Department of Dermatology, Hallym Institute for Translational Medicine, Anyang, Korea.
Department of Dermatology, Hallym University Sacred Heart Hospital, Anyang, Korea.
Skin Res Technol. 2023 Jan;29(1):e13275. doi: 10.1111/srt.13275.
Sensitive skin is a subjective cutaneous hyper-reactivity that occurs in response to various innocuous stimuli. Keratinocytes have recently been shown to participate in sensory transduction by releasing many neuroactive molecules that bind to intra-epidermal free nerve endings and modulate nociception. In the literature, the characterization of these interactions has been based on the co-culture of keratinocyte and mammalian-origin neuronal cell lines. In this study, we established an in vitro model based on a co-culture of primary human keratinocytes and differentiated SH-SY5Y cells, a human neuronal cell line.
Human epidermal keratinocytes and SH-SY5Y cells were monocultured and co-cultured. Changes in calcium influx, substance P, inflammatory cytokines, and neuropeptides between the monoculture and co-culture groups treated with capsaicin only and capsaicin with transient receptor potential channel vanilloid subfamily member 1 (TRPV1) antagonist, trans-4-tert-butylcyclohexanol (TTBC), together. In addition, the difference in stinging sensation was evaluated by applying it to the volunteers.
When SH-SY5Y cells were co-cultured with keratinocytes, they had no significant effect on axonal development. Substance P was also released after capsaicin treatment and reduced by TTBC under co-culture conditions. Moreover, the expression of inflammatory cytokines and neuropeptides was significantly increased in co-cultured keratinocytes compared to that under monoculture conditions. In addition, the stinging sensation was significantly induced after the application of capsaicin in vivo and was relieved after the application of the TRPV1 antagonist.
We demonstrated that the novel co-culture model is functionally valid through capsaicin and TRPV1 antagonist. We also confirmed that TTBC could be used for the treatment of sensitive skin through a co-culture model and in vivo tests. This co-culture model of keratinocytes and SH-SY5Y cells may be useful in vitro alternatives for studying the close communication between keratinocytes and neuronal cells and for screening therapeutic drugs for sensitive skin.
敏感皮肤是一种对各种无害刺激产生的主观皮肤超敏反应。最近发现角质形成细胞通过释放许多与表皮内游离神经末梢结合并调节伤害感受的神经活性分子参与感觉转导。在文献中，这些相互作用的特征是基于角质形成细胞和哺乳动物来源的神经元细胞系的共培养。在这项研究中，我们建立了一种基于原代人角质形成细胞和分化的 SH-SY5Y 细胞（一种人神经元细胞系）共培养的体外模型。
单独培养和共培养人表皮角质形成细胞和 SH-SY5Y 细胞。用辣椒素单独和辣椒素与瞬时受体电位通道香草素亚家族成员 1（TRPV1）拮抗剂 Trans-4-叔丁基环己醇（TTBC）处理后，比较单独培养和共培养组之间钙内流、P 物质、炎症细胞因子和神经肽的变化。此外，通过将其应用于志愿者来评估刺痛感的差异。
当 SH-SY5Y 细胞与角质形成细胞共培养时，它们对轴突发育没有显著影响。辣椒素处理后也释放 P 物质，共培养条件下用 TTBC 减少。此外，与单独培养条件相比，共培养角质形成细胞中炎症细胞因子和神经肽的表达显著增加。此外，体内应用辣椒素后明显引起刺痛感，应用 TRPV1 拮抗剂后缓解。
我们通过辣椒素和 TRPV1 拮抗剂证明了新型共培养模型在功能上是有效的。我们还通过共培养模型和体内试验证实 TTBC 可用于治疗敏感皮肤。这种角质形成细胞和 SH-SY5Y 细胞的共培养模型可能是研究角质形成细胞和神经元细胞之间密切通讯以及筛选敏感皮肤治疗药物的有用体外替代方法。
