Ultrastructure of periosteal cells induced by bone morphogenetic protein 7 in vitro

doi:10.3969/j.issn.2095-4344.2014.33.007

Abstract

Abstract:

BACKGROUND: Study confirms that bone morphogenetic protein can induce osteogenesis; however the ultrastructure of periosteal cells induced by bone morphogenetic protein-7 remains poorly reported.
OBJECTIVE: To study the bioactivity and ultrastructure of periosteal cells induced by bone morphogenetic protein-7 in vitro.
METHODS: The primary periosteal cells isolated from adult tibial bone were in vitro cultured, and then divided into experimental group and control group. In the experimental group, cells were cultured with bone morphogenetic protein-7 and culture adjuvant; while cells in the control group were only cultured with the adjuvant. Three samples in each group were tested at 5, 10, 15 days, respectively. The general structure of cultured cells was observed using von Kossa staining, and the ultrastructure was observed under transmission electron microscopy.
RESULTS AND CONCLUSION: The periosteal cells in the two groups grew well in vitro, showing uniform morphology. Early cells were spindle-shaped, with strong three-dimensional sense and full transparency; mitotic cells were short columnar or cubic shaped, there were a lot of rough endoplasmic reticulum and Golgi complex in osteoblasts under electron microscope. Later stage of cells developed from long fusiform into wide shuttle and irregular shape, there were a large number of matrix vesicles within the cells under the electron microscope. The membrane coating, alkaline phosphatase and calcium-binding protein in the cytoplasm, as well as calcium crystals were found. The osteogenesis basement and lateral sides appeared projections, which were connected with adjacent bone cells. Induction of bone morphogenetic protein-7 in vitro promotes the osteoblasts proliferation, division and bone formation speed. The results suggest that bone morphogenetic protein-7 can significantly enhance the proliferation ability of osteoblasts in vitro.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: bone morphogenetic protein-7, osteoblasts, cell structure

CLC Number:

R318

Liao Jia-cheng, Bei Kang-sheng, Lian Yin-chuan, Xu Jin-wen, Liu Yan-xiao, Huang Xiao-yu. Ultrastructure of periosteal cells induced by bone morphogenetic protein 7 in vitro[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(33): 5288-5292.

Figures/Tables 4

2.3 实验分组后传代细胞的形态学观察结果相差显微镜下观察，细胞培养初期(传代后第5天)：细胞数量较少，细胞形状多为梭形或三角形，略光滑，细胞突起较少。细胞数量明显增多，细胞形状多样，大量纤维样伪足于其他细胞伪足交织成三维网状。有少量细胞聚集，电镜下观察见细胞核规则，常位于细胞的一侧，常染色质丰富，核仁少见。胞质中可见散在的线粒体、内质网、高尔基复合体，可见有较多溶酶体，表现为多个高电了密度的小泡。相差显微镜观察细胞培养后期(传代后第15天)：细胞数多，排列紧密；胞体较大，细胞核核大，细胞核呈分叶状，电镜下可见一两个核仁，核仁致密，核膜光滑清晰，有凹陷。胞浆内细胞器明显增多，线粒体丰富，嵴少，电子密度高。胞浆内有丰富的粗面内质网，Golgi复合体,粗面内质网末端膨大成池，内充以低电子密度物质,表面有核糖体, 大量分泌小泡散在分布，少量溶酶体。出现基质小泡，基质小泡内充满中等电子密度物质，同时能见到部分衰老、凋亡的细胞，细胞内出现了大量空泡(可能来源于线粒体肿胀变性)，周围出现双层膜空泡结构，有些细胞完全崩解。 2.4 单纯应用成骨培养辅助剂的对照组体外骨膜细胞普通培养的形态学观察结果骨膜细胞24 h后贴壁，相差显微镜观察见早期呈梭形, 饱满透明, 立体感强；个别可见细胞两极有细丝状突起，随后开始增殖，72 h后细胞增殖分裂迅速，分裂期呈立方形或短柱状，体积逐渐增大，向成纤维细胞状改变逐渐变成宽梭和不规则形，后期细胞排列紧密，成漩涡状生长，细胞之间无接触抑制现象，呈典型的长梭形、平行或旋涡状排列，集落间出现重叠。 2.5 骨形态发生蛋白7诱导后的实验组细胞形态学观察结果加入骨形态发生蛋白7及辅助剂后，细胞生长速度明显增加，细胞初始仍为长梭形，随时间延长逐渐变成多边形和三角形，与成骨细胞形态相似，并能增殖形成细胞集落(图2)。"

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