Pinto Bronson I, Tabor Aaron J, Stearns Diane M, Diller Robert B, Kellar Robert S
Department of Biological Sciences, Northern Arizona University, Flagstaff, Arizona.
Department of Chemistry and Biochemistry, Northern Arizona University, Flagstaff, Arizona.
Appl In Vitro Toxicol. 2016 Sep 1;2(3):151-156. doi: 10.1089/aivt.2016.0001.
Cellular migration assays are useful tools to investigate physiologic events on the bench top. Furthermore, this migration assay can be utilized to investigate wound healing therapeutics (those that encourage or accelerate wound closure) as well as deleterious agents (ones that mitigate or slow wound closure). The current study used an scratch assay to measure the effects of platelet-rich plasma (PRP) and depleted uranium (DU) in the form of uranyl acetate on cellular migration of human neonatal dermal fibroblasts in an simulation of wound healing. Data analyses included percent wound closure measured as the distance between cell margins, and rates of wound closure versus untreated controls. The highest doses of PRP (0.063, 0.125%) resulted in 50-65% wound closure after 4-8 hours relative to 38-44% in controls and the low-dose treatment group (0.031%). The high-dose treatments of PRP (0.125, 0.063%) reached 100% wound closure at 12 hours postwound versus 16 hours for controls and the low-dose treatment group (0.031%). Conversely, the higher doses of DU treatments (50 and 100 μM) resulted in <80% closure versus 100% closure in controls after 16 hours, with full closure observed at 20 hours. The highest dose of DU (1,000 μM) resulted in <20% closure versus 100% closure in controls after 16 hours. The use of the described scratch assay serves as a translatable bench-top model that has the potential to predict outcomes, and in many early studies can help to demonstrate proof-of-concept before moving into complex biological systems.
细胞迁移实验是在实验台上研究生理事件的有用工具。此外,这种迁移实验可用于研究伤口愈合治疗方法(那些促进或加速伤口闭合的方法)以及有害因子(那些减轻或延缓伤口闭合的因子)。当前的研究使用划痕实验来测量富血小板血浆(PRP)和醋酸双氧铀形式的贫铀(DU)对人新生儿真皮成纤维细胞在伤口愈合模拟中的细胞迁移的影响。数据分析包括以细胞边缘之间的距离衡量的伤口闭合百分比,以及与未处理对照相比的伤口闭合速率。最高剂量的PRP(0.063%、0.125%)在4至8小时后导致50 - 65%的伤口闭合,而对照组和低剂量治疗组(0.031%)为38 - 44%。高剂量的PRP治疗(0.125%、0.063%)在伤口后12小时达到100%伤口闭合,而对照组和低剂量治疗组(0.031%)为16小时。相反,较高剂量的DU治疗(50和100 μM)在16小时后导致<80%的闭合,而对照组为100%闭合,在20小时观察到完全闭合。最高剂量的DU(1000 μM)在16小时后导致<20%的闭合,而对照组为100%闭合。所描述的划痕实验的使用作为一种可转化的实验台模型,有预测结果的潜力,并且在许多早期研究中可以在进入复杂生物系统之前帮助证明概念。