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人牙周膜细胞和牙囊祖细胞的分离、鉴定及其分化潜能的研究以及它们在体外对骨形态发生蛋白-7(BMP-7)的反应

Isolation, characterization and investigation of differentiation potential of human periodontal ligament cells and dental follicle progenitor cells and their response to BMP-7 in vitro.

作者信息

Açil Yahya, Yang Fan, Gulses Aydin, Ayna Mustafa, Wiltfang Jörg, Gierloff Matthias

机构信息

Department of Oral and Maxillofacial Surgery, Christian Albrechts University, Kiel, Germany.

Affiliated Hospital of Stomatology, Medical College, Zhejiang University, Hangzhou, China.

出版信息

Odontology. 2016 May;104(2):123-35. doi: 10.1007/s10266-015-0198-1. Epub 2015 Mar 11.

Abstract

The aim of this study was to assess the factors, mechanisms and the differences between periodontal ligament (PDL) cells and denta l follicle (DF) progenitor cells towards the osteoblastic/cementoblastic differentiation and to investigate the effects of BMP-7 on developmental (DF) and mature tissue-derived (PDL) cells, respectively. Primary cell culture of PDL cells and DF progenitor cells was performed. Osteogenic differentiation was evaluated using von Kossa, Alizarin Red S and immuno-histo-chemistry staining of osteocalcin. Gene expression pattern was evaluated via real-time PCR. A series of CD surface marks were tested using flow cytometry and fluorescence-activated cell-sorting analysis was performed. Real-time RT-PCR demonstrated similar gene expression pattern of PDL cells and DF progenitor cells: the expression of OPN and OCN significantly was elevated when incubated with osteogenic components, Runx2 was unaffected, and Osteorix was hardly expressed whether in basic medium or induction medium. In addition, BMP-7 induced osteoblast/cementoblast differentiation of PDLSCs and DF progenitor cells in a dose- and time-dependent manner, as reflected by enhanced Runx2 and (OCN) mRNA transcript expression. BMP-7 triggers PDL cells and DF progenitor cells to differentiate towards an osteoblast/cementoblast phenotype.

摘要

本研究旨在评估牙周膜(PDL)细胞和牙囊(DF)祖细胞向成骨细胞/成牙骨质细胞分化的影响因素、机制及差异,并分别研究骨形态发生蛋白-7(BMP-7)对发育性(DF)和成熟组织源性(PDL)细胞的作用。进行了PDL细胞和DF祖细胞的原代细胞培养。使用冯·科萨染色、茜素红S染色及骨钙素免疫组织化学染色评估成骨分化。通过实时定量PCR评估基因表达模式。使用流式细胞术检测一系列CD表面标志物并进行荧光激活细胞分选分析。实时逆转录PCR显示PDL细胞和DF祖细胞具有相似的基因表达模式:与成骨成分孵育时,骨桥蛋白(OPN)和骨钙素(OCN)的表达显著升高,Runx2不受影响,无论在基础培养基还是诱导培养基中,骨成骨因子(Osteorix)几乎不表达。此外,BMP-7以剂量和时间依赖性方式诱导PDLSCs和DF祖细胞向成骨细胞/成牙骨质细胞分化,这通过Runx2和(OCN)mRNA转录本表达增强得以体现。BMP-7触发PDL细胞和DF祖细胞向成骨细胞/成牙骨质细胞表型分化。

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