Center for Cancer Biomarkers CCBIO and Gade Laboratory for Pathology, Department of Clinical Medicine, Faculty of Medicine, University of Bergen, Bergen, Norway.
Department of Oral Rehabilitation and Department of Biochemistry, 'Carol Davila' University of Medicine and Pharmacy, Bucharest, Romania.
Eur J Oral Sci. 2022 Jun;130(3):e12867. doi: 10.1111/eos.12867. Epub 2022 Apr 22.
Oral epithelial differentiation is known to be directed by underlying fibroblasts, but the responsible factor(s) have not been identified. We aimed here to identify fibroblast-derived factors responsible for oral epithelial differentiation. Primary normal human oral keratinocytes and fibroblasts were isolated from healthy volunteers after informed consent (n = 5) and 3D-organotypic (3D-OT) cultures were constructed. Various growth factors were added at a range of 0.1-100 ng/ml. 3D-OTs were harvested after ten days and assessed histologically, by immunohistochemistry and the TUNEL method. Epithelium developed in 3D-OT without fibroblasts showed an undifferentiated phenotype. Addition of granulocyte macrophage-colony stimulating factor (GM-CSF) induced expression of cytokeratin 13 in suprabasal cell layers. Admixture of GM-CSF and keratinocyte growth factor (KGF) induced, in addition, polarization of epidermal growth factor (EGF) receptor and β1-integrin to basal cell layer and collagen IV deposition. Terminal differentiation with polarization of TUNEL-positive cells to superficial layers occurred only in the presence of fibroblasts in collagen gels either in direct contact or at distance from normal oral keratinocytes. Taken together, these results show that major aspects of oral epithelial differentiation are regulated by the synergic combination of GM-CSF and KGF. However, the terminal stage seems to be controlled by other yet unidentified fibroblast-derived diffusible factor(s).
已知口腔上皮的分化受下层成纤维细胞的调控，但尚未确定其相关的调节因子。我们的目的是确定与口腔上皮分化相关的成纤维细胞衍生因子。在获得知情同意后（n=5），从健康志愿者中分离出原代正常的人口腔角质形成细胞和成纤维细胞，并构建 3D 器官型（3D-OT）培养物。在 0.1-100ng/ml 的范围内添加各种生长因子。在十天后收获 3D-OT 并进行组织学评估，通过免疫组织化学和 TUNEL 方法进行评估。在没有成纤维细胞的情况下，3D-OT 中发育的上皮表现出未分化的表型。添加粒细胞巨噬细胞集落刺激因子（GM-CSF）可诱导细胞角蛋白 13 在基底上层细胞中表达。GM-CSF 和角质形成细胞生长因子（KGF）的混合物还诱导表皮生长因子（EGF）受体和β1-整合素向基底细胞层极化，并沉积胶原蛋白 IV。只有在胶原凝胶中成纤维细胞的直接或远距离接触下，或在正常口腔角质形成细胞存在下，才能发生具有极化的 TUNEL 阳性细胞的终末分化。总之，这些结果表明，口腔上皮分化的主要方面受 GM-CSF 和 KGF 的协同组合调控。然而，终末阶段似乎受其他尚未确定的成纤维细胞衍生的可扩散因子控制。
