不同切削深度兔PRK和去瓣EPI-LASIK术后早期角膜愈合反应的观察

发布时间：2018-04-18 12:44

本文选题：不同切削深度 + 准分子激光角膜切削术　；参考：《南华大学》2011年硕士论文

【摘要】：目的研究不同切削深度兔准分子激光屈光性角膜切削术(Photorefractive keratectomy,PRK)和去瓣EPI-LASIK( Epipolis laser in-situ keratomileusis,EPI-LASIK)术后角膜上皮和基质中细胞形态、炎症细胞、以及细胞因子碱性成纤维细胞生长因子(Basicity fibroblast growth factor,bFGF)、核转录因子(Nuclear transcription factors,NF-κB)的表达,探讨两种手术方式在不同切削深度时早期角膜愈合反应,为临床工作提供理论依据。方法将所选实验兔22只随机分为三个组,高度切削组10只(-10.00D)、低度切削组10只(-3.00D)、剩余2只为空白对照组。建立实验动物模型,右眼去瓣EPI-LASIK,左眼PRK,高度切削组和低度切削组于术后1d、3d、5d、7d空气栓塞法每天随机处死1只,共8只动物,剩余14只于术后7d一并处死,沿角膜缘取角膜组织。用荧光染色法每日观察实验动物术后眼部的基本状况、角膜上皮的愈合情况以及愈合的时间;石蜡切片行HE染色光镜观察角膜上皮与基质细胞的形态学变化和炎症反应情况,并在光镜下计数炎症细胞的数量;免疫组化方法检测早期角膜创伤愈合反应时细胞因子bFGF、NF-κB的表达,应用Image Pro Plus 5.0图像分析系统进行定量分析;TUNEL法检测角膜基质细胞的凋亡情况。所有数据应用SPSS17.0行统计学分析。结果1、眼部状况:PRK眼术后症状明显,3d时有所减轻,去瓣EPI-LASIK术后3d时症状基本消失,高度切削组较低度组症状明显。2、角膜上皮愈合情况:荧光染色观察,高度切削组PRK眼角膜上皮愈合时间最短3d,最长6d,平均为4.33d,去瓣EPI-LASIK眼角膜上皮愈合时间最短3d,最长5d,平均为3.83d,低度切削组愈合较快。3、角膜的形态学改变:在低度切削组,PRK术后角膜上皮过度增生,细胞排列紊乱,可见大量空泡细胞,基质层排列紊乱,炎症细胞浸润。去瓣EPI-LASIK术后角膜上皮未见明显增生,细胞和基质层排列稍紊乱,未见明显的空泡细胞和炎症细胞浸润。高度切削组形态学改变明显。4、炎症反应:低度切削组,PRK与去瓣EPI-LASIK角膜中炎症细胞数量相比,差异无统计学意义(t=2.20,P0.05)。高度切削组,PRK较去瓣EPI-LASIK角膜中炎症细胞的数量明显增加,差异有统计学意义(t=10.32,P0.01)。5、角膜基质细胞凋亡情况:低度切削组,PRK与去瓣EPI-LASIK相比,凋亡细胞的数量未见明显异常(t=2.45,P0.05),而高度切削组,PRK较去瓣EPI-LASIK凋亡细胞的数量明显增加,差异有统计学意义(t=6.03,P0.01)。6、角膜中bFGF、NF-κB的表达:PRK和去瓣EPI-LASIK术后角膜中bFGF、NF-κB的表达均较空白对照组增加,低度切削组,PRK与去瓣EPI-LASIK角膜中bFGF、NF-κB表达未见明显异常(t=2.55,t=2.13,t=1.02,t=2.22,P0.05),而高度切削组,PRK较去瓣EPI-LASIK角膜中bFGF的表达减少,NF-κB的表达增加,两者均有统计学意义(P0.01)。结论去瓣EPI-LASIK用微型角膜刀制作上皮瓣并去除,基底膜完整,可以阻挡泪液及损伤细胞所释放的各种因子,使术后角膜上皮愈合快,炎症反应轻,尤其在高度切削时,去瓣EPI-LASIK较PRK有显著的优越性。
[Abstract]:Objective to study the effect of different cutting depth in rabbits after excimer laser photorefractive keratectomy (Photorefractive keratectomy PRK) and EPI-LASIK (Epipolis laser in-situ flap keratomileusis, EPI-LASIK) after cell morphology of corneal epithelium and stromal inflammatory cells, and cytokine basic fibroblast growth factor (Basicity fibroblast, growth factor, bFGF), nuclear transcription factors (Nuclear transcription factors, NF- K B) the expression of two kinds of operation mode in different cutting depth of early corneal wound healing response, and provide a theoretical basis for clinical work.
Methods the selected 22 experimental rabbits were randomly divided into three groups, high cutting group 10 (-10.00D), low cutting group 10 (-3.00D), the remaining 2 mice as control group. The establishment of experimental animal model, the right to left PRK, EPI-LASIK valve, high cutting group and low cutting group after operation. 1D, 3D, 5D, 7d were sacrificed 1 day air embolism method, a total of 8 animal, the remaining 14 rats at 7d after operation. All were killed, along the limbal corneal tissue. The basic condition of staining observed daily experimental animal postoperative ocular fluorescence, corneal epithelial healing and healing time of paraffin; the slices were stained by HE staining and morphological changes of the inflammatory response of light microscope to observe the corneal epithelium and stromal cells, and in the light microscope to count the number of inflammatory cells; immunohistochemical method to detect early corneal wound healing response of cytokine bFGF, expression of NF- kappa B, Image Pro with Plus 5 image The analysis system was used for quantitative analysis. The apoptosis of corneal stromal cells was detected by TUNEL method. All data were statistically analyzed by SPSS17.0.
