Miyata K, Murao M, Sawa M, Tanishima T
Department of Ophthalmology, University of Tokyo School of Medicine, Japan.
Jpn J Ophthalmol. 1990;34(3):257-66.
A new in vitro model to evaluate the corneal endothelial wound-healing process was developed. Confluent monolayer cell sheets of either rabbit or bovine corneal endothelial cells were cultured on cover slips, the surface of which had been precoated with hydroxyethyl methacrylate (HEMA) except for the 8 mm-diameter area for cell culture. Wounds were made by scraping off cells in the center of the cell sheets using a rotating silicone tip. The areas of these wounds were measured by a method of computed planimetry. The average wound area was 3.00 +/- 0.21 mm2 in rabbit cell cultures and 3.18 +/- 0.28 mm2 in bovine. The healing process after the wounding was observed using an inverted phase contrast microscope and wound-healing rates (healed area per hour) were calculated. Wounded areas were recovered by 60 hours in rabbit experiments, and by 84 hours in bovine experiments. Wound-healing rates were different among the observation periods and showed differences between the two species. The healing rates in the rabbit were higher than those in the bovine in the early phase. This in vitro model can be used for quantitative evaluation of the wound-healing process of the corneal endothelium.
建立了一种用于评估角膜内皮伤口愈合过程的新型体外模型。将兔或牛角膜内皮细胞的汇合单层细胞片培养在盖玻片上，盖玻片表面除了用于细胞培养的直径8mm区域外，均预先用甲基丙烯酸羟乙酯（HEMA）包被。使用旋转硅胶尖端刮除细胞片中心的细胞来制造伤口。这些伤口的面积通过计算机平面测量法进行测量。兔细胞培养中平均伤口面积为3.00±0.21mm²，牛细胞培养中为3.18±0.28mm²。使用倒置相差显微镜观察伤口后的愈合过程，并计算伤口愈合率（每小时愈合面积）。在兔实验中，伤口面积在60小时内恢复，在牛实验中在84小时内恢复。伤口愈合率在观察期之间有所不同，并且在两个物种之间也存在差异。在早期阶段，兔的愈合率高于牛。这种体外模型可用于角膜内皮伤口愈合过程的定量评估。
