Mimura Tatsuya, Yamagami Satoru, Usui Tomohiko, Yokoo Seiichi, Ono Kyoko, Honda Norihiko, Kaneda Atsuko, Sugisaki Kenji, Sayegh Rony R, Amano Shiro
Department of Ophthalmology, University of Illinois at Chicago, 1855 W. Taylor Street, Chicago, IL 60612, USA.
Curr Eye Res. 2007 May;32(5):407-11. doi: 10.1080/02713680701299342.
Although sheet transplantation with cultured corneal limbal epithelium has been widely performed as a strategy for ocular surface reconstruction, there has been no optimal method for evaluating the morphology of these sheets prior to transplantation. We propose the use of in vivo confocal microscopy as a novel method for the evaluation of limbal corneal epithelium cultured on amniotic membrane.
Human limbal epithelial sheets were grown on amniotic membranes by following a standard protocol and were stained with hematoxylin and eosin. Morphology was studied using in vivo confocal microscopy for cultured corneal epithelium on amniotic membrane, human intact amniotic membranes, and epithelium-denuded human amniotic membranes.
Histologic examination showed a stratified corneal epithelium sheet by the fourth week of culture. The surface and basal layers of the cultured limbal epithelium and amniotic membrane were clearly distinguished by in vivo confocal microscopy. A monolayer of amniotic epithelial cells was observed on the intact amniotic membrane, but not on the epithelium-denuded human amniotic membrane.
Our findings support the use of in vivo confocal microscopy as a valid technique for the preoperative evaluation of cultured corneal limbal epithelial cell sheets on amniotic membrane.
尽管培养的角膜缘上皮片移植作为眼表重建的一种策略已被广泛应用，但在移植前尚无评估这些上皮片形态的最佳方法。我们提出将活体共聚焦显微镜检查作为评估羊膜上培养的角膜缘上皮的一种新方法。
按照标准方案在羊膜上培养人角膜缘上皮片，并用苏木精和伊红染色。使用活体共聚焦显微镜检查研究羊膜上培养的角膜上皮、完整的人羊膜和上皮剥脱的人羊膜的形态。
组织学检查显示培养至第4周时形成分层的角膜上皮片。通过活体共聚焦显微镜检查可清楚区分培养的角膜缘上皮和羊膜的表层及基底层。在完整羊膜上观察到单层羊膜上皮细胞，但在上皮剥脱的人羊膜上未观察到。
我们的研究结果支持将活体共聚焦显微镜检查作为术前评估羊膜上培养的角膜缘上皮细胞片的有效技术。
