Abstract:
Current modalities for periodontal regeneration provide modest success, however complete periodontal regeneration is still not achievable. Recently in vitro synthesized nucleoside-modified messenger RNA (mRNA) has emerged as a novel platform in regenerative medicine. The aims of this study are to investigate the ability of human periodontal ligament cells (PDLCs), clinically relevant target cells, to produce bone morphogenetic protein-2 (BMP-2), a significant protein for bone formation after transfected with modified mRNA that encode this protein. We investigated the ability of translated protein for enhancing PDLC proliferation and promotes endothelial cell tube formation, marker of angiogenesis. Isolated PDLCs from healthy periodontal tissue were transfected with N1-methylpseudouridine modified mRNA encoding BMP-2 complexed with transfecting agent, Lipofectamine 2000. Cell lysates and supernatants were collected at 24, 48 and 72 hours (h) after transfection for protein production by ELISA and cell viability by AlamarBlue assay.
High levels of BMP-2 production were detected intracellulary and extracellulary. Secreted BMP-2 gradually increased up to 72 h. Cell viability was maintained above 90% throughout the observation period. The post-transfection supernatants were able to promote PDLC proliferation and endothelial cell tube formation. In conclusion, transfection of PDLCs with N1-methylpseudouridine modified mRNA encoding BMP-2 in lipofectamine 2000 led to high levels of functional BMP-2 protein. Using the in vitro synthesized nucleoside-modified mRNA may allow future application as novel therapeutics platform for periodontal regeneration, however further researches are required.
