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脂多糖(LPS)诱导的牙髓炎小鼠模型

Mouse Model of Lipopolysaccharide (LPS)-Induced Pulpitis.

作者信息

Shao Lanting, Chen Baian, Zheng Ying

机构信息

School of Basic Medical Sciences, Capital Medical University, Beijing, China.

Department of Stomatology, Peking Union Medical College Hospital, Beijing, China.

出版信息

Bio Protoc. 2025 Jan 20;15(2):e5128. doi: 10.21769/BioProtoc.5128.

Abstract

Pulpitis is an important and prevalent disease within the oral cavity. Thus, animal models are necessary tools for basic research focused on pulpitis. Researchers worldwide often use dogs and miniature pigs to construct animal models of pulpitis. However, gene editing in miniature pigs is difficult, the surgical modeling process is complex, and tooth demineralization time is lengthy. Although some researchers have attempted to establish a mouse model of pulpitis, most models have involved direct exposure of dental pulp. However, the causes of pulpitis vary considerably among individuals, hindering effective research. In this study, we established a mouse model of pulpitis by accessing the pulp cavity, exposing the pulp to lipopolysaccharide (LPS), and then filling the tooth. One day after surgery, we observed many necrotic tissues and extensive inflammatory exudate, including neutrophils, around the coronal cavity preparation. Additionally, we noted many more neutrophils and a small amount of chronic inflammatory cell infiltrates at the junction between inflamed and normal tissue. These findings indicated that our model can be used to explore the early stage of pulpitis. Ten days after surgery, we observed vacuolar degeneration in some fibroblasts and proliferation in others at the distal end of the inflamed tissue. We also noted dilation and congestion of the pulp blood vessels. Therefore, our model can also be used to explore the middle and later stages of pulpitis. Thirty days after surgery, we observed necrosis in the coronal pulp cavity and upper half of the root pulp, indicating that our model can also be used to explore the end stage of pulpitis. This model is easy to establish, shows pulpitis progression in the dental pulp, exhibits a clear inflammatory phenotype, and can be readily combined with gene editing techniques. Accordingly, it is suitable for basic research focused on pulpitis and has substantial practical value. Key features • Lipopolysaccharide (LPS) can induce pulpitis in mice. • The mouse model of LPS-induced pulpitis can be used in basic studies of pulpitis. • After 1 day, the mouse model of LPS-induced pulpitis can demonstrate the main phenotypes of early-stage pulpitis. • After 10 days, the mouse model of LPS-induced pulpitis can display the main phenotypes of middle and late stage pulpitis. Graphical overview Figure 1.Graphical overview of the C57BL/6 mouse model of lipopolysaccharide (LPS)-induced pulpitis.A. Weigh the mouse. B. Anesthetize the mouse. C. Secure the mouse to the surgical pad and expose its oral cavity. D. Open the pulp chamber of the right maxillary first molar. E. Rinse the medullary foramen with 0.9% NaCl solution. Apply a small cotton ball saturated with 1 mg/mL LPS to the medullary foramen for 5 min, then cover the medullary foramen with Esthet-X flow and irradiate the site. F. Perform tissue decalcification and paraffin embedding (sample collection, decalcification, dehydration, wax embedding, and sectioning), followed by Histopathology staining, microscopy examination, image acquisition, and analysis.

摘要

牙髓炎是口腔内一种重要且常见的疾病。因此,动物模型是专注于牙髓炎基础研究的必要工具。世界各地的研究人员经常使用狗和小型猪来构建牙髓炎动物模型。然而,小型猪的基因编辑困难,手术建模过程复杂,牙齿脱矿时间长。尽管一些研究人员试图建立牙髓炎小鼠模型,但大多数模型都涉及直接暴露牙髓。然而,牙髓炎的病因在个体之间差异很大,阻碍了有效研究。在本研究中,我们通过进入牙髓腔,将牙髓暴露于脂多糖(LPS),然后填充牙齿,建立了一种牙髓炎小鼠模型。手术后一天,我们在冠腔制备周围观察到许多坏死组织和广泛的炎性渗出物,包括中性粒细胞。此外,我们在炎症组织与正常组织交界处注意到更多的中性粒细胞和少量慢性炎性细胞浸润。这些发现表明我们的模型可用于探索牙髓炎的早期阶段。手术后十天,我们在炎症组织远端观察到一些成纤维细胞出现空泡变性,另一些成纤维细胞增殖。我们还注意到牙髓血管扩张和充血。因此,我们的模型也可用于探索牙髓炎的中晚期阶段。手术后三十天,我们观察到冠髓腔和根髓上半部分坏死,表明我们的模型也可用于探索牙髓炎的终末期阶段。该模型易于建立,显示牙髓中牙髓炎的进展,表现出明显的炎症表型,并且可以很容易地与基因编辑技术相结合。因此,它适用于专注于牙髓炎的基础研究,具有重要的实用价值。关键特征 • 脂多糖(LPS)可诱导小鼠发生牙髓炎。 • LPS诱导的牙髓炎小鼠模型可用于牙髓炎的基础研究。 • 1天后,LPS诱导的牙髓炎小鼠模型可表现出早期牙髓炎的主要表型。 • 10天后,LPS诱导的牙髓炎小鼠模型可显示中晚期牙髓炎的主要表型。图形概述 图1.LPS诱导的C57BL/6小鼠牙髓炎模型的图形概述。A.称小鼠体重。B.麻醉小鼠。C.将小鼠固定在手术垫上并暴露其口腔。D.打开右上颌第一磨牙的髓腔。E.用0.9%氯化钠溶液冲洗髓孔。将浸有1mg/mL LPS的小棉球敷于髓孔5分钟,然后用Esthet-X flow覆盖髓孔并照射该部位。F.进行组织脱钙和石蜡包埋(样本采集、脱钙、脱水、浸蜡和切片),随后进行组织病理学染色、显微镜检查、图像采集和分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7cab/11769714/120a56f345ee/BioProtoc-15-2-5128-g001.jpg

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