目的比较DX保存液与甘油保存液对人角膜基质透镜的保存效果。
方法收集2019年2—5月于山东第一医科大学附属青岛眼科医院在飞秒激光小切口角膜基质透镜取出术中取出的中高度近视患者30例60眼的完整角膜基质透镜标本，采用随机数表法将标本分为正常对照组、DX保存液保存1 d组、DX保存液保存1周组、甘油保存1 d组、甘油保存1周组和甘油保存2周组，每组10个标本。采用锥虫蓝染色法检测不同保存方法角膜基质透镜细胞死亡数；光学显微镜下观察角膜基质透镜形态结构的完整性；透射电子显微镜下观察保存的角膜基质透镜超微结构。
结果DX保存液保存1 d组、DX保存液保存1周组、甘油保存1 d组、甘油保存1周组和甘油保存2周组细胞死亡数分别为(53.1±14.2)、(50.8±9.8)、(70.4±13.6)、(172.8±31.7)和(182.8±14.2)个/视野，总体比较差异有统计学意义( F＝16.37， P<0.05)；DX保存液保存1 d组与DX保存液保存1周组角膜基质透镜死亡细胞数量差异无统计学意义( P>0.05)；甘油保存1 d组细胞死亡数明显少于甘油保存1周组和甘油保存2周组，甘油保存1周组死亡细胞数明显多于DX保存液保存1周组，差异均有统计学意义(均 P<0.05)。正常对照组角膜基质透镜胶原纤维排列规则；DX保存液保存1 d组和DX保存液保存1周组角膜基质组织胶原纤维排列较整齐，细胞较完整，甘油保存1 d组角膜基质组织水肿，细胞核裸露，随着甘油保存时间的延长，组织内胶原纤维逐渐变疏松。透射电子显微镜下可见DX保存液保存1 d组和DX保存液保存1周组角膜基质组织紧密，胶原纤维致密，细胞核完整；甘油保存1周组细胞分布稀疏，细胞结构不完整，甘油保存2周组细胞分布明显稀疏，细胞结构破坏。
结论人角膜基质透镜用DX保存液保存1周可有效保存角膜基质组织和维持细胞结构。DX保存液对角膜基质透镜的保存效果优于甘油保存液。
ObjectiveTo compare the preservation effect of DX preservation solution and glycerin preservation solution on the corneal stromal lens.
MethodsSixty intact corneal stromal lens samples were collected during femtosecond small incision lenticule extraction (SMILE) from 60 myopic eyes of 30 subjects at Qingdao Eye Hospital of Shandong First Medical University from February 2019 to May 2019.The samples were randomized into DX preserved 1-day group, DX preserved 1-week group, glycerin preserved 1-day group, glycerin preserved 1-week group and glycerin preserved 2-week group according to the different preservation methods, with 10 samples in each group.No intervention was done in the samples of the normal control group.Trypan blue staining was used to count the number of dead cells in the corneal stromal lens.The morphological structure of the corneal stromal lens was examined with an optical microscope, and its ultrastructure was observed under the transmission electron microscope.This study adhered to the Declaration of Helsinki.Written informed consent was obtained from each patient prior to any medical intervention.The study protocol was approved by an Ethics Committee of Qingdao Eye Hospital of Shandong First Medical University (No.2019-30).
ResultsThe number of dead cells was (53.1±14.2), (50.8±9.8), (70.4±13.6) and (172.8±31.7) and (182.8±14.2) cells/field in the DX preserved 1-day group, DX preserved 1-week group, glycerin preserved 1-day group, glycerin preserved 1-week group and glycerin preserved 2-week group, respectively, showing a significant difference among the five groups ( F=16.37, P<0.05). There was no significant difference between the DX preserved 1-day group and 1-week group ( P>0.05). The number of dead cells was significantly less in the glycerin preserved 1-day group than that of the glycerin preserved 1-week group and glycerin preserved 2-week group, and the number of dead cells was significantly increased in the glycerin preserved 1-week group compared with the DX preserved 1-week group (all at P<0.05). The arrangement of collagen fibers of the corneal stromal lens was regular and the cells were intact in the normal control group, DX preserved 1-day group and DX preserved 1-week group.The tissue edema, bare cell nuclei and loose collagen fibers were found in the samples in the glycerin preserved 1-day group.The corneal stromal lens was compact and the collagen fibers were dense and the nuclei were intact in the DX preserved 1-day group and DX preserved 1-week group.The distribution of the cells was sparse and the cell structure was abnormal under the transmission electron microscope in various glycerin preserved groups.
ConclusionsThe structure of corneal stromal lens can be well preserved for one week by DX storage solution.The preservation effect of DX solution is better for fresh human corneal stromal lens than glycerin solution.
田乐,李德卫,彭予苏,等. DX保存液与甘油保存液对人角膜基质透镜保存效果的比较[J]. 中华实验眼科杂志,2022,40(02)：126-132.
DOI:10.3760/cma.j.cn115989-20200603-00393版权归中华医学会所有。
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田乐：酝酿和设计实验、实施研究、采集数据、分析/解释数据、统计分析、起草文章；李德卫、彭予苏、张飞飞：参与实验，采集数据；陈敏：酝酿和设计实验、实施研究、对文章的知识性内容作批评性审阅
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