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。

• 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
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06404063/
• 关键词: Hard tissue; Three dimensional morphometry; Bone Metabolism; Aging; CLSM; Madibleimage analysis; Image analysis; Immunohistochemical; 副甲状腺ホルモン関連ペプチド; DBAレクチン; カルシトニン遺伝子関連ペプチド; 共焦点レーザー顕微鏡; ジーンターゲッティング; 骨形態計測; トレーサー実験; 組織細胞化学; CD44; ヘパラン硫酸鎖

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.

本研究旨在使用三维组织形态计量学和细胞形态计量学检查骨代谢引起的形态学变化。结果如下：1）观察了骨的生理和病理变化，用激光共聚焦扫描显微镜观察下颌骨的老化和骨质疏松，并将观察结果存储为数字化显微图像。2)所存储的图像用于开发可变形的形态测量图像分析和数据管理软件，其使得能够用Luzex-F对以下参数进行半自动化分析：骨表面;类骨质表面;成骨细胞表面;侵蚀表面;破骨细胞表面;骨质疏松;类骨质体积;总组织体积;纤维化体积;单标记表面;双标记表面;标记厚度;类骨质厚度;壁厚;静止表面;低矿化骨体积。3)保存四环素和钙黄绿素标记的不脱钙标本的CLSM观察荧光图像，对二维图像进行分析，然后利用Micro Voxel将二维图像重建为标本的三维图像。结果表明，该程序能够重建距离样品表面100 μ m深度的四环素和钙黄绿素标记体积的3D图像。4)利用Luzex-III表面建模软件，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。上述程序用于：i）老化对大鼠下颌骨影响的形态学和免疫组织化学研究; ii）对大鼠牙槽骨的形态学和免疫组织化学研究; iii）根据骨的解剖区域研究PTH对骨代谢的差异作用; iv）卵巢切除大鼠骨吸收的超微结构研究。

项目成果

Yoshiba,N.他: "A confocal laser scanning microscopic study of the immunofluorescent localization of fibronectin in odontoblast layer of human teeth." Arch.Oral Biol.39. 395-400 (1994)

Yoshiba, N. 等人：“人类牙齿成牙本质细胞层中纤连蛋白免疫荧光定位的共焦激光扫描显微镜研究。”Arch.Oral Biol.395-400 (1994)。

Nakamura,H.他: "Immunohistochemical localization of vacuolar-typeH^+-ATPase in osteoclasts." Arch.Histol.Cytol.57. 535-539 (1994)

Nakamura, H. 等人：“破骨细胞中液泡型 H^+-ATP 酶的免疫组织化学定位。”Arch.Histol.Cytol.535-539 (1994)。

風間順一郎他: "レーザー顕微鏡の骨動態観察への応用" CLINICAL CALCIUM. 4. 64-66 (1994)

Junichiro Kazama 等人：“激光显微镜在骨动力学观察中的应用”临床钙。4. 64-66 (1994)

Kazama,J.J.他: "A new histochemical double-stain method using three-dimensional analysis with confocal laser scanning microscopy." Biotech & Histochem. 69. 324-328 (1994)

Kazama, J.J. 等人：“使用共焦激光扫描显微镜进行三维分析的新组织化学双染色方法。”69. 324-328 (1994)

Koike,H.他: "Age-related histological changes in rat mandibular condyle." J.Bone Miner.Metab.13. 10-16 (1995)

Koike, H. 等人：“大鼠下颌髁突的年龄相关组织学变化。”J.Bone Miner.Metab.10-16 (1995)。