College of Pharmacy, Rangsit University, Pathum Thani 12000, Thailand.
Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.
Mar Drugs. 2019 Jul 20;17(7):424. doi: 10.3390/md17070424.
Stem cell activities in human tissues are critical for tissue integrity and function. Maintaining keratinocyte stem cells (KSCs) stemness helps sustain healthy skin by supporting keratinocyte renewal, involving the formation of epidermal barriers. In this study, abalone collagen (AC) extracts with molecular weights of 3 kDa (AC 1) and 300 kDa (AC 2) were compared to the epidermal growth factor (EGF) for their effects on cell proliferation, cell migration (wound healing), spheroid formation, and the expression level of stem cell markers on human keratinocytes (HaCaT cells). Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cell proliferation was quantified by ATP and DNA content analysis and Sulforhodamine B (SRB) assays. Cell migration assay was determined using the scratch wound healing test. Spheroid formation was evaluated and the expression level of stem cell markers was investigated by western blot analysis. The results showed that AC 1 at the concentration of 100 µg/mL could stimulate HaCaT cell proliferation, migration, spheroid formation, and the expression level of stem cell markers (keratin 19, β-catenin, ALDH1A1) compared to the control. In conclusion, a smaller molecular weight of abalone collagen extract exhibits a better effect on keratinocytes proliferation, migration, and stemness, which could be a potential active ingredient in cosmeceutical products.
人体组织中的干细胞活动对于组织完整性和功能至关重要。维持角质形成细胞干细胞（KSCs）的干性有助于通过支持角质形成细胞更新来维持健康的皮肤，其中包括表皮屏障的形成。在这项研究中，将分子量为 3 kDa（AC1）和 300 kDa（AC2）的鲍鱼胶原（AC）提取物与表皮生长因子（EGF）进行比较，以研究它们对人角质形成细胞（HaCaT 细胞）增殖、细胞迁移（伤口愈合）、球体形成和干细胞标志物表达水平的影响。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法测量细胞活力，并通过 ATP 和 DNA 含量分析和磺酰罗丹明 B（SRB）测定法定量细胞增殖。使用划痕愈合试验测定细胞迁移试验。通过 Western blot 分析评估球体形成并研究干细胞标志物的表达水平。结果表明，与对照组相比，浓度为 100 µg/mL 的 AC1 可刺激 HaCaT 细胞增殖、迁移、球体形成和干细胞标志物（角蛋白 19、β-连环蛋白、ALDH1A1）的表达。总之，鲍鱼胶原提取物的较小分子量对角质形成细胞的增殖、迁移和干性表现出更好的效果，这可能是化妆品产品中的一种潜在活性成分。
