Abstract
Purpose :
Chemically burned cornea requires appropriate measures for preventing inflammation and tissue damage, as well as for being repaired. Here we investigated an artificial corneal graft (ACG) based on lower lip mucosa stem cells (LMSC). The aim of this study was to characterize cells derived from lower lip mucosa in two species, including human and rabbit, as well as to understand if ACG can promote corneal restoration.
Methods :
Both human and rabbit LMSCs were characterized by several criteria, especially in terms of their proliferation capacity, differentiation ability, genomic stability, and immunophenotypic characterization. The proliferation rate of LMSCs was evaluated by producing their growth curves. Osteogenic, chondrogenic, and adipogenic differentiation potential was examined by culturing LMSCs in differentiation media for several weeks and after that von Kossa, safranin-O, and Oil Red O staining, respectively. Genomic stability was evaluated by producing their karyotypes, and the percentage of chromosomal rearrangements was assessed. The abundance of ABCB5, ABCG2, ALDH3A1, CK3, CK14, and CK15 was examined by immunocytochemistry. Finally, a decellularized human amniotic membrane (HAM) was used as a scaffold, and rabbit LMSCs were seeded on it in a density of 100000 cells/cm2. In vivo, the ability of ACG to repair damaged cornea was evaluated in a limbal stem cell deficiency (LSCD) rabbit model. Inflammation and corneal wound healing were examined by histological analysis. Untreated cornea and cornea with total LSCD acted as a control.
Results :
LMSCs were characterized at 6 passages. Among both human and rabbit LMSCs grew at the highest speed. The population doubling time of human and rabbit LMSCs was 36.22±0.57 and 54.78±1.3 hours (p<0.05), respectively. It was found that LMSCs were able to differentiate toward osteogenic, chondrogenic, and adipogenic lineages. However, rabbit LMSCs differentiation potential was slightly lower. The genome of both human and rabbit LMSCs was stable with the chromosomal rearrangements number of 6.67% and 10%, respectively. Immunocytochemistry showed that both human and rabbit LMSCs expressed epithelial and stem cell markers. In vivo, it was found that the transplantation of ACG promoted corneal reepithelization via 1 month.
Conclusions :
Thus, our results suggest that ACG based on HAM and LMSCs are able to repair the corneal structure and decrease inflammation after being wounded.
This is a 2021 ARVO Annual Meeting abstract.