FOXO1 regulates wound-healing responses in human gingival fibroblasts

IF 3.4 3区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE Journal of periodontal research Pub Date : 2024-03-18 DOI:10.1111/jre.13257

Leticia Rojas, Nicolás Tobar, Javier Espinoza, Susana Ríos, Constanza Martínez, Jorge Martínez, Dana T. Graves, Patricio C. Smith

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Abstract

Background and objective

Forkhead box-O 1 (FOXO1) is a transcription factor actively involved in oral wound healing at the epithelial barrier. However, less is known regarding the role of FOXO1 during the tissue repair response in the connective tissue compartment. This study explored the involvement of FOXO1 in the modulation of fibroblast activity related to wound healing.

Methods

Primary cultures of human gingival fibroblasts were obtained from four healthy young donors. Myofibroblastic differentiation, collagen gel contraction, cell migration, cell spreading, and integrin activation were evaluated in the presence or absence of a FOXO1 inhibitor (AS1842856). Variations in mRNA and proteins of interest were evaluated through qRT-PCR and western blot, respectively. Distribution of actin, α-smooth muscle actin, and β1 integrin was evaluated using immunofluorescence. FOXO1 and TGF-β1 expression in gingival wound healing was assessed by immunohistochemistry in gingival wounds performed in C57BL/6 mice. Images were analyzed using ImageJ/Fiji. ANOVA or Kruskal-Wallis test followed by Tukey's or Dunn's post-hoc test was performed. All data are expressed as mean ± SD. p < .05 was considered statistically significant.

Results

FOXO1 inhibition caused a decrease in the expression of the myofibroblastic marker α-SMA along with a reduction in fibronectin, type I collagen, TGF-β1, and β1 integrin mRNA level. The FOXO1 inhibitor also caused decreases in cell migration, cell spreading, collagen gel contraction, and β1 integrin activation. FOXO1 and TGF-β1 were prominently expressed in gingival wounds in fibroblastic cells located at the wound bed.

Conclusion

The present study indicates that FOXO1 plays an important role in the modulation of several wound-healing functions in gingival fibroblast. Moreover, our findings reveal an important regulatory role for FOXO1 on the differentiation of gingival myofibroblasts, the regulation of cell migration, and collagen contraction, all these functions being critical during tissue repair and fibrosis.

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FOXO1 调节人类牙龈成纤维细胞的伤口愈合反应。

背景和目的：叉头盒-O 1（FOXO1）是一种转录因子，积极参与口腔上皮屏障的伤口愈合。然而，人们对 FOXO1 在结缔组织区组织修复反应中的作用知之甚少。本研究探讨了 FOXO1 参与调节与伤口愈合相关的成纤维细胞活性的情况：方法：从四名健康的年轻供体处获得人牙龈成纤维细胞的原代培养物。在 FOXO1 抑制剂（AS1842856）存在或不存在的情况下，对成纤维细胞的分化、胶原凝胶收缩、细胞迁移、细胞扩散和整合素活化进行了评估。相关 mRNA 和蛋白质的变化分别通过 qRT-PCR 和 Western 印迹进行评估。肌动蛋白、α-平滑肌肌动蛋白和β1整合素的分布通过免疫荧光进行评估。在 C57BL/6 小鼠的牙龈伤口中使用免疫组化方法评估 FOXO1 和 TGF-β1 在牙龈伤口愈合中的表达。使用 ImageJ/Fiji 对图像进行分析。进行方差分析或 Kruskal-Wallis 检验，然后进行 Tukey's 或 Dunn's 事后检验。p 结果：抑制 FOXO1 可降低肌成纤维细胞标记物 α-SMA 的表达，同时降低纤维连接蛋白、I 型胶原、TGF-β1 和 β1 整合素 mRNA 水平。FOXO1 抑制剂还导致细胞迁移、细胞扩散、胶原凝胶收缩和 β1 整合素活化的减少。FOXO1和TGF-β1在牙龈伤口处位于伤口床的成纤维细胞中显著表达：本研究表明，FOXO1 在调节牙龈成纤维细胞的多种伤口愈合功能中发挥着重要作用。此外，我们的研究结果还揭示了 FOXO1 在牙龈肌成纤维细胞分化、细胞迁移调控和胶原收缩中的重要调控作用，所有这些功能在组织修复和纤维化过程中都至关重要。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of periodontal research 医学-牙科与口腔外科

CiteScore

6.90

自引率

5.70%

发文量

103

审稿时长

6-12 weeks

期刊介绍： The Journal of Periodontal Research is an international research periodical the purpose of which is to publish original clinical and basic investigations and review articles concerned with every aspect of periodontology and related sciences. Brief communications (1-3 journal pages) are also accepted and a special effort is made to ensure their rapid publication. Reports of scientific meetings in periodontology and related fields are also published. One volume of six issues is published annually.