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成骨细胞诱导消融转基因小鼠牙槽骨成骨活性的组织学检查。

Histological examination on osteoblastic activities in the alveolar bone of transgenic mice with induced ablation of osteocytes.

机构信息

Department of Bone Metabolism, School of Stomatology, Shandong University, Jinan, China.

出版信息

Histol Histopathol. 2013 Mar;28(3):327-35. doi: 10.14670/HH-28.327.

Abstract

The purpose of this study was to examine histological alterations on osteoblasts from the alveolar bone of transgenic mice with targeted ablation of osteoctyes. Eighteen weeks-old transgenic mice based on the diphtheria toxin (DT) receptor-mediated cell knockout (TRECK) system were used in these experiments. Mice were injected intraperitoneally with 50 µg/kg of DT in PBS, or only PBS as control. Two weeks after injections, mice were subjected to transcardiac perfusion with 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), and the available alveolar bone was removed for histochemical analyses. Approximately 75% of osteocytes from alveolar bones became apoptotic after DT administration, and most osteocytic lacunae became empty. Osteoblastic numbers and alkaline phosphatase (ALP) activity were markedly reduced at the endosteum of alveolar bone after DT administration compared with the control. Osteoblastic ALP activity in the periodontal ligament region, on the other hand, hardly showed any differences between the two groups even though numbers were reduced in the experiment group. Silver impregnation showed a difference in the distribution of bone canaliculi between the portions near the endosteum and the periodontal ligament: the former appeared regularly arranged in contrast to the latter's irregular distribution. Under transmission electron microscopy (TEM), the osteoblasts in the periodontal ligament showed direct contact with the Sharpey's fibers. Thus, osteoblastic activity was affected by osteocyte ablation in general, but osteoblasts in contact with the periodontal ligament were less affected than endosteal osteoblasts.

摘要

本研究旨在研究靶向敲除破骨细胞的转基因小鼠牙槽骨成骨细胞的组织学变化。该实验使用了基于白喉毒素(DT)受体介导的细胞敲除(TRECK)系统的 18 周龄转基因小鼠。这些小鼠通过腹腔内注射 50μg/kg 的 DT 于 PBS 中,或仅注射 PBS 作为对照。注射后两周,通过心脏灌注用 4%多聚甲醛于 0.1M 磷酸缓冲液(pH7.4)对小鼠进行处理,去除可用的牙槽骨进行组织化学分析。在 DT 给药后,约 75%的牙槽骨成骨细胞发生凋亡,且大多数骨陷窝空化。与对照组相比,DT 给药后牙槽骨内骨表面的成骨细胞数量和碱性磷酸酶(ALP)活性明显减少。另一方面,牙周韧带区域的成骨细胞 ALP 活性在两组之间几乎没有差异,尽管实验组的数量减少。银染显示骨小管在靠近骨内膜和牙周韧带部分的分布存在差异:前者呈规则排列,而后者呈不规则分布。在透射电子显微镜(TEM)下,牙周韧带中的成骨细胞与 Sharpey 纤维直接接触。因此,成骨细胞活性普遍受到破骨细胞消融的影响,但与牙周韧带接触的成骨细胞比骨内膜的成骨细胞受影响较小。

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