The conjunctiva is a clear tissue covering the white part of the eye and lines the back of the eyelids. Conjunctival diseases, such as symblepharon, cause inflammation, discharges, and photophobia. The treatment often requires excision of large parts of conjunctiva. Tissue engineering of conjunctival cells using human amniotic membrane (HAM) denuded of its epithelium as a basement membrane scaffold has been shown to be effective for covering conjunctival defects. However, most epithelial denudation protocols are time‐consuming and expensive or compromise HAM's basement membrane structure and matrix components. We have previously described a method to de‐epithelialize HAM using ice‐cold urea (uHAM). In this report, we used this method to provide tissue‐engineered constructs with cultivated conjunctival epithelial cells on uHAM in two patients, one with a giant conjunctival nevus and the other with a large symblepharon. Autologous conjunctival epithelial cells harvested from incisional biopsies of these two patients were cultured on the uHAM scaffold. The transplantation of tissue‐engineered constructs to patients' ocular surface immediately after the removal of lesions showed successful reconstruction of the ocular surface. Postoperatively, there were neither recurrence of lesions nor epithelial defects throughout the follow‐up (up to 7 and 19 months, respectively). This report highlights the translational potential of an efficient and inexpensive method to prepare de‐epithelialized HAM as a basement membrane scaffold for cell‐based tissue‐engineered treatments of ocular surface disorders. stem cells translational medicine 2019;8:620&626
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