Nakayama Yohei, Igarashi Kazuma, Jin Zhenyu, Yamaguchi Arisa, Ganss Bernhard, Ogata Yorimasa
Department of Periodontology, Nihon University School of Dentistry at Matsudo, Chiba, Japan.
Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, Chiba, Japan.
J Periodontal Implant Sci. 2025 Aug;55(4):255-272. doi: 10.5051/jpis.2403080154. Epub 2025 Jan 30.
The junctional epithelium (JE) covers the cervical areas of developing or existing teeth. It can re-establish itself even after being removed during periodontal therapies, followed by wound healing. However, the mechanisms that can maintain this universally conserved structure are still unclear.
The molecular mechanisms of JE homeostasis were investigated by altering levels of JE-specific genes in a telomerase immortalized human gingival epithelial cell line (TIGKs) by exposing TIGKs to conditioned medium (C-CM) from cultivated human cementoblasts. The mRNA and protein levels of JE-associated genes in TIGKs were examined using real-time polymerase chain reaction (PCR) and immunocytochemistry (ICC) after treatment with C-CM. The candidate pathways related to differential mRNA and protein expression were analyzed with a DNA microarray and identified using Kyoto Encyclopedia of Genes and Genomes and WikiPathways. Real-time PCR and ICC were conducted to confirm the changes in the expressions of candidate genes.
mRNA levels and protein expressions of amelotin () were upregulated after treatment with C-CM for 48 hours. DNA microarray analyses identified 595 genes that were upregulated >2-fold, and 820 genes that were downregulated >2-fold. C-CM promoted the expression of suppressor of cytokine signaling 3 and reduced the expression of an inactivator of Janus kinase 2. Both signaling molecules were found, using siRNA technology, to mediate the increase of mRNA and protein expression levels.
The upregulation of in gingival epithelial cells by C-CM suggests that this regulatory pathway is associated with the homeostasis of JE structures by the cementum.
结合上皮（JE）覆盖发育中或已存在牙齿的颈部区域。即使在牙周治疗期间被去除后，它仍能在伤口愈合后自我重建。然而，维持这种普遍保守结构的机制仍不清楚。
通过将永生化人牙龈上皮细胞系（TIGKs）暴露于培养的人成牙骨质细胞的条件培养基（C-CM）中，改变JE特异性基因的水平，研究JE稳态的分子机制。用C-CM处理后，采用实时聚合酶链反应（PCR）和免疫细胞化学（ICC）检测TIGKs中JE相关基因的mRNA和蛋白质水平。用DNA微阵列分析与差异mRNA和蛋白质表达相关的候选途径，并使用京都基因与基因组百科全书和WikiPathways进行鉴定。进行实时PCR和ICC以确认候选基因表达的变化。
用C-CM处理48小时后，釉成熟蛋白（）的mRNA水平和蛋白质表达上调。DNA微阵列分析确定了595个上调>2倍的基因和820个下调>2倍的基因。C-CM促进细胞因子信号转导抑制因子3的表达，并降低Janus激酶2失活剂的表达。使用siRNA技术发现这两种信号分子都介导了mRNA和蛋白质表达水平的增加。
C-CM使牙龈上皮细胞中上调，表明该调节途径与牙骨质对JE结构的稳态有关。
