In-vivo Cultivation of Limbal Stem Cells for Ocular Surface Reconstruction

To describe a novel surgical technique for in vivo limbal stem cell cultivation for reconstruction of corneal epithelium in total limbal stem cell deficiency (LSCD). Patients suffering from chronic total unilateral LSCD due to a chemical injury were sequentially included. One small limbal block (2x1 mm) harvested from contralateral healthy eye sutured over the amniotic membrane (AM) to the superior limbal area of the injured eye. Amniotic membrane extract eye drop (AMEED) at the concentration of 0.1 mg/ml was regularly used until total corneal epithelialization was achieved. All eyes underwent sequential penetrating keratoplasty (PKP) for optical indication and the corneal button was sent for histopathologic examinations and molecular analysis for the detection of limbal stem cell markers. Five eyes of 5 patients with a mean age of 32 years were operated. Mean follow up period was 12 months. Total epithelialization of the corneal surface was achieved in 2 weeks in all eyes. Corneal conjunctivalization and vascularization regressed dramatically after 3 to 4 weeks. Transplanted corneas were clear at the last follow-up examination. No complication developed in donor and recipient eyes. Laboratory examinations showed limbal stem cell markers over the corneal buttons. In vivo cultivation of limbal stem cells over the AM using AMEED is an effective procedure for ocular surface reconstruction in patients with unilateral total LSCD. However, its long term results should be confirmed in studies with a longer follow-up period.