The Effect Of MiR-29a-3p On The Implant Osseointergration Of Rat With Hyperlipidemia

BackgroudsThe morbidity of hyperlipidemia is high,and it can influence bone metabolism.Hyperlipidemia can cause osteoporosis,and effect the implant osseointegration.Earlier study found that multiple molecules of Wnt pathway changed,and these changes in the molecules may also lead to poor bone integration.MicroRNA has a specific expression at different stages of cell differentiation and tissue development,and is mainly involved in the regulation of gene transcription.The microRNA-29 family is a very important gene family of microRNA and plays a role in a variety of diseases such as cancer,diabetes,osteoporosis,and cardiovascular disease.ObjectiveThe aim of this study is to investigate the effect of miR-29a-3p on implant osseointegration of rat with hyperlipidemia and the osteoblastic differentiation of BMSCs in hyperlipidemia environment.Materials and methods1.In vitro experiments1.1 Acquisition and culture of BMSCsBMSCs were cultured and passaged.The third generation BMSCs were used to the subsequent experiments.1.2 Changes of miR-29a-3p expression during osteogenic differentiation of BMSCs.The experiment was divided into two groups.The high fat group was induced by high-fat osteogenesic induction,and the normal group was induced by a-MEM medium osteogenesic induction.After 3?5?7?14 days,total RNA was extracted and RT-PCR was used to evaluate the expression level of miR-29a-3p.1.3 The effect of miR-29a-3p on the osteoblast differentiation of BMSCs in hyperlipidemia environmentThe expression of miR-29a-3p was overexpressed by miR-29a-3p mmics,and reduced expression by miR-29a-3p inhibitor in BMSCs.Then,the expression of ALP and Runx2 in BMSCs induced by osteogenic induction were detected by RT-PCR and Western Blotting.2.In vivo experiment2.1 Build the model of hyperlipidemia ratsAdult male Wistar rats were randomly divided into two.The high fat group were fed with high fat diet and normal group were fed with normal deit.After continuous feeding for 6 weeks,the hyperlipidemia rat model was established.2.2 MiR-29a-3p expression changeds in bone tissue around implant of rats.The animals were sacrificed at 5?10?15?20 days after the surgery.The expression of miR-29a-3p was detected by RT-PCR.2.3 The role of miR-29a-3p in the implant osseointergration of rats with hyperlipidemia.In the overexpress group,miR-29a-3p was overexpressed by lentiviral vector,and the negitive control group was injected with negitive control injection.RT-PCR and Western Blotting was used to detect the expression of ALP and Runx2 in bone tissue around implant.Results1.During osteogenic induction,the expression of miR-29a-3p of 5 days was higher than that of 3 days,and 7 days was higher than that of 5 days,but 14 days compared to 7 days decreased.The expression levels of miR-29a-3p in the high fat group(3 days,5 days,7 days and 14 days after the surgery)were lower than those in the normal group(p<0.05).2.Overexpressed miR-29a-3p in BMSCs by miR-29a-3p mmics,ALP and Runx2 was significantly higher than the control group.Lower the expression of miR-29a-3p in BMSCs by miR-29a-3p inhibitor,the expression of ALP and Runx2 decreased.3.The expression of miR-29a-3p in the bone tissue around the implant after implantation of 10 days was higher than that of 5 days,but 15 days was lower than that of 10 days,and 20 days was lower than that of 15 days.The expression levels of miR-29a-3p in the experimental group at 5 days,10 days,15 days were lower than those in the control group(p<0.05).4.The over expression of miR-29a-3p in hyperlipidemic rats by lentiviral vector.After the implanted into the bone tissue of 10 days,the expression of ALP and Runx2 increased(p<0.05).Conclusion1.MiR-29a-3p plays a positive role in the regulation of bone marrow mesenchymal stem cells in high-fat environment.2.MiR-29a-3p can promote expression of ALP?Runx2 in the bone tissue around the implant of rats with hyperlipidemia,maybe miR-29a-3p can promote implant osseointergration of hyperlipidemia rats.