Department of Dental Regenerative Medicine, National Center for Geriatrics and Gerontology, Research Institute, Aichi, Japan.
J Endod. 2012 Jul;38(7):920-6. doi: 10.1016/j.joen.2012.01.010. Epub 2012 Mar 15.
Pulp stem/progenitor cells have been successfully transplanted after pulpectomy in dogs and have led to pulp regeneration. The regenerated pulp tissue was investigated by the qualitative and quantitative protein expression patterns in comparison with those of normal pulp. There is an unmet need for a quality standard for regenerated pulp tissue.
Three distinct human CD105(+) stem/progenitor cells from dental pulp, bone marrow, and amnion were compared by 2-dimensional electrophoresis and the differential proteomic expression profiles. The protein identified with high confidence in pulp CD105(+) cells was examined by immunohistochemistry and real-time reverse-transcription polymerase chain reaction in regenerated pulp tissue compared with normal pulp. Its migration effect was further examined in pulp CD105(+) cells using small interfering RNA techniques to knock down the protein.
Nine protein spots were detected solely in pulp CD105(+) cells; one of these was identified as vimentin. The expression of vimentin messenger RNA was highest in pulp tissue among a variety of human tissues and higher in pulp CD105(+) cells compared with other CD105(+) cells and unfractionated total pulp cells. Pulp cells and endothelial cells were positively stained with vimentin in regenerated pulp tissue similarly as those in normal pulp tissue. The expression of vimentin in regenerated pulp was similar to normal pulp. RNA interference knock down of vimentin expression in pulp CD105(+) cells significantly reduced the migration activity.
The highest expression of vimentin in pulp tissue among other tissues and its migration effect in pulp stem cells suggest that it is a quality standard for pulp regeneration and pulp cell function.
牙髓干细胞/祖细胞经牙髓切除术后在犬体内成功移植,并能促进牙髓再生。再生牙髓组织的定性和定量蛋白质表达模式与正常牙髓进行了比较研究。再生牙髓组织需要一个质量标准。
通过二维电泳和差异蛋白质组表达谱比较了来自牙髓、骨髓和羊膜的 3 种不同的人 CD105(+)干细胞/祖细胞。用免疫组织化学和实时逆转录聚合酶链反应检测牙髓 CD105(+)细胞中高置信度鉴定的蛋白质,与正常牙髓进行比较。采用小干扰 RNA 技术敲低该蛋白,进一步检测其在牙髓 CD105(+)细胞中的迁移作用。
在牙髓 CD105(+)细胞中检测到 9 个仅存在的蛋白质斑点,其中一个被鉴定为波形蛋白。在各种人类组织中,波形蛋白信使 RNA 的表达在牙髓组织中最高,在牙髓 CD105(+)细胞中的表达高于其他 CD105(+)细胞和未分离的总牙髓细胞。再生牙髓组织中的牙髓细胞和内皮细胞与正常牙髓组织一样,用波形蛋白呈阳性染色。再生牙髓组织中波形蛋白的表达与正常牙髓相似。RNA 干扰敲低牙髓 CD105(+)细胞中波形蛋白的表达显著降低了迁移活性。
在其他组织中,牙髓组织中波形蛋白表达最高,以及其在牙髓干细胞中的迁移作用提示它是牙髓再生和牙髓细胞功能的质量标准。