Dreiss Alexandra K, Winkler von Mohrenfels Christoph, Gabler Bernhard, Kohnen Thomas, Marshall John, Lohmann Chris P
The Rayne Insitute, Departmen of Ophthalmology, St. Thomas' Hospital, London.
Klin Monbl Augenheilkd. 2002 May;219(5):365-9; discussion 369. doi: 10.1055/s-2002-32631.
Laser epithelial keratomileusis (LASEK) is a new surgical procedure to treat myopia. An epithelial flap is created after the exposure to 20 % alcohol and following the laser ablation the epithelium is repositioned to its original location. The advantage of LASEK is that the ablated corneal surface is covered by a full thickness epithelium immediately after surgery. It is hypothesised that this epithelial coverage inhibits the wound healing response of the cornea. However, this concept will only work if the epithelial cells are still vital after the exposure to alcohol.
The vitality of the corneal epithelial cells was investigated in 6 human cadaver eyes after the exposure to 20 % alcohol over 15 to 60 s. The vitality of the corneal epithelial cells was assessed by soaking the specimen in a 0.1 % trypan blue solution and incubated at 37 degrees C for 2 min. After a wash with BSS the specimen were re-incubated at 37 degrees C for 30 min in culture medium. After one more wash with BSS the cells were observed with a standard light microscope. Cells which retained the blue colour would be dead and vital cells would appear clear. Light microscopy and transmission electron microscopy was used to detect morphological changes.
Mainly vital corneal epithelial cells were seen for an alcohol exposure time of up to 30 s. In particular the basal epithelial cells appeared alive. With a exposition time of 60 s most cells were dead.
In LASEK the exposure time of 20 % alcohol is between 20 and 30 s. Based on our results we can conclude that after such exposure time most cells are alive, which is essential for the postoperative wound healing response.
准分子激光上皮下角膜磨镶术(LASEK)是一种治疗近视的新型手术方法。在暴露于20%酒精后制作上皮瓣,激光消融后上皮瓣重新复位到其原始位置。LASEK的优点是术后被消融的角膜表面立即被全层上皮覆盖。据推测,这种上皮覆盖可抑制角膜的伤口愈合反应。然而,只有上皮细胞在暴露于酒精后仍保持活力,这一概念才会起作用。
对6只人尸体眼在暴露于20%酒精15至60秒后角膜上皮细胞的活力进行了研究。将标本浸泡在0.1%台盼蓝溶液中,并在37℃孵育2分钟,以评估角膜上皮细胞的活力。用平衡盐溶液(BSS)冲洗后,将标本在培养基中于37℃再孵育30分钟。再用BSS冲洗一次后,用标准光学显微镜观察细胞。保留蓝色的细胞为死亡细胞,活细胞将呈现透明。用光镜和透射电子显微镜检测形态学变化。
酒精暴露时间长达30秒时,主要可见存活的角膜上皮细胞。特别是基底上皮细胞看起来是活的。暴露60秒时,大多数细胞死亡。
在LASEK中,20%酒精的暴露时间为20至30秒。根据我们的结果可以得出结论,在这样的暴露时间后,大多数细胞是存活的,这对术后伤口愈合反应至关重要。