Stem cells of the oral mucosa during reconstructive corneal surgery under conditions of experimental limbal insufficiency

Session Details

Session Title: Presented Poster Session: Surgical Cornea

Venue: Poster Village: Pod 2

First Author: : I.Gavrilyik RUSSIA

Co Author(s): :    A. Kulikov   S. Churashov   V. Chernysh   O. Aleksandrova   M. Blinova           

Abstract Details

Purpose:

Perform reconstruction of the limbus and Bowman membrane of the cornea with simultaneous re-epithelialization of its surface by transplantation of a bio-engineering structure based on an amniotic membrane with cultured stem cells of the oral mucosa with total experimental limbal insufficiency.

Setting:

1. Department of Ophthalmology. Military Medical Academy named after SMKirov, Ministry of Defense of the Russian Federation, st. Academician Lebedev 6, 194044, St. Petersburg, Russia 2. Institute of Cytology, Russian Academy of Sciences, Tikhoretsky ave. 4, 194064 St. Petersburg, Russia.

Methods:

The study was performed on 10 rabbits (20 eyes). Pre-simulated total limbal insufficiency. After isolating stem cells from biopsies of the oral mucosa, cultivation was performed at 37 ° C in an atmosphere of 5% CO2 for 7–14 days. The cell population was stained for stem markers (ABCB5, ABCG2, ALDH3A1, p63α) and differentiation markers (cytokeratins 3, 5, 12, 14, 15, 19). Cultured stem cells were placed on the amniotic membrane. After 1-3 days, the grafts obtained in this way were transplanted onto the corneal surface of the experienced eyes. In the control, only the amniotic membrane was transplanted.

Results:

Stem cells created multiple colonies that fused to form a monolayer by 7–10 days. The population was positively stained for ABCB5, ABCG2, ALDH3A1 and cytokeratins 3/12 and 15. The p63α factor was found in trace amounts. After surgery, active epithelization was observed in both groups. In the main group, by the 30th day, complete epithelialization of the corneal surface was observed. In the control group, even to 45 days epithelialization did not occur. Impression cytology of the cornea revealed differentiated epithelial cells of the oral mucosa in the first group, and conjunctivalization of the cornea in the second group.

Conclusions:

Trasplantation of the bioengineering construction on the basis of the amniotic membrane with cultured stem cells of the oral mucosa allows the reconstruction of the limb region and the Bowmen membrane of the cornea under conditions of total limbal insufficiency. Further research is needed to achieve a more rapid epithelialization of the corneal surface.

Financial Disclosure:

None