Pucinelli Carolina Maschietto, Nelson-Filho Paulo, Lucisano Marília Pacífico, León Jorge Esquiche, Faccioli Lúcia Helena, Sorgi Carlos Arterio, Silva Clara Marina Pereira Cavalcanti, Silva Lea Assed Bezerra da, Silva Raquel Assed Bezerra da
Universidade de São Paulo, Faculdade de Odontologia de Ribeirão Preto, Departamento de Clínica Infantil, Ribeirão Preto, SP, Brasil.
Universidade de São Paulo, Faculdade de Odontologia de Ribeirão Preto, Departamento de Estomatologia, Saúde Coletiva e Odontologia Legal, Ribeirão Preto, SP, Brasil.
J Appl Oral Sci. 2025 Jun 27;33:e20240579. doi: 10.1590/1678-7757-2024-0579. eCollection 2025.
This study evaluated the altered expression levels of genes and cytokines associated with M1 and M2 macrophages during the development of periapical lesion (PL).
PL was induced in the lower first molars of 96 mice. After the experimental periods of two, seven, 14, 21, and 42 days, the animals were euthanized and their jaws were dissected and submitted to the following analyses: microscopic descriptive analysis and fluorescence microscopy morphometry of PL size (mm2); quantitative gene expression analysis by qRT-PCR for M1 (Cxcl10, Cxcl9, and Nos2) and M2 phenotypes (Arg1, Fizz1, Ym1, and Mrc1); and M1- (GM-CSF, IFN-γ, IL-6, IL-1β, TNF-α) and M2- (IL-4, IL-13, and IL- 10) related cytokines quantification by Luminex. Data were statistically compared by ANOVA, Tukey post-test, Kruskal-Wallis, and Dunn post-test (α=5%).
PL area and inflammatory infiltrate increased over experimental periods. From a contextual view, a pro-inflammatory profile and a higher activation of M1 phenotype markers in the initial periods of two and seven days could be observed. On day 21, microscopic features and M2 subtype predominance indicated a repair attempt. However, on day 42, an acute exacerbation of immunoinflammatory process and return to the M1 macrophage profile were evidenced.
M1 and M2 macrophage polarization-related markers were expressed alternately throughout the experimental periods, according to the stage of PL progression.
本研究评估了根尖周病变(PL)发展过程中与M1和M2巨噬细胞相关的基因和细胞因子表达水平的变化。
在96只小鼠的下颌第一磨牙中诱导PL。在实验的2、7、14、21和42天结束后,对动物实施安乐死,解剖其颌骨并进行以下分析:PL大小(mm²)的显微镜描述性分析和荧光显微镜形态测量;通过qRT-PCR对M1(Cxcl10、Cxcl9和Nos2)和M2表型(Arg1、Fizz1、Ym1和Mrc1)进行定量基因表达分析;以及通过Luminex对与M1(GM-CSF、IFN-γ、IL-6、IL-1β、TNF-α)和M2(IL-4、IL-十三、和IL-10)相关的细胞因子进行定量。数据通过方差分析、Tukey事后检验、Kruskal-Wallis检验和Dunn事后检验进行统计学比较(α=5%)。
在实验期间,PL面积和炎症浸润增加。从背景来看,在第2天和第7天的初始阶段,可以观察到促炎特征和M1表型标志物的更高激活。在第21天,微观特征和M2亚型占主导地位表明有修复尝试。然而,在第42天,免疫炎症过程急性加重并恢复到M1巨噬细胞特征得到证实。
根据PL进展阶段,M1和M2巨噬细胞极化相关标志物在整个实验期间交替表达。