Star volume in bone research. A histomorphometric analysis of trabecular bone structure using vertical sections

Conventional bone histomorphometry performed on iliac crest biopsies does not generally provide unbiased stereological estimates of parameters related to bone structure due to the anisotropy of trabecular bone; this, however, can be obtained with vertical sections, which are anisotropic sections, in combination with an anisotropic test system. A practical procedure for obtaining vertical sections from bone is described. The new stereological parameter, the star volume, can provide an unbiased estimation of the absolute mean size of the marrow space and thus give an indirect estimate of the connectivity of trabecular bone structure. The marrow space star volume of vertebral bodies and iliac crest increases with age in both sexes demonstrating that the structural bone changes which occur with age is a topological one with changes in trabecular connectivity. The practical procedure including sampling efficiency for marrow space star volume is described. © 1993 Wiley-Liss, Inc.     

The three-dimensional microstructure of the trabecular bone in the mandible

This study investigated the three dimensional (3D) trabecular microstructure of the alveolar and basal bone in the mandible using micro-CT and compared the morphometric values of the different sites. Ten specimens were prepared and scanned using a micro-CT system. Both the alveolar and basal trabecular bone of the premolar region in the mandible were measured for the structural analysis. Cross-sectional 1024×1024 pixel images were created. From the two-dimensional (2D) images produced, 3D structural images were reconstructed. After scanning the specimen, the volumes of interest (VOI) of the alveolar and basal bone regions were selected from the 3D reconstruction images, and the structural parameters such as bone volume fraction, bone surface density, trabecular thickness, trabecular separation, trabecular number and structural model index were analyzed. The trabecular structure showed a marked variation within the sites of the specimen, especially in the basal trabecular bone inferior to the mandibular canal. In both the alveolar and basal bone regions, a mixture of both plate-like and rod-like structures was observed. The alveolar region showed a more compact, plate-type trabecular structure than the basal regions. In parametric comparison with the basal bone, the alveolar bone generally had a higher bone volume fraction, bone trabecular thickness and trabecular number, and lower bone surface density, trabecular separation and structural model index. The alveolar bone consisted of a compact bone structure with a large amount of thick plate-type trabecular bone, which was effectively resistant to the masticatory forces. As the measurements were made closer to the basal bone, a loose structure was observed with lower bone volume and fewer, thin, rod-like trabeculae.     

Trabecular bone structure in patients with primary hyperparathyroidism

To evaluate the effects of increased parathyroid hormone (PTH)-secretion on trabecular bone structure in patients with primary hyperparathyroidism (PHPT) we have analysed iliac crest biopsies from 84 patients (38 male, 46 female, age 20–85 years) with surgically proven PHPT by using quantitative histomorphometry. As there might be an influence of age or sex, subjects were divided into 4 subgroups according to age (younger than 50 years, older than 50 years) and sex. Eighty four age and sex matched autopsy cases of accidental death served as controls. When compared with age matched controls, trabecular bone volume (BV/TV) and trabecular diameter in PHPT were increased. Trabecular density in PHPT was elevated significantly in the older age-groups. These results suggest a preservation of intertrabecular connectivity, which we were able to confirm by measurement of the number of intertrabecular nodes and trabecular bone pattern factor (TBPf). No differences were demonstrated between male and female subjects. Furthermore, no variations according to the histological type (with or without endosteal fibrosis) could be demonstrated in our material. We conclude that increased trabecular bone volume in PHPT is the result not only of thicker trabecula but also of a reduction of the age-dependent loss of complete trabecular plates due to perforations. Thus PHPT leads to a substantial preservation of intertrabecular connectivity.     