The results of 1 eye condition: PRK eye postoperative symptoms, 3D decrease to EPI-LASIK flap 3D after the symptoms disappeared, high cutting group less obvious symptoms of group.2, corneal epithelial healing: fluorescent staining, the shortest 3D height of cornea ablation group PRK epithelial healing time, the longest 6D that was an average of 4.33d to EPI-LASIK corneal epithelial flap healing time 3D, the longest 5D, average 3.83d, low cutting group heal faster.3, the change of corneal morphology in low cutting group, PRK corneal epithelial hyperplasia, cell arrangement disorder, vacuoles cells, stromal layer derangement the infiltration of inflammatory cells. The postoperative corneal epithelial flap EPI-LASIK no obvious hyperplasia cells and stroma arranged a little disorder. No obvious vacuolar degeneration and inflammatory cell infiltration. Change the high cutting group obvious morphological.4, inflammatory reaction, low cutting group, PRK Compared with the number of inflammatory cells in the corneal flap EPI-LASIK, there was no statistically significant difference (t=2.20, P0.05). High cutting group, PRK compared to the number of inflammatory cells in the corneal flap EPI-LASIK increased significantly, the difference was statistically significant (t=10.32, P0.01).5, corneal stromal cell apoptosis: low cutting group, compared with PRK to flap EPI-LASIK, number of apoptotic cells was significantly abnormal (t=2.45, P0.05), and high cutting group, the number of apoptotic cells compared to the PRK EPI-LASIK flap was significantly increased, the difference was statistically significant (t=6.03, P0.01).6, corneal bFGF, expression of NF- kappa B: bFGF PRK and EPI-LASIK corneal flap in the expression of NF- kappa B increased significantly compared with the blank control group, low cutting group, PRK and bFGF to EPI-LASIK in NF- corneal flap, kappa B expression had no obvious abnormalities (t=2.55, t=2.13, t=1.02, t=2.22, P0.05), and high cutting group, compared to the PRK bFGF EPI-LASIK in the form of corneal flap The expression of NF- kappa B increased, and both had statistical significance (P0.01).
Conclusion EPI-LASIK flap made by microkeratome flap and removal of basement membrane integrity, various factors can stop the tear and damage cells release, the postoperative corneal epithelium healing, less inflammatory reaction, especially in high cutting, to flap EPI-LASIK than PRK has obvious advantages.

【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R779.63

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