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• W3048663854 abstract "Background: Stem cells isolated from dental pulp (DPSCs) are characterized by a high rate of proliferation, low immunogenicity and a high ability to differentiate in different lineages (i.e. osteogenic, chondrogenic, adipogenic, myogenic and neural commitment). Their multipotency can be attributed to the peculiar embryological origin from the neural crest. DPSCs represent a promising stem cell resource since they hold a low ethical impact and can be easily isolated through routine dental procedures. These cells own immuno-modulatory properties, exerted through the activation of different mechanisms, including the Fas / FasL pathway, as well as through the release of soluble factors. Currently, other molecular mechanisms are under consideration such as PD-1 / PD-L1 (Programmed Death 1 and its Ligand) which are supposed to be involved in the induction and / or maintenance of immune tolerance. Objectives: The aim of this research was to investigate whether the stimulation of PD-L1 in DPSCs can affect the immunomodulatory effects of these stem cells on peripheral blood mononuclear cells (PBMCs). Furthermore, the expression of PD-L1 was also assayed after the induction of osteogenic differentiation of DPSCs in order to evaluate a possible application of DPSCs in autoimmune inflammatory osteo-erosive diseases. Methods: Immuno-selection was performed on DPSCs, isolated from waste material, against the stemness markers c-Kit and STRO-1, to obtain a pure stem cell population. Then, STRO-1+/c-Kit+ DPSCs, were co-cultured either directly and indirectly with peripheral blood mononuclear cells (PBMCs) from healthy adult donors, previously activated by anti-CD3 and anti-CD28 antibodies. Co-cultures of PBMCs with amniotic fluid stem cells (AFSCs) and bone marrow mesenchymal stem cells (BM-MSCs) were also set up. The expression of PD-1 in PBMCs as well as of PD-L1 in DPSCs, AFSCs, BM-MSCs and PBMCs, was evaluated by Western Blot (WB) and immunofluorescence (IF) analyses, before and after osteogenic differentiation. Osteogenic differentiation of DPSCs, after 30 days of induction, was verified by IF and WB, of osteopontin, osteocalcin and RUNX2 markers. Interleukin-2 (IL-2) expression levels in PBMCs were analyzed by Real-Time PCR analysis. Results: Our data highlight that, after direct and indirect co-culture with activated PBMCs, PD-L1 expression was up-regulated not only in DPSCs, but also in BM-MSCs and AFSCs (Figure 1), thus suggesting that 1) this is a common ability of mesenchymal stem cells and 2) this event can be also mediated by soluble factors release. Moreover, when evaluating the effects of DPSCs co-culture on PBMCs an increased expression of cleaved caspase 3 was observed, together with a decreased expression of IL-2 - a growth factor essential for the proliferation and survival of T cells (Figure 2). These findings showed how DPSCs can modulate the immune system by PD-L1 up-regulation. On the other hand, it is noteworthy that, after reaching osteogenic commitment, DPSCs down-regulated the expression of PD-L1, allowing to hypothesize that PD-L1 expression is strictly related to the maintenance of stemness. Figure 1. Figure 2. Conclusion: Taken together, our findings suggest that the expression of PD-L1 in DPSCs is involved in the modulation of immune response and pave the way for further investigations on the role of PD-1/PD-L1 pathway in controlling inflammation and immune response when applied to the treatment of autoimmune inflammatory diseases. References: [1]Keir ME, et al. PD-1 and its ligands in tolerance and immunity. Annu Rev Immunol. 2008 Disclosure of Interests: None declared" @default.
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• W3048663854 date "2020-06-01" @default.
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• W3048663854 title "AB0039 ROLE OF MESENCHYMAL STEM CELLS ISOLATED FROM DENTAL PULP (DPSCS) IN IMMUNOREGULATION PROCESSES MEDIATED BY PROGRAMMED DEATH-LIGAND 1 (PD-L1)" @default.
• W3048663854 doi "https://doi.org/10.1136/annrheumdis-2020-eular.3696" @default.
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