adult (4-month), middle-aged (12-month), and old (24-month) mice were analyzed for trabecular bone volume fraction (BV/TV), volumetric bone mineral density (vBMD), cortical thickness (Ct.Th), and tissue mineral density (TMD). Parameters were measured for full vertebrae, cranial, middle, and caudal vertebral regions, comparing both 1/3-vertebral and standardized 30-slice reconstruction approaches.</div></div><div><h3>Results</h3><div>Age was associated with a reduction in vBMD and BV/TV in both sexes, with bone volume, structure, and density most significantly reduced at L1 and cranial vertebral regions. Females exhibited greater declines than males, particularly in BV/TV and vBMD. Ct.Th was the greatest mid-spine but declined with age at L1 and L5. Regional differences were observed for both cortical parameters.</div></div><div><h3>Conclusions</h3><div>Trabecular bone is affected by age, sex, lumbar level, and vertebral region in mice. The greatest sensitivity to age-related bone loss was detected in the L1 lumbar level and cranial vertebral regions in female mice. These findings highlight the need for region- and method- specific analysis in bone research and exhibit age-related changes in vertebrate by level and region that exhibit similarities to humans.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101879"},"PeriodicalIF":2.6,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145263211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-02DOI: 10.1016/j.bonr.2025.101881
Allison L. Horenberg , Eric Z. Zeng , Yunke Ren , Tanishk Sinha , Emith Uyanwatte , Melia Matthews , Erica L. Scheller , Karl J. Lewis , Arvind P. Pathak , Warren L. Grayson
Peripheral sensory and sympathetic nerves innervating bony tissues are of interest for their role in modulating bone function and responding to physiologic stimuli. However, due to their filamentous structure and small diameter, peripheral nerves are difficult to visualize and characterize in dense tissues such as bone. Recent advances in imaging have resulted in novel techniques to reconstruct the 3D bone microenvironment following fluorescent staining, using methods such as whole-mount immunostaining, tissue clearing, and lightsheet microscopy (LSM). While these methods have significantly improved visualization of peripheral nerves within the bony skeleton, current practices in skeletal nerve segmentation rely on manual tracing methods that are both subjective and time consuming. Further, due to excess background signal and moderate off-target staining of skeletal nerves, traditional methods for nerve segmentation remain inadequate for effective quantification. Here, we describe a methods-focused workflow for 3D imaging and spatial analysis of nerves in bone that consists of combining LSM with nerve segmentation using the Ilastik® open-source software. First, LSM was used to acquire high-resolution 3D images of nerves in uninjured and injured murine calvaria. Next, we employed Ilastik® to generate accurate segmentations of peripheral nerves in bone. This approach reduced the need for manual segmentation and overcame the limitations of conventional segmentation methods. We then demonstrated the utility of this workflow as a broadly applicable tool, by performing Ilastik®-based segmentation for three distinct skeletal bone contexts (i.e. use cases) with varied bone samples, tissue clearing, processing protocols, inflammatory states, and imaging modalities. Collectively, this workflow enabled the characterization of spatial nerve patterning in bone and has the potential to provide novel insights into spatially regulated phenomena in bone homeostasis, injury, and disease.
