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应用原子力显微镜研究淫羊霍苷诱导人脐带间充质干细胞成骨分化

ISSN号：1004-4957
期刊名称：分析测试学报
时间：0
页码：478-483
语言：中文
分类：O766.1[理学—晶体学]
作者机构：[1]暨南大学生命科学技术学院化学系,广东广州510632, [2]暨南大学附属第一医院骨科,广东广州510632
相关基金：国家自然科学基金面上项目资助（30872404）; 海外及港澳学者合作研究基金资助项目（30828028）; 973重大项目子项目资助（2010CB833603）
相关项目：近场光学研究γδT细胞免疫的分子机制

关键词：原子力显微镜, 淫羊霍苷, 人脐带间充质干细胞, 成骨分化, atomic force microscope, icariin, human umbilical cord mesenchymal stem cells, osteogenic differentiation

中文摘要：

利用碱性磷酸酶（ALP）染色和钙结节（Vonkossa）染色的方法对诱导21 d的淫羊霍苷诱导人脐带间充质干细胞进行鉴定;应用原子力显微镜（AFM）观察淫羊霍苷的形貌和人脐带间充质干细胞诱导0、5、10、15、21 d后的细胞形貌。结果表明,经成骨诱导分化21 d后,ALP染色呈强阳性,Vonkossa染色可见明显钙结节。AFM分析表明,淫羊霍苷在盖玻片上呈分散状分布,在细胞表面上聚集并呈微米域分布。实验发现,由于吸附在细胞表面时,被细胞膜分子包裹,更有利于在细胞表面的吸附,进入细胞内部,细胞表面的淫羊霍苷颗粒较在盖玻片上时增大,由淫羊霍苷颗粒进入细胞后在细胞表面留下一些小孔,可知其通过进入细胞内部诱导成骨分化。分化后,细胞表面有小突触,是由成骨分化后细胞内形成钙结节造成。

英文摘要：

Atomic force microscope was used to study the induction of human umbilical cord mesenchymal（hUCM） stem cells into osteogenic differentiation by icariin in vitro at different inducing periods（0,5,10,15,21 days）.After 21 days of induction,the induced osteogenic differentiation of hUCM stem cells were evaluated by alkaline phosphatase（ALP）staining and calcium nodules（Vonkossa）staining methods.Results showed that the alkaline phosphate staining for hUCM stem cells after 21 days of osteogenic differentiation was strongly positive and calcium nodules could be evidently observed on the Vonkossa staning.AFM results showed that the icariin particles were dispersively distributed on the coverslip,but aggregated on the cell surface in micron-domain.It was found that the icariin particles on the cell surface were larger than those on the coverslip,since the adsorbed particles on the cell surface were enwrapped by the cell membrane molecules.Some holes were left on the cell surface as the icariin particles entered into the interior of cells,thus the induced osteogenic differentiation of hUCM stem cells were obtained.After differentiation,some small synapses,formed by calcium nodules,were observed on the cell surface.

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