【 abstract 】 objective comparative study to disc type excimer laser epithelial keratomileusis (under (epi-polis laser in-situ keratomileusis, epi-LASIK) and excimer laser corneal refractile cutting technique (photorefractive keratectomy, PRK) to the rabbit corneal pathological effects. Methods the New Zealand white rabbit only 34, random take of 32 rabbit right eye line to disc type Epi-LASIK (group A), left eye line PRK (group B) and the other two rabbit (4 only not surgery eyes) for normal control group (group C).Postoperative 4 h, 1 d, 3 d, 5 d, 7 d, 14 d, 1 month, 3 months take corneal specimens, HE dyeing observation before the number of corneal stromal cell changes, immune cells chemical dyeing observation central corneal Ki-67, alpha SMA positive cells quantity and cutting edge area PCNA positive cornea cell count, and transmission electron microscope corneal ultrastructural changes. Results the cornea stromal cell number before normal for every 400 times the light (88. 00 ± 5. 38); Postoperative different time points before the group B cells are A number of matrix group A little, differences were significantly statistical significance (P < 0, are 01).
Normal corneal epithelial cells basal PCNA positive cells for every 400 times the number of light (15. 94 + 3. 02); After one d, 3 d, 5 d, 7 d, 14 d group A PC making positive cells are less than the number of group B, A statistically significant difference were (both for P < 0. 05 or 0. 01). Normal almost do not express the stromal layer cornea Ki-67; Postoperative 4 h, 1 d, 3 d, 5 d, 7 d, 14 d group A Ki-67 positive cells which is less number of group B, differences were significantly statistical significance (P < 0 are.01). Normal corneal almost don't express alpha SMA; Postoperative 5 d, 7 d, 14 d, 1 month, 3 months group A number of alpha SMA positive cells which is less group B, differences were significantly statistical significance (P < 0 are.01). Transmission electron microscopy (sem) examination showed that the 7 d group A combination between the cells within more closely, 3 months collagen arrangement in the substrate is neat.Conclusion to disc type Epi with a more than PRK-LASIK light corneal epithelium damage responses, less corneal epithelial and stromal cell proliferation cornea reaction and lower to muscle into fiber mother cell transformation potential, so under the corneal epithelium mist cloudy (haze) the likelihood of lower.
【 key words 】 excimer laser corneal refractile cutting art; To disc type excimer laser epithelial keratomileusis (under; Corneal pathology
【摘要】 目的 对比研究去瓣型准分子激光上皮下角膜磨镶术(epi-polis laser in-situ keratomileusis, Epi-LASIK)和准分子激光屈光性角膜切削术(photorefractive keratectomy, PRK)对兔角膜病理的影响。方法 新西兰白兔34只,随机取其中32只兔的右眼行去瓣型Epi-LASIK(A组),左眼行PRK(B组),另外2只兔(4只未手术眼)为正常对照组(C组)。术后4 h、1 d、3 d、5 d、7 d、14 d、1个月、3个月取角膜标本,HE染色观察角膜前基质细胞的数量变化,免疫细胞化学染色观察角膜中央区Ki-67、α-SMA阳性细胞数量及角膜切削缘区PCNA阳性细胞数量,同时透射电镜观察角膜超微结构的变化。结果 正常角膜前基质细胞数量为每400倍光镜(88. 00±5. 38)个;术后各时间点B组前基质细胞数量均较A组少,差异均有显著统计学意义(均为P<0•01)。正常角膜上皮细胞基底层PCNA阳性细胞数量为每400倍光镜(15. 94±3. 02)个;术后1 d、3 d、5 d、7 d、14 d A组PC-NA阳性细胞数量均较B组少,差异均有统计学意义(均为P<0. 05或0. 01)。正常角膜基质层几乎不表达Ki-67;术后4 h、1 d、3 d、5 d、7 d、14 d A组Ki-67阳性细胞数均较B组少,差异均有显著统计学意义(均为P<0. 01)。正常角膜几乎不表达α-SMA;术后5 d、7 d、14 d、1个月、3个月A组α-SMA阳性细胞数均较B组少,差异均有显著统计学意义(均为P<0. 01)。透射电镜检查显示,术后7 d之内A组细胞之间结合更紧密, 3个月时基质内胶原纤维排列较整齐。结论 去瓣型Epi-LASIK比PRK有着更轻的角膜上皮损伤反应,更少的角膜上皮细胞和角膜基质细胞增殖反应以及更低的向肌成纤维母细胞转化潜能,所以角膜上皮下雾状混浊(haze)发生的可能性更低。
【关键词】 准分子激光屈光性角膜切削术;去瓣型准分子激光上皮下角膜磨镶术;角膜病理
1988年,Marshall等[1]发明了准分子激光屈光性角膜切削术(photorefractive keratectomy, PRK),这是第一种用于临床的准分子激光屈光手术术式。2001年,为防止酒精浸泡破坏上皮细胞基底膜和前弹力层的完整性, Pallikaris等又提出了机械法准分子激光上皮下角膜磨镶术(epi-polis laser in-situ ker-atomileusis,Epi-LASIK)[2]。PRK和Epi-LASIK都没有保留角膜前弹力层,但术后角膜上皮下雾状混浊(haze)的发生率PRK远高于Epi-LASIK[3-4]。临床发现,Epi-LASIK去瓣后虽然和PRK同样没有上皮瓣,但前者上皮生长更快,反应更轻, haze发生率更低,为此,我们采用光镜、电镜和免疫细胞化学染色对PRK和去瓣型Epi-LASIK术后的角膜病理和超微结构、增殖细胞核抗原(proliferation cell nuclear anti-gen,PCNA)阳性细胞、增殖蛋白Ki-67阳性细胞、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)阳性细胞等进行研
