Synergy effect of SMILE lenticule and conditional medium to induce the differentiation of mesenchymal stem cells into keratocytes
Mohsen Ghiasi1 , Mehrdad Hashmi2 , Khosrow Jadidi3 , Hossein Aghamollaei4 *
- Department of Molecular and Cellular Sciences, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
- Department of Genetics, Faculty of Advanced Science and Technology, Islamic Azad University, Tehran Medical Sciences, Tehran, Iran.
- Vision Health research Center, Semnan university of medical sciences, Semnan, Iran
- Chemical Injures Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Abstract: Keratocytes are quiescent cells that are not easily cultured. Differentiation of stem cells into keratocytes has been proposed as an alternative method. The aim of this study was to investigate the differentiation potential of adipose derived mesenchymal stem (ADMSC)cells into corneal keratocyte cells by combination of SMILE lenticule scaffold and conditional medium
Methods: conditioned medium was collected from keratocytes culture media. (ADMSC) were cultured on the decellularized human SMILE lenticule (SL), and were exposed to keratocyte-conditioned medium for 21 days. Differentiation was evaluated using Real-time PCR and immunocytochemistry (ICC). ADMSC were cultured on the SL scaffolds and after 21 days of induction of differentiation, they were implanted in the corneal stroma of 8 New Zealand male rabbits. Rabbits were followed for 3 months and, the safety was evaluated by clinical and histological variables.
Results: After culture ADMSC on the surface of SL in the presence of keratocyte-conditioned medium, Real-time PCR and ICC analysis showed a significant increase in the expression of keratocytes specific markers including keratocan, lumican, ALDH3A1, and CD34 compared to the control group (p<0.05). After implantation of SL containing differentiated cells in the animal cornea, no serious complications including angiogenesis, corneal opacity, inflammation, or signs of tissue rejection were observed.
Conclusion: The use of cells differentiated by combination of SL and conditional medium in the stroma of the cornea of the animal is safe and has no serious side effects. After more evaluations, this cells can be used in corneal stromal regeneration.
