胶原交联对兔角膜穿通伤瘢痕愈合的生物力学影响

发布时间：2018-07-04 16:20

本文选题：胶原交联 + 角膜　；参考：《遵义医学院》2017年硕士论文

【摘要】：目的:探讨核黄素联合紫外光A(Ultraviolet-A,UVA)诱导兔眼角膜穿通伤胶原交联(corneal collagen crosslinking,CXL)后对角膜瘢痕的生物力学的影响,为预防外伤性角膜葡萄肿的形成探索新方法。方法:选86只健康家兔。80只随机分为正常对照组(A组)和实验组(B、C、D 3组,均制作角膜全层穿通伤模型),B组(单纯缝合组)、C组(缝合+术后即刻CXL)、D组(缝合+第7天CXL),B、C、D每组又根据不同处死时间(4周、6周、8周)分为a、b、c3组,即Ba、Bb、Bc;Ca、Cb、Cc;Da、Db、Dc组,每组8只。在相应的时间统一取4mm×10mm角膜试件在生物力学实验机上进行角膜拉伸,记录每个角膜瘢痕试件所拉伸的应力、应变,然后计算在10%应变时的杨氏模量(Young's modulus,E),绘制应力-应变曲线。根据所测出的CXL组及单纯缝合组中生物力学特性最强组,按相应的处理方法再次建模,每组3只,通过蛋白免疫印迹(Western Boltting,WB)检测α-平滑肌肌动蛋白(Alpha smooth muscle action,α-SMA)表达情况。结果:(1)角膜瘢痕最大的应力:A组为5.71±1.29MPa,B组:Ba、Bb、Bc组分别为1.37±0.62MPa,2.21±0.30MPa,3.14±0.74MPa;C组:Ca、Cb、Cc组分别为1.87±0.56MPa,2.83±0.54MPa,3.26±0.83MPa;D组:Da、Db、Dc组分别为2.17±0.52MPa,2.92±0.63MPa,4.21±0.68MPa。与A组比较,各组均有统计学差异(P=0.00),即穿通伤后所形成的瘢痕的应力较正常角膜明显降低。杨氏模量:A组为:16.37±4.06MPa;B组:Ba、Bb、Bc组分别为4.64±1.69MPa,7.75±0.82MPa,9.02±0.87MPa;C组:Ca、Cb、Cc组分别为9.67±2.42MPa,10.67±2.07MPa,11.08±1.61MPa;D组:Da、Db、Dc组分别为10.94±1.57MPa,11.16±2.50MPa,13.36±2.10MPa。除了Dc组与A组织比较(P=0.007),其余与A组比较,均有统计学差异(P=0.00),即穿通伤后所形成的瘢痕的杨氏模量明显降低。(2)组内最大应力比较:随时间推移,应力明显增加。Ba与Bb相比,具有统计学差异(P=0.026),Bb与Bc 3组相比,有统计学差异(P=0.015);Ca与Cb相比有统计学差异(P=0.010),而Cb与Cc组无明显差异(P=0.235);Da与Db相比具有统计学差异(P=0.042),Db与Dc相比有统计学差异(P=0.01)。(3)组内杨氏模量比较:随时间推移,单纯缝合组杨氏模量在增加,在第6周后增加缓慢,缝合后第7天CXL在第8周最强,而缝合后即刻CXL虽呈增加趋势,但变化较为缓慢。Ba、Bb、Bc 3组相比,Ba与Bb有统计学差异(P=0.008),Bb相比Bc相比无统计学差异(P=0.264);Ca、Cb、Cc 3组相比,Ca与Cb相比,无统计学差异(P=0.364),Cb与Cc相比无统计学差异(P=0.707);Da与Db相比无统计学差异(P=0.842),Db与Dc相比有统计学差异(P=0.047)。(4)组间最大应力比较:总体上看,单纯缝合后形成的瘢痕的应力低于缝合后第7天CXL后的应力。Ba、Ca、Da 3组相比,Ba与Ca比较无统计学差异(P=0.181),Ca与Da相比无统计学差异(P=0.405),Ba与Da相比有统计学差异(P=0.034);Bb、Cb、Db 3组相比,Bb与Cb相比,无统计学差异(P=0.094),Cb与Db相比无统计学差异(P=0.805);Bc、Cc、Dc 3组相比,除Bc与Cc组对比无统计学差异(P=0.764),Cc与Dc相比,有统计学差异(P=0.010),Bc与Dc相比有统计学差异(P=0.005)。(5)组间杨氏模量比较:总体上看,单纯缝合后所形成的瘢痕的抗变形能力低于缝合后第7天CXL的。Ba、Ca、Da 3组相比,Ba与Ca及Da相比有统计学差异(P值均=0.000),Ca与Da组对比无统计学差异(P=0.249);Bb、Cb、Db 3组相比,Bb与Cb及Da对比均有统计学差异(P分别为0.009、0.003),Cb与Db对比无统计学差异(P=0.654);Bc、Cc、Dc 3组相比,Bc与Cc组对比无明显统计学差异(P=0.072),Cc及Dc相比有统计学差异(P=0.040)。(6)WB的结果显示:兔角膜穿通伤单纯缝合组与缝合+第7天CXL的α-SMA的表达比率分别为0.28±0.11和0.65±0.20,差异有统计学意义(P=0.048)。结论:(1)兔角膜穿通伤缝合术后所形成的瘢痕的生物力学特性随时间延长而增强,本次实验在第8周最强。(2)CXL可以增强兔角膜穿通伤形成瘢痕的生物力学特性,随时间推移抗生物力学特性逐渐增强,在兔角膜穿通伤缝合后第7天进行CXL要好于即刻CXL,可能可以预防角膜葡萄肿发生。(3)在角膜穿通伤后CXL可以增加α-SMA的分泌。
[Abstract]:Objective: To explore the effect of collagen cross-linking (corneal collagen crosslinking, CXL) on corneal scar after corneal perforating injury in rabbits induced by riboflavin combined with A (Ultraviolet-A, UVA), and to explore a new method to prevent the formation of traumatic corneal staphyloma. Methods: 86 healthy rabbits were selected to be randomly divided into normal control group (A group). And the experimental group (group B, C, D 3, all corneal penetrating injury models), group B (simple suture group), group C (suture + immediate CXL), D group (suture + seventh days CXL), B, C, D each group were divided into a, i.e., 4 weeks, 6 weeks, 8 weeks. The tensile stress and strain of each corneal scar specimen were recorded on the biomechanical experiment machine, and then the young modulus (Young's modulus, E) at the 10% strain was calculated and the stress strain curve was drawn. According to the strongest group of biological mechanics characteristics in the CXL group and the simple suture group, the model was modeled again according to the corresponding treatment method. The expression of alpha smooth muscle actin (Alpha smooth muscle action, alpha -SMA) was detected by Western Boltting (Western, WB) in each group. Results: (1) the maximum stress of corneal scar: A group was 5.71 + 1.29MPa, B group was 1.37 +, 2.21 +, 3.14 +. 