Potenziali applicazioni delle DPSC nella medicina rigenerativa: capacità di differenziamento e supporto nel processo di angiogenesi

Dental pulp has been revealed as an accessible and rich source of mesenchymal stem cells (MSCs), and its biological potential is currently under intense investigation. MSCs from dental pulp stem/stromal cells (DPSCs) have been indicated as a heterogeneous population, originating from the neural crest, and physiologically involved in dentin homeostasis; moreover, they contribute to bone remodeling and differentiation into several tissues including cartilage, bone, adipose, hepatic, and nervous tissues. DPSCs have also been shown to influence the angiogenesis process, for example through the release of secretory factors or by differentiating into vascular and/or perivascular cells. This multifaceted nature underscores the potential therapeutic applications of DPSCs across various fields, ranging from tissue regeneration to vascular pathology. Based on this information, the primary goal of this study was to investigate the capacity of DPSCs to differentiate into various cell types, including osteo-odontogenic, hepatic, neurogenic, and perivascular phenotypes. This investigation utilized both traditional two-dimensional (2D) and advanced three-dimensional (3D) culture methods. The ultimate aim of this research is to showcase the potential versatility of DPSCs for future applications in regenerative medicine research. For the experiments, DPSCs were subjected to various treatments (osteo-odontogenic and hepatic differentiation media, hypoxia at 1%, and MACS separation). At the same time, a stationary technique was employed to generate organoids. The results revealed a significant ability of DPSCs to differentiate into the osteo-odontogenic phenotype. Notably, these cells exhibited high expression levels of genes characteristic of osteo-odontoblasts and produced a matrix containing dentin and calcium phosphates, both in 2D and 3D environments. DPSCs possess the capacity to differentiate in neuronal cells by specific media. Additionally, they can be influenced by the environment to differentiate into specific neuronal cells of the nervous system, such as under hypoxic conditions. In light of this, DPSCs were exposed to hypoxia (1% O2) for 5 and 16 days, revealing that hypoxia-induced DPSCs differentiation was time-dependent. Moreover, conditioned media (CM) derived from DPSCs stimulated by hypoxia had the ability to induce neural differentiation in both SH-SY5Y neuroblastoma cells and undifferentiated DPSCs. This suggests that the differentiation of DPSCs mediated by hypoxia is likely to occur through an autocrine/paracrine mechanism. To investigate the potential to differentiate into the peri-vascular cells were subjected to immunomagnetic separation (MACS) targeting the pericytic marker NG2, resulting in the isolation of three subpopulations of interest (Total, NG2-, and NG2+ DPSCs). The data showed that NG2+ DPSCs, displaying a pericyte-like phenotype, were able to stabilize tubules in vitro for 14 days by directly interacting with endothelial cells. This suggests that DPSCs have the capability to differentiate into different cell types, making them a valuable resource for regenerative medicine. Their potential offers hope for the treatment of various medical conditions.