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一种用于基因改造小鼠骨骼三维微观结构表型特征分析的无损技术:生长激素转基因动物和正常对照的初步数据。

A non-destructive technique for 3-D microstructural phenotypic characterisation of bones in genetically altered mice: preliminary data in growth hormone transgenic animals and normal controls.

作者信息

Graichen H, Lochmüller E M, Wolf E, Langkabel B, Stammberger T, Haubner M, Renner-Müller I, Englmeier K H, Eckstein F

机构信息

Anatomische Anstalt, Ludwig-Maximilians-Universität, München, Germany.

出版信息

Anat Embryol (Berl). 1999 Mar;199(3):239-48. doi: 10.1007/s004290050225.

Abstract

A non-destructive, three-dimensional technique for microstructural phenotypic characterisation of skeletal elements in genetically altered mice is presented. Preliminary data in bovine growth-hormone transgenic animals and control littermates are shown. The technique is based on microcomputed tomography (microCT) and digital postprocessing and allows for a differential quantitative analysis of the cortical and trabecular bone compartments in the axial and peripheral skeleton. The distal femora and the first lumbar vertebral bodies of six animals were CT scanned in the axial plane with an isotropic resolution of 20 microm. The periostal surface and the marrow spaces were segmented fully automatically, and the trabecular and cortical compartments were separated interactively. After 3-D reconstruction, various regions of interest (diaphyseal, metaphyseal and epiphyseal) were selected for the analysis. The femora and vertebrae of the transgenic animals showed obvious differences in size, shape, and trabecular arrangement compared with the control animals. The total bone mass was increased by a factor of two to three, but the trabecular bone was increased much more (up to 12 times) than the cortical bone. The transgenic animals showed an increased ratio of trabecular vs cortical bone (0.90 to 1.27 vs 0.14 to 0.36 in the femoral diaphysis) and an elevated trabecular bone volume fraction (49% to 73% vs 18% to 43% in the femoral metaphysis). The mean 3-D cortical thickness was similar in the normal and transgenic animals (values between 93 microm and 232 microm in the dia- and metaphyses), but the minimal cortical thickness was lower in the transgenic animals (22 to 31 microm vs 54 microm to 110 microm in the diaphysis). The technique presented is suitable for phenotypic characterisation of bone structure in genetically altered mice.

摘要

本文介绍了一种用于对基因工程小鼠骨骼元素进行微观结构表型特征分析的非破坏性三维技术。展示了在牛生长激素转基因动物及其对照同窝仔鼠中的初步数据。该技术基于微计算机断层扫描(microCT)和数字后处理,可对轴向和外周骨骼中的皮质骨和小梁骨部分进行差异定量分析。对六只动物的股骨远端和第一腰椎椎体进行了轴向平面的CT扫描,各向同性分辨率为20微米。骨膜表面和骨髓腔完全自动分割,小梁骨和皮质骨部分通过交互式分离。三维重建后,选择不同的感兴趣区域(骨干、干骺端和骨骺)进行分析。与对照动物相比,转基因动物的股骨和椎骨在大小、形状和小梁排列上有明显差异。总骨量增加了两到三倍,但小梁骨的增加幅度(高达12倍)远大于皮质骨。转基因动物的小梁骨与皮质骨的比例增加(股骨干中为0.90至1.27,而对照为0.14至0.36),小梁骨体积分数升高(股骨干骺端中为49%至73%,而对照为18%至43%)。正常动物和转基因动物的平均三维皮质厚度相似(骨干和干骺端的值在93微米至232微米之间),但转基因动物的最小皮质厚度较低(骨干中为22至31微米,而对照为54微米至110微米)。所介绍的技术适用于对基因工程小鼠的骨结构进行表型特征分析。

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