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However, little work to date has focused on the emerging market of aerosolized nicotine delivery known as electronic nicotine delivery systems (ENDS) or electronic cigarettes, and their potential for new effects on human health. Challenges studying these devices include heterogeneity in the formulation of the common components of most available ENDS, including nicotine and a carrier (commonly composed of propylene glycol and vegetable glycerin, or PG\/VG). In the present study, we report on experiments interrogating the effects of major identified components in e\u2010cigarettes. Specifically, the potential concomitant effects of nicotine and common carrier ingredients in commercial \u201cvape\u201d products are explored in\u00a0vitro to inform the potential health effects on the craniofacial skeleton through novel vectors as compared to traditional tobacco products. MC3T3\u2010E1 murine pre\u2010osteoblast cells were cultured in\u00a0vitro with clinically relevant liquid concentrations of nicotine, propylene glycol (PG), vegetable glycerin (VG), Nicotine+PG\/VG, and the vape liquid of a commercial product (\n                    <jats:italic>Juul<\/jats:italic>\n                    ). Cells were treated acutely for 24\u2009h and RNA\u2010Seq was utilized to determine segregating alteration in mRNA signaling. Influential gene targets identified with sparse partial least squares discriminant analysis (sPLS\u2010DA) implemented in mixOmics were assessed using the PANTHER Classification system for molecular functions, biological processes, cellular components, and pathways of effect. Additional endpoint functional analyses were used to confirm cell cycle changes. The initial excitatory concentration (EC50) studied defined a target concentration of carrier PG\/VG liquid that altered the cell cycle of the calvarial cells. Initial sPLS\u2010DA analysis demonstrated the segregation of nicotine and non\u2010nicotine exposures utilized in our in\u00a0vitro modeling. Pathway analysis suggests a strong influence of nicotine exposures on cellular processes including metabolic processes and response to stimuli including autophagic flux. Further interrogation of the individual treatment conditions demonstrated segregation by treatment modality (Control, Nicotine, Carrier (PG+VG), Nicotine+PG\/VG) along three dimensions best characterized by: latent variable 1 (PLSDA\u20101) showing strong segregation based on nicotine influence on cellular processes associated with cellular adhesion to collagen, osteoblast differentiation, and calcium binding and metabolism; latent variable 2 (PLSDA\u20102) showing strong segregation of influence based on PG+VG and Control influence on cell migration, survival, and cycle regulation; and latent variable 3 (PLSDA\u20103) showing strong segregation based on Nicotine and Control exposure influence on cell activity and growth and developmental processes. Further, gene co\u2010expression network analysis implicates targets of the major pathway genes associated with bone growth and development, particularly craniofacial (FGF, Notch, TGF\u03b2, WNT) and analysis of active subnetwork pathways found these additionally overrepresented in the Juul exposure relative to Nicotine+PG\/VG. Finally, experimentation confirmed alterations in cell count, and increased evidence of cell stress (markers of autophagy), but no alteration in apoptosis. These data suggest concomitant treatment with Nicotine+PG\/VG drives alterations in pre\u2010osteoblast cell cycle signaling, specifically transcriptomic targets related to cell cycle and potentially cell stress. Although we suspected cell stress and well as cytotoxic effects of Nicotine+PG\/VG, no great influence on apoptotic factors was observed. Further RNA\u2010Seq analysis allowed for the direct interrogation of molecular targets of major pathways involved in bone and craniofacial development, each demonstrating segregation (altered signaling) due to e\u2010cigarette\u2010type exposure. These data have implications directed toward ENDS formulation as synergistic effects of Nicotine+PG\/VG are evidenced here. Thus, future research will continue to interrogate how varied formulation of Nicotine+PG\/VG affects overall cell functions in multiple vital systems.\n                  <\/jats:p>","DOI":"10.1096\/fj.202302014rrr","type":"journal-article","created":{"date-parts":[[2024,8,30]],"date-time":"2024-08-30T11:58:55Z","timestamp":1725019135000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Acute exposure to electronic cigarette components alters\n                    <scp>mRNA<\/scp>\n                    expression of pre\u2010osteoblasts"],"prefix":"10.1096","volume":"38","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1374-8347","authenticated-orcid":false,"given":"Shareef M.","family":"Dabdoub","sequence":"first","affiliation":[{"name":"Division of Biostatistics and Computational Biology College of Dentistry, University of Iowa  Iowa City Iowa USA"},{"name":"Department of Periodontics College of Dentistry, 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