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Jing Jie, Jie Yang, Hui He, Hua Q. Xiao, Zhi Y. Li, Jing Y. Chen, Nuo Dong, Hu P. Wu, Zu G. Liu, Wei Li; Tissue Remodelization After Ocular Surface Reconstruction With Intact Or Denuded Amniotic Membrane. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5163.
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© ARVO (1962-2015); The Authors (2016-present)
Intact amniotic membrane (iAM), which contains amniotic epithelial cells, was widely used as permanent graft in ocular surface reconstruction surgery. Previous studies showed that corneal epithelial cells cultured on iAM exhibited different biological characteristics compared with those cultured on denuded amniotic membrane (dAM). However, there was no study investigating the efficacy of dAM on ocular surface reconstruction and the tissue remodelization after AM transplantation.
Eighteen New Zealand rabbits were performed partial limbal and conjunctival tissue removal (5×5mm in size) in both eyes. The identical size of cryo-preserved human dAM and iAM were then transplanted in hibateral eyes respectively. The epithelial growth on the amniotic membrane was evaluated by fluorescent staining. The expression of basement membrane components type VII collagen and Laminin 5, epithelial differentiation markers such as K19 and K12 cytokeratins, and infiltration of macrophages were investigated by immunostaining. PAS staining was performed to determine goblet cell differentiation. Clinically, twenty pterygium patients were performed with pterygium excision and AM transplantation, 10 of them were transplanted with iAM, while other 10 with dAM. The epithelial growth on the amniotic membrane was also evaluated by fluorescent staining.
The ocular surface was successfully reconstructed 2 weeks after AM transplantation both in rabbit model and pterygium patients. The growth rate of limbal and conjunctival epithelium was significantly faster on dAM than that on iAM both in rabbit and human. H&E staining of rabbit corneal and conjunctival tissue after AM transplantation showed prominent stratification of epithelium on dAM. PAS staining showed more goblet cells on dAM at 2 weeks after surgery. Basement membrane components collagen VII and Laminin 5 were degraded in dAM within 2 weeks post-surgery, while still remained in iAM. There was less macrophage infiltration in dAM transplantation group. K12 and K19 expression was present in epithelial cells spread on iAM or dAM, and showed no significant difference.
dAM facilitates epithelial repopulation and goblet cell differentiation during ocular surface reconstruction.
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