脊髓损伤早期及制动大鼠股骨干骺端的显微CT观察

背景：脊髓损伤后可引起损伤平面以下骨量大量丢失,导致骨质疏松。 目的：观察比较脊髓损伤及失用性制动模型大鼠股骨远端骨密度及骨微观结构的改变。 方法：将SD大鼠随机分为3组：对照组,切除T10椎板,不损伤硬膜及脊髓;脊髓损伤组,切除T10椎板后行Allen's法造成脊髓损伤;制动组,以大鼠双侧腿－尾缝合造成双下肢制动。10 d后取一侧尺、桡骨及股骨行骨密度检测,另一侧股骨行显微CT扫描。 结果与结论：脊髓损伤组与制动组大鼠股骨远端骨密度、骨矿物质含量、骨体积分数表、骨小梁厚度、骨皮质面积及厚度、骨小梁数量均低于对照组(P < 0.05 ),骨小梁结构模型指数、骨表面积体积比、骨小梁分离度均高于对照组;脊髓损伤组上述指标较制动组变化程度更显著(P < 0.05)。3组尺、桡骨密度差异无显著性意义。说明脊髓损伤及制动均可导致骨量丢失,在脊髓损伤早期损伤平面以下部位骨微观结构呈现骨质疏松明显改变,且程度比失用性因素严重。     

骨小梁Micro-CT图像形态计量学参数计算方法综述

骨质疏松症是骨科领域的研究热点之一,研究发现,除骨密度(骨量)改变的因素外,骨小梁的结构变化也是患病的重要影响因素。骨小梁形态计量学分析是研究骨小梁结构形态变化的一种重要方法。基于骨小梁的Micro-CT图像介绍部分骨小梁形态学参数的计算方法,主要包括骨小梁各向异性、连通性、结构模型指数以及纹理等不同特征,同时列举部分相关研究实例,总结上述形态学参数的适用性以及优缺点,为更加有效地评价骨质疏松状态以及药物治疗效果提供依据。     

Measures of complexity for cancellous bone.

The problem of quantifying the structure of cancellous bone has been addressed in the past by histomorphometry and more recently by imaging techniques using X-ray attenuation. The current approaches compute and describe parts of the construction of the trabecular net. We developed a new technique which quantifies cancellous bone of human lumbar vertebrae as a whole. The interactions, transactions, and interrelationships of all parts of the structural composition of the trabeculae are accounted for and quantified. The method is based on the concept of structural complexity within the framework of nonlinear dynamics. The methodology was developed by using axial high resolution computed tomography images. The technique was transferred to quantitative computed tomography images and is based on the non-invasive assessment of 50 human L3 specimens. The value of Houndsfield units per pixel representing trabecular bone of the vertebrae was transformed into color-encoded and alphabet-encoded symbols. The procedure of transformation of the X-ray attenuation pixels into symbols was necessary as a basis on which measures of complexity were introduced to assess the composition of symbols within the images. The development of a generalization of symbolic dynamics, a mathematical method, to work with two-dimensional images was a prerequisite. The results of this study demonstrate that the structural composition of cancellous bone declines more rapidly than bone mineral density during the loss of bone. This outcome strongly suggests an exponential relationship between bone mineral density and the architectural composition of cancellous bone. Normal trabecular bone has a complex ordered structure. The structural composition during the osteopenic phase of bone loss is characterized by lower structural complexity and a significantly higher level of architectural disorder. A high grade of osteoporosis leads again to an ordered structure, although its structural complexity is minimal.     

Fractal analysis of radiographs: assessment of trabecular bone structure and prediction of elastic modulus and strength.

The purpose of this study was to determine whether fractal dimension of radiographs provide measures of trabecular bone structure which correlate with bone mineral density (BMD) and bone biomechanics, and whether these relationships depend on the technique used to calculate the fractal dimension. Eighty seven cubic specimen of human trabecular bone were obtained from the vertebrae and femur. The cubes were radiographed along all three orientations--superior-inferior (SI), medial-lateral (ML), and anterior-posterior (AP), digitized, corrected for background variations, and fractal based techniques were applied to quantify trabecular structure. Three different techniques namely, semivariance, surface area, and power spectral methods were used. The specimens were tested in compression along three orientations and the Young's modulus (YM) was determined. Compressive strength was measured along the SI direction. Quantitative computed tomography was used to measure trabecular BMD. High-resolution magnetic-resonance images were used to obtain three-dimensional measures of trabecular architecture such as the apparent bone volume fraction, trabecular thickness, spacing, and number. The measures of trabecular structure computed in the different directions showed significant differences (p<0.05). The correlation between BMD, YM, strength, and the fractal dimension were direction and technique dependent. The trends of variation of the fractal dimension with BMD and biomechanical properties also depended on the technique and the range of resolutions over which the data was analyzed. The fractal dimension showed varying trends with bone mineral density changes, and these trends also depended on the range of frequencies over which the fractal dimension was measured. For example, using the power spectral method the fractal dimension increased with BMD when computed over a lower range of spatial frequencies and decreased for higher ranges. However, for the surface area technique the fractal dimension increased with increasing BMD. Fractal measures showed better correlation with trabecular spacing and number, compared to trabecular thickness. In a multivariate regression model inclusion of some of the fractal measures in addition to BMD improved the prediction of strength and elastic modulus. Thus, fractal based texture analysis of radiographs are technique dependent, but may be used to quantify trabecular structure and have a potentially valuable impact in the study of osteoporosis.     

