Comparison Of The Osteogenesis Of Bone Marrow-and Adipose Tissue-derived Mesenchymal Stem Cells Under Noggin Suppression

Objectives:This study attempts to observe the effect and compare of osteogenesis abilitybetween adipose mesenchymal stem cells (ADSCs, Adipose Derived Stem Cells) andbone marrow mesenchymal stem cells (BMSCs, Bone Mesenchymal Stem Cells) withknockdowe Noggin and increasing BMP2in vitro.Methods:BMSCs and ADSCs from the same SD rat are transfected by BMP2and nogginshRNA with lipidosome in vitro so that osteogenesis can be induced. In3,7,14d threetime points， we detect osteogenesis related gene expression with Realtime-PCRtechnique,And detect the osteogenetic differentiation ability through the alizarin redstaining method。Results:1. ADSCs and BMSCs cell morphological observation showd that：we achievedADSCs and BMSCs from the same SD rats successfully,and observed its alizarin redstaining and oil red O staining after osteogenesis and adipogenesis induction.2. Realtime-PCR results showd that：3,7,14days ADSCs and BMSCsBMP2geneexpression trend is the same，the expression of BMP2gene of BMP2group wasobviously higher than that of EGFP group（control group）（P<0.01）,and expression ofBMP2gene of BMP2+nogginshRNA group was obviously higher than that of BMP2group（P<0.01）.3. Realtime-PCR results showd that：after transfection3,7,14days ADSCsexperimental group (BMP2, BMP2+NogginshRNA) Runx2, Col1, ALP, Sp7osteogenesis gene expression were significantly increased compared with control group (EGFP（)P<0.01）。BMP2+NogginshRNA Group Runx2, Col1, ALP, Sp7osteogenesisgene expression compared with BMP2group were significantly higher (P <0.01).Likewise, after transfection3,7,14days BMSCs experimental group (BMP2, BMP2+NogginshRNA) Runx2, Col1, ALP, Sp7osteogenesis gene expression significantlyincreased compared with control group (EGFP)（P<0.05）.BMP2+NogginshRNAGroup Runx2, Col1, ALP, Sp7osteogenesis gene expression compared with BMP2group increased significantly（P<0.05）.4. Alizarin red staining results showed that：EGFP group, BMP2group, BMP2+NogginshRNA group the amount of calcium nodules obviously showed a trend ofincreasing. Quantitative analysis results showed increasing trend.There are statisticallysignificant（P<0.01）the calcium nodules of BMSCs are significantly greater than that ofADSCs.There are statistically significant（P<0.01）.5.Realtime-PCR results showd that：after BMP2gene and noggin gene silencingcommon modified BMSCs, the expression of osteogenesis gene Runx2, Col1, ALP andSp7compared with ADSCs in the same condition were significantly up-regulation（P<0.01）. on3,7,14days,BMP2gene and noggin gene silencing common modifiedADSCs BMP2gene expression compared with control group (EGFP) BMSCs weresignificantly up-regulation.（P<0.01），osteogenesis gene Runx2, Col1, ALP, Sp7expression is significantly down-regulation（P<0.01）.Conclusions:1. Under BMP2gene and NogginshRNA modified, the expression of BMP2andosteogenesis differentiation ability of two kinds of stem cells is better than that of BMP2genetic modification.2. Under the same genetic modification, osteogenesis ability of BMSCs is betterthan that of ADSCs.3. Without genetic modification, osteogenesis ability of BMSCs is superior to thatof the genetic modification ADSCs.