羊膜移植于角膜基质内的转归及其影响因素
Outcome and influencing factors of amniotic membrane transplanted into corneal stroma

摘要：目的 研究羊膜移植于角膜基质内的转归并探讨其影响因素。方法 取15只新西兰大白兔，将二氯三苯胺荧光素(dichlorotriazinyl aminofluoresclin, DTAF)标记的羊膜埋植于角膜基质内。术后进行临床观察，并于4、8、13、17、22周取材做HE染色观察组织病理学改变,激光共聚焦显微镜检查追踪羊膜踪迹，以及22周取材做角膜透射电镜检查，观察超微结构。 结果 HE染色术后4周可见多层均质红染的羊膜组织弯曲排列，结构保留完整，基底膜和实质层未溶解。术后8周，羊膜组织仍呈均质红染的条状结构，实质层较完整。术后13周羊膜形态渐显模糊，但仍呈条带状，少量角膜细胞聚集在羊膜周围。术后17周明显可见碎片状的羊膜组织。术后22周角膜基质内存留小片状羊膜组织。激光共聚焦显微镜发现羊膜在角膜基质内可以存留22周以上。术后22周角膜透射电镜可见整合于角膜基质中的羊膜基质胶原纤维成分较不规则，羊膜周围角膜基质超微结构无明显改变。结论 羊膜移植于角膜基质内存留时间可长达22周以上,羊膜可以融合于角膜组织中，作为替代材料为行角膜移植手术争取了时间。
ABSTRACT: Objective To investigate the turnover and factors influencing amniotic membrane transplantation into corneal stroma. Methods Amniotic membrane stained with dichlorotriazinyl aminofluoresclin (DTAF) was implanted into the corneal stroma of 15 New Zealand white rabbits. After transplantation, we observed the clinical changes. The tissue samples from grafted area were observed with HE staining and laser confocal microscope to trace amniotic membrane at 4, 8, 13, 17 and 22 week after surgery. And the tissue samples at week 22 after surgery from grafted area were also observed by transmission electron microscopy. Results After 4 weeks of operation, amniotic membrane tissue bent with visible multi-layer homogeneous red dye. The structure remained intact; basement membrane and stroma were no dissolved. After 8 weeks, amniotic tissue was a strip structure of homogeneous red dye, and the substance layer was complete. After 13 weeks, amniotic membrane morphology appeared fuzzy, but still took the shape of strips; a small number of corneal cells accumulated around the amniotic membrane. At 17 weeks after operation, fragmented amnion was evident. At 22 weeks after operation, small pieces of amnion tissue remained in the corneal stroma. Laser scanning confocal microscopy revealed that amniotic membrane could retain for over 22 weeks in the corneal stroma. After 22 weeks, amnion stroma collagen fibers that were integrated into the corneal stroma were irregular. Corneal stromal ultrastructure surrounding the amniotic membrane did not obviously change. Conclusion Retention time of amniotic membrane transplanted into corneal stroma can be as long as over 22 weeks and amniotic membrane can be merged into the corneal tissue as a substitute, which wins time for corneal transplantation operation