{"title":"A novel workflow for 3D imaging and spatial analysis of nerves in bone","authors":"Allison L. Horenberg , Eric Z. Zeng , Yunke Ren , Tanishk Sinha , Emith Uyanwatte , Melia Matthews , Erica L. Scheller , Karl J. Lewis , Arvind P. Pathak , Warren L. Grayson","doi":"10.1016/j.bonr.2025.101881","DOIUrl":"10.1016/j.bonr.2025.101881","url":null,"abstract":"<div><div>Peripheral sensory and sympathetic nerves innervating bony tissues are of interest for their role in modulating bone function and responding to physiologic stimuli. However, due to their filamentous structure and small diameter, peripheral nerves are difficult to visualize and characterize in dense tissues such as bone. Recent advances in imaging have resulted in novel techniques to reconstruct the 3D bone microenvironment following fluorescent staining, using methods such as whole-mount immunostaining, tissue clearing, and lightsheet microscopy (LSM). While these methods have significantly improved visualization of peripheral nerves within the bony skeleton, current practices in skeletal nerve segmentation rely on manual tracing methods that are both subjective and time consuming. Further, due to excess background signal and moderate off-target staining of skeletal nerves, traditional methods for nerve segmentation remain inadequate for effective quantification. Here, we describe a methods-focused workflow for 3D imaging and spatial analysis of nerves in bone that consists of combining LSM with nerve segmentation using the Ilastik® open-source software. First, LSM was used to acquire high-resolution 3D images of nerves in uninjured and injured murine calvaria. Next, we employed Ilastik® to generate accurate segmentations of peripheral nerves in bone. This approach reduced the need for manual segmentation and overcame the limitations of conventional segmentation methods. We then demonstrated the utility of this workflow as a broadly applicable tool, by performing Ilastik®-based segmentation for three distinct skeletal bone contexts (i.e. use cases) with varied bone samples, tissue clearing, processing protocols, inflammatory states, and imaging modalities. Collectively, this workflow enabled the characterization of spatial nerve patterning in bone and has the potential to provide novel insights into spatially regulated phenomena in bone homeostasis, injury, and disease.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101881"},"PeriodicalIF":2.6,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145263212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01DOI: 10.1016/j.bonr.2025.101880
E. Montagnino , W. Bush , J. Bustamante , W. Bandara , P. Jalaie , M.R. Allen , J.M. Wallace , T. Siegmund , R.K. Surowiec , J.A. Howarter
Mineral imbalances in the body from chronic kidney disease can impact bone turnover and cause cortical bone loss. Synthetic salmon calcitonin is an FDA-approved treatment for bone fragility observed in diseases such as osteoporosis, and clinical trials have demonstrated a reduction in fractures compared to untreated individuals. This study documents the effects of calcitonin on cortical bone using an in vivo mouse model of chronic kidney disease. Serum BUN and PTH are reported. Calcitonin was found to impact at a dose of 50/IU/kg/day five times a week for five weeks. MicroCT was used to evaluate bone quantity measures, such as cortical porosity, thickness, bone area, and long bone structural geometric parameters. It was found that porosity, thickness, and bone geometry are affected by disease, but not by treatment at the specified regime. Small and wide-angle x-ray scattering (SAXS and WAXS) was used to obtain the nanostructure of the mineral-collagen-water composite, including mineral dimensions, -periodicity and collagen spacing. Data from thermogravimetric analysis (TgA) were used to quantify wt.% of the mineral, collagen, and bound water of each sample. Chronic kidney disease was found to decrease collagen spacing to increase mineral weight fractions, and to reduce loosely bound water. There were no changes from chronic kidney disease on the -Periodicity. Treatment increased the weight percent of collagen, with no effect on other bone quality parameters.
{"title":"Nanostructural changes in bone quality in a mouse model of chronic kidney disease and treatment with calcitonin","authors":"E. Montagnino , W. Bush , J. Bustamante , W. Bandara , P. Jalaie , M.R. Allen , J.M. Wallace , T. Siegmund , R.K. Surowiec , J.A. Howarter","doi":"10.1016/j.bonr.2025.101880","DOIUrl":"10.1016/j.bonr.2025.101880","url":null,"abstract":"<div><div>Mineral imbalances in the body from chronic kidney disease can impact bone turnover and cause cortical bone loss. Synthetic salmon calcitonin is an FDA-approved treatment for bone fragility observed in diseases such as osteoporosis, and clinical trials have demonstrated a reduction in fractures compared to untreated individuals. This study documents the effects of calcitonin on cortical bone using an in vivo mouse model of chronic kidney disease. Serum BUN and PTH are reported. Calcitonin was found to impact at a dose of 50/IU/kg/day five times a week for five weeks. MicroCT was used to evaluate bone quantity measures, such as cortical porosity, thickness, bone area, and long bone structural geometric parameters. It was found that porosity, thickness, and bone geometry are affected by disease, but not by treatment at the specified regime. Small and wide-angle x-ray scattering (SAXS and WAXS) was used to obtain the nanostructure of the mineral-collagen-water composite, including mineral dimensions, <span><math><mi>D</mi></math></span>-periodicity and collagen spacing. Data from thermogravimetric analysis (TgA) were used to quantify wt.% of the mineral, collagen, and bound water of each sample. Chronic kidney disease was found to decrease collagen spacing to increase mineral weight fractions, and to reduce loosely bound water. There were no changes from chronic kidney disease on the <span><math><mi>D</mi></math></span>-Periodicity. Treatment increased the weight percent of collagen, with no effect on other bone quality parameters.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101880"},"PeriodicalIF":2.6,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145358576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-19DOI: 10.1016/j.bonr.2025.101878
Frederik Duch Bromer , Andreas Lodberg , Lykke Sylow , Michala Carlsson , Christian Brix Folsted Andersen , Jesper Skovhus Thomsen , Annemarie Brüel
Aim
Weight-loss therapy often results in an unintended loss of muscle and bone mass. Inhibitors of the activin receptor signaling pathway, such as bimagrumab, an anti-activin receptor antibody (αActRIIA/IIB ab), are under investigation to counteract weight-loss induced muscle loss, but their skeletal effects in obesity remain unclear. This study investigates αActRIIA/IIB ab on bone in mice exposed to a high-fat diet (HFD) model of obesity or standard chow.
