Netto Marcelo V, Mohan Rajiv R, Medeiros Fabricio W, Dupps William J, Sinha Sunilima, Krueger Ronald R, Stapleton W Michael, Rayborn Mary, Suto Chikako, Wilson Steven E
The Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio 44195, USA.
J Refract Surg. 2007 Sep;23(7):667-76. doi: 10.3928/1081-597X-20070901-05.
To examine early postoperative wound healing in rabbit corneas that had LASIK flaps formed with three different models (15 KHz, 30 KhZ, and 60 KHz) of a femtosecond laser compared with flaps formed with a microkeratome.
Thirty-nine rabbit eyes were randomized to receive either no surgery or corneal flaps formed with one of the lasers or the microkeratome. Sixteen eyes also had lamellar cuts with no side cuts with the 30 KHz laser. Animals were sacrificed and corneas processed as frozen sections or fixed for transmission electron microscopy. Frozen sections were evaluated with the TUNEL assay to detect apoptosis, immunocytochemistry for Ki67 to detect cell mitosis, and immunocytochemistry for CD11b to detect mononuclear cells.
Rabbit corneas that had flaps formed with the 15 KHz laser had significantly more stromal cell death, greater stromal cell proliferation, and greater monocyte influx in the central and peripheral comea at 24 hours after surgery than corneas that had flaps formed with the 30 KHz or 60 KHz laser or the microkeratome. Results of the 60 KHz laser and microkeratome were not significantly different for any of the parameters at 24 hours, except for mitotic stromal cells at the flap margin. Transmission electron microscopy revealed that the primary mode of stromal cell death at 24 hours after laser ablation was necrosis.
Stromal cell necrosis associated with femtosecond laser flap formation likely contributes to greater inflammation after LASIK performed with the femtosecond laser, especially with higher energy levels that result in greater keratocyte cell death.
比较用飞秒激光的三种不同模式(15千赫兹、30千赫兹和60千赫兹)制作准分子原位角膜磨镶术(LASIK)瓣的兔角膜与用微型角膜刀制作瓣的兔角膜术后早期伤口愈合情况。
39只兔眼随机分为不做手术组,或接受用其中一种激光或微型角膜刀制作角膜瓣组。16只眼还用30千赫兹激光进行了无侧切的板层切割。处死动物,将角膜制成冰冻切片或固定用于透射电子显微镜检查。对冰冻切片进行末端脱氧核苷酸转移酶介导的缺口末端标记(TUNEL)分析以检测细胞凋亡,进行Ki67免疫细胞化学检测以检测细胞有丝分裂,进行CD11b免疫细胞化学检测以检测单核细胞。
与用30千赫兹或60千赫兹激光或微型角膜刀制作瓣的角膜相比,用15千赫兹激光制作瓣的兔角膜在术后24小时中央和周边角膜有更多的基质细胞死亡、更强的基质细胞增殖和更多的单核细胞流入。除瓣边缘的有丝分裂基质细胞外,60千赫兹激光和微型角膜刀在24小时时的任何参数结果均无显著差异。透射电子显微镜显示,激光消融后24小时基质细胞死亡的主要方式是坏死。
飞秒激光瓣形成相关的基质细胞坏死可能导致飞秒激光LASIK术后炎症反应更强,尤其是能量水平较高导致角膜细胞死亡更多时。