微創(chuàng)核黃素—紫外線A兔鞏膜膠原交聯(lián)的實(shí)驗(yàn)研究
發(fā)布時(shí)間：2018-08-26 14:44
【摘要】：目的核黃素-紫外線A誘導(dǎo)的鞏膜膠原交聯(lián)療法已在體外及活體動(dòng)物實(shí)驗(yàn)中證明了其增強(qiáng)鞏膜生物力學(xué)強(qiáng)度的長(zhǎng)期有效性與安全性,但既往手術(shù)操作創(chuàng)傷大,且無法暴露解剖上較為靠后極的鞏膜區(qū)域,本文旨在研究一種操作創(chuàng)傷小、可有效照射至后鞏膜區(qū)域的微創(chuàng)核黃素-紫外線A鞏膜膠原交聯(lián)法,通過設(shè)計(jì)合適的實(shí)驗(yàn)參數(shù),在活體兔上展開一系列的實(shí)驗(yàn)研究,以驗(yàn)證其有效性與安全性。方法選用70只體重為2.0Kg~2.5Kg的成年清潔級(jí)新西蘭大白兔,雌雄各半,無任何眼部疾病。隨機(jī)分為7組,每組各10只,分別為非交聯(lián)對(duì)照組(Ⅰ)、交聯(lián)后1天組(Ⅱ)、交聯(lián)后7天組(Ⅲ)、交聯(lián)后15天組(Ⅳ)、交聯(lián)后1月組(Ⅴ)、交聯(lián)后2月組(Ⅵ)、交聯(lián)后3月組(Ⅶ)。交聯(lián)組(Ⅱ至Ⅶ組)和非交聯(lián)對(duì)照組皆選擇右眼作為實(shí)驗(yàn)眼,左眼作為對(duì)照眼。應(yīng)用微創(chuàng)核黃素-紫外線A鞏膜交聯(lián)儀器對(duì)交聯(lián)組行鞏膜膠原交聯(lián)實(shí)驗(yàn),具體實(shí)驗(yàn)參數(shù)設(shè)置如下:紫外線波長(zhǎng)為370nm,能量密度設(shè)置為3m W/cm2,照射時(shí)間為30分鐘;通過微創(chuàng)結(jié)膜切口插入特制LED光源并貼近鞏膜照射;每隔2分鐘滴加1次濃度為0.1%的核黃素溶液(不含右旋糖酐)作為光敏感劑。分別在各交聯(lián)組所規(guī)定的術(shù)后時(shí)間處死對(duì)應(yīng)各交聯(lián)組的兔子,摘取眼球,每組隨機(jī)選取6只用于生物力學(xué)檢測(cè)以比較各組鞏膜生物力學(xué)指標(biāo)變化(極限應(yīng)力、極限應(yīng)變和8%彈性模量),剩余4只用于HE染色、TUNEL細(xì)胞凋亡檢測(cè),以分析其組織病理學(xué)變化。結(jié)果1.各組(Ⅰ至Ⅶ組)左右眼鞏膜厚度比較,組別差異無統(tǒng)計(jì)學(xué)意義(P0.05),眼別差異無統(tǒng)計(jì)學(xué)意義(P0.05),組別和眼別間不存在交互作用(P0.05)。2.各組(Ⅰ至Ⅶ組)鞏膜生物力學(xué)指標(biāo)比較:組別差異均有統(tǒng)計(jì)學(xué)意義(均P0.05),眼別差異均有統(tǒng)計(jì)學(xué)意義(均P0.01),組別和眼別間均存在交互作用(均P0.01);Ⅱ至Ⅶ組各組分別與Ⅰ組比較,左眼生物力學(xué)指標(biāo)差異均無統(tǒng)計(jì)學(xué)意義(均P0.05),右眼生物力學(xué)指標(biāo)差異均有統(tǒng)計(jì)學(xué)意義(均P0.01);Ⅱ至Ⅵ組各組的組間比較,左眼生物力學(xué)指標(biāo)差異均無統(tǒng)計(jì)學(xué)意義(均P0.05),右眼生物力學(xué)指標(biāo)差異均無統(tǒng)計(jì)學(xué)意義(均P0.05);Ⅱ至Ⅵ組各組分別與Ⅶ組比較,左眼生物力學(xué)指標(biāo)差異均無統(tǒng)計(jì)學(xué)意義(均P0.05),右眼生物力學(xué)指標(biāo)差異均有統(tǒng)計(jì)學(xué)意義(均P0.05);Ⅰ組組內(nèi)比較,左右眼的生物力學(xué)指標(biāo)差異無統(tǒng)計(jì)學(xué)意義(P0.05);Ⅱ至Ⅶ組各組組內(nèi)比較,左右眼的生物力學(xué)指標(biāo)差異均有統(tǒng)計(jì)學(xué)意義(均P0.01)。