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Three-dimensional structural studies of bone tissues by ion-etching-scanning electron microscopy.

通过离子蚀刻扫描电子显微镜研究骨组织的三维结构。

基本信息

批准号：
16591852
负责人：
NAGATO Toshikazu
金额：
$ 1.98万
依托单位：
Fukuoka Dental College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

Three studies were performed in this research project; (1) Three-dimensional structures of the bone tissues of fumerus and alveolar bone and periodontium of mandible of rats were studied with the resin section-ion etching (IE)-scanning electron microscopy (SEM) and energy filter (EF) transmission electron microscope (TEM)-TEM tomography. (2) Immuno histo and cyto-chemistry of rat salivary glands including parotid and von Ebner's gland was performed by using IE-SEM method. (3) Stem cell factors on cultured bone marrow cells which were harvested from mouse tibiae were also examined.In IE-SEM studies, it was suggested that the structure of inner circumferential lamellae was clear, bat arrangement of some collagen fibrils were parallel, some oblique or some crossing as well as osteocytes. On the other hand, in alveolar bone, lamellar structure was absent, and osteocyte arrangement were irregular. In FETEM-TEM tomography study, lamellar structure were clearly observed in the semi-thin secti … More ons of the inner circumferential lamellae in fumerus, but not observed in the alveolar bone. Complicated arrangement of collagen fibrils were alse observed in the semi-thin sections of the periodontium.The ability to apply both light and scanning electron microscopy to immunostained specimens was examined with rat salivary gland. Immunoscanning electron micrographs of the sections which were immunohistochemically stained with anti-amylase and immunogold in association with silver enhancement techniques showed that secretory granules in the secretory terminal portion and intercalated duct cell of the rat parotid gland were immunoreaction positive.In bone marrow microenvironment, various cells are expected to interact each other revulating their functions through mediators and cell-contact Then, the effect of TGF-β1, bFGF, TNF-α, IL-4, and LIF on the stem cell factor (SCF) splice variants expression of bone marrow cells were studied by using flow cytometry and quantitative RT-PCR. Flow cytometry analysis demonstrated that the number of cells expressing membrane-bound SCF increased when the cells were treated with TGF-β1 or bFGF. Quantitative RT-PCR analysis demonstrated that TGF-β 1 recuced both SCF-1 and SCF-2 mRNAs synthesis in a dose dependent manner These results suggest that expressions of stem cell factor splice variants in mouse bone marrow cells were regulated individually under co-stimulation of TGF-β1, bFGF, TNF-α, IL-4, and LIF. Less

本研究项目共进行了三项研究；(1)采用树脂切片离子刻蚀(IE)-扫描电子显微镜（SEM）和能量滤波（EF）透射电子显微镜(TEM)-透射电子显微镜（TEM）断层扫描技术，研究了大鼠下颌骨股骨、牙槽骨和牙周组织的三维结构。(2)采用IE-SEM法对大鼠腮腺、艾氏腺等唾液腺进行免疫组织和细胞化学检测。(3)对小鼠胫骨培养的骨髓细胞的干细胞因子进行了检测。电镜观察发现，骨细胞内周片状结构清晰，部分胶原原纤维平行排列，部分呈斜向排列，部分呈交叉排列。另一方面，牙槽骨没有板层结构，骨细胞排列不规则。在FETEM-TEM扫描研究中，在半薄切片上可以清晰地观察到板层结构，在股骨内侧有较多的周向板层结构，但在牙槽骨中未观察到。牙周组织半薄切片中胶原原纤维排列复杂。用大鼠唾液腺检查了对免疫染色标本应用光镜和扫描电镜的能力。经抗淀粉酶和免疫金联合银增强技术免疫组织化学染色切片的免疫扫描电镜显示，大鼠腮腺分泌末端和插层管细胞的分泌颗粒呈免疫反应阳性。在骨髓微环境中，各种细胞之间通过介质和细胞接触相互作用，调节其功能。我们利用流式细胞术和定量RT-PCR技术研究了TGF-β1、bFGF、TNF-α、IL-4和LIF对骨髓细胞干细胞因子（SCF）剪接变异体表达的影响。流式细胞术分析显示，TGF-β1或bFGF处理后，表达膜结合SCF的细胞数量增加。定量RT-PCR分析表明，TGF-β1以剂量依赖的方式减少了SCF-1和SCF-2 mrna的合成。这些结果表明，在TGF-β1、bFGF、TNF-α、IL-4和LIF的共同刺激下，小鼠骨髓细胞中干细胞因子剪接变异体的表达被单独调节。少

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

エポン切片-イオンエッチング-SEM法の骨組織三次元構造解析への応用。

Epon截面-离子刻蚀-SEM方法在骨组织三维结构分析中的应用

DOI：
发表时间：
2005
期刊：
福岡歯大学会雑誌 31・4
影响因子：
0
作者：
小崎健一;他;井上庸子
通讯作者：
井上庸子

エポン切片-イオンエッチング-SEM法の骨組織三次元構造解析への応用

Epon截面-离子刻蚀-SEM方法在骨组织三维结构分析中的应用

DOI：
发表时间：
2005
期刊：
福岡歯科大学学会雑誌 31・4
影响因子：
0
作者：
I Sasagawa;M Ishiyama;井上庸子
通讯作者：
井上庸子

Morphological analysis of the three-dimensional structure of bone tissues by the epoxy resin section-ion etching-SEM method.

环氧树脂切片-离子刻蚀-SEM法对骨组织三维结构进行形态分析

DOI：
发表时间：
2005
期刊：
The journal of Fukuoka Dental College. Vol.31, No.4
影响因子：
0
作者：
Uchida;K.;Akita;Y.;Matsuo;K.;Fujiwara;S.;Nakagawa;A.;Kazaoka;Y.;Hachiya;H.;Naganawa;Y.;Oh-iwa;I.;Ohura;K.;Saga;S.;Kawai;T.;Matsumoto;Y.;Shimozato;K.;Kozaki;K.;Yoko INOUE
通讯作者：
Yoko INOUE

Application of ion etching to immunoscanning electron microscopy.

离子蚀刻在免疫扫描电子显微镜中的应用。