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In vitro and in vivo study on the osseointegration of magnesium and strontium ion with two different proportions of mineralized collagen and its mechanism

To explore the optimal combination of Mg2+, Sr2+ and mineralized collagen (nHAC) with two different proportions of hydroxyapatite (HA) and collagen (COL) on differentiation of MC3T3-E1 and the underlying mechanism, as well as achieve bone osseointegration. MC3T3-E1 cells were cultured in a complete medium with Mg2+ at the concentration of 0, 4, 8, 12, 16, 20 mmol/L, Sr2+ at the concentration of 0, 3, 6, 12 mmol/L, and the impregnation solution of 3:7 and 5:5nHAC. The differentiation of MC3T3-E1 was measured by expression of osteogenic genes and proteins including Runx-2, BMP-2 and OCN and determined the activation of PI3K/AKT/GSK3β/β-catenin signaling pathway in 12 mmol/LMg2++3 mmol/LSr2++3:7nHAC group. Osteoporosis was induced in 18 female rats by means of ovariectomy, the implants were immersed in 60 mmol/LMg2++15 mmol/LSr2++3:7nHAC impregnation solution and implanted into the mesial alveolar fossa for immediate implantation. The osseointegration of the implants was observed by Confocal laser scanning microscopy (CLSM) and histology at 4 and 8 weeks. The groups cultured with 12 mmol/LMg2+, 3 mmol/LSr2+ and 3:7nHAC impregnation solution showed the osteogenic genes and proteins were significantly higher respectively (P < 0.05), as well as p-Akt, p-GSK3β and β-catenin proteins (P < 0.05). CLSM and histology showed that the implant surface was surrounded by thick lamellar bone plate, and the trabecular bone were dense and continuous in the impregnation solution. These results found that magnesium and strontium ion-loaded mineralized collagen play an critical role in up-regulating the cells activity through PI3K/AKT/GSK3β/β-catenin signaling pathway and could be promote the formation of osseointegration.