Materials and methods
Male C57BL/6 J mice were stratified into four groups (n = 10/group, standard chow or HFD for 10 weeks ± αActRIIA/IIB ab). αActRIIA/IIB ab (10 mg/kg) was administered twice weekly during the final three weeks. The femur and vertebral body were assessed using DEXA, μCT, mechanical testing, and histomorphometry.
Results
HFD did not affect bone density, microstructure, or strength but reduced histological bone formation markers. In standard chow mice, αActRIIA/IIB ab increased trabecular bone volume fraction (BV/TV) and volumetric bone mineral density (vBMD) by 36 %. In HFD mice, the effect of αActRIIA/IIB ab was less pronounced but still increased BV/TV (+16 %) and vBMD (+13 %). For cortical bone, μCT parameters remained largely unaffected by αActRIIA/IIB ab, while the treatment increased periosteal mineralizing bone surfaces in standard chow mice (+217 %), but not in HFD mice.
Conclusions
αActRIIA/IIB ab enhanced trabecular bone properties in standard chow-fed mice, but its anabolic effects were blunted in HFD-fed mice. Furthermore, αActRIIA/IIB ab improved cortical histological bone formation markers, while morphology remained unaffected, suggesting a site- or time-specific difference. Thus, αActRIIA/IIB ab holds potential for mitigating weight-loss-associated bone deterioration.
{"title":"High-fat diet induced obesity and anti-activin receptor antibody: Effects on bone properties in mice","authors":"Frederik Duch Bromer , Andreas Lodberg , Lykke Sylow , Michala Carlsson , Christian Brix Folsted Andersen , Jesper Skovhus Thomsen , Annemarie Brüel","doi":"10.1016/j.bonr.2025.101878","DOIUrl":"10.1016/j.bonr.2025.101878","url":null,"abstract":"<div><h3>Aim</h3><div>Weight-loss therapy often results in an unintended loss of muscle and bone mass. Inhibitors of the activin receptor signaling pathway, such as bimagrumab, an anti-activin receptor antibody (αActRIIA/IIB ab), are under investigation to counteract weight-loss induced muscle loss, but their skeletal effects in obesity remain unclear. This study investigates αActRIIA/IIB ab on bone in mice exposed to a high-fat diet (HFD) model of obesity or standard chow.</div></div><div><h3>Materials and methods</h3><div>Male C57BL/6 J mice were stratified into four groups (<em>n</em> = 10/group, standard chow or HFD for 10 weeks ± αActRIIA/IIB ab). αActRIIA/IIB ab (10 mg/kg) was administered twice weekly during the final three weeks. The femur and vertebral body were assessed using DEXA, μCT, mechanical testing, and histomorphometry.</div></div><div><h3>Results</h3><div>HFD did not affect bone density, microstructure, or strength but reduced histological bone formation markers. In standard chow mice, αActRIIA/IIB ab increased trabecular bone volume fraction (BV/TV) and volumetric bone mineral density (vBMD) by 36 %. In HFD mice, the effect of αActRIIA/IIB ab was less pronounced but still increased BV/TV (+16 %) and vBMD (+13 %). For cortical bone, μCT parameters remained largely unaffected by αActRIIA/IIB ab, while the treatment increased periosteal mineralizing bone surfaces in standard chow mice (+217 %), but not in HFD mice.</div></div><div><h3>Conclusions</h3><div>αActRIIA/IIB ab enhanced trabecular bone properties in standard chow-fed mice, but its anabolic effects were blunted in HFD-fed mice. Furthermore, αActRIIA/IIB ab improved cortical histological bone formation markers, while morphology remained unaffected, suggesting a site- or time-specific difference. Thus, αActRIIA/IIB ab holds potential for mitigating weight-loss-associated bone deterioration.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101878"},"PeriodicalIF":2.6,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145118986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-10DOI: 10.1016/j.bonr.2025.101876
Honey Hendesi , Dana A. Godfrey , Ana Ferreira Ruble , Aaron M. Tran , David A. Villani , Samantha H. Landgrave , Nur A. Hasan , Douglas J. Adams , Michael J. Zuscik
