Trejo-Remigio David A, Jacinto-Alemán Luis F, Leyva-Huerta Elba R, Navarro-Bustos Bogdan R, Portilla-Robertson Javier
Department of Oral Medicine and Pathology, Graduate Dental School, National Autonomous Mexico University, Mexico City, Mexico.
Laboratory of Cell Culture and Immunohistochemistry, Department of Oral Medicine and Pathology, Graduate Dental School, National Autonomous Mexico University, Mexico City, Mexico.
Minerva Stomatol. 2019 Jun;68(3):132-141. doi: 10.23736/S0026-4970.19.04166-9.
Odontomas are odontogenic tumors with hamartoma features that are classified as compound or complex. Our objective was to characterize the proliferation of ectodermal and ectomesenchymal profile markers of primary cell cultures of complex and compound odontomas.
Four samples of compound odontomas (OdCm) and three of complex odontomas (OdCx) were obtained from patients attending the Oral Pathology and Medicine Clinic of the Graduate Dental School, National Autonomous University of Mexico for primary culture generation. MTT, immunocytochemistry and RT-PCR assays of CD34, Sox2, Amel, Ambn, p21, EDAR, Msx1, Msx2, Pax9, RUNX2, BSP, OPN, Barx1 and GAPDH (control) were performed. Additionally, six paraffin-embedded odontomas were obtained for immunocytochemistry and RT-PCR validation assays. The mean and standard deviation were determined, and ANOVA and Kruskall-Wallis tests were performed.
Cultured compound odontoma exhibited higher proliferation, and an ectomesenchymal immunocytochemistry profile with predominant expression of Amel, BSP, Pax9, EDAR, Barx and Msx2; in complex cultured odontoma Sox2, CD34, RUNX2 and OPN predominated. Our statistical analysis showed a significant difference in PCR analysis (P<0.05) for OPN and CD34. Paraffin-embedded odontomas showed similar pattern with difference for NGFR and Sox2 for immunohistochemistry and EDAR, BARX1 and PAX9 for RT-PCR assays.
The results suggested heterogeneous behavior for both odontoma cell lines, because in compound odontomas predominant biomarkers are related to the enamel knot, late-stage odontogenesis and ectomesenchymal interactions; and in complex odontoma the significant expression of CD34 and OPN could be responsible for the difference behavior and mineralized amorphous structure.
牙瘤是具有错构瘤特征的牙源性肿瘤，分为复合性或复杂性。我们的目的是表征复合性和复杂性牙瘤原代细胞培养物中外胚层和外间充质特征标志物的增殖情况。
从墨西哥国立自治大学牙科学院口腔病理学与医学诊所的患者中获取4份复合性牙瘤样本（OdCm）和3份复杂性牙瘤样本（OdCx），用于原代培养。对CD34、Sox2、Amel、Ambn、p21、EDAR、Msx1、Msx2、Pax9、RUNX2、BSP、OPN、Barx1和GAPDH（对照）进行MTT、免疫细胞化学和RT-PCR检测。此外，获取6份石蜡包埋的牙瘤用于免疫细胞化学和RT-PCR验证检测。确定均值和标准差，并进行方差分析和克鲁斯卡尔-沃利斯检验。
培养的复合性牙瘤表现出更高的增殖率，以及外间充质免疫细胞化学特征，主要表达Amel、BSP、Pax9、EDAR、Barx和Msx2；在培养的复杂性牙瘤中，Sox2、CD34、RUNX2和OPN占主导。我们的统计分析显示，OPN和CD34的PCR分析存在显著差异（P<0.05）。石蜡包埋的牙瘤显示出相似的模式，免疫组化中NGFR和Sox2存在差异，RT-PCR检测中EDAR、BARX1和PAX9存在差异。
结果表明两种牙瘤细胞系存在异质性行为，因为在复合性牙瘤中，主要生物标志物与釉结、牙发育后期和外间充质相互作用有关；而在复杂性牙瘤中，CD34和OPN的显著表达可能导致不同的行为和矿化无定形结构。
