Department of Ophthalmology, Osaka Medical College, 2-7 Daigaku-machi, 569-8686, Takatsuki, Osaka, Japan.
Graefes Arch Clin Exp Ophthalmol. 2012 Apr;250(4):557-63. doi: 10.1007/s00417-011-1849-8. Epub 2011 Nov 3.
We have developed inorganically-coated all-trans retinoic acid (atRA) nanoparticles, nano-sized egg-like particles of atRA (NANOEGG®-atRA). The purpose of this study was to determine the effects of NANOEGG®-atRA on corneal wound healing in vivo and in vitro.
A rabbit corneal epithelial wound healing model was exposed to different concentrations of NANOEGG®-atRA. Wound healing was serially quantified as the ratio of fluorescein-stained area at the selected times to that at baseline. After wound closure, the barrier function of the cornea was determined using low concentrations of tropicamide. At the completion of the experiments, the corneal epithelium was histologically examined. For the in vitro studies, linear scratch wounds were made on cultured SV40-immortalized human corneal epithelial cells (HCE-T). Then, the cells were exposed to different concentrations of NANOEGG®-atRA, and wound healing was determined by the degree of closure of the scratch wound. In addition, the effects of NANOEGG®-atRA on the proliferation of HCE-T cells were determined by WST-8 assays.
Exposure to NANOEGG®-atRA decreased the injured area 24 hrs after the ablation. The maximum effect of NANOEGG®-atRA was observed at a concentration of 33 mM. Histologically, no abnormal or differentiated corneal epithelial cells were observed in the histological sections treated with NANOEGG®-atRA. The tropicamide-induced pupillary dilation was significantly slowed in the eyes treated with NANOEGG®-atRA. NANOEGG®-atRA at concentrations of 3.3 and 33 nM induced earlier wound closure in vitro, but did not induce proliferation of HCE-T cells.
NANOEGG®-atRA promotes wound healing and should be considered for the treatment of wounds of the corneal epithelium.
我们开发了无机包裹的全反式视黄酸（atRA）纳米颗粒，即纳米大小的 atRA 卵状颗粒（NANOEGG®-atRA）。本研究旨在确定 NANOEGG®-atRA 对体内和体外角膜伤口愈合的影响。
采用不同浓度的 NANOEGG®-atRA 暴露于兔角膜上皮伤口愈合模型。在选定时间点通过荧光素染色面积与基线面积的比值连续定量评估伤口愈合情况。在伤口闭合后，用低浓度的托吡卡胺测定角膜的屏障功能。实验完成后，对角膜上皮进行组织学检查。对于体外研究，在培养的 SV40 永生化人角膜上皮细胞（HCE-T）上制作线性划痕伤口。然后，将细胞暴露于不同浓度的 NANOEGG®-atRA，通过划痕伤口的闭合程度来确定伤口愈合情况。此外，通过 WST-8 测定法确定 NANOEGG®-atRA 对 HCE-T 细胞增殖的影响。
暴露于 NANOEGG®-atRA 可减少消融后 24 小时的受伤面积。在 33mM 浓度下观察到 NANOEGG®-atRA 的最大效果。组织学上，用 NANOEGG®-atRA 处理的组织切片中未观察到异常或分化的角膜上皮细胞。用 NANOEGG®-atRA 处理的眼睛中，托吡卡胺诱导的瞳孔扩张明显减慢。浓度为 3.3 和 33nM 的 NANOEGG®-atRA 可在体外更早地诱导伤口闭合，但不会诱导 HCE-T 细胞增殖。
NANOEGG®-atRA 促进伤口愈合，可考虑用于治疗角膜上皮伤口。
