摘要
目的明确果蝇裸露角蛋白2(Nkd2)在大鼠下颌第一磨牙牙本质发育过程中表达。方法取出生后1d、3d、5d、7d、9d的SD乳鼠下颌骨制作石蜡切片免疫组化染色,倒置显微镜观测Nkd2在大鼠牙胚中的表达;体外培养大鼠牙乳头细胞并采用细胞免疫组化染色进行组织来源鉴定,诱导其成牙本质向分化后茜素红染色,荧光定量RT-qPCR和Western blot检测Nkd2在此过程中的表达变化趋势。结果免疫组织化学染色显示Nkd2在牙乳头、成牙本质细胞层和成釉细胞和牙囊中均阳性,在牙本质形成早期的成牙本质细胞层中强表达,随后减弱。Nkd2在大鼠牙乳头细胞中的表达呈阳性,矿化诱导4周后茜素红染色显示有矿化结节形成。荧光定量RT-qPCR和Western blot检测显示矿化诱导培养过程中Nkd2的表达呈先上调后下调趋势。结论Nkd2在大鼠牙胚组织中的表达分布具有时空特异性,体外大鼠牙乳头细胞诱导成牙本质细胞向分化过程中,Nkd2表达呈先上调后下调的趋势,推测Nkd2可能在牙本质形成早期发挥作用。
Objective To detect the expression of Nkd2 in the development of dentin.Methods Immunohistochemistry was performed on the mandibles of SD rats at 1,3,5,7 and 9 days after birth.The expression of Nkd2 in rat dental germ was observed by inverted microscopy.Rat dental papilla cells were cultured in vitro and identified by immunohistochemistry.After inducing odontoblast differentiation of dental papilla cells,alizarin red staining was performed,and RT-qPCR and western blotting were used to detect the expression of Nkd2 in the process.Results Immunohistochemical staining showed positivity for Nkd2 in the dental papilla,odontoblast layer,ameloblasts and dental follicles,and Nkd2 was strongly expressed in the early stage of dentinogenesis and then weakened.The expression of Nkd2 was positive in rat dental papilla cells.Alizarin red staining showed that mineralized nodules formed 4 weeks after mineralization induction.RT-qPCR and western blotting showed that the expression of Nkd2 increased first and then decreased during mineralization induction culture.Conclusion The expression and distribution of Nkd2 in rat dental germ tissues are time-and spacespecific.During the odontoblast differentiation of rat dental papilla cells in vitro,the expression of Nkd2 increased first and then decreased.It is suggested that Nkd2 may be involved in the regulation of dentin development and play a role in the early stage of dentin formation.
作者
陈婵婵
廖志清
王勤
张叶
丁桂聪
CHEN Chanchan;LIAO Zhiqing;WANG Qin;ZHANG Ye;DING Guicong(Shenzhen Children's Hospital,Gongdong,Shenzhen 518026)
出处
《现代口腔医学杂志》
CAS
2020年第4期197-201,共5页
Journal of Modern Stomatology
基金
广东省自然科学基金(2017A030310595)
深圳市知识创新计划基础研究项目(JCYJ20170303155435885)。
