D-M@Mir-21 Effect On Osteogenic Differentiation

DPS-MEVs-miR-21(D-M@miR-21) was constructed. The effect of D-M@miR-21 on osteogenic differentiation of MC3T3-E1 cells was investigated by adding D-M@miR-21 to MC3T3-E1 cells. MEVs were separated from milk by means of ultrafast centrifugation. DPS-MEVs was obtained by co-incubation. The D-M@miR-21 was obtained by transfection. MEVs and D-M@miR-21 were identified by transmission electron microscopy, nanoflow detection, and Western blot. The experiments were divided into Control group, miR-21 group, MEVs group, DPS group, and D-M@miR-21 group. Cell proliferation was detected by CCK-8. The gene and protein levels of ALP, Runx2 and OPN were detected by Real-time PCR and Western blot. The results of transmission electron microscopy, nanoflow, Western blot and immunomagnetic beaded capture showed that MEVs and D-M@miR-21 were successfully prepared. The results of CCK-8 showed that compared with the Control group, miR-21 group, MEVs group and DPS group, D-M@miR-21 group could promote the proliferation of these cells at 24 h and 48 h. Real-time PCR and Western blot results showed that the expressions of related factors ALP, Runx2 and OPN genes and proteins in D-M@miR-21 group were the most significantly increased, and the results were statistically different (P<0.05). D-M@miR-21 successfully prepared; D-M@miR-21 can promote the proliferation and osteogenic differentiation of MC3T3-E1 cells.