Intermittent Fasting (IF) is a dietary strategy with metabolic benefits that can reverse certain obesity-related pathologies. This study aimed to investigate whether IF can mitigate delayed bone fracture healing associated with obesity. Using cohorts of mice on high-fat or control diets, we applied either an ad libitum feeding or an alternate-day fasting regimen to animals from both diet groups. We assessed bone healing outcomes by evaluating callus mineralization and adipocyte accumulation within the callus through micro computed tomography (micro-CT), histology, and immunohistochemical analyses. Since IF is known to modulate gut microbiome composition, often associated with improvement in various metabolic and inflammatory processes, particularly in high-fat-fed mice, we also explored the microbial community changes in IF mice through 16S rRNA sequencing of cecal samples. Metabolically, IF led to reduced body weight and improved glucose tolerance in obese mice. Regarding fracture healing outcomes, reduced/delayed mineralization and adipocyte accumulation in fracture callus tissue in the high-fat-fed cohort were significantly attenuated when the high-fat-fed mice were subjected to alternate-day fasting. These benefits of IF were not observed in lean mice fed a control diet. Furthermore, IF significantly altered the gut microbiota of mice on a high-fat diet, including an increased abundance of short-chain fatty acid producing bacteria, known for their positive effect on bone density, and a reduction in various pro-inflammatory taxa. While the mechanistic role remains unknown, these findings suggest that the improved fracture healing observed in obese mice following IF may be associated with alterations in gut microbiome composition and function.
{"title":"Intermittent fasting alleviates obesity-associated impairments in bone fracture healing: Exploring the role of gut microbiome","authors":"Honey Hendesi , Dana A. Godfrey , Ana Ferreira Ruble , Aaron M. Tran , David A. Villani , Samantha H. Landgrave , Nur A. Hasan , Douglas J. Adams , Michael J. Zuscik","doi":"10.1016/j.bonr.2025.101876","DOIUrl":"10.1016/j.bonr.2025.101876","url":null,"abstract":"<div><div>Intermittent Fasting (IF) is a dietary strategy with metabolic benefits that can reverse certain obesity-related pathologies. This study aimed to investigate whether IF can mitigate delayed bone fracture healing associated with obesity. Using cohorts of mice on high-fat or control diets, we applied either an ad libitum feeding or an alternate-day fasting regimen to animals from both diet groups. We assessed bone healing outcomes by evaluating callus mineralization and adipocyte accumulation within the callus through micro computed tomography (micro-CT), histology, and immunohistochemical analyses. Since IF is known to modulate gut microbiome composition, often associated with improvement in various metabolic and inflammatory processes, particularly in high-fat-fed mice, we also explored the microbial community changes in IF mice through 16S rRNA sequencing of cecal samples. Metabolically, IF led to reduced body weight and improved glucose tolerance in obese mice. Regarding fracture healing outcomes, reduced/delayed mineralization and adipocyte accumulation in fracture callus tissue in the high-fat-fed cohort were significantly attenuated when the high-fat-fed mice were subjected to alternate-day fasting. These benefits of IF were not observed in lean mice fed a control diet. Furthermore, IF significantly altered the gut microbiota of mice on a high-fat diet, including an increased abundance of short-chain fatty acid producing bacteria, known for their positive effect on bone density, and a reduction in various pro-inflammatory taxa. While the mechanistic role remains unknown, these findings suggest that the improved fracture healing observed in obese mice following IF may be associated with alterations in gut microbiome composition and function.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101876"},"PeriodicalIF":2.6,"publicationDate":"2025-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145047410","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-06DOI: 10.1016/j.bonr.2025.101877
Guido Filler , Harry Chandrakumaran , Funmbi Babalola , Dougenie Emile , Shih-Han Susan Huang , Robert Stein
Background
Burosumab, a monoclonal antibody to fibroblast growth factor 23 (FGF23), is effective for X-linked hypophosphatemic rickets (XLH). Renal effects, particularly calciuria and nephrocalcinosis, remain incompletely characterized.
