![]() ![]() ![]() Further investigation revealed that the NF-κB/NLRP3/procaspase-1/IL-1β signaling pathway played a key role in LPS pre-Exo-mediated regulation of macrophage polarization. Flow cytometric analysis of BMDMs showed that LPS pre-Exo were involved in the regulation of macrophage polarization and immune homeostasis during inflammation. In vivo, LPS pre-Exo significantly attenuated inflammatory infiltration, thus improving the survival of allogeneic skin graft. These loaded factors inhibited the expressions of inflammatory factors via a negative feedback mechanism. LPS pre-Exo could better ablate inflammation compared to untreated MSC-derived exosomes (BM-Exo). The expressions of downstream inflammatory cytokines were determined by Elisa, and the polarization of macrophages was analyzed by flow cytometry. The molecular signaling pathway responsible for modulating inflammation was examined by Western blotting. Specifically, bone marrow-derived macrophages (BMDMs) were isolated and cultured with LPS (100 ng/ml) for 3 h, and were further treated with LPS pre-Exo for 24 h or 48 h. For in vitro, an inflammatory model was established. The differentiation of macrophages in the spleen was analyzed by flow cytometry. The accumulation and polarization of macrophages were examined by immunohistochemistry. LPS pre-Exo and rapamycin were injected via the tail vein into C57BL/6 mice allografted with BALB/c skin graft survival was observed and evaluated. The exosomes were isolated from the supernatant of MSCs pretreated with LPS. This study investigated whether exosomes from LPS pretreated bone marrow mesenchymal stem cells (LPS pre-MSCs) could prolong skin graft survival.
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