Changes in the cortical and trabecular bone compartments with different types of menopause measured by peripheral quantitative computed tomography

Our objective was to study the changes in the bone mineral density of the cortical and trabecular compartments with different types of menopause. A total of 153 normal postmenopausal women (mean age 48 ± 5 years) were studied. The women were divided into three groups based on mean age at menopause: early menopause (menopause before 43 years), normal menopause (menopause at 44–52 years), and late menopause (menopause after 52 years). The number of years since menopause was similar in all three groups (±5 years). Cortical and trabecular bone mineral density was determined in all the women using peripheral quantitative computed tomography. Our results show that only the trabecular bone mineral density differed significantly among the groups (Kruskal-Wallis: P = 0.0029). The women with early menopause bad a lower trabecular bone density than the women with normal and late menopause (P = 0.0019 and P < 0.0001, respectively). Among the women with early menopause, 22 had experienced menopause before the age of 40 and 25 after the age of 40; there were significant differences in trabecular bone mineral density between these two subgroups (P < 0.05). Trabecular bone mineral density, the only variable studied that varied among the groups, correlated significantly with the duration of reproductive life (simple linear regression: r = 0.340, P < 0.0001). In conclusion, these findings emphasize the importance of the duration of reproductive life as a determinant of bone mass in women.     

Osseous microarchitecture of the scaphoid: Cadaveric study of regional variations and clinical implications

Bone strength and structure are closely associated with fracture and screw fixation, however osseous micro architecture on scaphoid has not been clearly addressed. We conducted histomorphometric study of the scaphoid using micro CT to find regional variations and differences in the scaphoid to provide better understanding of fracture mechanism and suggest optimal screw position. We divided scaphoid into eight regions and collected regional data from eleven different cadaveric scaphoids. A computer program was used to measure parameters, which includes mean subchondral bone thickness, bone mineral density for bone density parameters, and tissue mineral density, trabecular thickness, trabecular spacing, trabecular number and bone volume fraction for bone quality parameters. All bone strength parameters were measured the maximum value in the regions where scaphoid articulates with radius. Articular regions presented higher bone strength parameters and thicker subchondral bone. The minimum value of trabecular number was in midcarpal side of waist portion. There was trend of higher subchondral bone thickness in the scaphoid which articulates with capitate and radius. This histomorphometric study showed regional variation of the scaphoid in terms of bone density and quality parameters. Waist portion presented thick subchondral and trabecular bone for high cross section moment of inertia against bending. Three point bending for scaphoid fracture and vertical screw placement are suggested based on these variations.     

