Franco-Arellanes María Cristina, Toledo-Valdes Perla Xóchitl, Díaz-Hernández Cynthia, Díaz-Castillejos Risk, García-Reyes Eunice Daysi, Ramírez-Thomé Saira Karina, Ávila-Curiel Beatriz Xóchitl, Castañeda-Patlán María Cristina, Zenteno Edgar, Solórzano-Mata Carlos Josué
Faculty of Medicine and Surgery, Universidad Autónoma Benito Juárez de Oaxaca, Oaxaca, México.
Faculty of Dentistry, Universidad Autónoma Benito Juárez de Oaxaca, Oaxaca, México.
Folia Histochem Cytobiol. 2025;63(1):1-10. doi: 10.5603/fhc.103756. Epub 2025 Mar 28.
O-GlcNAcylation is a post-translational modification in which a single N-Acetyl-D-Glucosamine (GlcNAc) molecule is added to Ser or Thr residues of proteins. The O-N-acetylglucosaminyl transferase (OGT) enzyme is responsible for adding GlcNAc to the target proteins and N-acetyl-β-D-glucosaminidase (OGA) that removes the GlcNAc residue. O-GlcNAcylation has been described in the pathophysiology of several diseases; however, little has been studied in dental tissue. The aim of the present work is to characterise the product of O-GlcNAcylation and its enzymes at the tissue level in the dental pulp, as well as its expression in dental pulp stem cells (DPSCs) both in situ and in vitro. This enables the recognition of the behaviour of O-GlcNAcylation in pulp tissue without pathology.
Pulp tissue was obtained from 10 healthy donors, and the expression of O-GlcNAc, OGT, and OGA was analysed using immunofluorescence with specific antibodies in different regions of the dental pulp. DPSCs were isolated, cultured, and identified with anti-STRO1 (antibody specific for human CD34+ cells, useful for DPSC identification). The expression of O-GlcNAc in DPSCs was confirmed in vitro through Western blot.
Different regions of the dental pulp and DPSCs express O-GlcNAc and the enzymes OGT and OGA. O-GlcNAc and OGT expression was more prominent in the odontoblastic layer, cell-rich zone, and in the central core. OGA was distributed throughout the pulp tissue with lower immunoreactivity compared to OGT.
Our results suggest that O-GlcNAcylation may play a relevant role in human dental pulp homeostasis and in physiology of DPSCs.
O-连接的N-乙酰葡糖胺化（O-GlcNAcylation）是一种翻译后修饰，其中单个N-乙酰-D-葡糖胺（GlcNAc）分子被添加到蛋白质的丝氨酸或苏氨酸残基上。O-N-乙酰葡糖胺基转移酶（OGT）负责将GlcNAc添加到靶蛋白上，而N-乙酰-β-D-葡糖胺酶（OGA）则去除GlcNAc残基。O-GlcNAcylation已在多种疾病的病理生理学中被描述；然而，在牙组织中的研究较少。本研究的目的是在组织水平上表征牙髓中O-GlcNAcylation的产物及其酶，以及其在牙髓干细胞（DPSC）原位和体外的表达。这有助于识别牙髓组织中无病理状态下O-GlcNAcylation的行为。
从10名健康供体获取牙髓组织，使用特异性抗体通过免疫荧光分析牙髓不同区域中O-GlcNAc、OGT和OGA的表达。分离、培养DPSC并用抗STRO1（对人CD34+细胞特异的抗体，可用于DPSC鉴定）进行鉴定。通过蛋白质印迹在体外证实DPSC中O-GlcNAc的表达。
牙髓的不同区域和DPSC表达O-GlcNAc以及OGT和OGA酶。O-GlcNAc和OGT的表达在成牙本质细胞层、细胞丰富区和中央核心更为突出。与OGT相比，OGA分布于整个牙髓组织，免疫反应性较低。
我们的结果表明，O-GlcNAcylation可能在人牙髓稳态和DPSC生理学中发挥相关作用。
