Simultaneous utilizing lenticule SMILE and keratocytes-conditioned medium to prompt human adipose-derived mesenchymal stem cells to transform into keratocyte-like cells, and assessing its safety in the rabbit's cornea
Mohsen Ghiasi1 , Khosrow Jadidi 2 , Ali Salimi3 , Hossein Aghamollaei4 *
- Rajaie Cardiovascular Medical and Research Center, Iran University of Medical Sciences, Tehran, Iran.
- Vision Health Research Center, Semnan University of Medical Sciences, Semnan, Iran.
- Tissue Engineering and Regenerative Medicine Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
- Chemical Injures Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Abstract: Background: Keratocytes are important for maintaining the structure and transparency of the cornea, but they are stationary and difficult to culture. The study aimed to investigate the differentiation of human adipose mesenchymal stem cells (hADSCs) into corneal keratocyte cells using SMILE scaffolds and conditioned medium (CM) and evaluate the safety of cell-containing scaffolds in rabbit cornea.
Methods: Method and Material: The hADSCs were cultured on different SMILE and exposed to keratocyte-CM (KCM) for 7, 14, and 21 days. Real-time PCR and immunocytochemistry (ICC) were used to evaluate differentiation. hADSCs were differentiated on the SL scaffolds and inserted into the corneal stroma of 8 male New Zealand rabbits. The rabbits were monitored for 3 months, and safety was assessed using clinical and histological measures.
Results: Results: The study showed that the combination of SMILE and KCM can induce the differentiation of hADSC keratocytes. The safety of the cell-containing scaffolds was confirmed by clinical and histological variables. In addition, Real-time PCR and immunohistochemistry (IHC) analysis confirmed the assessment of keratocyte-like cells' existence in the rabbit stroma after three months. The results of the present study showed that the constructed scaffolds do not cause any serious side effects such as inflammation, neovascularization, corneal opacity, infection or signs of tissue rejection in the cornea of rabbits.
Conclusion: Conclusion: In conclusion, the combined application of corneal extracellular matrix and KCM has the potential to induce the differentiation of hADSC keratocytes. These results suggest a potential alternative approach for generating necessary keratocytes in corneal tissue engineering.
