Seo Seung Beom, Dananjaya S H S, Nikapitiya Chamilani, Park Bae Keun, Gooneratne Ravi, Kim Tae-Yoon, Lee Jehee, Kim Cheol-Hee, De Zoysa Mahanama
College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungnam National University, Yuseong-gu, Daejeon, 34134, Republic of Korea.
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea.
Fish Shellfish Immunol. 2017 Sep;68:536-545. doi: 10.1016/j.fsi.2017.07.057. Epub 2017 Jul 27.
Silver nanoparticles (AgNPs) were successfully synthesized by a chemical reduction method, physico-chemically characterized and their effect on wound-healing activity in zebrafish was investigated. The prepared AgNPs were circular-shaped, water soluble with average diameter and zeta potential of 72.66 nm and -0.45 mv, respectively. Following the creation of a laser skin wound on zebrafish, the effect of AgNPs on wound-healing activity was tested by two methods, direct skin application (2 μg/wound) and immersion in a solution of AgNPs and water (50 μg/L). The zebrafish were followed for 20 days post-wounding (dpw) by visual observation of wound size, calculating wound healing percentage (WHP), and histological examination. Visually, both direct skin application and immersion AgNPs treatments displayed clear and faster wound closure at 5, 10 and 20 dpw compared to the controls, which was confirmed by 5 dpw histology data. At 5 dpw, WHP was highest in the AgNPs immersion group (36.6%) > AgNPs direct application group (23.7%) > controls (18.2%), showing that WHP was most effective in fish immersed in AgNPs solution. In general, exposure to AgNPs induced gene expression of selected wound-healing-related genes, namely, transforming growth factor (TGF-β), matrix metalloproteinase (MMP) -9 and -13, pro-inflammatory cytokines (IL-1β and TNF-α) and antioxidant enzymes (superoxide dismutase and catalase), which observed differentiation at 12 and 24 h against the control; but the results were not consistently significant, and many either reached basal levels or were down regulated at 5 dpw in the wounded muscle. These results suggest that AgNPs are effective in acceleration of wound healing and altered the expression of some wound-healing-related genes. However, the detailed mechanism of enhanced wound healing remains to be investigated in fish.
通过化学还原法成功合成了银纳米颗粒(AgNPs),对其进行了物理化学表征,并研究了它们对斑马鱼伤口愈合活性的影响。制备的AgNPs呈圆形,水溶性良好,平均直径和zeta电位分别为72.66 nm和-0.45 mv。在斑马鱼身上造成激光皮肤伤口后,通过两种方法测试了AgNPs对伤口愈合活性的影响,即直接皮肤应用(2 μg/伤口)和浸入AgNPs与水的溶液(50 μg/L)。通过肉眼观察伤口大小、计算伤口愈合百分比(WHP)和组织学检查,对斑马鱼伤口后20天进行跟踪观察。肉眼观察发现,与对照组相比,在伤口后5、10和20天,直接皮肤应用和浸入AgNPs处理均显示出明显更快的伤口闭合,这在伤口后5天的组织学数据中得到了证实。在伤口后5天,AgNPs浸入组的WHP最高(36.6%)>AgNPs直接应用组(23.7%)>对照组(18.2%),表明浸入AgNPs溶液的鱼中WHP最有效。总体而言,暴露于AgNPs会诱导选定的伤口愈合相关基因的表达,即转化生长因子(TGF-β)、基质金属蛋白酶(MMP)-9和-13、促炎细胞因子(IL-1β和TNF-α)和抗氧化酶(超氧化物歧化酶和过氧化氢酶),在12和24小时观察到与对照组的差异;但结果并不一致显著,许多基因在伤口后5天在受伤肌肉中要么达到基础水平,要么下调。这些结果表明,AgNPs在加速伤口愈合方面是有效的,并改变了一些伤口愈合相关基因的表达。然而,在鱼类中增强伤口愈合的详细机制仍有待研究。