Espana E M, Ti S-E, Grueterich M, Touhami A, Tseng S C G
TissueTech, Inc. and Ocular Surface Center, Miami, FL 33173, USA.
Br J Ophthalmol. 2003 Dec;87(12):1509-14. doi: 10.1136/bjo.87.12.1509.
To study corneal stromal changes and the presence of myofibroblasts after transplantation of ex vivo expanded limbal epithelium.
A state of limbal deficiency was induced in 16 rabbits. After transplantation with autologous ex vivo expanded limbal epithelium on amniotic membrane (AM), their clinical outcomes were classified as success, partial success or failure according to surface smoothness, stromal clarity, and vascularisation. Clinical outcomes were correlated with phenotypic outcomes of corneal, conjunctival, or mixed epithelium, defined by expression of K3 keratin or MUC5AC. Immunostaining was performed with antibodies against collagen IV, fibronectin, and alpha-smooth muscle actin (alpha-SMA) to assess stromal wound remodelling.
Rabbits were sacrificed after a mean follow up of 10 (SD 3.3) months. Collagen IV, expressed in the basement membrane of all three groups, was found in the stroma of the partial success, but not in that of the success or the failure. Fibronectin was absent in the success and the failure, but expressed in the stroma of the partial success. Alpha-SMA was expressed in superficial stroma of the partial success, but suppressed in areas with AM remnants.
Restoration of a clear and transparent cornea is associated with a normal corneal epithelium and complete wound remodelling. In contrast, wound healing remains active and incomplete in conjunctivalised corneas, which remain opaque with myofibroblasts.
研究体外扩增的角膜缘上皮移植后角膜基质的变化及肌成纤维细胞的存在情况。
诱导16只兔出现角膜缘缺陷状态。在羊膜（AM）上移植自体体外扩增的角膜缘上皮后，根据表面光滑度、基质清晰度和血管化情况将其临床结果分为成功、部分成功或失败。临床结果与由K3角蛋白或MUC5AC表达定义的角膜、结膜或混合上皮的表型结果相关。用抗IV型胶原、纤连蛋白和α-平滑肌肌动蛋白（α-SMA）的抗体进行免疫染色，以评估基质伤口重塑情况。
平均随访10（标准差3.3）个月后处死兔子。IV型胶原在所有三组的基底膜中均有表达，在部分成功组的基质中也有发现，但在成功组或失败组的基质中未发现。纤连蛋白在成功组和失败组中不存在，但在部分成功组的基质中表达。α-SMA在部分成功组的浅层基质中表达，但在有AM残余物的区域受到抑制。
透明角膜的恢复与正常角膜上皮和完全的伤口重塑有关。相比之下，结膜化角膜中的伤口愈合仍然活跃且不完全，角膜仍不透明且存在肌成纤维细胞。
