Aqueous extract of tobacco induces mitochondrial potential dependent cell death and epithelial-mesenchymal transition in gingival epithelial cells

Abstract

Smokeless tobacco habits are detrimental to oral health. A correlation between tobacco use and local epithelial tissue damage exists. Yet, the underlying cellular mechanism is not precisely characterized. This study assessed the dose-dependent action of Smokeless tobacco extract on gingival epithelial cells. Gingival tissue was taken from 5 healthy donors. Gingival epithelial cells were isolated by an enzymatic method and cultured up to passage 2. The cultured cells were treated with smokeless tobacco extract at 10%, 25%, 50%, and 75% volume concentration. After 48 h of incubation, MTT assay, Annexin V/PI assay, and DiIC1(5) assay were used to evaluate viability, apoptosis, and mitochondrial potential of the cells. RT-qPCR was used to determine the expression of BAX, BCL2, ECAD, NCAD, and TWIST. The Smokeless tobacco extract reduced cell viability by disrupting the mitochondrial potential and inducing apoptosis. Further, the Smokeless tobacco extract induced a dose-dependent epithelial-mesenchymal-transition in gingival epithelial cells. Apoptotic cellular death caused by tobacco extract on the gingival epithelial system was dependant on the mitochondrial potential of the cell. The results demonstrate that smokeless tobacco causes detrimental metabolic alterations of the periodontium. Featured application This study elucidates the mechanism by which Smokeless tobacco products cause cellular damage to the gingival epithelium. The use of Smokeless tobacco products can lead to major cellular and surface changes in the gingiva and its appearance. The consequences of these changes are not limited to oral cancer but also increases a person’s risk for dental and periodontal disease.

Oxidative Damage Induced by Phototoxic Pheophorbide a 17-Diethylene Glycol Ester Encapsulated in PLGA Nanoparticles

“…Changes in mitochondrial membrane potential (MMP) may evidence the organelle failure and lead to apoptosis [ 53 ]. We applied a permeable lipophilic fluorescent cationic dye DiIC1(5) as a marker of mitochondrial membrane depolarization (∆Ψm) [ 54 ]. Viable mitochondria with high membrane potential actively accumulated DiIC1(5), demonstrating a strong fluorescence, while in cells treated with XL-8 and XL-8-NPs mitochondria displayed dose-dependent fluorescence intensity decrease ( Figure 6 F).…”

The oral bioaccessibility and gingival cytotoxicity of metal(loid)s in wild vegetables from mining areas: Implication for human oral health

“…In this study, the extracts with high Ni contents (exclude PAL ) only slightly exhibited DNA damage on HGEC ( Figures 2B , 5F , L , O ). These inconsistent results could be attributed to the differences in cellular physiology as well as the constituents of the exposure and exposed systems ( 46 , 47 ). As such, these comparisons further demonstrated that the cell apoptosis and DNA damages may not be correlated with the levels of heavy metal(loid)s in edible vegetables, but are also impacted by the other factors in each specific exposure scenario.…”

Mitochondrial Dysfunction in Periodontitis and Associated Systemic Diseases: Implications for Pathomechanisms and Therapeutic Strategies

“…Volatile sulfur compounds and tobacco extract can induce mitochondrial MMP loss and apoptosis in GECs, leading to local epithelial damage [57,58]. The effect of cyclosporine A, an immunosuppressant that induces gingival hyperplasia, on GEC apoptosis is still debated.…”