2.83 + 0.54MPa, 3.26 + 0.83MPa, group D: Da, Db, Dc group were 2.17 + 0.52MPa, 2.92 + 0.63MPa and 4.21 + 0.68MPa. compared with A group. All groups were statistically different (P=0.00), that is, the stress of scar formed after penetrating injury was significantly lower than that of normal cornea. Young's modulus: A group was 16.37 + 4.64, 7.75 + 0. respectively. 82MPa, 9.02 + 0.87MPa, group C: Ca, Cb, Cc group were 9.67 + 2.42MPa, 10.67 + 2.07MPa, 11.08 + 1.61MPa, D group: Da, Db, 11.16 +, 13.36 + and 13.36. (2) the maximum stress comparison in the group: compared with Bb, the stress increased significantly (P=0.026) compared with Bb (P=0.026). Compared with the Bc 3 groups, there were statistical differences (P=0.015); Ca and Cb had statistical differences (P=0.010), while Cb and Cc group had no significant difference (P=0.235). There was a statistical difference (P=0.01). (3) the young modulus of the group was compared: the young's modulus of the simple suture group increased as time went on, increased slowly after sixth weeks, and the CXL was the strongest in seventh days after the suture, while the CXL after the suture increased, but the change was slow.Ba, Bb, and Bc 3 groups, Ba and Bb were statistically different (P=0.008), Bb B compared to B. There was no statistical difference in C (P=0.264); there was no statistical difference between Ca and Cb (P=0.364) compared with Cb in Ca, Cb and Cc 3 groups, and there was no statistical difference between Cb and Cc (P=0.707). (4) the maximum stress comparison between groups: in general, the scar formation after simple suture should be compared. There was no statistical difference between Ba and Ca (P=0.181) compared to the stress.Ba, Ca and Da after CXL seventh days after suture (P=0.181), Ca and Da had no statistical difference compared with Da (P=0.405), Ba compared with Da, and there were no statistical differences compared with those of the 3 groups. Compared to group Bc and Cc, there was no statistical difference (P=0.764), compared with Dc (P=0.010), there was a statistical difference (P=0.010), and there was a statistical difference between Bc and Dc (P=0.005). (5) the young modulus of the group was compared: in general, the anti deformation ability of the scar formed after the simple suture was lower than that of the.Ba on the seventh day CXL after the suture, and compared with the 3 groups. There was no statistical difference (P value =0.000), and there was no statistical difference between Ca and Da group (P=0.249), and there was no statistical difference between Bb and Cb and Da compared with the 3 groups of Bb, Cb, Db. The difference (P=0.040). (6) the results of WB showed that the ratio of the expression of the simple suture group and the suture + seventh day CXL of the rabbit cornea was 0.28 + 0.11 and 0.65 + 0.20, respectively, and the difference was statistically significant (P=0.048). Conclusion: (1) the biomechanical properties of the scar formed after corneal perforation suture in rabbits increased with time, this experiment The strongest in the eighth week. (2) CXL can enhance the biomechanical properties of the rabbit corneal perforation scar formation scar. The anti biomechanical properties gradually increase with time, and CXL is better than immediate CXL at seventh days after the wound suture of the rabbit cornea. It may prevent the occurrence of corneal staphyloma. (3) CXL can increase the secretion of alpha -SMA after the penetrating injury of the cornea.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R779.1

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