PDF(Pubmed)
Abstract：
Scientific progress and ethical considerations are increasingly shifting the toxicological focus from in vivo animal models to in vitro studies utilizing physiologically relevant cell cultures. Consequently, we evaluated and validated a three-dimensional (3D) model of the human lung using Calu-3 cells cultured at an air-liquid interface (ALI) for 28 days. Assessment of seven essential genes of differentiation and transepithelial electrical resistance (TEER) measurements, in conjunction with mucin (MUC5AC) staining, validated the model. We observed a time-dependent increase in TEER, genetic markers of mucus-producing cells (muc5ac, muc5b), basal cells (trp63), ciliated cells (foxj1), and tight junctions (tjp1). A decrease in basal cell marker krt5 levels was observed. Subsequently, we utilized this validated ALI-cultured Calu-3 model to investigate the adversity of the aerosols generated from three flavored electronic cigarette (EC) e-liquids: cinnamon, vanilla tobacco, and hazelnut. These aerosols were compared against traditional cigarette smoke (3R4F) to assess their relative toxicity. The aerosols generated from PG/VG vehicle control, hazelnut and cinnamon e-liquids, but not vanilla tobacco, significantly decreased TEER and increased lactate dehydrogenase (LDH) release compared to the incubator and air-only controls. Compared to 3R4F, there were no significant differences in TEER or LDH with the tested flavored EC aerosols other than vanilla tobacco. This starkly contrasted our expectations, given the common perception of e-liquids as a safer alternative to cigarettes. Our study suggests that these results depend on flavor type. Therefore, we strongly advocate for further research, increased user awareness regarding flavors in ECs, and rigorous regulatory scrutiny to protect public health.
摘要：
科学进步和伦理考虑越来越多地将毒理学重点从体内动物模型转移到利用生理相关细胞培养物的体外研究。因此,我们使用在气-液界面(ALI)培养28天的Calu-3细胞评估并验证了人肺的三维(3D)模型.评估分化和跨上皮电阻（TEER）测量的七个必需基因，结合粘蛋白（MUC5AC）染色，对模型进行了验证。我们观察到TEER的时间依赖性增加，粘液产生细胞的遗传标记(muc5ac，muc5b),基底细胞（trp63），纤毛细胞（foxj1），和紧密连接（TJP1）。观察到基底细胞标志物krt5水平降低。随后，我们利用这个经过验证的ALI培养的Calu-3模型来研究三种调味电子烟（EC）电子液体产生的气溶胶的逆境：肉桂，香草烟草,还有榛子.将这些气溶胶与传统香烟烟雾（3R4F）进行比较，以评估其相对毒性。PG/VG车辆控制产生的气溶胶，榛子和肉桂电子液体，但不是香草烟草，与培养箱和仅空气对照相比，TEER显着降低，乳酸脱氢酶（LDH）释放增加。与3R4F相比，除香草烟草外，测试的调味EC气溶胶在TEER或LDH方面没有显着差异。这与我们的期望形成鲜明对比，考虑到电子液体作为香烟更安全的替代品的普遍看法。我们的研究表明，这些结果取决于风味类型。因此,我们强烈主张进一步研究，提高用户对EC口味的认识，和严格的监管审查，以保护公众健康。
