Raffaele Nuzzi1, Lola Buono2, Simona Scalabrin1, Marco De Iuliis1, Benedetta Bussolati2. 1. Eye Clinic, Department of Surgical Sciences, University of Turin, AOU Città della Salute e della Scienza, Turin, Italy. 2. Department of Biotechnology and Health Sciences, University of Turin, Turin, Italy.
Abstract
PURPOSE: Human corneal endothelial cells (HCECs) are essential to visual function; however, since they have limited proliferative capacity in vivo, they are prone to corneal endothelial dysfunction. At present, the only treatment is a corneal transplantation from donor cadavers. Also, due to a global shortage of donor corneas, it is important to find alternative strategies. Recent studies highlight that stem cell-derived extracellular vesicles (EVs) play a relevant role in stem cell-induced regeneration by reprogramming injured cells and inducing proregenerative pathways. The aim of this work is to evaluate whether EVs derived from mesenchymal stem cells (MSC-EVs) are able to promote regeneration of damaged HCECs. METHODS: We isolated HCECs from discarded corneas in patients undergoing corneal transplantation or enucleation (N = 23 patients). Bone marrow mesenchymal stem cells (MSCs) were obtained from Lonza, cultured, and characterized. MSC-EVs were obtained from supernatants of MSCs. In order to establish a valid in vitro damage model to test the regenerative potential of EVs on HCECs, we evaluated the proliferation rate and the apoptosis after exposing the cells to serum-deprived medium at different concentrations for 24 hours. We then evaluated the HCEC migration through a wound healing assay. RESULTS: In the selected serum deprivation damage conditions, the treatment with different doses of MSC-EVs resulted in a significantly higher proliferation rate of HCECs at all the tested concentrations of EVs (5-20 × 103 MSC-EV/cell). MSC-EVs/cell induced a significant decrease in number of total apoptotic cells after 24 hours of serum deprivation. Finally, the wound healing assay showed a significantly faster repair of the wound after HCEC treatment with MSC-EVs. CONCLUSIONS: Results highlight the already well-known proregenerative potential of MSC-EVs in a totally new biological model, the endothelium of the cornea. MSC-EVs, indeed, induced proliferation and survival of HCECs, promoting the migration of HCECs in vitro.
PURPOSE: Human corneal endothelial cells (HCECs) are essential to visual function; however, since they have limited proliferative capacity in vivo, they are prone to corneal endothelial dysfunction. At present, the only treatment is a corneal transplantation from donor cadavers. Also, due to a global shortage of donor corneas, it is important to find alternative strategies. Recent studies highlight that stem cell-derived extracellular vesicles (EVs) play a relevant role in stem cell-induced regeneration by reprogramming injured cells and inducing proregenerative pathways. The aim of this work is to evaluate whether EVs derived from mesenchymal stem cells (MSC-EVs) are able to promote regeneration of damaged HCECs. METHODS: We isolated HCECs from discarded corneas in patients undergoing corneal transplantation or enucleation (N = 23 patients). Bone marrow mesenchymal stem cells (MSCs) were obtained from Lonza, cultured, and characterized. MSC-EVs were obtained from supernatants of MSCs. In order to establish a valid in vitro damage model to test the regenerative potential of EVs on HCECs, we evaluated the proliferation rate and the apoptosis after exposing the cells to serum-deprived medium at different concentrations for 24 hours. We then evaluated the HCEC migration through a wound healing assay. RESULTS: In the selected serum deprivation damage conditions, the treatment with different doses of MSC-EVs resulted in a significantly higher proliferation rate of HCECs at all the tested concentrations of EVs (5-20 × 103 MSC-EV/cell). MSC-EVs/cell induced a significant decrease in number of total apoptotic cells after 24 hours of serum deprivation. Finally, the wound healing assay showed a significantly faster repair of the wound after HCEC treatment with MSC-EVs. CONCLUSIONS: Results highlight the already well-known proregenerative potential of MSC-EVs in a totally new biological model, the endothelium of the cornea. MSC-EVs, indeed, induced proliferation and survival of HCECs, promoting the migration of HCECs in vitro.
Authors: Rebeca Blazquez; Francisco Miguel Sanchez-Margallo; Olga de la Rosa; Wilfried Dalemans; Verónica Alvarez; Raquel Tarazona; Javier G Casado Journal: Front Immunol Date: 2014-11-04 Impact factor: 7.561
Authors: Stefania Bruno; Marta Tapparo; Federica Collino; Giulia Chiabotto; Maria Chiara Deregibus; Rafael Soares Lindoso; Francesco Neri; Sharad Kholia; Sara Giunti; Sicheng Wen; Peter Quesenberry; Giovanni Camussi Journal: Tissue Eng Part A Date: 2017-06-13 Impact factor: 3.845
Authors: Fatma Y Meligy; Hanan Sharaf El-Deen Mohammed; Tarek M Mostafa; Mohamed M Elfiky; Israa El-Sayed Mohamed Ashry; Ahmed M Abd-Eldayem; Nermin I Rizk; Dina Sabry; Eman S H Abd Allah; Salwa Fares Ahmed Journal: Pharmaceutics Date: 2022-06-22 Impact factor: 6.525