Prediction of mechanical properties of trabecular bone using quantitative MRI

Techniques for quantitative magnetic resonance imaging (MRI) have been developed for non-invasive estimation of the mineral density and structure of trabecular bone. The R*(2) relaxation rate (i.e. 1/T*(2)) is sensitive to bone mineral density (BMD) via susceptibility differences between trabeculae and bone marrow, and by binarizing MRI images, structural variables, such as apparent bone volume fraction, can be assessed. In the present study, trabecular bone samples of human patellae were investigated in vitro at 1.5 T to determine the ability of MRI-derived variables (R*(2) and bone volume fraction) to predict the mechanical properties (Young's modulus, yield stress and ultimate strength). Further, the MRI variables were correlated with reference measurements of volumetric BMD and bone area fraction as determined with a clinical pQCT system. The MRI variables correlated significantly (p < 0.01) with the mechanical variables (r = 0.32-0.46), BMD (r = 0.56) and bone structure (r = 0.51). A combination of R*(2) and MRI-derived bone volume fraction further improved the prediction of yield stress and ultimate strength. Although pQCT showed a trend towards better prediction of the mechanical properties, current results demonstrate the feasibility of combined MR imaging of marrow susceptibility and bone volume fraction in predicting the mechanical strength of trabecular bone and bone mineral density.     

雷公藤甲素通过抑制破骨细胞生成预防骨丢失的研究

目的 探讨雷公藤甲素抗骨质疏松的作用及机制。方法 建立大鼠老年性骨质疏松模型。40只22月龄雄性SD大鼠随机分为雷公藤甲素（每天15μg/kg腹腔注射）治疗组和生理盐水对照组（每天15μg/kg腹腔注射）,连续8周。采用显微CT分析胫骨近端骨松质的骨密度(BMD)和骨显微结构。WB检测成骨相关蛋白表达水平。TRACP-5b染色法测定破骨细胞数,同时检测骨吸收标志物表达水平。结果 显微CT结果显示,雷公藤甲素治疗组大鼠骨密度、骨体积/总体积比值(Bv/Tv)、骨小梁厚度(Tb.Th)、骨小梁数目(Tb.N)、骨小梁间距(Tb.Sp)均显著高于对照组（P<0.05）。两组的成骨相关蛋白表达水平无明显差异,TRACP-5b染色显示雷公藤甲素减少了体内破骨细胞的数量（P<0.05）,同时血液骨吸收标志物水平也明显降低（P<0.05）。结论 雷公藤甲素通过抑制破骨细胞生成进而对老年性骨质疏松有保护作用。雷公藤甲素可能是治疗老年性骨质疏松症的一种可行方案。     

Development of the trabecular structure within the ulnar medial coronoid process of young dogs

This study describes the timing of development of the trabecular structure of the ulnar medial coronoid process (MCP) in the dog. The right MCPs of nine healthy golden retrievers, aged 4 to 24 weeks, without signs of secondary joint disease were dissected and scanned with microcomputed tomography (micro-CT) at a voxel size of 34 microm to determine histomorphometric parameters. Bone volume fraction and mean trabecular separation show a reciprocal pattern in time, reflecting an initial high bone density (and low trabecular separation), and then a sharp drop in density at 8-10 weeks, followed by a gradual increase to high values at 24 weeks. With a similar bone volume fraction as in young bone, the older bone shows thicker trabeculae and a more plate-like structure. This is reflected in the much smaller number of trabeculae and the lower surface/volume ratio at higher age. An anisotropic structure of the trabeculae with an orientation in the direction of the proximodistal axis of the ulna is already present at 6 weeks after birth. This primary alignment was perpendicular to the humeroulnar articular surface, matching the direction of the compressive forces applied to the MCP by the humeral trochlea. The secondary alignment appeared at 13 weeks after birth and was directed along the craniocaudal axis of the MCP, toward the attachment of the anular ligament. In comparison with data from long bones and vertebrae, the findings of a high bone volume fraction and a well-defined trabecular alignment at a very early age are remarkable. The high bone volume fraction is possibly a remnant of the fetal trabecular structure, as dogs are relatively immature at birth compared to other animals. Soon after the start of steady locomotion, the trabecular structure changes into a more mature-like structure. The early trabecular alignment is possibly a reflection of the early load-bearing function of the MCP in the elbow joint.     

A review of magnetic resonance (MR) imaging of trabecular bone micro-architecture: contribution to the prediction of biomechanical properties and fracture prevalence.

