A new method of comprehensive static histomorphometry applied on human lumbar vertebral cancellous bone.

The aim of the present study was to assess age-related changes in the human spine by use of established static histomorphometry, and to determine how these static histomorphometric measures are interrelated in human cancellous bone tissue. The material comprised normal human lumbar vertebral bodies (L-2) from 12 women (19-96 years) and 12 men (23-91 years) selected from a larger autopsy material to give an even age and gender distribution. In addition, L-2 from three female subjects (80, 88, and 90 years) with a known vertebral fracture of L-2 were considered. Approximately 9-mm-thick frontal (mediolateral) slices were embedded in methylmetacrylate, stained with aniline blue, and scanned into a computer with a flatbed image scanner at a high resolution (2400 dpi). With a custom-made computer program the following static histomorphometric measures were determined: trabecular bone volume; marrow space star volume; bone space star volume; anisotropy of bone and marrow phase (star length distribution method); node-strut analysis (node:terminus ratio); trabecular thickness; trabecular number; trabecular separation; and trabecular bone pattern factor. In addition, connectivity density was determined (by the ConnEulor method). All 11 histomorphometric measures, except bone space star volume and the two measures of anisotropy, showed a significant correlation with age. Marrow space star volume (r = 0.82) and trabecular bone volume (r = -0.81) showed the highest correlation with age. Furthermore, it was found that all of the histomorphometric measures were correlated, to different degrees. Trabecular bone volume correlated significantly with all ten histomorphometric measures, whereas the two anisotropy measures were poorly correlated to the other measures. Finally, we found the histomorphometric values in this study to be in excellent accordance with various previously published results from studies of human trabecular vertebral bone, the sole exception being marrow space star volume, which was probably due to the small (artificial) region of interest (ROI) that was used in the earlier studies. In conclusion, the new method applied herein allows for easy assessment of age-related changes and also for assessment of relationships between histomorphometric measures in human vertebral cancellous bone.

[1]  S A Goldstein,et al.  The relationship between the structural and orthogonal compressive properties of trabecular bone. , 1994, Journal of biomechanics.

[2]  J. Compston,et al.  A new method for the two‐dimensional analysis of bone structure in human iliac crest biopsies , 1986, Journal of microscopy.

[3]  L. Mosekilde,et al.  Sex differences in age-related loss of vertebral trabecular bone mass and structure--biomechanical consequences. , 1989, Bone.

[4]  W. J. Whitehouse The quantitative morphology of anisotropic trabecular bone , 1974, Journal of microscopy.

[5]  D. C. Sterio The unbiased estimation of number and sizes of arbitrary particles using the disector , 1984, Journal of microscopy.

[6]  G V Cochran,et al.  A new manual method for assessing two‐dimensional cancellous bone structure: Comparison between iliac crest and lumbar vertebra , 1991, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[7]  J S Thomsen,et al.  Age‐ and Gender‐Related Differences in Vertebral Bone Mass, Density, and Strength , 1999, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[8]  D. Chappard,et al.  Altered trabecular architecture induced by Corticosteroids: A Bone Histomorphometric Study , 1996, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[9]  H. Gundersen,et al.  Biologically meaningful determinants of the in vitro strength of lumbar vertebrae. , 1991, Bone.

[10]  Keiichi Abe,et al.  Thinning of Gray-Scale Images with Combined Sequential and Parallel Conditions for Pixel Removal , 1992, IEEE Trans. Syst. Man Cybern. Syst..

[11]  A Odgaard,et al.  Three-dimensional methods for quantification of cancellous bone architecture. , 1997, Bone.

[12]  M Vogel,et al.  Trabecular bone pattern factor--a new parameter for simple quantification of bone microarchitecture. , 1992, Bone.

[13]  Schneider,et al.  Star length distribution: a volume‐based concept for the characterization of structural anisotropy , 1998, Journal of microscopy.

[14]  R. Huiskes,et al.  Relationships between bone morphology and bone elastic properties can be accurately quantified using high‐resolution computer reconstructions , 1998, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[15]  M. Hahn,et al.  Polyostotic heterogeneity of the spine in osteoporosis. Quantitative analysis and three-dimensional morphology. , 1994, Bone and mineral.

[16]  H. Gundersen,et al.  Direct stereological estimation of 3-D connectivity density in human iliac cancellous bone: the effect of age and sex , 1994 .

[17]  J R Nyengaard,et al.  The Conneulor: unbiased estimation of connectivity using physical disectors under projection. , 1993, Bone.

