PMID- 25994935 OWN - NLM STAT- MEDLINE DCOM- 20160217 LR - 20181113 IS - 1475-925X (Electronic) IS - 1475-925X (Linking) VI - 14 DP - 2015 May 21 TI - A comparative study of mechanical strain, icariin and combination stimulations on improving osteoinductive potential via NF-kappaB activation in osteoblast-like cells. PG - 46 LID - 10.1186/s12938-015-0039-z [doi] LID - 46 AB - BACKGROUND: The combination of drugs and exercise was the effective treatment in bone injure and rebuilding in clinic. As mechanical strain has potential in inducing the differentiation of osteoblasts in our previous study, the further research to investigate the combination of mechanical strain and icariin stimulation on inducing osteoblast proliferation, differentiation and the possible mechanism in MC3T3-E1 cell line. METHODS: A whole cell enzyme-linked immunosorbent assay that detects the bromodeoxyuridine incorporation during DNA synthesis was applied to evaluate the proliferation. The mRNA expression of alkaline phosphatase (ALP), osteocalcin (OCN), type I collagen (Col I), bone morphogenetic protein-2 (BMP-2) and BMP-4 was detected by real-time reverse-transcription polymerase chain reaction. The activity of ALP was analyzed by ELISA and the protein expression of OCN, Col I and BMP-2 was assessed by western blot. Moreover, the activity of nuclear transcription factor kappa-B (NF-kappaB) signaling pathway was investigated with the expression of inhibitor of kappaB (IkappaB) alpha, phosphorylation of IkappaB-alpha (P-IkappaB-alpha), p65, P-p65 by western blot. RESULTS: We observed that compared to single mechanical strain or icariin stimulation, the mRNA and protein expressions of ALP (P < 0.05 or P < 0.01), OCN (P < 0.01) and Col I (P < 0.05 or P < 0.01) were increased significantly by the combination of mechanical strain and icariin stimulation. Moreover, the combination of mechanical strain and icariin stimulation could up-regulate the expression of BMP-2 (P < 0.01) and BMP-4 compared to single mechanical strain or icariin stimulation. The combination of mechanical strain and icariin stimulation could activate NF-kappaB signaling pathway by increasing the expression of IkappaB alpha, P-IkappaB-alpha, p65, P-p65 (P < 0.01). CONCLUSION: The combination of mechanical strain and icariin stimulation could activate the NF-kappaB pathway to improve the proliferation, differentiation of osteoblast-like cells. FAU - Wang, Qiang-Song AU - Wang QS AD - Tianjin Institute of Medical Equipment, Academy of Military Medical Sciences, No. 106 Wandong Road, Hedong District, Tianjin, 300162, People's Republic of China. wangqiangsong2008@163.com. FAU - Zhang, Xin-Chang AU - Zhang XC AD - Tianjin Institute of Medical Equipment, Academy of Military Medical Sciences, No. 106 Wandong Road, Hedong District, Tianjin, 300162, People's Republic of China. xinchangzhang@163.com. FAU - Li, Rui-Xin AU - Li RX AD - Tianjin Institute of Medical Equipment, Academy of Military Medical Sciences, No. 106 Wandong Road, Hedong District, Tianjin, 300162, People's Republic of China. limxinxin@163.com. FAU - Sun, Jing-Gong AU - Sun JG AD - Tianjin Institute of Medical Equipment, Academy of Military Medical Sciences, No. 106 Wandong Road, Hedong District, Tianjin, 300162, People's Republic of China. sunjg@sina.vip.com. FAU - Su, Wei-Hua AU - Su WH AD - Tianjin Institute of Medical Equipment, Academy of Military Medical Sciences, No. 106 Wandong Road, Hedong District, Tianjin, 300162, People's Republic of China. 13512062095@126.com. FAU - Guo, Yong AU - Guo Y AD - Tianjin Institute of Medical Equipment, Academy of Military Medical Sciences, No. 106 Wandong Road, Hedong District, Tianjin, 300162, People's Republic of China. guoyong74@163.com. FAU - Li, Hao AU - Li H AD - Tianjin Institute of Medical Equipment, Academy of Military Medical Sciences, No. 106 Wandong Road, Hedong District, Tianjin, 300162, People's Republic of China. lihao2006011653@sina.com. FAU - Zhang, Xi-Zheng AU - Zhang XZ AD - Tianjin Institute of Medical Equipment, Academy of Military Medical Sciences, No. 106 Wandong Road, Hedong District, Tianjin, 300162, People's Republic of China. z56717@sohu.com. LA - eng PT - Comparative Study PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150521 PL - England TA - Biomed Eng Online JT - Biomedical engineering online JID - 101147518 RN - 0 (Bmp2 protein, mouse) RN - 0 (Bmp4 protein, mouse) RN - 0 (Bone Morphogenetic Protein 2) RN - 0 (Bone Morphogenetic Protein 4) RN - 0 (Collagen Type I) RN - 0 (Flavonoids) RN - 0 (NF-kappa B) RN - 104982-03-8 (Osteocalcin) RN - EC 3.1.3.1 (Alkaline Phosphatase) RN - VNM47R2QSQ (icariin) SB - IM MH - 3T3 Cells MH - Alkaline Phosphatase/metabolism MH - Animals MH - Bone Morphogenetic Protein 2/genetics/metabolism MH - Bone Morphogenetic Protein 4/genetics/metabolism MH - Bone Remodeling/drug effects MH - Cell Differentiation/drug effects MH - Cell Proliferation/drug effects MH - Collagen Type I/genetics/metabolism MH - Flavonoids/*pharmacology MH - Gene Expression Regulation/drug effects MH - Mice MH - NF-kappa B/*metabolism MH - Osteoblasts/*cytology/*drug effects/metabolism MH - Osteocalcin/genetics/metabolism MH - Signal Transduction/drug effects MH - *Stress, Mechanical PMC - PMC4455701 EDAT- 2015/05/23 06:00 MHDA- 2016/02/18 06:00 PMCR- 2015/05/21 CRDT- 2015/05/22 06:00 PHST- 2015/01/08 00:00 [received] PHST- 2015/04/16 00:00 [accepted] PHST- 2015/05/22 06:00 [entrez] PHST- 2015/05/23 06:00 [pubmed] PHST- 2016/02/18 06:00 [medline] PHST- 2015/05/21 00:00 [pmc-release] AID - 10.1186/s12938-015-0039-z [pii] AID - 39 [pii] AID - 10.1186/s12938-015-0039-z [doi] PST - epublish SO - Biomed Eng Online. 2015 May 21;14:46. doi: 10.1186/s12938-015-0039-z.