Bone mineral density and three-dimensional trabecular structure play a significant role in predicting bone strength and biomechanical properties. MR is a non-invasive technique for determining trabecular architecture both in vivo and in vitro. In this paper we review the use of magnetic resonance imaging to obtain high resolution images of trabecular bone structure and quantify the three-dimensional architecture of the trabecular bone network. Studies assessing the anisotropy of the trabecular architecture in human cadaveric specimens from the distal and proximal femur, and the thoracic and lumbar vertebrae, are reviewed. The contributions of the MR derived measures of 3D trabecular bone structure to the biomechanical strength of the specimen are presented. In vivo, the relationship between the high resolution MR derived trabecular bone structure parameters in the distal radius and calcaneus in patients with hip fractures, are compared to age matched normal controls. MR derived measures are compared to measures of trabecular bone mineral density (BMD) in the hip using dual X-ray absorptiometry (DXA).     

超重环境对骨组织生物力学性能和形态结构的影响

目的 探讨超重环境对骨组织生物力学性能及骨组织形态结构的影响,为航天员和飞行员安全飞行训练及 最大耐受高重力值的确定提供理论支撑。 方法 采用本课题组自主设计的超重加载平台构建超重动物模型,加载 方式为腹背方向承受向心加速度,加速度分别为 5、10、20 g,离心 45 s,1 次/ d,5 d / 周,连续 4 周。 使用 Instron 5865 材料力学试验机对离体骨组织进行三点弯曲试验,通过测量力学参数的变化,分析超重环境对骨组织生物力学性 能的影响;通过双能 X 线骨密度仪扫描、Micro-CT 扫描、骨组织染色等技术,检测骨矿物质含量、骨密度及骨小梁等 形态学参数的变化,观察超重环境对骨组织形态结构的影响。 结果 超重加载可显著降低小鼠胫骨的断裂应力及 弹性模量。 超重环境导致骨组织中骨体积分数下降,骨小梁厚度以及骨小梁数量显著降低,骨小梁排列紊乱。 结 论 超重环境对骨组织生物力学性能及骨组织形态结构的影响与超重载荷的强度密切相关。 低(5 g)强度超重载 荷能够降低骨小梁厚度,但对其力学性能的影响很小。 中(10 g)、高(20 g)强度超重载荷能够显著触发骨流失以 及骨力学性能的下降。 随着超重载荷的增加,机体的生理调节无法阻止骨量和力学性能下降的趋势     

Long-term osteopenic changes in cancellous bone structure in ovariectomized rats

Cancellous bone mass decreases following ovariectomy in rodents, providing a useful model for post-menopausal bone loss in humans. This study describes and quantifies the longer-term changes in cancellous bone structure in the ovariectomized (OVX) rat. Rats were OVX or sham-OVX at 100 days of age and bones were collected 540 days later. Lumbar vertebral bodies were prepared for microradiography and structural analyses (nodal analyses and star volume analyses) of cancellous bone. Proximal humerii were prepared for scanning electron microscopy (SEM). Microradiography confirmed the loss of cancellous bone from the central spongiosa regions of the vertebral bodies and the humerii in the OVX rats. Changes in trabecular structural elements included relative increases in the number of free to free, cortical to free, cortical to node struts and decreases in the node to node struts in the OVX animals compared with controls. There were increases in average lengths of the node to free, node to node, and free to free trabecular struts in the OVX animals. The marrow star volume was increased in the OVX animals indicating a greater trabecular separation in these animals compared with controls. Viewed by SEM, metaphyseal trabeculae in the controls consisted of rods and plates but in the OVX animals the remaining trabeculae were mostly longitudinal rods with smaller transverse connecting rods. The remaining bone in the OVX animals was found in the lateral metaphyseal areas and is consistent with maintenance of the structural capacity of the bone. These long-term changes in cancellous bone structure are likely due to the continuation of functional skeletal loading but a decrease in gonadal hormones resulting in a decreased necessity to maintain a skeletal mineral store for reproduction (e.g., pregnancy and lactation). © 1993 Wiley-Liss, Inc.     

