Mechano-growth factor regulated cyclic stretch-induced osteogenic differentiation and MMP-1, MMP-2 expression in human periodontal ligament cells by activating the MEK/ERK1/2 pathway

doi:10.19439/j.sjos.2019.01.002

Abstract

Abstract: 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.

Key words: Orthodontics, MGF, Human periodontal ligament cells, Cyclic stretch, Osteogenic differentiation, MMPs

CLC Number:

R783.5

CHEN Jing-tao, WANG Yan, ZHOU Zhi-fei, WEI Ke-wen. Mechano-growth factor regulated cyclic stretch-induced osteogenic differentiation and MMP-1, MMP-2 expression in human periodontal ligament cells by activating the MEK/ERK1/2 pathway[J]. Shanghai Journal of Stomatology, 2019, 28(1): 6-12.

References

[1] 张晓东, 宋九余, 李响, 等. 周期性牵张力对人牙周膜细胞MMP-1 mRNA表达的影响[J]. 临床口腔医学杂志, 2009, 25(1): 16-19.
[2] Chen YJ, Jeng JH, Chang HH, et al.Differential regulation of collagen, lysyl oxidase and MMP-2 in human periodontal ligament cells by low- and high-level mechanical stretching[J]. J Periodontal Res, 2013, 48(4): 466-474.
[3] Tang JJ, Podratz JL, Lange M, et al.Mechano growth factor, a splice variant of IGF-1, promotes neurogenesis in the aging mouse brain[J]. Mol Brain, 2017, 10(1): 23.
[4] Matheny RW Jr, Nindl BC, Adamo ML.Minireview: Mechano-growth factor: a putative product of IGF-I gene expression involved in tissue repair and regeneration[J]. Endocrinology, 2010, 151(3): 865-875.
[5] Sha Y, Lv Y, Xu Z, et al.MGF E peptide pretreatment improves the proliferation and osteogenic differentiation of BMSCs via MEK-ERK1/2 and PI3K-Akt pathway under severe hypoxia[J]. Life Sci, 2017, 189: 52-62.
[6] Tong Y, Feng W, Wu Y, et al.Mechano-growth factor accelerates the proliferation and osteogenic differentiation of rabbit mesenchymal stem cells through the PI3K/AKT pathway[J]. BMC Biochem, 2015, 16: 1.
[7] Zhang B, Luo Q, Sun J, et al.MGF enhances tenocyte invasion through MMP-2 activity via the FAK-ERK1/2 pathway[J]. Wound Repair Regen, 2015, 23(3): 394-402.
[8] 邹德慧, 赵莹, 赵寅华, 等. 周期性动态压力对人牙周膜干细胞中力生长因子表达的影响[J]. 牙体牙髓牙周病学杂志, 2016, 26(4): 191-196.
[9] 高锦瑜, 余小琴, 王军强. 周期性牵张应力作用下KLF5对人牙周膜细胞增殖和成骨分化的影响[J]. 上海口腔医学, 2018, 27(1): 28-33.
[10] 李彦, 李石, 牛忠英, 等. 微重力环境下Smads信号通路对人牙周膜干细胞成骨向分化的影响[J]. 上海口腔医学, 2012, 21(3): 246-250.
[11] Komori T.Runx2, an inducer of osteoblast and chondrocyte differentiation[J]. Histochem Cell Biol, 2018, 149(4): 313-323.
[12] 宋京, 任大鹏, 颜世果, 等. 细胞外信号调节蛋白激酶1/2在介导周期性牵张力对牙周膜细胞成骨分化中的作用[J]. 华西口腔医学杂志, 2017, 35(5): 520-526.
[13] Sha Y, Afandi R, Zhang B, et al.MGF E peptide pretreatment improves collagen synthesis and cell proliferation of injured human ACL fibroblasts via MEK-ERK1/2 signaling pathway[J]. Growth Factors, 2017, 35(1): 29-38.
[14] 余小琴, 高锦瑜, 刘华. 机械应力诱导HMGB1通过金属基质蛋白酶促进人牙周膜细胞的迁移[J]. 牙体牙髓牙周病学杂志, 2017, 27(9): 499-505.
[15] 沙永强, 吕永钢. 力生长因子E肽对前交叉韧带成纤维细胞活力、迁移与侵袭的影响[J]. 医用生物力学, 2016, 31(3): 227-234.
[16] 任大鹏. ERK1/2-Runx2信号通路介导周期性张应力作用下牙周膜成纤维细胞成骨分化的机制研究[D]. 济南: 山东大学, 2016.