IADR Abstract Archives
Bone Regeneration Induced by microRNA-200c and microRNA-200a Inhibitor
Objectives: To test the effectiveness of miR-200c and miR-200a inhibition on osteogenic differentiation and bone regeneration in vitro and in vivo.
Methods: Plasmid DNA encoding miR-200c and plasmid-based miR-200a inhibitor (PMIS-200a) were transfected into human bone marrow MSCs in vitro. The cells with miR-200c overexpression and miR-200a inhibition were subsequently exposed to osteogenic differentiation media for up to 4 weeks. The biomarkers of osteogenic differentiation were quantitatively measured at different time points. In addition, plasmid miR-200c and PMIS-200a were loaded into collagen sponge at different doses and implanted into calvarial defects at a rat model. The rats were euthanized after 4 weeks and the bone regeneration with different treatments were analyzed using µCT and histology.
Results: Overexpression of miR-200c and PMIS-200a in human bone marrow MSCs were confirmed after the cells were treatd with plasmid miR-200c and PMIS-200a in vitro. The biomarkers of osteogenic differentiation, including ALP, Runx2, and OCN, were up-regulated in the cells , compared to the non-treated cells and the cells treated with empty vectors. Bone formation quantitatively measured using µCT and histology in the defects treated with miR-200c and PMIS-200a are significantly higher than those receiving collagen sponge alone or empty vectors. The newly formed bone volume treated with miR-200c and PMIS-200a are twice higher than the controls.
Conclusions: These data demonstrated that miR-200c and miR-200a inhibition potently improve osteogenic differentiation of human MSCs and promote bone formation, which may potentially be used to restore oral and craniofacial bone defects clinically.
Methods: Plasmid DNA encoding miR-200c and plasmid-based miR-200a inhibitor (PMIS-200a) were transfected into human bone marrow MSCs in vitro. The cells with miR-200c overexpression and miR-200a inhibition were subsequently exposed to osteogenic differentiation media for up to 4 weeks. The biomarkers of osteogenic differentiation were quantitatively measured at different time points. In addition, plasmid miR-200c and PMIS-200a were loaded into collagen sponge at different doses and implanted into calvarial defects at a rat model. The rats were euthanized after 4 weeks and the bone regeneration with different treatments were analyzed using µCT and histology.
Results: Overexpression of miR-200c and PMIS-200a in human bone marrow MSCs were confirmed after the cells were treatd with plasmid miR-200c and PMIS-200a in vitro. The biomarkers of osteogenic differentiation, including ALP, Runx2, and OCN, were up-regulated in the cells , compared to the non-treated cells and the cells treated with empty vectors. Bone formation quantitatively measured using µCT and histology in the defects treated with miR-200c and PMIS-200a are significantly higher than those receiving collagen sponge alone or empty vectors. The newly formed bone volume treated with miR-200c and PMIS-200a are twice higher than the controls.
Conclusions: These data demonstrated that miR-200c and miR-200a inhibition potently improve osteogenic differentiation of human MSCs and promote bone formation, which may potentially be used to restore oral and craniofacial bone defects clinically.