Li Danfeng, Feng Yi, Tang Hang, Huang Lijia, Tong Zhongchun, Hu Cheng, Chen Xiaodan, Tan Jiali
Department of Orthodontics, Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, China.
Front Bioeng Biotechnol. 2020 May 25;8:444. doi: 10.3389/fbioe.2020.00444. eCollection 2020.
Periodontitis, a common disease that can lead to bone destruction, periodontal attachment loss, and tooth loss, is the major cause for oral tissue engineering. Experimental periodontitis is a suitable disease-model for studying bone regeneration and the potential therapeutic role of biomaterials on periodontal tissue engineering, as this model could be employed to mimic the natural host response under bacteria-caused oral pathological environments. Although large animals with ligature-induced periodontitis have mostly been used for experiments, a mouse model is a better choice for several reasons. Inserting ligature threads through the interproximal space between the teeth is the key step in establishing a periodontitis model, and it is easy to achieve in large animals, but difficult in mice due to the limited operating space. In this work, we provide a new and proven approach for periodontitis induction in mice using C+ nickel-titanium root canal files and stainless-steel ligature wires. The validity of this method was assessed by evaluating alveolar bone loss via micro-CT and detecting periodontal inflammation by histological staining and qPCR after the treatments. Progressive alveolar bone loss was observed from day 3 after the ligature-placement. Infiltration and accumulation of F4/80+ macrophage was also detected. In accordance with the histological results, there was upregulation of the expression levels of the inflammatory genes β, α, and in gingival tissues isolated from the ligation sites. Our results suggest that this novel method could resolve the difficulty of ligature-placement in mice and consequently contribute to further use of mouse models for studying the pathological mechanisms of periodontitis and developing potential periodontal tissue regeneration strategies. C+ files, which are made of nickel-titanium, are tough, elastic, and sufficiently thin to pass through the interproximal space between the teeth after pre-bending to form an appropriate angle, thus providing an access for ligature wire insertion. As a common tool in the dental clinic, it is familiar to researchers of oral biology, and can provide the feasibility for wide application of our method.
牙周炎是一种常见疾病，可导致骨质破坏、牙周附着丧失和牙齿脱落，是口腔组织工程的主要诱因。实验性牙周炎是研究骨再生以及生物材料在牙周组织工程中的潜在治疗作用的合适疾病模型，因为该模型可用于模拟细菌引起的口腔病理环境下的天然宿主反应。尽管大多使用结扎诱导牙周炎的大型动物进行实验，但出于几个原因，小鼠模型是更好的选择。将结扎丝穿过牙齿间的邻间隙是建立牙周炎模型的关键步骤，这在大型动物中很容易实现，但在小鼠中由于操作空间有限而困难。在这项工作中，我们提供了一种新的且经过验证的方法，使用C +镍钛根管锉和不锈钢结扎丝在小鼠中诱导牙周炎。通过微计算机断层扫描（micro-CT）评估牙槽骨丢失，并在处理后通过组织学染色和定量聚合酶链反应（qPCR）检测牙周炎症，以此评估该方法的有效性。在结扎放置后第3天观察到牙槽骨逐渐丢失。还检测到F4/80 +巨噬细胞的浸润和积聚。根据组织学结果，从结扎部位分离的牙龈组织中炎症基因β、α和的表达水平上调。我们的结果表明，这种新方法可以解决小鼠结扎放置的困难，从而有助于进一步利用小鼠模型研究牙周炎的病理机制并制定潜在的牙周组织再生策略。由镍钛制成的C +锉坚韧、有弹性且足够细，在预先弯曲形成合适角度后能够穿过牙齿间的邻间隙，从而为插入结扎丝提供通道。作为牙科诊所的常用工具，口腔生物学研究人员对其很熟悉，可为我们方法的广泛应用提供可行性。
