Ha Hyo Shin, Song Kye Yong, Kim Jae Chan
Department of Ophthalmology, Chung-Ang University College of Medicine, Seoul, Korea.
J Korean Med Sci. 2006 Jun;21(3):544-9. doi: 10.3346/jkms.2006.21.3.544.
The purpose of this study is to characterize and compare the ultrastructural changes occurring during the in vivo cultivation of corneal epithelium on amniotic membrane (AM) at several different time points. Corneal burn patients (n=7) with a corneal epithelial defect and severe limbal damage were selected. Initially, AM transplantation with limbal autograft was performed at the acute stage of corneal burn to reconstruct the damaged ocular surface. One to six (mean interval; 3.3+/-1.2) months later, the central part of AM containing an in vivo expanded corneal epithelium was excised and retransplanted in adjacent lesions. The excised epithelium with AM was examined by electron microscopy and immunohistochemical study. By electron microscopy, one and two months after expansion, cultivated epithelium on AM showed an undifferentiated epithelium and an incomplete basement membrane (BM). But, after three months, the cultivated epithelium began to differentiate into a multilayered epithelium with a continuous BM with increased hemidesmosomes. These findings were further confirmed by immunohistochemical study, that cytokeratin K3 was expressed in the cultivated corneal epithelium and newly formed BM was partially positive of collagen IV at three months. At least 3 months may be needed for the proliferation and differentiation of in vivo cultivated corneal epithelium on AM.
本研究的目的是在几个不同时间点对羊膜（AM）上角膜上皮细胞体内培养过程中发生的超微结构变化进行表征和比较。选择了角膜上皮缺损且严重角膜缘损伤的角膜烧伤患者（n = 7）。最初，在角膜烧伤急性期进行带角膜缘自体移植的羊膜移植，以重建受损的眼表。1至6个月（平均间隔；3.3±1.2）后，切除含有体内扩增角膜上皮的羊膜中央部分，并重新移植到相邻病变处。对切除的带有羊膜的上皮进行电子显微镜和免疫组织化学研究。通过电子显微镜观察，扩增1个月和2个月后，羊膜上培养的上皮显示为未分化上皮和不完整的基底膜（BM）。但是，3个月后，培养的上皮开始分化为多层上皮，基底膜连续，半桥粒增加。免疫组织化学研究进一步证实了这些发现，即细胞角蛋白K3在培养的角膜上皮中表达，并且在3个月时新形成的基底膜部分呈IV型胶原阳性。羊膜上体内培养的角膜上皮细胞增殖和分化可能至少需要3个月。