Methods
In this retrospective cohort, 13 children with genetically confirmed XLH (7 females, 6 males; 0.6–16.3 years) were evaluated after conversion from conventional therapy to burosumab. Longitudinal changes in serum phosphate, TmP/GFR, TRP, 1,25(OH)₂D, intact parathyroid hormone (PTH), and urinary calcium/creatinine (Ca:Cr) were assessed. Hypercalciuria was defined using age-specific SI thresholds; Ca:Cr was also expressed as ×ULN (Ca:Cr divided by the age-specific upper limit).
Results
Burosumab improved serum phosphate (p < 0.001), TmP/GFR (p < 0.001), and TRP (p = 0.007). Despite normalized phosphate handling, two patients developed de novo nephrocalcinosis. No child was hypercalciuric at washout or Day 14; two had ≥1 episode later. A repeated-measures analysis of log10(×ULN) showed no overall time effect from washout to Day 56 (F(4,29) = 1.66, p = 0.185). A larger decline in PTH correlated with higher Ca:Cr (p < 0.05), whereas 1,25(OH)₂D did not.
Conclusion
Burosumab improves phosphate homeostasis in children with XLH, but a minority may develop hypercalciuria and nephrocalcinosis, potentially linked to PTH suppression. Vigilant biochemical and ultrasound monitoring—particularly early after conversion—and consideration of prophylaxis in high-risk cases are advisable.
{"title":"Effect of burosumab conversion on calciuria and nephrocalcinosis in children with XLH: A real-world cohort study","authors":"Guido Filler , Harry Chandrakumaran , Funmbi Babalola , Dougenie Emile , Shih-Han Susan Huang , Robert Stein","doi":"10.1016/j.bonr.2025.101877","DOIUrl":"10.1016/j.bonr.2025.101877","url":null,"abstract":"<div><h3>Background</h3><div>Burosumab, a monoclonal antibody to fibroblast growth factor 23 (FGF23), is effective for X-linked hypophosphatemic rickets (XLH). Renal effects, particularly calciuria and nephrocalcinosis, remain incompletely characterized.</div></div><div><h3>Methods</h3><div>In this retrospective cohort, 13 children with genetically confirmed XLH (7 females, 6 males; 0.6–16.3 years) were evaluated after conversion from conventional therapy to burosumab. Longitudinal changes in serum phosphate, TmP/GFR, TRP, 1,25(OH)₂D, intact parathyroid hormone (PTH), and urinary calcium/creatinine (Ca:Cr) were assessed. Hypercalciuria was defined using age-specific SI thresholds; Ca:Cr was also expressed as ×ULN (Ca:Cr divided by the age-specific upper limit).</div></div><div><h3>Results</h3><div>Burosumab improved serum phosphate (<em>p</em> < 0.001), TmP/GFR (p < 0.001), and TRP (<em>p</em> = 0.007). Despite normalized phosphate handling, two patients developed de novo nephrocalcinosis. No child was hypercalciuric at washout or Day 14; two had ≥1 episode later. A repeated-measures analysis of log10(×ULN) showed no overall time effect from washout to Day 56 (F(4,29) = 1.66, <em>p</em> = 0.185). A larger decline in PTH correlated with higher Ca:Cr (<em>p</em> < 0.05), whereas 1,25(OH)₂D did not.</div></div><div><h3>Conclusion</h3><div>Burosumab improves phosphate homeostasis in children with XLH, but a minority may develop hypercalciuria and nephrocalcinosis, potentially linked to PTH suppression. Vigilant biochemical and ultrasound monitoring—particularly early after conversion—and consideration of prophylaxis in high-risk cases are advisable.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101877"},"PeriodicalIF":2.6,"publicationDate":"2025-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145107001","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-03DOI: 10.1016/j.bonr.2025.101874
Molly E. Muehlebach , Sarah A. Holstein
