Study on Odaph Promoting Osteoblasts Adhesion and Mineralization

doi:10.13701/j.cnki.kqyxyj.2023.09.009

Abstract

Abstract: Objective: To explore the influence of odontogenesis-associated phosphoprotein (Odaph) on osteoblast adhesion and mineralization. Methods: The expression of Odaph in mouse bone tissue was detected by quantitative real-time PCR (qRT-PCR). The effect of Odaph overexpression on MC3T3-E1 was detected by qRT-PCR. qRT-PCR, Western blot, and immunocytochemistry were used to detect the effect of Odaph overexpression on the expression of Lamininγ2 and Integrinβ6. Odaph and Lamininγ2 plasmid were cotransfected into human embryonic kidney cells-293T(HEK-293T), and co-immunoprecipitation was used to verify the interaction between Odaph and Lamininγ2. After mineralization induction of MC3T3-E1, the expressions of mineralization-related markers ALP, OCN, and Runx2 were detected by qRT-PCR, and ALP activity was observed by alkaline phosphatase (ALP) staining. Results: Odaph was expressed in mouse bone tissue and MC3T3-E1. Odaph overexpression increased the expression of Lamininγ2 and Integrinβ6, and Odaph interacted with Lamininγ2. After mineralization induction, Odaph overexpression promoted the expression of ALP, OCN, and Runx2, and enhanced ALP activity. Conclusion: Odaph promoted osteoblast adhesion and mineralization.

Key words: odontogenesis-associated phosphoprotein, MC3T3-E1, adhesion, mineralization, alkaline phosphatase

LI Mingyue, XIAO Shuang, LUAN Linqun, ZHANG Jie, GAO Yuguang, ZHANG Li. Study on Odaph Promoting Osteoblasts Adhesion and Mineralization[J]. Journal of Oral Science Research, 2023, 39(9): 810-814.

References

[1] Parry DA, Brookes SJ, Logan CV, et al. Mutations in C4orf26, encoding a peptide with in vitro hydroxyapatite crystal nucleation and growth activity, cause amelogenesis imperfecta [J]. Am J Hum Genet, 2012, 91(3):565-571.
[2] Govitvattana N, Kaku M, Ohyama Y, et al. Molecular cloning of mouse homologue of enamel protein C4orf26 and its phosphorylation by FAM20C [J]. Calcif Tissue Int, 2021, 109(4):445-454.
[3] Ji YK, Li C, Tian Y, et al. Maturation stage enamel defects in Odontogenesis-associated phosphoprotein (Odaph) deficient mice [J]. Dev Dyn, 2021, 250(10):1505-1517.
[4] Mu HY, Dong ZH, Wang YM, et al. Odontogenesis-associated phosphoprotein (Odaph) overexpression in ameloblasts disrupts enamel formation via inducing abnormal mineralization of enamel in secretory stage [J]. Calcif Tissue Int, 2022, 111(6):611-621.
[5] Liu SD, Li ZR, Wang QX, et al. Graphene oxide/chitosan/hydroxyapatite composite membranes enhance osteoblast adhesion and guided bone regeneration [J]. ACS Appl Bio Mater, 2021, 4(11):8049-8059.
[6] Kim JM, Lin C, Stavre Z, et al. Osteoblast-osteoclast communication and bone homeostasis [J]. Cells, 2020, 9(9):2073.
[7] Yao Y. Laminin: loss-of-function studies [J]. Cell Mol Life Sci, 2017, 74(6):1095-1115.
[8] 禹翠翠,叶潇元,董志恒,等.miR-450b-5p调控IL-1β诱导人牙龈上皮细胞粘附和迁移[J].口腔医学研究,2022,38(6):572-577.
[9] Campbell ID, Humphries MJ. Integrin structure, activation, and interactions [J]. Cold Spring Harb Perspect Biol, 2011, 3(3):a004994.
[10] Koivisto L, Bi J, Hakkinen L, et al. Integrin alphavbeta6: Structure, function and role in health and disease [J]. Int J Biochem Cell Biol, 2018, 99:186-196.
[11] Ido H, Nakamura A, Kobayashi R, et al. The requirement of the glutamic acid residue at the third position from the carboxyl termini of the laminin gamma chains in integrin binding by laminins [J]. J Biol Chem, 2007, 282(15):11144-11154.
[12] Yamada M, Sekiguchi K. Molecular basis of laminin-integrin interactions [J]. Curr Top Membr, 2015, 76:197-229.
[13] Vimalraj S. Alkaline phosphatase: Structure, expression and its function in bone mineralization [J]. Gene, 2020, 754:144855.
[14] Kinoshita Y, Mohamed FF, Amadeu de Oliveira F, et al. Gene therapy using adeno-associated virus serotype 8 encoding TNAP-D10 improves the skeletal and dentoalveolar phenotypes in Alpl^-/- mice [J]. J Bone Miner Res, 2021, 36(9):1835-1849.
[15] Pang Y, Liu LL, Mu H, et al. Nobiletin promotes osteogenic differentiation of human osteoblastic cell line (MG-63) through activating the BMP-2/RUNX-2 signaling pathway [J]. Saudi J Biol Sci, 2021, 28(9):4916-4920.
[16] Tosa I, Yamada D, Yasumatsu M, et al. Postnatal Runx2 deletion leads to low bone mass and adipocyte accumulation in mice bone tissues [J]. Biochem Biophys Res Commun, 2019, 516(4):1229-1233.