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Experimental Bone Biopsies Using Two Bone Biopsy Needles

Rationale and Objectives

The aim of this study was to investigate whether samples obtained using two kinds of small trephines, 2.4 and 1.8 mm in inner diameter, are sufficient for the quantitative evaluation of metabolic bone disease using micro–computed tomographic (CT) three-dimensional parameter data sets.

Materials and Methods

A total of 19 porcine lumbar vertebrae prior to biopsy and biopsy samples from the use of 2.4- and 1.8-mm trephines were examined using micro–CT imaging. For quantitative analysis, seven three-dimensional structural parameters, including trabecular bone volume, trabecular number, trabecular thickness, trabecular separation, the structure model index, the degree of anisotropy, and the trabecular bone pattern factor, were measured using CtAn software. The difference and agreement between the biopsy samples and the baseline vertebrae specimens before biopsy were assessed using paired t tests and Bland-Altman anaylsis, respectively.

Results

There were no significant differences between the 2.4-mm samples and the baseline vertebrae specimens for trabecular bone volume, trabecular thickness, and trabecular number, with mean differences of −0.9%, 2.3%, and −3.1%, respectively; there was no significant difference between the 1.8-mm samples and the baseline vertebrae specimens only for trabecular thickness, with a mean difference of 1.9%.

Conclusion

Samples taken from the use of the 2.4-mm trephine were better for quantitative analysis than those from the use of the 1.8-mm trephine and were acceptable for the quantitative evaluation of trabecular bone volume, trabecular thickness, and trabecular number.

Bone biopsies have been used with increasing frequency for the diagnosis of metabolic bone diseases and for research purposes . In terms of microstructure analysis from a bone biopsy, an appropriate sample size is a major determinant for the accurate qualitative and quantitative histologic diagnosis of metabolic bone disease . Crushing artifacts that occur during the retrieval of samples can cause morphometric differences by resulting in structural damage and compression. Therefore, a biopsy resulting in a bone core with diameter ≥ 5 mm is recommended . More invasive techniques are required for bone specimens of increasing size; thus, it is important to establish a sample size that is sufficient for accurate and representative histologic results. To identify the smallest sample size that can be used with consistency and reliability, several investigations have been undertaken and have reached differing conclusions .

Standard bone histomorphometry has been used as the gold standard to assess trabecular microstructure, but it is time consuming, invasive, and subject to artifacts during preparation. Shrinkage during the cutting process will inevitably lead to the distortion of specimens . Recent advances in the use of micro–computed tomographic (CT) imaging have made it feasible to perform three-dimensional bone structural evaluations nondestructively, quickly, and more simply because of the absence of preparation and may provide high reproducibility and accuracy . Therefore, micro-CT three-dimensional data sets can be used as a substitute for conventional histologic sections for bone structural evaluations . To the best of our knowledge, there has been no report on the smallest sample size that is sufficient for the quantitative diagnosis of metabolic bone disease by using micro-CT imaging instead of histomorphometry.

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Materials and methods

Specimens

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Figure 1, Three- and two-dimensional micro–computed tomographic images of a porcine vertebral body before biopsy. (a) Three-dimensional reconstruction image shows two holes (arrows) scratched at the cortex. Each hole located at the upper and lower cortex indicates the subsequent biopsy site for the use of the 2.4-mm trephine and 1.8-mm trephine, respectively. (b) Axial reconstruction image of the baseline vertebra before biopsy at the level of the lower hole (arrow) . Biopsy was performed in a direction perpendicular to the midline sagittal cut (arrowheads) . (c) Two-dimensional image of the biopsy sample obtained from the 2.4-mm trephine shows a small degree of artifact at the peripheral portion (arrowheads) . This artifact seems to have occurred during the retrieval of the sample, indicating a crushing artifact. (d) Two-dimensional image of the biopsy sample obtained from the 1.8-mm trephine shows a broken and shorter specimen and a generalized moderate degree of artifact in the periphery and outer layer (arrowheads) , indicating crushing artifacts and accumulated drilling residue.

