Biological Impact of the Ratio of E-Cigarette Liquid Base Constituents, Propylene Glycol and Vegetable Glycerin, on Primary Human Melanocytes

Abstract

Oral cavity is the first site to encounter e-cigarette (EC) or tobacco smoke. Increased gingival pigmentation can lead to aesthetic concerns and hinder successful outcomes of gingival depigmentation procedures as well as lead to color alterations in patients with dental restorations. While the effects of tobacco smoke and nicotine in increasing pigmentation in the gingiva of the smoker have been well-documented, the effects of EC on pigmentation have not been explored. Due to large variations in e-liquids from different sources, this study focused on the effects of EC liquid base constituents, propylene glycol (PG) and vegetable glycerin (VG), which are a universal constituent of all e-liquids. Effects of PG and VG solutions mixed at different ratios (0/100, 20/80, 55/45, 80/20, and 100/0 % v/v) were examined using primary human melanocytes obtained from neonatal foreskin; this cell model is representative of the physiological model of gingival melanocytes and has been used in our previous study. Results showed significant concentration-dependent cytotoxicity for all groups, although mixtures with higher PG content showed higher cytotoxicity to cells as compared to those with VG. Melanogenesis was robustly activated by PG-containing mixtures with the greatest effect obtained for 80/20 PG/VG mixture as compared to other ratios, while VG by itself did not activate melanogenesis. The activation of melanin synthesis within cells was not correlated to intracellular tyrosinase activity as that was suppressed by PG at higher ratios. Morphological changes of a multidendritic phenotype were observed in cells exposed to all PG/VG mixtures, with markedly greater effects for groups with higher PG content. Taken together, the results of this pilot study demonstrate for the first time that EC base constituents possess the capacity to significantly activate melanogenesis in human melanocytes at nontoxic concentrations, with the dominant effect obtained at a PG/VG ratio of 80/20, indicative of a nonlinear response with increasing concentrations of PG. Moreover, further studies to address the impact of PG/VG with the addition of nicotine and the effects of different EC flavors are underway. Future studies to elucidate mechanisms of increased pigmentation as well as further investigate effects in melanocytes with the presence of other oral cell types and other components of the oral microenvironment such as saliva and bacterial flora are warranted. This research emphasizes the need to reconsider the regulation of EC base constituents PG and VG as different ratios of these compounds can cause differential effects.