Purpose: To explore the role and mechanism of mechano-growth factor (MGF) in cyclic stretch (CS)-induced osteogenic differentiation and MMP-1, MMP-2 expression in human periodontal ligament cells (hPDLCs).
Methods: HPDLCs were isolated and transfected with si-MGF, or stimulated with MGF or MEK/ERK pathway inhibitor U0126. Cells were cultured in Flexercell system with 10% elongation at 0.1 Hz. An alkaline phosphatase (ALP) kit was used to detect ALP activity. QRT-PCR assay was performed to determine the transcript levels of MGF and osteogenic genes, including ALP, runt-related transcription factor 2 (Runx2) and osteopontin (OPN). Western bot was used to evaluate the effect on MEK/EKR1/2 signaling. Statistical analysis was performed using SPSS 19.0 software package.
Results: CS induced the expression of MGF in hPDLCs in a time-dependent manner (P＜0.05). In contrast to the control group, transfection with si-MGF inhibited the expression of MGF in hPDLCs (P＜0.05). Moreover, cessation of MGF dramatically suppressed ALP activity (P＜0.05) and the expression of osteogenic gene ALP, Runx2 and OPN (P＜0.05) in hPDLCs. Furthermore, down-regulation of MGF restrained the expression of MMP-1 and MMP-2, in contrast to CS group (P＜0.05). Conversely, stimulation with MGF further enhanced the effects of CS on osteogenic differentiation of hPDLCs and MMP-1, MMP-2 expression (P＜0.05). Additionally, MGC silencing abrogated CS-induced expression of p-ERK (P＜0.05), which was further enhanced following MGF treatment (P＜0.05). Simultaneously, precondition with U0126 antagonized MGF-enhanced effects on CS-triggered osteogenic differentiation and MMP-1, MMP-2 expression (P＜0.05).
Conclusions: Mechano-growth factor regulates cyclic stretch-induced osteogenic differentiation and MMP-1, MMP-2 expression in human periodontal ligament cells by activating MEK/ERK1/2 signaling pathway.
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