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Dose-dependent green tea effect on decrease of inflammation in human oral gingival epithelial keratinocytes: in vitro study.

Clinical oral investigations (2019-10-28)
Ana Hagiu, Thomas Attin, Patrick R Schmidlin, Liza L Ramenzoni
ABSTRACT

This in vitro study aimed to analyze the anti-inflammatory and wound healing potential of green tea extract (GTE) in human gingival epithelial keratinocytes (HGEK) treated with lipopolysaccharides (LPS). A cell viability assay was conducted using MTT to determine nontoxic levels of GTE on immortalized HGEK. Cells were concomitantly treated with LPS (1 μg/ml) and GTE (1 mg/ml, 2.5 mg/ml, 5 mg/ml, and 10 mg/ml) to assess inflammation. Gene expression levels of inflammatory markers IL-β1, IL-6, and TNFα were measured by RT-PCR and their protein production was assessed by ELISA. The scratch wound healing assay was used to investigate the effects of different concentrations of GTE on cell migration. We also explored the effect of GTE on the induction of the Nrf2/HO-1 pathway in the cells with or without LPS. GTE at concentrations of 2.5 mg/ml, 5 mg/ml, and 10 mg/ml significantly enhanced cell viability (p < 0.05). And IL-β1, IL-6, and TNFα gene expression presented up to 10-fold decrease compared with LPS-treated cells, which was also similarly found on the protein levels. At the same concentrations, cell migration increased. The mechanism results showed that GTE produced the anti-inflammatory response by activating the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway and increasing the level of anti-oxidant protein heme oxygenase-1 (HO-1). GTE may be potentially used as oral rinse anti-inflammatory drug for treatment and prevention of oral inflammatory diseases, which is shown here by the ability to reduce the inflammation and increase in cell migration in a dose-dependent manner.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Glutamine solution, 200 mM, solution, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
Human IL-6 ELISA Kit, for serum, plasma, cell culture supernatant and urine
Sigma-Aldrich
Human Tumor Necrosis Factor α ELISA Kit, for serum, plasma, cell culture supernatants and urine
Sigma-Aldrich
Human IL-1 β ELISA Kit, for serum, plasma, cell culture supernatant and urine
Sigma-Aldrich
Bis(2-oxo-3-oxazolidinyl)phosphinic chloride, ≥97.0% (AT)

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