Project description:The present study shows by in vivo experiments, the impact of RhoA inhibitor treatment using a functionalized alginate scaffold and the beneficial results at the early stage by increasing neurites outgrowth and synaptic contact. However, 7 days after treatment a plateau is obtained in BBB score. We thus performed an in vitro and proteomic associated to systemic biology analyses of secreted factors from ND7/23 DRG cell line incubated with secreted factors from different segments after spinal cord lesion treated wih RhoA inhibitor. We then performed a time course proteomic study from the DRG cells in same conditions as for the secretome. The results taken together allow a better understanding of the mechanisms occurring in both side of the lesion and led a positive impact of the treatment on growth cone guidance in both sides of the lesion (Rostral 1 and Caudal 1). We identified the transcription factors involved in time course at the beginning of the treatment and the growth cone factors (receptors, ligands, signaling pathways) implicated in this time course. We also demonstrate the involvement of immunoglobulins (IgG1, IgG2) in this process. Anti-GFAP seem to bind to its own receptor in DRG cells when neurite outgrowth is initiated and thus modulate the process like in traumatic brain injury. These results open the door of a concomitant treatment at both sides of the lesion with RhoA inhibitor. However, we also pointed that RhoA inhibitor treatment led to an inhibition of several factors implicated in neurogenesis reflecting a global inhibition at term as detected in vivo that reached 2 weeks after treatment. RhoA inhibitor treatment is useful to initiate the neurogenesis process but needs to be supplied by a second treatment for increasing synaptic connections and the neuron reconnections.