微创核黄素—紫外线A兔巩膜胶原交联的实验研究
发布时间：2018-08-26 14:44
【摘要】：目的核黄素-紫外线A诱导的巩膜胶原交联疗法已在体外及活体动物实验中证明了其增强巩膜生物力学强度的长期有效性与安全性,但既往手术操作创伤大,且无法暴露解剖上较为靠后极的巩膜区域,本文旨在研究一种操作创伤小、可有效照射至后巩膜区域的微创核黄素-紫外线A巩膜胶原交联法,通过设计合适的实验参数,在活体兔上展开一系列的实验研究,以验证其有效性与安全性。方法选用70只体重为2.0Kg~2.5Kg的成年清洁级新西兰大白兔,雌雄各半,无任何眼部疾病。随机分为7组,每组各10只,分别为非交联对照组(Ⅰ)、交联后1天组(Ⅱ)、交联后7天组(Ⅲ)、交联后15天组(Ⅳ)、交联后1月组(Ⅴ)、交联后2月组(Ⅵ)、交联后3月组(Ⅶ)。交联组(Ⅱ至Ⅶ组)和非交联对照组皆选择右眼作为实验眼,左眼作为对照眼。应用微创核黄素-紫外线A巩膜交联仪器对交联组行巩膜胶原交联实验,具体实验参数设置如下:紫外线波长为370nm,能量密度设置为3m W/cm2,照射时间为30分钟;通过微创结膜切口插入特制LED光源并贴近巩膜照射;每隔2分钟滴加1次浓度为0.1%的核黄素溶液(不含右旋糖酐)作为光敏感剂。分别在各交联组所规定的术后时间处死对应各交联组的兔子,摘取眼球,每组随机选取6只用于生物力学检测以比较各组巩膜生物力学指标变化(极限应力、极限应变和8%弹性模量),剩余4只用于HE染色、TUNEL细胞凋亡检测,以分析其组织病理学变化。结果1.各组(Ⅰ至Ⅶ组)左右眼巩膜厚度比较,组别差异无统计学意义(P0.05),眼别差异无统计学意义(P0.05),组别和眼别间不存在交互作用(P0.05)。2.各组(Ⅰ至Ⅶ组)巩膜生物力学指标比较:组别差异均有统计学意义(均P0.05),眼别差异均有统计学意义(均P0.01),组别和眼别间均存在交互作用(均P0.01);Ⅱ至Ⅶ组各组分别与Ⅰ组比较,左眼生物力学指标差异均无统计学意义(均P0.05),右眼生物力学指标差异均有统计学意义(均P0.01);Ⅱ至Ⅵ组各组的组间比较,左眼生物力学指标差异均无统计学意义(均P0.05),右眼生物力学指标差异均无统计学意义(均P0.05);Ⅱ至Ⅵ组各组分别与Ⅶ组比较,左眼生物力学指标差异均无统计学意义(均P0.05),右眼生物力学指标差异均有统计学意义(均P0.05);Ⅰ组组内比较,左右眼的生物力学指标差异无统计学意义(P0.05);Ⅱ至Ⅶ组各组组内比较,左右眼的生物力学指标差异均有统计学意义(均P0.01)。3.光镜下观察并比较Ⅰ至Ⅶ组各组的巩膜HE染色石蜡切片,结果显示:Ⅱ至Ⅶ组各交联组的角膜、巩膜、虹膜、睫状体和脉络膜等均未发生细胞坏死和水肿等异常的形态学改变,视网膜无变性、内外核层细胞数减少等改变。4.TUNEL细胞凋亡检测结果显示:交联组(Ⅱ至Ⅶ组)右眼的细胞凋亡率平均值为12.13%,非交联组(Ⅰ组)细胞凋亡率为11.00%,组间细胞凋亡率比较均无统计学差异(均P0.05)。结论交联组各组的兔巩膜生物力学强度均得到有效提高,眼球各组织无明显病理损害,微创核黄素-紫外线A巩膜胶原交联法可安全、有效地提高兔巩膜的生物力学强度,此外,交联组各组兔巩膜的交联效果会随交联后时间的增加而出现减弱。
[Abstract]:Objective Riboflavin-ultraviolet A-induced scleral collagen cross-linking therapy has proved its long-term efficacy and safety in enhancing the biomechanical strength of the sclera in vitro and in vivo. However, previous surgical procedures were traumatic and could not expose the anatomically more posterior scleral areas. This study was designed to study a less traumatic operation. Methods Seventy clean adult New Zealand white rabbits weighing 2.0Kg~2.5Kg were selected, half male and half female, without any eye. Diseases were randomly divided into 7 groups, 10 in each group, which were non-crosslinking control group (I), 1 day after crosslinking group (II), 7 days after crosslinking group (III), 15 days after crosslinking group (IV), 1 month after crosslinking group (V), 2 months after crosslinking group (VI), 3 months after crosslinking group (_). Cross-linking group (group II to_) and non-crosslinking control group all chose right eye as experimental eyes, left eye as control. Eye. Scleral collagen cross-linking test was performed with minimally invasive riboflavin-ultraviolet A scleral cross-linking instrument in the cross-linking group. The specific experimental parameters were as follows: UV wavelength was 370 nm, energy density was 3 m W/cm 2, irradiation time was 30 minutes; special LED light source was inserted through minimally invasive conjunctival incision and irradiated close to the sclera; the concentration was added once every 2 minutes. 0.1% riboflavin solution (without dextran) was used as photosensitizer. The rabbits in each cross-linking group were sacrificed and their eyeballs were taken out. Six rabbits in each group were randomly selected for biomechanical testing to compare the changes of scleral biomechanical indices (ultimate stress, ultimate strain and 8% elastic modulus) and the remaining 4. Results 1. There was no significant difference in scleral thickness between the two groups (P 0.05). There was no significant difference between the two groups (P 0.05). There was no significant difference between the two groups (P 0.05). There was no interaction between the two groups (P 0.05). 2. Standard comparison: There were statistically significant differences between groups (all P 0.05), eye differences were statistically significant (all P 0.01), there were interaction between groups and eye groups (all P 0.01); Group II to_compared with group I, there were no significant differences in left eye biomechanical indicators (all P 0.05), right eye biomechanical indicators were statistically significant differences. Meaning (all P 0.01); There was no significant difference in left eye biomechanical indexes between groups II to VI (all P 0.05), and no significant difference in right eye biomechanical indexes (all P 0.05); there was no significant difference in left eye biomechanical indexes between groups II to VI and_, respectively (all P 0.05). The differences were statistically significant (all P 0.05); there was no significant difference in the biomechanical indexes between the left and right eyes in group I (P 0.05); there was significant difference in the biomechanical indexes between the left and right eyes in group II to_ (all P 0.01). 3. The scleral HE stained paraffin sections in group I to_were observed and compared under light microscope. The results showed that there were no abnormal morphological changes in cornea, sclera, iris, ciliary body and choroid, no degeneration of retina, and no decrease in the number of cells in the inner and outer nuclear layers. 4. The results of TUNEL apoptosis detection showed that the average apoptosis rate of right eye in the cross-linked group (group II to_) was 12.13%. Conclusion The biomechanical strength of the rabbit sclera in the cross-linking group was improved effectively, and there was no obvious pathological damage in the eyeball tissues. The minimally invasive riboflavin-ultraviolet A scleral collagen cross-linking method was safe and effective to improve the rabbit sclera. In addition, the cross-linking effect of each group of rabbits sclera decreased with the increase of post-crosslinking time.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R778.11
