Expression of bone associated markers by tooth root lining cells, in situ and in vitro
Expression of bone associated markers by tooth root lining cells, in situ and in vitro
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DOI：
10.1016/s8756-3282(96)00348-1
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发表时间：
1997-02-01
期刊：
影响因子：
4.1
通讯作者：
Somerman, MJ
中科院分区：
文献类型：
--
作者：
DErrico, JA;Macneil, RL;Somerman, MJ
牙周病的特征是炎症和随后牙齿支持组织的丢失和/或损害，包括骨、牙骨质和牙周膜。牙周疾病后牙周发育和再生的初始过程中的一个关键组织是牙骨质。由于无法分离和培养参与牙骨质形成的细胞(即成牙骨质细胞)，旨在了解牙周发育和再生机制，特别是牙根骨质形成机制的研究工作一直受到阻碍。通过使用骨细胞培养的实验，已经了解了许多涉及骨形成和功能的过程和机制。因此，这项研究的目的是开发一种策略，从而可以分离、培养和鉴定成牙骨质细胞。作为第一步，利用原位杂交，我们确定了矿物相关蛋白在CD-1小鼠第一磨牙根发育过程中的时间和空间表达。这些蛋白包括牙本质唾液蛋白(DSP)、骨桥蛋白(OPN)、骨涎蛋白(BSP)、骨钙素(OCN)和I型胶原。在小鼠的牙根发育过程中，BSP、OPN和OCN的mRNAs由牙根表面的细胞-成牙骨质细胞-选择性地表达，在第41天表达水平很高。重要的是，在这个时间点，BSP、OPN和OCN的mRNAs没有在整个牙周膜中表达。这些发现为我们提供了对牙根衬里细胞或成牙骨质细胞有选择性的标记，并确定了分离细胞用于体外研究的时间(第41天)。为了从附着在牙根表面的组织中分离细胞，使用了酶消化法，类似于现在被认为是分离成骨细胞的经典技术。为了确定体外培养的细胞是否含有牙根衬里细胞和成牙骨质细胞，对培养的细胞进行了矿物质相关蛋白的表达分析。通过原位杂交检测，异质原代群体中的细胞表达I型胶原、BSP、OPN和OCN。相比之下，该人群中的细胞不表达牙本质唾液蛋白，这是一种成牙本质细胞特有的蛋白。这些程序提供了一种在体外获得牙根衬里细胞的方法，现在可以克隆这些细胞，并用于研究体外确定牙根衬里细胞或成牙骨质细胞的特性。(C)1997年，爱思唯尔科学公司。
Periodontal disease is marked by inflammation and subsequent loss and/or damage to tooth-supporting tissues including bone, cementum, and periodontal ligament. A key tissue in the initial process of periodontal development as well as regeneration following periodontal disease is cementum. Research efforts aimed toward understanding mechanisms involved in periodontal development and regeneration, and in particular the formation of root cementum, have,been hampered by an inability to isolate and culture cells involved in cementum production (i.e., cementoblasts). Much has been learned regarding the processes and mechanisms involved in bone formation and function from experiments using bone cell cultures. Therefore, the purpose of this study was to develop a strategy whereby cementoblasts could be isolated, cultured, and characterized. As a first step, using in situ hybridization, we determined the timed and spatial expression of mineral-associated proteins during first molar root development in CD-1 mice. These proteins included dentin sialoprotein (DSP), osteopontin (OPN), bone sialoprotein (BSP), osteocalcin (OCN), and type I collagen. During root development in mice BSP, OPN, and OCN mRNAs were expressed selectively by cells lining the tooth root surface-cementoblasts-with high levels of expression at day 41. Importantly, at this time point BSP, OPN, and OCN mRNAs were not expressed throughout the periodontal ligament. These findings provided us with markers selective to root-lining cells, or cementoblasts, in situ, and established the time (day 41) for isolating cells for in vitro studies. To isolate cells from tissues adherent to the root surface, enzymatic digestion was used, similar to what are now considered classical techniques for isolation of osteoblasts. To determine whether cells in vitro contained root-lining cells and cementoblasts, cultured cells were analyzed for expression of mineral-associated proteins. Cells within this heterogeneous primary population expressed type I collagen, BSP, OPN, and OCN as determined by in situ hybridization. In contrast, cells within this population did not express dentin sialoprotein, an odontoblast-specific protein. These procedures have provided a means to obtain root-lining cells in vitro that can now be cloned and used for studies directed at determining the properties of root-lining cells, or cementoblasts, in vitro. (C) 1997 by Elsevier Science Inc.