3.光鏡下觀察并比較Ⅰ至Ⅶ組各組的鞏膜HE染色石蠟切片,結(jié)果顯示:Ⅱ至Ⅶ組各交聯(lián)組的角膜、鞏膜、虹膜、睫狀體和脈絡(luò)膜等均未發(fā)生細(xì)胞壞死和水腫等異常的形態(tài)學(xué)改變,視網(wǎng)膜無變性、內(nèi)外核層細(xì)胞數(shù)減少等改變。4.TUNEL細(xì)胞凋亡檢測(cè)結(jié)果顯示:交聯(lián)組(Ⅱ至Ⅶ組)右眼的細(xì)胞凋亡率平均值為12.13%,非交聯(lián)組(Ⅰ組)細(xì)胞凋亡率為11.00%,組間細(xì)胞凋亡率比較均無統(tǒng)計(jì)學(xué)差異(均P0.05)。結(jié)論交聯(lián)組各組的兔鞏膜生物力學(xué)強(qiáng)度均得到有效提高,眼球各組織無明顯病理?yè)p害,微創(chuàng)核黃素-紫外線A鞏膜膠原交聯(lián)法可安全、有效地提高兔鞏膜的生物力學(xué)強(qiáng)度,此外,交聯(lián)組各組兔鞏膜的交聯(lián)效果會(huì)隨交聯(lián)后時(shí)間的增加而出現(xiàn)減弱。
[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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R778.11
本文編號(hào)：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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R778.11
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
1 張學(xué)敏;趙旭;張豐菊;張淼;;核黃素在鞏膜組織內(nèi)滲透性的實(shí)驗(yàn)研究[J];中華眼科雜志;2015年06期
2 張亞麗;李志偉;牟國(guó)營(yíng);劉蕾;;核黃素/紫外線A誘導(dǎo)的膠原交聯(lián)對(duì)兔和人鞏膜生物力學(xué)性能的影響[J];中華眼視光學(xué)與視覺科學(xué)雜志;2014年05期
3 田恬;;活體材料的納米生物力學(xué)[J];科技導(dǎo)報(bào);2014年08期
4 王瑩;韓泉洪;韓風(fēng)梅;楚艷華;趙堪興;;甘油醛后鞏膜交聯(lián)治療豚鼠形覺剝奪性近視眼的研究[J];中華眼科雜志;2014年01期
5 趙旭;王萌萌;張豐菊;;紫外線A-核黃素交聯(lián)加固角鞏膜安全性的研究進(jìn)展[J];中華眼科雜志;2013年02期
6 呂雅平;周浩;夏文濤;褚仁遠(yuǎn);周行濤;戴錦暉;;紫外光-核黃素交聯(lián)法對(duì)豚鼠鞏膜生物力學(xué)特性的影響[J];中國(guó)實(shí)驗(yàn)動(dòng)物學(xué)報(bào);2012年04期
7 王萌萌;張豐菊;;膠原交聯(lián)在眼科的應(yīng)用進(jìn)展[J];中國(guó)實(shí)用眼科雜志;2011年10期
8 蒲玲玲;韋京豫;余奇;;維生素B_2檢測(cè)方法的研究進(jìn)展[J];中國(guó)老年學(xué)雜志;2009年11期
9 汪多仁;;維生素B_2的開發(fā)與應(yīng)用[J];發(fā)酵科技通訊;2008年03期
10 靳韜;王超英;;不同應(yīng)力水平對(duì)幼兔鞏膜生物力學(xué)特性的影響[J];眼科研究;2007年09期
相關(guān)博士學(xué)位論文 前2條
1 張亞麗;鞏膜核黃素/紫外線A膠原交聯(lián)的實(shí)驗(yàn)研究[D];山東大學(xué);2013年
2 王萌萌;鞏膜紫外光—核黃素交聯(lián)術(shù)的有效性和安全性研究[D];首都醫(yī)科大學(xué);2013年
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