Effects of prolonged stimulation with heated tobacco products (Ploom TECH+ ) on gingival epithelial cells

Osamu Uehara; Norihiro Nakamoto; Daichi Hiraki; Durga Paudel; Nodoka Sugiyama; Tetsuro Morikawa; Koki Yoshida; Yutaka Kawano; Tsuyoshi Shimo; Yasushi Furuichi; Hiroko Miura; Yoshihiro Abiko

doi:10.1111/jre.13123

Effects of prolonged stimulation with heated tobacco products (Ploom TECH⁺ ) on gingival epithelial cells

J Periodontal Res. 2023 Jun;58(3):553-563. doi: 10.1111/jre.13123. Epub 2023 Mar 27.

Authors

Osamu Uehara^{1

2}, Norihiro Nakamoto³, Daichi Hiraki⁴, Durga Paudel², Nodoka Sugiyama³, Tetsuro Morikawa⁵, Koki Yoshida^{2

5}, Yutaka Kawano^{2

6}, Tsuyoshi Shimo⁴, Yasushi Furuichi^{2

3}, Hiroko Miura¹, Yoshihiro Abiko^{2

5}

Affiliations

¹ Division of Disease Control and Molecular Epidemiology, Department of Oral Growth and Development, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, 061-0293, Japan.
² Advanced Research Promotion Center, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, 061-0293, Japan.
³ Division of Periodontology and Endodontology, Department of Oral Rehabilitation, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, 061-0293, Japan.
⁴ Division of Reconstructive Surgery for Oral and Maxillofacial Region, Department of Human Biology and Pathophysiology, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, 061-0293, Japan.
⁵ Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, 061-0293, Japan.
⁶ Department of Gastroenterology and Oncology, Tokushima University Graduate School of Biomedical Sciences Tokushima, 3-18-15, Kuramoto-cho, Tokushima, 770-8503, Japan.

PMID: 36974375
DOI: 10.1111/jre.13123

Abstract

Objective and background: Heated tobacco products have recently become commercially available. These products, as well as combustible cigarettes, produce aerosols; the risk of various diseases associated with heated tobacco products may be the same or higher than that with combustible cigarettes. In this study, we examined the effect of Ploom TECH⁺ extract on gingival epithelial cells.

Methods: Tobacco leaves from Ploom TECH⁺ tobacco capsules and water were mixed and heated; the supernatant subsequently collected was the heated tobacco product (HTP; control: HTP not added). Normal human gingival epithelial progenitors were cultured alternately with or without HTP for a total of 1 month. Subsequently, RNA, DNA, and proteins were isolated from these samples and comprehensively analyzed using RNA sequencing (RNA-seq), reduced representation bisulfite sequencing (RRBS), and western blotting, respectively.

Results: RNA-seq revealed that 284 genes showed a twofold increase and 145 genes showed a twofold decrease in gene expression. A heat map showed genetic differences between the control and HTP groups. A principal component analysis plot showed a clear genetic distribution between the control and HTP. Gene Ontology (GO) analysis showed that genes related to seven GO terms, including cornification and keratinization, were induced by long-term HTP stimulation. By contrast, GO pathways with a significant decrease in component expression were not detected. RRBS revealed that CpG island methylation increased more than twofold in 158 genes and decreased to less than twofold in 171 genes. Methylation of these CpG islands was not correlated with changes in gene expression levels. HTP treatment increased S100A7 expression.

Conclusion: Long-term HTP stimulation affected epithelial differentiation and keratinization of gingival epithelial cells. Thus, habitual use of Ploom TECH⁺ may be a risk factor for tobacco-related oral mucosal diseases.

Keywords: DNA methylation; RNA sequencing; gingival epithelial cell; heated tobacco product; reduced representation bisulfite sequencing.

MeSH terms

Epithelial Cells
Hot Temperature
Humans
Risk Factors
Tobacco Products*