纤维粘连蛋白在牙本质形成中的作用

纤维粘连蛋白(fibronectin,Fn),是一种多功能高分子量糖蛋白,广泛存在于胞外基质中,在发育和损伤修复中调节细胞附着、移行和分化。近年来在牙齿发育和牙髓修复过程中,Fn在诱导成牙本质细胞及成牙本质细胞样细胞分化,促进牙本质形成过程中发挥重要的调控作用,本文就Fn在牙本质形成中的作用研究进展作一综述。     

人牙髓干细胞向成牙本质细胞定向分化过程中miRNAs表达谱筛选及鉴定

目的:筛选人牙髓干细胞向成牙本质细胞定向分化过程中差异表达的microRNAs(miRNAs)并进行初步鉴定。方法:体外分离、培养和鉴定人牙髓干细胞,miRNAs芯片筛选牙髓干细胞向成牙本质细胞分化过程中差异表达的miRNAs,并通过TargetScan数据库预测靶基因,实时定量PCR法对结果进行初步鉴定。结果:人牙髓干细胞vimentin、nestin、GFAP和I型胶原4种细胞表型均表达;成脂诱导3周后细胞内有油红O染色阳性脂滴出现;成牙本质诱导可见钙结节形成;基因芯片结果显示,牙髓干细胞向成牙本质细胞分化过程中,发生2倍以上表达变化的miRNAs有6条,其中上调3条,下调3条。通过miRNA靶标预测工具预测靶基因,发现hsa-miR-633和hsa-miR-210有与牙髓干细胞分化相关的靶基因;real time-PCR验证hsa-miR-633和hsa-miR-210表达变化与基因芯片结果相符。结论:人牙髓干细胞经诱导向成牙本质细胞分化过程中miRNAs表达谱具有显著变化,为牙髓干细胞分化机制研究提供了新的提示。     

纤维粘蛋白在牙本质形成中的作用

纤维粘连蛋白是一种多功能高分子量糖蛋白，广泛存在于胞牙基质中，在发育和损伤修复中调节细胞附着，移行和分化。近年来在牙齿发育和牙髓修复过程中，Ｆｎ在诱导成牙本质细胞及成本成本质细胞样细胞分化，促进本质形成过程中发挥重要的调控作用。本文就Ｆｎ在牙本质形成中的作用研究进展作了一综述。     

小整联蛋白结合配体N-连结糖蛋白家族成员在牙发育中的作用

小整联蛋白结合配体N-连结糖蛋白家族是一组主要在牙和骨组织中表达的蛋白质，其基因具有通用的外显子内含子结构，影响细胞的增殖和分化。目前已知其包括骨桥蛋白、骨涎蛋白、牙本质基质蛋白-1、牙本质涎磷蛋白、细胞外基质磷酸糖蛋白和釉蛋白等6个成员，本文就其在牙发育过程中的作用作一综述。     

人牙髓细胞在离体牙髓腔内向成牙本质细胞样细胞分化的研究

目的观察人牙髓细胞在离体牙髓腔内向成牙本质细胞样细胞分化的潜能。方法将原代培养的人牙髓细胞接种至处理过的离体牙髓腔内,2周后固定、脱钙、包埋、切片、染色,显微镜下观察其生长及分化情况。结果牙髓细胞在牙本质表面生长良好,部分细胞伸出胞质突伸入牙本质小管中,表现出成牙本质细胞样细胞的形态。结论牙本质可以诱导牙髓细胞向成牙本质细胞样细胞分化,可为组织工程化牙髓的研制提供实验依据。     

成牙本质细胞特异分子标志物的研究进展

牙髓干细胞是一类存在于牙髓组织中,保持着高度的增殖和分化潜能,受到刺激后能向终末细胞分化的细胞。在牙髓干细胞的研究过程中,常需要根据成牙本质细胞分子标记物的表达来判断牙髓干细胞的分化进程。随着研究的进展,可供选择的成牙本质细胞分子标记物也越来越广泛。下面就成牙本质细胞分化的特异分子标记物的研究进展作一综述。     

牙髓干细胞与牙本质再生相关性的研究进展

牙髓中存在具备自我更新和多向分化潜能的牙髓干细胞。这些细胞经诱导可向成牙本质细胞分化并形成牙本质样结构，有望成为牙本质再生的种子细胞。本文就牙髓干细胞的来源、生物学特性及其在牙本质再生中的应用作一综述。     

牙髓干细胞与牙本质再生相关性的研究进展

牙髓中存在具备自我更新和多向分化潜能的牙髓干细胞.这些细胞经诱导可向成牙本质细胞分化并形成牙本质样结构,有望成为牙本质再生的种子细胞.本文就牙髓干细胞的来源、生物学特性及其在牙本质再生中的应用作一综述.     

