Cell biological studies of hard tissue using three dimensional morphometry.

本研究旨在利用三维组织形态学和细胞形态学检测骨代谢引起的形态学变化。结果如下：1)利用CLSM检测下颌骨老化、骨质疏松等骨的生理病理变化，并将观察结果存储为数字化显微图像。2)将存储的图像用于开发可使用的形态测量图像分析和数据管理软件，该软件可以使用Luzex-F对以下参数进行半自动分析：骨表面；骨样表面；成骨细胞表面；侵蚀面；破骨表面；Bonevolume;骨样体积；总组织体积；纤维化体积；单标签表面；双标签表面；标签厚度；类骨厚度；壁厚；静态表面；低矿化骨体积。3)保存四环素和钙黄素标记的未钙化标本CLSM观察所得的荧光图像，分析二维图像，利用Micro Voxel将二维图像重建为标本的三维图像。结果表明，该程序能够重建四环素和钙绿素标记体积的三维图像，距离样品表面深度可达100微米。4)利用表面建模软件Luzex-III，成功地同时重建了破骨细胞、成骨细胞与骨表面之间的三维空间关系。采用上述方法：1)对衰老对大鼠下颌骨的影响进行形态学和免疫组化研究；Ii)大鼠牙槽骨的形态学和免疫组织化学研究；iii)研究甲状旁腺激素对骨代谢的不同影响，根据骨的解剖区域；Iv)去卵巢大鼠骨吸收的超微结构研究。

This study aimed to examine morphological changes caused by bone metabolism using a 3-dimensional histomorphometry and cytomorphometry . The results obtained were as follows :1)Examined physiologic and pathologic changes of bone, e.g., aging of mandible and osteoporosis with CLSM and storing the views as digitized microscopic images . 2)The stored images were used for developing a madible morphometric image analysis and data management software, which enabled a semi-automated analysis with a Luzex-F for the following parameters : Bone surface ; Osteoid surface ; Osteoblast surface ; Eroded surface ; Psteoclast surface ; Bonevolume ; Osteoid volume ; Total tissue volume ; Fibrosis volume ; Single labeled surface ; Double labeled surface ; Label thickness ; Osteoid thickness ; Wall thickness ; Quiescent surface ; Low mineralizing bone volume . 3)Store fluorescence images obtained by CLSM observation of tetracyclin-and calcein-labeled undecalcified specimens, analyze 2-dimensional images, then use the 2-D images to reconstitute into 3-Dimages of the specimens by Micro Voxel. The results showed that the procedure enables the reconstitution of 3-D images of tetracyclin-and calcein-labeled volume up to a 100 um depth from the sample surface. 4)Theuse of Luzex-III,a surface modeling software, succeeded in simultaneously reconstitute the 3-D spatial relationship between osteoclasts, osteoblasts and the bone surface. The above-described procedures were used for : i)a morphological and immunohistochemical study on the effect of aging on rat mandibular bone ; ii)a morphological and immunohistochemical study on rat alveolar bone ; iii)study on the differential effect of PTH on bone metabolism according to the anatomical regions of bone ; iv)ultrastructural study on bone resorption in ovarectomized rats.