新鲜和深低温保存人羊膜重建兔眼结膜的研究

The study on fresh or preserved human amniotic membrane transplantation for rabbit conjunctival reconstruction

目的对比新鲜与深低温保存人类羊膜重建兔眼结膜后的组织改变,分析新鲜羊膜在重建结膜中的应用价值。方法 24只新西兰白兔等分成两组。剪去双眼上方球结膜形成结膜缺损,一组右眼行深低温保存人羊膜移植,另一组右眼行新鲜人羊膜移植,两组的左眼行自体球结膜移植。于术后1、3、5、12周处死动物,每组3只,连带巩膜切除术区,进行组织学(HE和PAS)检查,12周标本行透射电镜检查。光镜下对术区炎症细胞计数,数据统计学处理。结果新鲜羊膜植片在术后早期(最少1周)自身羊膜上皮细胞存活;3周后,新鲜或保存羊膜植片皆被有杯状细胞分化的结膜上皮覆盖;12周时羊膜被吸收,植片成为正常眼表的一部分;新鲜和保存羊膜移植在术后早期炎症反应高于球结膜移植眼(p<0.05),又以新鲜羊膜移植者略强烈(p<0.05),但无任何治疗的情况下,皆可自行平息。结论人羊膜作为一种异种异体移植材料,可成功重建兔眼结膜。新鲜羊膜在正确取材、清洗彻底的条件下,可以与深低温保存羊膜同样应用于结膜重建。

Purpose To observe the difference of histological reaction between fresh and preserved human amniotic membrane (HAM) transplantation, and analyze the application value of the fresh amniotic membrane for conjunctival reconstruction. Methods 24 New Zealand albino rabbits were divided into two groups. The preserved HAM were transplanted on the sclera of right eyes as group A, and the fresh HAM were transplanted on the right eyes as group B. The left eyes of both groups were transplanted autogenic conjunctiva.Eyes were enucleated 1 week, 3 weeks,5 weeks and 12 weeks (n= 3 for each) after surgery respectively,and they were examined histologically with staining of HE and PAS. The specimens of 12 weeks after surgery were examined with transmission electronic microscopy .The inflammatory cells were calculated with light microscopy. All the data were analyzed wity SPSS. Results 1 week after transplantation, the HAM firmly attached on the sclera in group A, with the dead epithelial cells remaining on its surface. While the amniotic epithelium was alive in group B. 3 weeks later, conjunctiva! epithelization was completed on the HAM in both groups. Abundant PAS - positive goblets were recognized. After 12 weeks, the amnion dissolved in both groups. On transmission electronic microscopy, the cell polarity was prominent. The epithelial cells contained a large number of mitochondria and endoplasmic reticulum. Microvilli were pronounced on the apical cell membrane. There were many intracellular desmosomes and hemidesmosomes. In a short time after surgery, the inflammation of all the right eyes were more apparent than controls (p < 0.05) .During the third week to the fifth week after surgery,the inflammation of the right eyes in group B was more intense than that of the same eyes in group A (p < 0.05) . Conclusion As a kind of xenograft, human amniotic membrane can reconstruct the rabbit eye surface successfully. If the HAM is cleaned and prepared carefully, the fresh amniotic membrane can be used for conjunctival reconstruction.