Human gingival epithelial cells stimulate proliferation, migration, and tube formation of lymphatic endothelial cells in vitro

J Periodontal Res. 2023 Jun;58(3):596-606. doi: 10.1111/jre.13110. Epub 2023 Feb 26.


Objective: The aim of this study was to investigate the response of gingival epithelial cells to microbial and inflammatory signals.

Background: The gingival epithelial barrier provides the first line of defense and supports tissue homeostasis by maintaining the cross-talk between gingival epithelium, oral microbiota, and immune cells. Lymphatic vessels are essential to sustaining this homeostasis. The gingival epithelial cells have been shown to produce prolymphangiogenic factors during physiologic conditions, but their role in response to microbial and inflammatory signals is unknown.

Methods: Immortalized human gingival epithelial cells (HGEC) and human dermal lymphatic microvascular endothelial cells (LEC) were cultured. HGEC were exposed to Porphyromonas gingivalis derived-LPS, human IL-1 beta/IL-1F2 protein, or recombinant human IL-6/IL-6R. Levels of vascular growth factors (VEGF-A, VEGF-C, and VEGF-D) in cell supernatants were determined by ELISA. LEC were grown to confluence, and a scratch was induced in the monolayer. Uncovered area was measured up to 48 h after exposure to conditioned medium (CM) from HGEC. Tube formation assays were performed with LEC cocultured with labelled HGEC or exposed to CM.

Results: VEGF-A, VEGF-C, and low levels of VEGF-D were constitutively expressed by HGEC. The expression of VEGF-C and VEGF-D, but not VEGF-A, was upregulated in response to proinflammatory mediators. VEGF-C was upregulated in response to P. gingivalis LPS, but not to Escherichia coli LPS. A scratch migration assay showed that LEC migration was significantly increased by CM from HGEC. Both the CM and coculture with HGEC induced significant tube formation of LEC.

Conclusions: HGEC can regulate production of lymphangiogenic/angiogenic factors during inflammatory insults and can stimulate proliferation, migration, and tube formation of LEC in vitro in a paracrine manner.

Keywords: conditioned medium; cytokines; keratinocytes; lymphatic endothelial cells; vascular endothelial growth factors.

MeSH terms

  • Cell Proliferation
  • Cells, Cultured
  • Endothelial Cells* / metabolism
  • Epithelial Cells
  • Humans
  • Lipopolysaccharides / pharmacology
  • Vascular Endothelial Growth Factor C* / metabolism
  • Vascular Endothelial Growth Factor C* / pharmacology
  • Vascular Endothelial Growth Factor D / metabolism


  • Vascular Endothelial Growth Factor C
  • Vascular Endothelial Growth Factor D
  • Lipopolysaccharides