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目的应用比格犬建立年轻恒牙根尖周炎动物模型,并进行再生性牙髓治疗(regenerative endodontic treatment,RET),探究参与RET的间充质干细胞(mesenchymal stem cells,MSCs)的可能来源。方法本研究于2013年12月至2014年7月在中国医科大学口腔医学院中心实验室完成。选择3只6月龄的比格犬,每只犬双根前磨牙6颗,共18颗牙,随机分为正常对照组、根尖周炎组、RET组(分为RET-1、RET-2组)以及明胶海绵组(分为Gelfoam-1、Gelfoam-2组),每组3颗牙。RET-1、RET-2组分别在封抗生素糊剂炎症控制后,刺破根尖周组织出血,常规行RET 1、2周后拔牙,劈牙,取根管内容物。明胶海绵组,不刺破根尖周组织,而是在根管内放明胶海绵,使之接触到根尖部;Gelfoam-1、Gelfoam-2组分别于放明胶海绵1、2周后拔牙,劈牙,取根管内容物。正常对照组与根尖周炎组牙齿于建立根尖周炎模型4周后拔除,劈牙,取根管内容物。内容物的PBS冲洗液经甩片后,用免疫荧光标记MSCs标记物STRO-1和CD105。结果根尖周炎组根管内容物中STRO-1和CD105的阳性表达高于正常对照组。RET组和明胶海绵组,根管内容物中都可检测到MSCs标记物的表达。结论进行RET时,根管内的MSCs可来源于刺破根尖周组织出血,也可能由内源性干细胞向根管内“归巢”而来。
Objective To establish the animal model of periapical periodontitis of young permanent teeth in beagle dogs and to perform regenerative endodontic treatment (RET) to explore the possible source of mesenchymal stem cells (MSCs) involved in RET. Methods The study was performed at the Central Laboratory of Stomatology, China Medical University from December 2013 to July 2014. Three beagle dogs aged 6 months and 6 pairs of premolar teeth per dog were randomly divided into normal control group, apical periodontitis group and RET group (divided into RET-1 and RET- 2 groups) and gelatin sponge group (divided into Gelfoam-1, Gelfoam-2 group) with 3 teeth in each group. The RET-1 and RET-2 groups respectively punctured the periapical tissue after the antibiotic paste was inflamed, and routinely extracted the tooth root and split the teeth and took root canal contents after 1 and 2 weeks. Gelatin sponge group, do not puncture the periapical tissue, but put gelatin sponge in the root canal to make contact with the root tip; Gelfoam-1, Gelfoam-2 group were placed gelatin sponge 1, 2 weeks after tooth extraction, Cleave teeth, take root canal content. The normal control group and the periapical periodontitis group established the periapical periodontal model 4 weeks after the removal, chopped teeth, take root canal contents. The contents of the PBS rinse after rejection throne, immunofluorescence labeled MSCs markers STRO-1 and CD105. Results The positive expression of STRO-1 and CD105 in the root canal of the periapical periodontitis group was higher than that of the normal control group. RET group and gelatin sponge group, root canal content MSCs marker expression can be detected. Conclusions For RET, MSCs in the root canal may originate from punctured periapical tissue hemorrhage, or from endogenous stem cells to the root canal.