CD146在人牙髓干细胞中表达的研究

目的:研究CD146在人牙髓干细胞及其诱导分化过程中的表达情况。方法:体外培养人牙髓干细胞,免疫荧光及流式细胞术检测CD146的表达。矿化诱导人牙髓干细胞分化,检测牙本质唾蛋白的表达,从mRNA及蛋白水平检测诱导过程中CD146的表达。结果:免疫荧光及流式细胞术证明人牙髓干细胞中CD146表达阳性。使用矿化诱导液培养人牙髓干细胞,通过检测到牙本质唾蛋白的表达,证明细胞已向成牙本质细胞方向分化;在此诱导过程中,CD146在人牙髓干细胞中的表达逐渐下调。CD146在人牙髓干细胞中有较特异性的表达,有可能作为其特异性标志物。     

转化生长因子β信号转导与成牙本质细胞分化

转化生长因子β在牙胚发育，成牙本质细胞分化和牙髓损伤修复过程中均发挥着重要的作用，本文就TGF－β近年来的研究进展，尤其是TGF－β－Smad信号途径与成牙本质细胞分化机制的相互关系作一综述。     

Expression of mineralization markers in dental pulp cells

There is an increasing interest in the utility of dental pulp stem cells (DPSCs) for dentin regeneration. The mechanisms involved in DPSC differentiation remain poorly understood. The purpose of the study was to investigate the mineralization capacity of human dental pulp cells (DPCs) and identify potential markers for odontoblast differentiation. The isolated DPCs expressed mesenchymal stem-cell markers as shown by flow cytometry and could differentiate in vitro into odontogenic, adipogenic, and chondrogenic lineages. Alkaline phosphatase activity of DPCs elevated over time, with significant upregulation on day 21 in odontogenic induction. Quantitative RT-PCR revealed that osteocalcin, dentin sialophosphoprotein (DSPP), and matrix extracellular phosphoglycoprotein (MEPE) expression also increased time dependently in the induction cultures. In conclusion, we isolated DPCs with stem cell characteristics. MEPE and DSPP showed a similar regulatory pattern of DPCs mineralization. MEPE along with DSPP may be potential odontogenetic differentiation markers.     

Autophagy in the dentin‐pulp complex against inflammation

The dentin‐pulp complex is a highly specialized tissue for protecting the dental pulp. Odontoblasts are long‐lived, hard‐tissue‐forming cells in the dentin‐pulp complex and critically involved in inflammatory responses against invading pathogens. Autophagy is a highly conserved homeostasis mechanism of living cells under various stress conditions. Growing evidence in the literature addresses the role of autophagy in odontoblast differentiation and aging. This review summarizes the current knowledge about autophagy for the dentin‐pulp complex in resisting inflammation.     

Dentin induction by implants of autolyzed antigen-extracted allogeneic dentin on amputated pulps of dogs

Abstract Implantation of autolyzed antigen-extracted allogeneic (AAA) dentin matrix gelatin powder caused homeoinduction on amputated dental pulps. This event began with migration of spindle-shaped mesenchymal cells into the cavity on the amputated pulp. This was followed by proliferation of undifferentiated large cells concomitantly with vascular invasion and attachment of spindle-shaped cells and large cells to the AAA dentin. The adhering cells differentiated into osteodentinoblasts and/or preodontoblasts to form osteodentin matrix and predentin respectively. Osteodentinocytes and odontoblasts, then, formed osteodentin and tubular dentin. The healing of pulp dressed with inactivated AAA dentin using guanidine HCl was delayed. These results suggested that AAA dentin matrix powder may have chemotactic and mitogenic activity for undifferentiated mesenchymal cells and may provide a suitable scaffolding for fixation of these cells. Compartments of microenvironment formed by AAA dentin and the enveloping hard substances may play some role in differentiation into odontoblasts.     

Dental pulp stem cells, niches, and notch signaling in tooth injury

Stem cells guarantee tissue repair and regeneration throughout life. The decision between cell self-renewal and differentiation is influenced by a specialized microenvironment called the 'stem cell niche'. In the tooth, stem cell niches are formed at specific anatomic locations of the dental pulp. The microenvironment of these niches regulates how dental pulp stem cell populations participate in tissue maintenance, repair, and regeneration. Signaling molecules such as Notch proteins are important regulators of stem cell function, with various capacities to induce proliferation or differentiation. Dental injuries often lead to odontoblast apoptosis, which triggers activation of dental pulp stem cells followed by their proliferation, migration, and differentiation into odontoblast-like cells, which elaborate a reparative dentin. Better knowledge of the regulation of dental pulp stem cells within their niches in pathological conditions will aid in the development of novel treatments for dental tissue repair and regeneration.     

