Lee Y-L, Liu J, Clarkson B H, Lin C-P, Godovikova V, Ritchie H H
Department of Cariology, Restorative Sciences and Endodontics, University of Michigan School of Dentistry, Ann Arbor, MI 48109-1078, USA.
Caries Res. 2006;40(3):256-64. doi: 10.1159/000092235.
To understand the molecular events underlying the dentin-pulp complex responses to carious progression, we systematically analyzed tissue morphology and dentin matrix protein distribution in non-carious teeth and in teeth with enamel and dentin caries. Dentin matrix proteins analyzed included collagen type I, phosphophoryn (PP) and dentin sialoprotein (DSP), all of which play decisive roles in the dentin mineralization process. Human non-carious and carious third molar teeth were freshly collected, demineralized, and processed for hematoxylin and eosin staining. The ABC-peroxidase method was used for immunohistochemical staining of collagen type I, PP and DSP proteins using specific antibodies. In situ hybridization was also performed. In contrast to elongated odontoblasts in non-carious teeth, odontoblasts subjacent to dentin caries were cuboidal and fewer in number. The predentin zone was also dramatically reduced in teeth with dentin caries. The staining intensity for collagen type I, PP and DSP in the dentin-pulp complex increased progressively from non-carious teeth, to teeth with enamel and dentin caries. In situ hybridization studies showed DSP-PP mRNA expression in odontoblasts and dental pulp that was consistent with our immunohistochemical results. These results suggest that carious lesions stimulate the dentin-pulp complex to actively synthesize collagen type I, PP and DSP proteins. This response to carious lesions is likely to provide a basis for reparative and/or reactionary dentin formation.
为了解牙本质-牙髓复合体对龋病进展反应的分子机制,我们系统分析了非龋牙以及患有釉质龋和牙本质龋牙齿的组织形态和牙本质基质蛋白分布。所分析的牙本质基质蛋白包括I型胶原、磷酸磷蛋白(PP)和牙本质涎蛋白(DSP),它们在牙本质矿化过程中均起决定性作用。新鲜采集人类非龋和龋坏的第三磨牙,进行脱矿处理,并进行苏木精-伊红染色。采用ABC过氧化物酶法,使用特异性抗体对I型胶原、PP和DSP蛋白进行免疫组织化学染色。同时还进行了原位杂交。与非龋牙中细长的成牙本质细胞不同,牙本质龋下方的成牙本质细胞呈立方形,数量较少。牙本质龋患牙的前期牙本质区也显著减少。牙本质-牙髓复合体中I型胶原、PP和DSP的染色强度从非龋牙到患有釉质龋和牙本质龋的牙齿逐渐增加。原位杂交研究显示,成牙本质细胞和牙髓中DSP-PP mRNA的表达与我们的免疫组织化学结果一致。这些结果表明,龋损刺激牙本质-牙髓复合体积极合成I型胶原、PP和DSP蛋白。这种对龋损的反应可能为修复性和/或反应性牙本质的形成提供基础。