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

(1)采用树脂切片-离子刻蚀(IE)-扫描电子显微镜(SEM)和能量过滤(EF)-透射电子显微镜(TEM)断层扫描技术，研究了大鼠下颌骨穹隆、牙槽骨和牙周组织的三维结构。(2)用IE-SEM方法对大鼠唾液腺包括腮腺和von Ebner‘s腺进行免疫组织化学和细胞化学研究。(3)对培养的小鼠胫骨骨髓细胞进行了干细胞因子的检测，IE-SEM显示内周板结构清晰，部分胶原纤维平行排列，有的倾斜或交叉排列，有的呈交错排列。而在牙槽骨中，板层结构缺失，骨细胞排列不规则。在FETEM-TEMCT研究中，在半薄层…中可以清楚地观察到片层结构内周板在发孔较多，但在牙槽骨中未见。在牙周组织的半薄切片中，还观察到复杂的胶原纤维排列。用大鼠唾液腺对免疫染色标本进行光镜和扫描电子显微镜观察。免疫扫描电子显微镜观察大鼠腮腺分泌终末和间质导管细胞的分泌颗粒呈免疫反应阳性。在骨髓微环境中，各种细胞可能通过中介和细胞接触而相互作用，发挥各自的功能。应用流式细胞术和定量RT-β技术研究转化生长因子-α1、碱性成纤维细胞生长因子、肿瘤坏死因子-α、白介素4和激光诱导因子对骨髓细胞干细胞因子剪接变异体表达的影响。流式细胞仪分析显示，经转化生长因子-β-1或碱性成纤维细胞生长因子处理后，表达膜结合性干细胞因子的细胞数增加。定量RT-β分析显示，转化生长因子-β-1以剂量依赖方式抑制干细胞因子-1和干细胞因子-2的合成。这些结果表明，在转化生长因子-碱性成纤维细胞生长因子1、碱性成纤维细胞生长因子、肿瘤坏死因子-α、IL-4和激光诱导因子的共同刺激下，小鼠骨髓细胞中干细胞因子剪接变异体的表达受到单独的调控。较少

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