[18]  A. Bailey,et al.  Age-Related Changes in the Biochemical Properties of Human Cancellous Bone Collagen: Relationship to Bone Strength , 1999, Calcified Tissue International.

[19]  P. Croucher,et al.  Assessment of cancellous bone structure: Comparison of strut analysis, trabecular bone pattern factor, and marrow space star volume , 1996, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[20]  H J Gundersen,et al.  Estimation of structural anisotropy based on volume orientation. A new concept , 1990, Journal of microscopy.

[21]  R. T. DeHoff,et al.  Experimental determination of the topological properties of three‐dimensional microstructures , 1972 .

[22]  H J Gundersen,et al.  Star volume of marrow space and trabeculae of the first lumbar vertebra: sampling efficiency and biological variation. , 1989, Bone.

[23]  M. Kleerekoper,et al.  Relationships between surface, volume, and thickness of iliac trabecular bone in aging and in osteoporosis. Implications for the microanatomic and cellular mechanisms of bone loss. , 1983, The Journal of clinical investigation.

[24]  A. Vesterby,et al.  Star volume of marrow space and trabeculae in iliac crest: sampling procedure and correlation to star volume of first lumbar vertebra. , 1990, Bone.

[25]  D. Chappard,et al.  Comparison of eight histomorphometric methods for measuring trabecular bone architecture by image analysis on histological sections , 1999, Microscopy research and technique.

[26]  A. Boyde,et al.  Cancellous Bone Structure in the Growing and Aging Lumbar Spine in a Historic Nubian Population , 1997, Calcified Tissue International.

[27]  P. Croucher,et al.  The effects of gonadotrophin-releasing hormone agonists on iliac crest cancellous bone structure in women with endometriosis. , 1995, Bone.

[28]  M Vogel,et al.  Intervertebral variation in trabecular microarchitecture throughout the normal spine in relation to age. , 1995, Bone.

[29]  Søren E. Larsen,et al.  Characterizing anisotropy: A new concept☆ , 1992 .

[30]  J S Thomsen,et al.  Lumbar vertebral body compressive strength evaluated by dual-energy X-ray absorptiometry, quantitative computed tomography, and ashing. , 1999, Bone.

[31]  R. Huiskes,et al.  Fabric and elastic principal directions of cancellous bone are closely related. , 1997, Journal of biomechanics.

[32]  O. Boachie-Adjei,et al.  Histomorphometric Analysis of Vertebral and Iliac Crest Bone Samples A Correlated Study , 1990, Spine.

[33]  L. Mosekilde,et al.  Nondestructive determination of iliac crest cancellous bone strength by pQCT. , 1997, Bone.

[34]  Ching Y. Suen,et al.  Thinning Methodologies - A Comprehensive Survey , 1992, IEEE Trans. Pattern Anal. Mach. Intell..

[35]  L. Mosekilde Age-related changes in vertebral trabecular bone architecture--assessed by a new method. , 1988, Bone.

[36]  S. Cowin,et al.  Wolff's law of trabecular architecture at remodeling equilibrium. , 1986, Journal of biomechanical engineering.

[37]  L. Mosekilde,et al.  Consequences of the remodelling process for vertebral trabecular bone structure: a scanning electron microscopy study (uncoupling of unloaded structures). , 1990, Bone and mineral.

[38]  E Schneider,et al.  Structure and Function of Vertebral Trabecular Bone , 1997, Spine.

[39]  J S Thomsen,et al.  Relationships between static histomorphometry and bone strength measurements in human iliac crest bone biopsies. , 1998, Bone.

[40]  S C Cowin,et al.  The fabric dependence of the orthotropic elastic constants of cancellous bone. , 1990, Journal of biomechanics.

[41]  M H Bartley,et al.  Skeletal Changes in Aging and Disease , 1966, Clinical orthopaedics and related research.

[42]  D. Mitton,et al.  High-Resolution Computed Tomography for Architectural Characterization of Human Lumbar Cancellous Bone: Relationships with Histomorphometry and Biomechanics , 1999, Osteoporosis International.

[43]  R. Mann,et al.  Characterization of microstructural anisotropy in orthotropic materials using a second rank tensor , 1984 .

[44]  R. Bromley,et al.  Quantitative histological study of human lumbar vertebrae. , 1966, Journal of gerontology.

[45]  H. Beck-Nielsen,et al.  Vertebral bone density evaluated by dual-energy X-ray absorptiometry and quantitative computed tomography in vitro. , 1998, Bone.