To describe a novel surgical technique of in vivo limbal stem cell cultivation for reconstruction of corneal epithelium in unilateral total limbal stem cell deficiency (LSCD).

Patients suffered of chronic total unilateral LSCD due to a chemical injury were sequentially included. One small limbal block (2x1mm) harvested from contralateral healthy eye sutured over the amniotic membrane (AM) on the superior limbal area of the injured eye. Amniotic membrane extract eye drop (AMEED) at the dose of 0.1 mg/ml was regularly used until cell proliferation was started. The frequency of drop decreased until the total corneal epithelialization was achieved. All eyes underwent sequential penetrating keratoplasty (PKP) for optical results and the corneal button analyzed for histopathologic examinations and molecular analysis for detection of limbal stem cells.

: Five eyes of 5 patients with a mean age of 32 years were operated. Mean follow up period was 12 months. In all eyes total epithelialization of the corneal surface was occurred in 2 weeks. Corneal conjunctivalization and vascularisation decreased dramatically 3 to 4 weeks after surgery. Sequential PKP was performed in all eyes. At the last follow up visit all transplanted corneas were clear. No complication developed in donor and recipient eyes. Labarotary 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, it’s long term results should be confirmed in studies with longer follow ups .