Influence of pattern of menopausal transition on the amount of trabecular bone loss. Results from a 6-year prospective longitudinal study

INTRODUCTION: Bone density is lower in postmenopausal than in premenopausal women. Recent findings have suggested that accelerated bone loss already begins before menopause. Despite numerous cross-sectional studies on menopause-related bone density, longitudinal data on perimenopausal bone density changes are scarce. This study sought to characterize the dynamics of changes leading to postmenopausal osteopenia and to possibly find the time point at which accelerated bone loss begins. METHODS: We prospectively followed 34 pre-, peri- and early postmenopausal women without prior external hormone use, measuring their lumbar spine trabecular bone density with quantitative computer tomography at 0, 2 and 6 years. The analysis of the changes over time was done in a tri-parted fashion, since menopausal status changed variably for individual subjects: we grouped the participants according to their currently valid menopausal classification for prospective (baseline classification), interim (2 years) and retrospective (6-year classification) analysis. RESULTS: Six different patterns of menopausal transition were identified in our sample. Bone loss in the groups not reaching postmenopause during 6 years of observation was >50% of the maximum bone loss observed during the study period. Invariably for all analyses, the perimenopausal phase with estrogen levels still adequate was associated with the greatest reduction of trabecular bone mineral density, reaching 6.3% loss annually in the lumbar spine. By comparison, the average rate of loss was slower in the early postmenopause; total bone loss differed by pattern of menopausal transition (one-way ANOVA p<0.05). CONCLUSION: The presented data for the first time show the perimenopausal course of trabecular bone loss (as measured by QCT of the lumbar spine). Acceleration of bone loss during perimenopause reached half-maximal values of the total bone loss measured around menopause, despite adequate serum estradiol levels.     

bBMP异位诱导成骨的Micro—CT评价

借助Micro—CT评价单纯牛骨形态发生蛋白（bovine bone morphogenetic protein,bBMP）异位诱导成骨的长期三维影像学及骨质变化。（20±2）g昆明小鼠21只,麻醉后于双侧股部肌肉中植入bBMP各2mg,分别于1、2、4、6、8、10、12周各处死3只,切取诱导分化组织,5％戊二醛固定,行Micro—CT扫描和三维重建,运用ABA专用骨骼分析软件测定组织矿含量（tissue mineral content,TMC）,组织骨密度（tissue mineral density,TUB）,骨体积分数（bone volume fraction,BVF）,结构模型指数（structure model index,SMI）,骨小梁厚度（trabecular thickness,Tb．Th）,骨小梁数量（trabecular number,Tb．N）及皮质骨骨密度（bone mineral density,BMD）等参数,运用SPSS10．0统计软件进行统计学分析。bBMP从植入2周开始逐渐形成一椭圆形骨组织块,2～4周,异位生成骨呈疏松的新生骨,4周时组织矿含量达第一个峰值,骨小梁数量最多;随着观察时间的延长（6-12周）,异位诱导生成的椭圆形骨组织内部骨小梁逐渐吸收,数量减少,12周时骨小梁数量最少;而外层骨组织逐渐塑形成为皮质骨,12周时骨矿含量值、骨小梁厚度、组织骨密度和皮质骨骨密度均达最大值。说明bBMP具有强大的异位骨诱导能力,血供不足时,骨质降解吸收;血供充足时,骨质逐渐成熟改建。     

Architecture and mineralization of developing cortical and trabecular bone of the mandible

Ossification of the presumptive trabecular bone in the mandibular condyle and the presumptive cortical bone in the mandibular corpus of the pig mandible was investigated during development, using micro-computed tomography (microCT). Three-dimensional architecture and mineralization characteristics were assessed from ten pigs of different developmental ages. In the condyle, increases in trabecular thickness and separation and a decrease in the trabecular number, led to an unchanged bone volume fraction. A conversion from rod-like into plate-like trabeculae was observed. Bone volume and trabecular thickness were always higher in the corpus, where an increase in bone volume fraction was caused by an increase in the trabecular thickness and a decrease in separation. A transition from a plate-like structure into a more compact structure took place. The average degree of mineralization in the condyle and the corpus increased with age. In the corpus, the degrees of mineralization were higher than in the condyle. The differences between the condyle and corpus and the changes with age could be explained by differences in the distribution of mineralization within the trabecular elements. Generally, the degrees of mineralization increased from the surface toward the centers of the trabecular elements, indicating growth of the trabecular elements by the surface apposition of new mineral.