Effect of calcium on proliferation, migration and osteogenic differentiation of human dental follicle cells

doi:10.19439/j.sjos.2019.06.003

Abstract

Abstract: PURPOSE: To determine the role of Ca²⁺ in proliferation,migration and osteogenic differentiation of human dental follicle cells(hDFCs). METHODS: hDFCs were isolated and cultured. The source of hDFCs was detected by immunofluorescence staining. Osteogenesis and adipogenic differentiation of hDFCs was detected by alizarin red staining and oil red O staining, to identify its multi-directional differentiation ability. A series of Ca²⁺ solutions with different concentrations was prepared, CCK8 assay was used to detect the proliferative abilities at 1, 3, 5, and 7 d;migratory ability of 24 h was detected by Transwell assay. Calcium nodules were detected by semiquantitative analysis of alizarin red staining. mRNA expression of osteogenic differentiation related genes was examined by real-time quantitative polymerase chain reaction (RT-qPCR).Statistical analysis was performed using SPSS 17.0 software package. RESULTS: Compared with the control group, 3,4 and 5 mmol/L Ca²⁺ significantly promoted proliferation of hDFCs at 3, 5 and 7 d (P<0.05). 3, 4, 5 and 6 mmol/L Ca²⁺ significantly promoted the migration of hDFCs at 24 h(P<0.01). High concentration of Ca²⁺ had no significant effect on its proliferation and migration. The results of alizarin red staining showed that when Ca²⁺concentration reached 4 mmol/L, formation of mineralized nodules were increased(P<0.01), and Ca²⁺ concentration-dependent. RT-qPCR results showed that Ca²⁺ up-regulated the expression of RUNX2 and OCN in osteogenic differentiation genes (P<0.01). CONCLUSIONS: Low Ca²⁺ concentration is beneficial to proliferation and migration, and high Ca²⁺ concentration is beneficial to osteogenic differentiation of human dental follicle cells.

Key words: Human dental follicle cells, Calcium, Migration, Proliferation, Osteogenic differentiat

CLC Number:

R783.5

ZHAO Xian, ZENG Jin, ZUO Dong-chuan, XU Jie, YU Jing-hong, XU Xiao-mei. Effect of calcium on proliferation, migration and osteogenic differentiation of human dental follicle cells[J]. Shanghai Journal of Stomatology, 2019, 28(6): 572-577.

References

[1] Guo S, Guo W, Ding Y, et al.Comparative study of human dental follicle cell sheets and periodontal ligament cell sheets for periodontal tissue regeneration[J]. Cell Transplantat, 2013, 22(6): 1061-1073.
[2] Guo S, Kang J, Ji B, et al.Periodontal derived mesenchymal cell sheets promote periodontal regeneration in inflammatory microenvironment[J]. Tissue Eng Part A, 2017, 23(13-14): 585-596.
[3] Yildirim S, Zibandeh N, Genc D, et al.The Comparison of the immunologic properties of stem cells isolated from human exfoliated deciduous teeth, dental pulp, and dental follicles[J]. Stem Cells Int, 2016, 2016: 4682875.
[4] Silver IA, Murrills RJ, Etherington DJ.Microelectrode studies on the acid microenvironment beneath adherent macrophages and osteoclasts[J]. Exp Cell Res, 1988, 175(2): 266-276.
[5] Lee MN, Hwang HS, Oh SH, et al.Elevated extracellular calcium ions promote proliferation and migration of mesenchymal stem cells via increasing osteopontin expression[J]. Exp Mol Med, 2018, 50(11): 142.
[6] Nakashima T, Hayashi M, Takayanagi H.New insights into osteoclastogenic signaling mechanisms[J]. Trends Endocrinol Metab, 2012, 23(11): 582-590.
[7] Hu F, Pan L, Zhang K, et al.Elevation of extracellular Ca²+ induces store-operated calcium entry via calcium-sensing receptors: a pathway contributes to the proliferation of osteoblasts[J]. PLoS One, 2014, 9(9): e107217.
[8] Capiod T.Extracellular calcium has multiple targets to control cell proliferation[J]. Adv Exp Med Biol, 2016, 898: 133-156.
[9] Lange I, Koster J, Koomoa DT.Calcium signaling regulates fundamental processes involved in neuroblastoma progression[J]. Cell Calcium, 2019, 82: 102052.
[10] Dvorak MM, Siddiqua A, Ward DT, et al.Physiological changes in extracellular calcium concentration directly control osteoblast function in the absence of calciotropic hormones[J]. Proc Natl Acad Sci USA, 2004, 101(14): 5140-5145.
[11] Maeno S, Niki Y, Matsumoto H, et al.The effect of calcium ion concentration on osteoblast viability, proliferation and differentiation in monolayer and 3D culture[J]. Biomaterials, 2005, 26(23): 4847-4855.
[12] Kruppke B, Heinemann C, Wagner AS, et al.Strontium ions promote in vitro human bone marrow stromal cell proliferation and differentiation in calcium- lacking media[J]. Dev Growth Differ, 2018, 61(2): 166-175.
[13] Cheng S, Wang W, Lin Z, et al.Effects of extracellular calcium on viability and osteogenic differentiation of bone marrow stromal cells in vitro[J]. Human Cell, 2013, 26(3): 114-120.
[14] Zhang Y, Xu G, Lee RM, et al.Collective cell migration has distinct directionality and speed dynamics[J]. Cell Mol Life Sci, 2017, 74(20): 3841-3850.
[15] Komori T.Runx2, an inducer of osteoblast and chondrocyte differentiation[J]. Histochem Cell Biol, 2018, 149(4): 313-323.
[16] Koori K, Maeda H, Fujii S, et al.The roles of calcium-sensing receptor and calcium channel in osteogenic differentiation of undifferentiated periodontal ligament cells[J]. Cell Tissue Res, 2014, 357(3): 707-718.
[17] Lei Q, Jiang C, Huang W, et al.Proteomic analysis of the effect of extracellular calcium ions on human mesenchymal stem cells: Implications for bone tissue engineering[J]. Chem Biol Interact, 2015, 233: 139-146.
[18] Cui L, Xu SM, Ma DD, et al.The effect of TRPM7 suppression on the proliferation,migration and osteogenic differentiation of human dental pulp stem cells[J]. Int Endod J, 2014, 47(6): 583-593.
[19] Castiglioni S, Romeo V, Locatelli L, et al.The simultaneous downregulation of TRPM7 and MagT1 in human mesenchymal stem cells in vitro: Effects on growth and osteogenic differentiation[J]. Biochem Biophys Res Commun, 2019, 513(1): 159-165.
[20] Castiglioni S, Romeo V, Locatelli L, et al.TRPM7 and MagT1 in the osteogenic differentiation of human mesenchymal stem cells in vitro[J]. Sci Rep, 2018, 8(1): 16195.