壳聚糖包裹siRNA在小鼠牙胚发育研究中的应用  

Application of chitosan encapsulated siRNA on the development of mouse tooth germs

作　　者：倪锦雯 高山[2] 段小红[1] 

机构地区：[1]第四军医大学口腔医学院,陕西西安710032 [2]中南大学口腔医学院,湖南长沙410078

出　　处：《临床口腔医学杂志》2014年第3期146-148,共3页Journal of Clinical Stomatology

基　　金：国家自然科学基金(81271116)

摘　　要：目的:探索局部注射siRNA对小鼠牙胚生长发育产生影响的实验方法。方法:选择健康新生昆明小鼠作为实验对象,将阴性对照siRNA进行壳聚糖(chitosan,CS)包裹,分别将其注入小鼠的两个不同部位:①.腹腔注射组,将Cy5.5标记的CS包裹的siRNA纳米粒注射于新生昆明小鼠腹腔,于不同时间点在动物活体成像仪下观察荧光分布情况。②.颌骨局部注射组,将Cy5.5标记的siRNA/CS纳米粒局部直接注射于新生小鼠左上颌第一磨牙牙胚,通过动物活体成像结合冰冻切片观察荧光分布情况,验证局部注射方法的可行性。结果:在腹腔注射组,随着时间延长,荧光开始在腹腔内扩散,随后在腹腔内淬灭,荧光始终局限于腹腔,腹腔外没有观察到荧光;颌骨局部注射组通过活体动物成像显示注射的牙胚处有荧光,进一步的冰冻切片验证在牙胚处有荧光,且牙胚结构无明显损伤。结论:本实验将CS包裹siRNA纳米粒技术应用于牙胚发育的相关研究,首次成功构建了新生昆明小鼠牙胚局部注射siRNA的方法,为研究牙胚发育相关基因的作用开辟了新的途径。Objective： To explore the effect of local injected siRNA on the development of neonatal mice tooth germs. Method： Newborn Kunming mice were injected with chitosan encapsulated negative control siRNA through two different lo- cations respectively： （1）.Intraperitoneal injection group, Cy5.5-chitosan-siRNA was intraperitoneal injected in neonatal mice, then we observed fluorescence distribution by in vivo fluorescence imaging at different time points. （2）.Local maxillary bone injection group, Cy5.5-chitosan-siRNA was directly injected into tooth germs Of left maxillary first molars in neonatal mice, in order to verify the feasibility of this method by observing fluorescence distribution through in vivo fluorescence imaging and frozen section. Result：In intraperitoneal injection group,there was no fluorescence signal in tooth germ, since fluores- cence imaging confines to peritoneal cavity as time went on. Local maxillary bone injection group showed obvious fluores- cence distribution at the injected tooth germ sites via in vivo fluorescence imaging as well as frozen section tooth germ sites. Conclusion： A successful siRNA local injection method in neonatal mice left maxillary first molar tooth germs was estab- lished in this experiment, opening up a new way in the study of tooth germ development related genes.

关 键 词：腹腔注射 局部注射 牙胚 壳聚糖 小干扰RNA 动物活体成像 

分 类 号：R780.2[医药卫生—口腔医学]