Myeloma bone disease (MBD) is characterized by tumor-induced osteoclast activation with concomitant suppression of osteoblast activity. Nitrogen bisphosphonates (NBPs), including zoledronic acid (ZA), are a mainstay of MBD treatment, due to their anti-osteoclastic effects secondary to high bone affinity and indirect disruption of protein geranylgeranylation through inhibition of farnesyl diphosphate synthase. The development of geranylgeranyl diphosphate synthase (GGDPS) inhibitors (GGSIs) represents a more selective means of targeting geranylgeranylation. The GGSI RAM2061 has direct anti-myeloma activity in vitro and in vivo, achieves both systemic and skeletal distribution, and has anti-osteoclastic activity. However, the effects of this novel therapy on osteoblast activity or in a setting that recapitulates the MBD milieu have yet to be explored. Exposure to RAM2061 or ZA during MC3T3-E1 differentiation resulted in impairment in osteoblast function, including alkaline phosphatase and mineralization activity, however minimal effects were observed in differentiated MC3T3-E1 cells that were subsequently exposed to drug. To evaluate the impact of RAM2061 on osteoblast or osteoclast activity under MBD-like conditions, JJN3 myeloma cell conditioned media (CM) was collected and added to bone cell cultures in order to simulate osteoclast-activating or osteoblast-inhibitory MBD microenvironments. These studies determined that RAM2061 maintains anti-resorptive effects and osteoblast inhibitory effects in undifferentiated precursors while in the presence of JJN3 CM. However, no appreciable effects were detected in osteoblasts exposed to drug post-differentiation. Overall, these studies contribute to the mechanistic understanding of NBP and GGSI effects in the bone and provide support for the continued investigation of GGSIs in MBD.
{"title":"The effects of geranylgeranyl diphosphate synthase inhibitor treatment on osteoblast biology and application in a conditioned media model of myeloma bone disease","authors":"Molly E. Muehlebach , Sarah A. Holstein","doi":"10.1016/j.bonr.2025.101874","DOIUrl":"10.1016/j.bonr.2025.101874","url":null,"abstract":"<div><div>Myeloma bone disease (MBD) is characterized by tumor-induced osteoclast activation with concomitant suppression of osteoblast activity. Nitrogen bisphosphonates (NBPs), including zoledronic acid (ZA), are a mainstay of MBD treatment, due to their anti-osteoclastic effects secondary to high bone affinity and indirect disruption of protein geranylgeranylation through inhibition of farnesyl diphosphate synthase. The development of geranylgeranyl diphosphate synthase (GGDPS) inhibitors (GGSIs) represents a more selective means of targeting geranylgeranylation. The GGSI RAM2061 has direct anti-myeloma activity in vitro and in vivo, achieves both systemic and skeletal distribution, and has anti-osteoclastic activity. However, the effects of this novel therapy on osteoblast activity or in a setting that recapitulates the MBD milieu have yet to be explored. Exposure to RAM2061 or ZA during MC3T3-E1 differentiation resulted in impairment in osteoblast function, including alkaline phosphatase and mineralization activity, however minimal effects were observed in differentiated MC3T3-E1 cells that were subsequently exposed to drug. To evaluate the impact of RAM2061 on osteoblast or osteoclast activity under MBD-like conditions, JJN3 myeloma cell conditioned media (CM) was collected and added to bone cell cultures in order to simulate osteoclast-activating or osteoblast-inhibitory MBD microenvironments. These studies determined that RAM2061 maintains anti-resorptive effects and osteoblast inhibitory effects in undifferentiated precursors while in the presence of JJN3 CM. However, no appreciable effects were detected in osteoblasts exposed to drug post-differentiation. Overall, these studies contribute to the mechanistic understanding of NBP and GGSI effects in the bone and provide support for the continued investigation of GGSIs in MBD.</div></div>","PeriodicalId":9043,"journal":{"name":"Bone Reports","volume":"27 ","pages":"Article 101874"},"PeriodicalIF":2.6,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145010897","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号