IKVAV regulates ERK1/2 and Akt signalling pathways in BMMSC population growth and proliferation.
Bottom Line: IKVAV peptides were synthesized by the solid-phase method.Meanwhile, phosphorylation levels of ERK1/2 and Akt were partially blocked by ERK1/2 inhibitor (PD98059) and Akt inhibitor (wortmannin), respectively.This study is the first to reveal an enhancement effect of IKVAV peptide on BMMSC at the signal transduction level, and the outcome could provide experimental evidence for application of IKVAV-grafted scaffolds in the field of BMMSC-based tissue engineering.
Affiliation: State Key Laboratory of Advanced Technology for Materials Synthesis and Processing, and Biomaterials Science and Engineering Research Center, Wuhan University of Technology, Wuhan, 430070, China.Show MeSH
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Mentions: Effects of IKVAV at various concentrations (0, 0.004, 0.02, 0.1, 0.5 and 2.5 mm) treating BMMSC for different time intervals (0, 24, 48, 72 h) on their viability were determined by CCK-8 assay. OD values were tested every 24 h for 72 h after co-culture. As shown in Fig.3a, cell viability increased gradually at concentrations from 0 to 0.5 mm, peaked at 0.5 mm and then declined. Highest OD value was observed at 72 h when treated with IKVAV at 0.5 mm (Fig.3b) (*P < 0.05). These results demonstrated that IKVAV promoted BMMSC proliferation in a dose- and time-dependent manner. Apoptosis was tested for by FCM analysis of annexin V FITC/propidium iodide (PI) staining of BMMSC treated with IKVAV at 0.5 mm, after 24 h. Figure3c and 3d indicate that there was no clear reduction in live cell percentage in the IKVAV-treated group (93.25%) compared to the control group (94.34%). Only a low apoptotic fraction was observed after 24-h treatment. Apoptotic level of the 0.5 mm IKVAV-treated group (2.37%) was almost the same as that of the control group (2.35%).
Affiliation: State Key Laboratory of Advanced Technology for Materials Synthesis and Processing, and Biomaterials Science and Engineering Research Center, Wuhan University of Technology, Wuhan, 430070, China.