Microtophographies and phosphodiesterase inhibitors to improve the regenerative capabilities of a chitosan membrane. an in-depth study on nervous cells
Authors:
Federica Fregnan (1), Luisa Muratori (1), Federica Zen (1), Marwa El Soury (1), Ilaria Tonazzini (3), Luca Scaccini (3), Francesco Porpiglia (2), Matteo Manfredi (2), Cristian Fiori (2), Stefano Geuna (1), Stefania Raimondo (1)
Organisations:
1: Department of Clinical and Biological Sciences, and Cavalieri Ottolenghi Neuroscience Institute, University of Turin.
2: Division of Urology, Department of Oncology, and Department of Clinical and Biological Sciences, San Luigi Gonzaga Hospital, University of Turin
3: NEST (National Enterprise for nanoScience and nanoTechnology), Istituto Nanoscienze-CNR & Scuola Normale Superiore, Pisa
Presenting author:
Federica Fregnan
Exposition date:
Thursday, September 14, 2023 (Satellite Events Day)
Exposition position:
96
The treatment of localized prostate cancer is radical prostatectomy with frequent iatrogenic damages to the periprostatic neurovascular bundles (NVB) responsible of erectile dysfunctions. Our team previously demonstrated the neuro-regenerative effect of chitosan membrane on ex vivo cultures of autonomic ganglia and its anti-proliferative effect on metastatic prostatic cancer cells.
The aim of the present study was to test in vitro and to develop a functionalized microstructured chitosan membrane to support and promote the main regenerative mechanisms, underlying nerve outgrowth and glial cell survival and proliferation for an in vivo use, to repair a lesion affecting the cavernous nerve. The chitosan was blended with 5-15% glycerol and micropatterned to obtain the well-established gratings (GR) and the asymmetric pattern with scalene triangles (SCA). Moreover, the controlled release of phosphodiesterase inhibitors (PDEI) was designed to chemically promote nerve regeneration and functional recovery.
The results of in vitro and ex vivo direct cultures on microstructured chitosan membranes demonstrated the oriented growth of neurons, a very important step in making regeneration more effective.
The in vitro protocol for the administration of PDEI (sildenafil-PDE5I and rolipram-PDE4I) was developed and for both stimulations an interesting gene regulation linked to the neuroprotective brain-derived neurotrophic factor (BDNF) in immortalized cultures of sensory and motor neurons was observed. In glial cell cultures, the administration of PDEI resulted in up-regulation of the transcription factor Krox20, which can positively influence the expression of myelin genes, of the proangiogenic Vascular endothelial growth factor (VEGF) and in a decrease of cell migration. Furthermore, the administration of Sildenafil and Rolipram increased the neuritic extension in neuronal populations. Further investigations are underway to deepen the study of the effect of PDEI administration on organotypic cultures of somatic and autonomic ganglia, where neuronal and glial cells co-exist, similarly to what happens in vivo.
