In the process of orthodontic tooth movement (OTM), periodontal ligament fibroblasts (PDLFs) must undergo osteogenic differentiation. OTM increased the expression of Zinc finger and BTB domain-containing 16 (ZBTB16), which is implicated in osteogenic differentiation. Our goal was to investigate the mechanism of PDLF osteogenic differentiation mediated by ZBTB16. The OTM rat model was established, and PDLFs were isolated and exposed to mechanical force. Hematoxylin-eosin staining, Alizarin Red staining, immunofluorescence, and immunohistochemistry were carried out. The alkaline phosphatase (ALP) activity was measured. Dual-luciferase reporter gene assay and chromatin immunoprecipitation assay were conducted. In OTM models, ZBTB16 was significantly expressed. Additionally, there was an uneven distribution of PDLFs in the OTM group, as well as an increase in fibroblasts and inflammatory infiltration. ZBTB16 interference hindered PDLF osteogenic differentiation and decreased Wnt and β-catenin levels. Meanwhile, ZBTB16 activated the Wnt/β-catenin pathway. ZBTB16 also enhanced the expression of the osteogenic molecules osterix, osteocalcin (OCN), osteopontin (OPN), and bone sialo protein (BSP) at mRNA and protein levels. The interactions between Wnt1 and ZBTB16, as well as GCN5 and ZBTB16, were also verified. The adeno-associated virus-shZBTB16 injection also proved to inhibit osteogenic differentiation and reduce tooth movement distance in in vivo tests. ZBTB16 was up-regulated in OTM. Through acetylation modification of ZBTB16, GCN5 regulated the Wnt/β-catenin signaling pathway and further mediated PDLF osteogenic differentiation.

中文翻译：

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GCN5通过乙酰化调控ZBTB16，介导成骨分化，影响正畸牙移动。

在正畸牙齿移动（OTM）过程中，牙周韧带成纤维细胞（PDLFs）必须经历成骨分化。OTM 增加了锌指和含有 BTB 结构域的 16 (ZBTB16) 的表达，这与成骨分化有关。我们的目标是研究 ZBTB16 介导的 PDLF 成骨分化的机制。建立OTM大鼠模型，分离PDLFs并施加机械力。进行苏木精-伊红染色、茜素红染色、免疫荧光和免疫组化。测量碱性磷酸酶 (ALP) 活性。进行了双荧光素酶报告基因测定和染色质免疫沉淀测定。在 OTM 模型中，ZBTB16 显着表达。此外，OTM 组中的 PDLF 分布不均，以及成纤维细胞和炎症浸润的增加。ZBTB16 干扰阻碍了 PDLF 成骨分化并降低了 Wnt 和 β-catenin 水平。同时，ZBTB16 激活了 Wnt/β-catenin 通路。ZBTB16 还在 mRNA 和蛋白质水平上增强了成骨分子 osterix、骨钙素 (OCN)、骨桥蛋白 (OPN) 和骨唾液酸蛋白 (BSP) 的表达。还验证了 Wnt1 和 ZBTB16 以及 GCN5 和 ZBTB16 之间的相互作用。腺相关病毒-shZBTB16 注射液在体内试验中也证明可以抑制成骨分化并减少牙齿移动距离。ZBTB16 在 OTM 中被上调。GCN5通过对ZBTB16进行乙酰化修饰，调控Wnt/β-catenin信号通路，进一步介导PDLF成骨分化。