本文编号：2205192
[Abstract]:Objective Riboflavin-ultraviolet A-induced scleral collagen cross-linking therapy has proved its long-term efficacy and safety in enhancing the biomechanical strength of the sclera in vitro and in vivo. However, previous surgical procedures were traumatic and could not expose the anatomically more posterior scleral areas. This study was designed to study a less traumatic operation. Methods Seventy clean adult New Zealand white rabbits weighing 2.0Kg~2.5Kg were selected, half male and half female, without any eye. Diseases were randomly divided into 7 groups, 10 in each group, which were non-crosslinking control group (I), 1 day after crosslinking group (II), 7 days after crosslinking group (III), 15 days after crosslinking group (IV), 1 month after crosslinking group (V), 2 months after crosslinking group (VI), 3 months after crosslinking group (_). Cross-linking group (group II to_) and non-crosslinking control group all chose right eye as experimental eyes, left eye as control. Eye. Scleral collagen cross-linking test was performed with minimally invasive riboflavin-ultraviolet A scleral cross-linking instrument in the cross-linking group. The specific experimental parameters were as follows: UV wavelength was 370 nm, energy density was 3 m W/cm 2, irradiation time was 30 minutes; special LED light source was inserted through minimally invasive conjunctival incision and irradiated close to the sclera; the concentration was added once every 2 minutes. 0.1% riboflavin solution (without dextran) was used as photosensitizer. The rabbits in each cross-linking group were sacrificed and their eyeballs were taken out. Six rabbits in each group were randomly selected for biomechanical testing to compare the changes of scleral biomechanical indices (ultimate stress, ultimate strain and 8% elastic modulus) and the remaining 4. Results 1. There was no significant difference in scleral thickness between the two groups (P 0.05). There was no significant difference between the two groups (P 0.05). There was no significant difference between the two groups (P 0.05). There was no interaction between the two groups (P 0.05). 2. Standard comparison: There were statistically significant differences between groups (all P 0.05), eye differences were statistically significant (all P 0.01), there were interaction between groups and eye groups (all P 0.01); Group II to_compared with group I, there were no significant differences in left eye biomechanical indicators (all P 0.05), right eye biomechanical indicators were statistically significant differences. Meaning (all P 0.01); There was no significant difference in left eye biomechanical indexes between groups II to VI (all P 0.05), and no significant difference in right eye biomechanical indexes (all P 0.05); there was no significant difference in left eye biomechanical indexes between groups II to VI and_, respectively (all P 0.05). The differences were statistically significant (all P 0.05); there was no significant difference in the biomechanical indexes between the left and right eyes in group I (P 0.05); there was significant difference in the biomechanical indexes between the left and right eyes in group II to_ (all P 0.01). 3. The scleral HE stained paraffin sections in group I to_were observed and compared under light microscope. The results showed that there were no abnormal morphological changes in cornea, sclera, iris, ciliary body and choroid, no degeneration of retina, and no decrease in the number of cells in the inner and outer nuclear layers. 4. The results of TUNEL apoptosis detection showed that the average apoptosis rate of right eye in the cross-linked group (group II to_) was 12.13%. Conclusion The biomechanical strength of the rabbit sclera in the cross-linking group was improved effectively, and there was no obvious pathological damage in the eyeball tissues. The minimally invasive riboflavin-ultraviolet A scleral collagen cross-linking method was safe and effective to improve the rabbit sclera. In addition, the cross-linking effect of each group of rabbits sclera decreased with the increase of post-crosslinking time.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R778.11
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