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Micro-CT Protocol

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Micro-CT Scanning

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Quantitative Analysis Using Micro-CT Three-dimensional Trabecular Parameters

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Statistical Analysis

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Results

Quantitative Analysis Using Micro-CT Three-dimensional Trabecular Parameters

Agreement and Difference Between the Baseline Vertebrae Specimens and Samples Taken Using the 2.4-mm Trephine

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Table 1

Agreement and Comparison Between the 3D Micro-CT Parameters of Baseline Vertebrae Specimens and the 2.4-mm Trephine Biopsy Samples Using Bland-Altman Methods and the Paired t -Test

Baseline Specimen 2.4-mm Sample Agreement ∗ Comparison † Measurement Mean ± SD Mean ± SD Mean Difference (%) 95% Confidence Interval_P_ Value BV/TV 43.38 ± 5.21 43.72 ± 4.87 −0.9 −18.0 to 16.2 .69 Tb. Th 12.28 ± 0.78 12.04 ± 1.14 2.2 −22.8 to 27.2 .49 Tb. N 0.035 ± 0.0043 0.036 ± 0.0040 −3.1 −29.1 to 22.9 .34 Tb. Sp 22.39 ± 3.13 18.02 ± 2.31 22.7 −35.4 to 80.8 .003 Tb. Pf −0.036 ± 0.022 0.073 ± 0.067 388.8 −6201.7 to 6979.2 <.0001 SMI 0.44 ± 0.37 1.51 ± 0.51 −121.4 −249.0 to 6.2 <.0001 DA 20.31 ± 27.39 10.16 ± 19.01 63.1 −137.6 to 263.8 .23

3D, three-dimensional; BV/TV, trabecular bone volume; CT, computed tomography; DA, degree of anisotropy; SMI, structural model index; Tb. N, trabecular number; Tb. Pf: trabecular bone fraction factor; Tb.Sp: trabecular separation; Tb.Th, trabecular thickness.

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Figure 2, Bland-Altman analysis of the differences of each three-dimensional parameter between the baseline vertebrae specimens and the 2.4-mm biopsy samples compared to the average of each three-dimensional parameter. The differences are expressed as a percentage of the averages between two paired data sets. (a) Trabecular bone volume (BV/TV), (b) trabecular thickness (Tb. Th), (c) trabecular number (Tb. N), (d) trabecular separation (TB. Sp), (e) trabecular bone pattern factor (Tb. Pf), (f) the structure model index (SMI), and (g) the degree of anisotropy (DA).

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Agreement and Difference Between the Baseline Vertebrae Specimens and Samples Taken Using the 1.8-mm Trephine

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Table 2

Agreement and Comparison Between the 3D Micro-CT Parameters of the Baseline Vertebrae Specimens and the 1.8-mm Trephine Biopsy Samples Using Bland-Altman Methods and the Paired t -Test

Baseline Specimen 1.8-mm Sample Agreement ∗ Comparison † Measurement Mean ± SD Mean ± SD Mean Difference (%) 95% Confidence Interval_P_ Value BV/TV 42.81 ± 5.15 47.81 ± 7.02 −10.7 −31.6 to 10.2 .0003 Tb. Th 12.38 ± 0.81 12.17 ± 1.20 1.9 −18.7 to 22.6 .49 Tb. N 0.035 ± 0.0043 0.039 ± 0.0048 −12.6 −34.9 to 9.7 .0001 Tb. Sp 23.02 ± 3.78 14.39 ± 2.62 45.8 5.0 to 86.5 <.0001 Tb. Pf −0.031 ± 0.025 0.099 ± 0.053 −273.7 −2378.6 to 1831.1 <.0001 SMI 0.59 ± 0.34 1.88 ± 0.43 −111.1 −197.6 to −24.6 <.0001 DA 14.27 ± 12.53 5.84 ± 3.00 55.8 −105.5 to 217.1 .0135

3D, three-dimensional; BV/TV, trabecular bone volume; CT, computed tomography; DA, degree of anisotropy; SMI, structural model index; Tb. N, trabecular number; Tb. Pf: trabecular bone fraction factor; Tb.Sp: trabecular separation; Tb.Th, trabecular thickness.

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Figure 3, Bland-Altman analysis of the differences of each three-dimensional parameter between the baseline vertebrae specimens and the 1.8-mm biopsy samples compared to the average of each three-dimensional parameter. The differences are expressed as a percentage of the averages between two paired data sets. (a) Trabecular bone volume (BV/TV), (b) trabecular thickness (Tb. Th), (c) trabecular number (Tb. N), (d) trabecular separation (TB. Sp), (e) trabecular bone pattern factor (Tb. Pf), (f) the structure model index (SMI), and (g) the degree of anisotropy (DA).

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Discussion

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Acknowledgment

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