Dentin regeneration by dental pulp stem cell therapy with recombinant human bone morphogenetic protein 2

Regenerative medicine is based on stem cells, signals, and scaffolds. Dental pulp tissue has the potential to regenerate dentin in response to noxious stimuli, such as caries. The progenitor/stem cells are responsible for this regeneration. Thus, stem cell therapy has considerable promise in dentin regeneration. Culture of porcine pulp cells, as a three-dimensional pellet, promoted odontoblast differentiation compared with monolayers. The expression of dentin sialophosphoprotein (Dspp) and enamelysin/matrix metalloproteinase 20 (MMP20) mRNA confirmed the differentiation of pulp cells into odontoblasts and was stimulated by the morphogenetic signal, bone morphogenetic protein 2 (BMP2). Based on the in vitro experiments, an in vivo evaluation of pulp progenitor/stem cells in the dog was performed. The autogenous transplantation of the BMP2-treated pellet culture onto the amputated pulp stimulated reparative dentin formation. In conclusion, BMP2 can direct pulp progenitor/stem cell differentiation into odontoblasts and result in dentin formation.     

基质金属蛋白酶-8在人和大鼠牙胚发育钟状期的表达

目的　研究基质金属蛋白酶-8(MMP-8)在牙胚发育牙本质形成过程中的表达。方法　免疫组织化学方法检测MMP-8在人牙胚发育钟状期的表达;原位杂交方法检测MMP-8 mRNA在大鼠牙胚发育钟状期的表达。结果　 MMP-8在人牙胚钟状晚期硬组织形成期成牙本质细胞和牙本质基质阳性表达,近髓腔处,牙本质染色更深;MMP-8 mRNA在大鼠牙胚发育钟状早期个别分化的成牙本质细胞表达阳性,钟状晚期,即硬组织形成期,MMP-8mRNA在成牙本质细胞表达强阳性。结论　MMP-8可能参与了人和大鼠牙胚发育过程牙本质基质改建。     

人前磨牙牙髓牙本质复合体层粘连蛋白受体的免疫组织化学研究

目的研究层粘连蛋白受体在牙髓牙本质复合体内的表达,探讨其分布与功能的关系.方法收集新鲜健康人前磨牙60个,固定,脱钙,石蜡包埋,层粘连蛋白受体SABC免疫组织化学染色.结果层粘连蛋白受体免疫反应物(LNR-IR)普遍存在于牙髓牙本质复合体的神经纤维膜、血管内皮细胞、成牙本质细胞及成纤维细胞上,以牙髓的神经纤维膜、血管内皮细胞最为丰富.结论层粘连蛋白受体一般见于牙髓牙本质复合体的各种细胞膜上,它与层粘连蛋白一起在支持、固定牙髓牙本质复合体各结构的正常位置、形态和功能的发挥上可能起了重要作用.     

MEPE is downregulated as dental pulp stem cells differentiate

Previous studies on dental pulp cell culture have described heterogenous mixtures of cells that differentiate into odontoblasts and form mineralized dentin. OBJECTIVE: The aim of this study was to characterize the matrix extracellular phosphoglycoprotein (MEPE) expression by dental pulp stem cells (DPSC), related to cell differentiation. DESIGN: DPSC differentiation to form mineralized nodules was characterized by Alizarin red staining and micro-Raman spectroscopy. Osteogenesis SuperArray analysis was used to broadly screen for osteogenesis-related genes altered by DPSC differentiation. Relative levels of expression of MEPE and DSP were determined by semiquantitative RT-PCR and Western blot. RESULTS: Mineral analysis showed that as DPSC differentiated, they formed a carbonated hydroxyapatite mineral. Differentiation was initially marked by upregulation by Runx2, TGFbeta-related genes, EGFR and genes involved in collagen metabolism. ALP activity first increased, as DPSCs reached confluence but later decreased when cells further differentiated three weeks after confluence. MEPE was the only marker that was downregulated as DPSCs differentiated. CONCLUSION: DPSC differentiation can be characterized by downregulation of MEPE as other markers of DPSC differentiation, such as DSP, are upregulated. Expression of MEPE related to DSP and can be used to monitor DPSC as they are used for studies of odontoblast differentiation, tissue engineering or vital pulp therapy. The downregulation of MEPE as DPSC differentiate, suggests that MEPE is an inhibitor of mineralization.