摘要
目的用Shh反义寡脱氧核苷酸(ODN)抑制体外培养小鼠牙胚中Shh的翻译,观察该抑制反应对牙胚生长及矿化的影响。方法培养牙胚取自胎龄为E17.5d昆明小鼠下颌第一磨牙,利用简易Trowell支架,建立小鼠牙胚体外培养模型,实验共分为三组:反义ODN组;正义ODN组及空白对照组,核酸浓度为30μmol/L。体外分别培养10d后,进行HE染色、von Kossa染色。结果蛋白印迹结果显示所应用的反义核酸阻断了牙胚中Shh的表达。HE、von Kossa染色结果显示:培养10d,S-ODN组与空白对照牙胚进入钟状晚期,细胞分化,并可见分泌的基质带;钙磷酸盐在成牙本质细胞与成釉细胞间的基质中沉积。AS-ODN组牙胚培养10d,牙胚进入钟状期,但牙胚体积明显变小,牙尖形态发育不良,成牙本质细胞排列紊乱,但仍可见高柱状分泌期细胞;von Kossa染色未见明显钙磷酸盐沉积。结论Shh反义核酸减缓体外培养牙胚的发育,可能通过对成牙本质细胞分泌功能的影响而影响牙胚的矿化。
Objective To understand the role of Sonic hedgehog （Shh） during tooth develolpment. Methods Shh antisense oligodeoxynucleotide （ODN） was designed, to specifically inhibit the translation of Shh mRNA. Mouse first molar germ was obtained from 17.5d embryo and cultured with Shh antisense ODN or Shh sense ODN or nothing as the controls for 10 days. The tooth germ was stained with HE and yon Kossa staining. Shh expression in the tooth germ was examined with Western blot. Results Shh expression was significantly decreased in Shh ODN treated group than that in the controls groups. The tooth germs treated with the Shh ODN were smaller than the controls. Von Kossa staining showed a dark staining band between enamel and predentin in control germs but not in the AS - ODN germs. Conclusion Shh played a role in the development and mineralization of cultured tooth germ.
出处
《现代口腔医学杂志》
CAS
CSCD
北大核心
2007年第2期169-172,共4页
Journal of Modern Stomatology
关键词
SHH
牙胚发育
Sonic hedgehog Tooth germ development
