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Cell biological studies of hard tissue using three dimensional morphometry.

使用三维形态测定法对硬组织进行细胞生物学研究。

基本信息

批准号：
06404063
负责人：
OZAWA Hidehiro
金额：
$ 16.64万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

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.

本研究旨在利用三维组织形态计量学和细胞形态计量学研究骨代谢引起的形态变化。结果如下：1)用激光共聚焦显微镜观察了下颌骨老化、骨质疏松等骨的生理和病理变化，并将图像存储为数字化显微图像。2)将存储的图像用于开发可加工的形态测量图像分析和数据管理软件，该软件能够使用Luzex-F对以下参数进行半自动分析：骨面、骨样表面、成骨细胞表面、侵蚀表面、破骨细胞表面、骨核、类骨质体积、总组织体积、纤维化体积、单标记面、双标记面、标记厚度、类骨质厚度、壁厚、静止表面、低矿化骨体积。3)保存四环素和钙黄素标记的不脱钙样品的激光共聚焦显微镜观察获得的荧光图像，进行二维图像分析，然后利用Micro Voxel将二维图像重建为样品的三维图像。结果表明，该方法能够从样品表面重建深达100微米的四环素和钙黄绿素标记的体积的三维图像。4)利用表面造型软件Luzex-III成功地同时重建了破骨细胞、成骨细胞和骨表面之间的三维空间关系。上述方法用于：1)衰老对大鼠下颌骨影响的形态和免疫组织化学研究；2)大鼠牙槽骨的形态和免疫组织化学研究；3)甲状旁腺素对骨代谢的不同部位影响的研究；4)去卵巢大鼠骨吸收的超微结构研究。