Department of Physiology and Pathology, São Paulo State University (UNESP), Araraquara School of Dentistry, Araraquara, Brazil.
Department of Dentistry, Universidade de Ribeirão Preto, UNAERP, Ribeirão Preto, Brazil.
J Periodontal Res. 2023 Aug;58(4):791-799. doi: 10.1111/jre.13138. Epub 2023 May 24.
This study assessed the metabolism of oral mucosal cells cultured on titanium discs (Ti) coated (or not) with epidermal growth factor (EGF) and exposed to tumor necrosis factor alpha (TNF-α).
Fibroblasts or keratinocytes were seeded on Ti coated or not with EGF, and then exposed to 100 ng/mL of TNF-α for 24 h. Groups were established: G1: Ti (control); G2: Ti + TNF-α; G3: Ti + EGF; and G4: Ti + EGF + TNF-α. Both cell lines were evaluated for: viability (AlamarBlue®, n = 8); interleukin 6 and 8 (IL-6, IL-8) gene expression (qPCR, n = 5), and protein synthesis (ELISA, n = 6). For keratinocytes cells, the matrix metalloproteinase type 3 (MMP-3) was evaluated by qPCR (n = 5) and ELISA (n = 6). A 3-D culture of fibroblasts was analyzed by confocal microscopy. The data were subjected to ANOVA analysis, α = 5%.
Increased cell viability was observed in all groups compared with G1. Enhanced gene expression and synthesis of IL-6 and IL-8 by fibroblasts and keratinocytes in G2 and modulation of hIL-6 gene expression in G4 was noted. Modulation of IL-8 synthesis occurred in keratinocytes in G3 and G4. Keratinocytes in G2 showed enhanced gene expression of hMMP-3. A 3-D culture showed more cells in G3. Fibroblasts in G2 exhibited disrupted cytoplasmic membrane. Cells in G4 showed elongated morphology with intact cytoplasm.
EGF coating increases cell viability and modulates the response of oral cells exposed to an inflammatory stimulus.
本研究评估了表皮生长因子（EGF）涂层或未涂层的钛盘（Ti）上培养的口腔黏膜细胞在肿瘤坏死因子α（TNF-α）作用下的代谢情况。
将成纤维细胞或角质形成细胞接种在涂有或未涂有 EGF 的 Ti 上，然后用 100ng/ml TNF-α孵育 24 小时。建立以下实验组：G1：Ti（对照）；G2：Ti+TNF-α；G3：Ti+EGF；G4：Ti+EGF+TNF-α。用 AlamarBlue®评估两种细胞系的活力（n=8）；用 qPCR（n=5）和 ELISA（n=6）评估白细胞介素 6 和 8（IL-6、IL-8）的基因表达和蛋白合成。对于角质形成细胞，通过 qPCR（n=5）和 ELISA（n=6）评估基质金属蛋白酶 3（MMP-3）。用共聚焦显微镜分析成纤维细胞的 3D 培养。对数据进行方差分析，α=5%。
与 G1 相比，所有组的细胞活力均增加。G2 中观察到成纤维细胞和角质形成细胞的 IL-6 和 IL-8 基因表达增强和蛋白合成增强，以及 G4 中 hIL-6 基因表达的调节。G3 和 G4 中的角质形成细胞的 IL-8 合成受到调节。G2 中的角质形成细胞表现出 hMMP-3 的基因表达增强。3D 培养显示 G3 中细胞数量增加。G2 中的成纤维细胞表现出细胞质膜破裂。G4 中的细胞表现出伸长的形态，细胞质完整。
EGF 涂层增加了细胞活力，并调节了暴露于炎症刺激的口腔细胞的反应。
