Role of neuropeptide substance P and the bone morphogenetic protein signaling pathway in osteogenic differentiation of ST2 cells

doi:10.7518/hxkq.2018.04.006

Abstract

Abstract:

Objective This study aimed to investigate the role and mechanism of neuropeptide substance P (SP) in ST2 cell (bone mesenchymal stem cells of mice) osteogenic differentiation to provide a basis for the treatment of temporomandibular joint osteoarthritis. Methods Third-generation ST2 cells were cultured with different concentrations of SP (0, 10^-10, 10^-8, 10^-6, and 10^-5mol·L^-1). After 24, 48, and 72 h, cell proliferation was detected by CCK-8. The ST2 cells were cultured with 10^-6mol·L^-1 SP for 1, 3, 5, and 7 days. Subsequently, the expression of alkaline phosphatase (ALP), collagen typeⅠ(CollaⅠ), and osteocalcin (OCN) in the culture supernatant was tested by enzyme-linked immunosorbent assay (ELISA). ALP activity was detected by immunofluorescence staining. The ST2 cells were cultured with SP, Noggin (inhibitor of the bone morphogenetic protein signaling pathway), SP+Noggin, and 2% fetal bovine serum, respectively. Finally, the expression of ALP, CollaⅠ, and OCN in the culture supernatant was tested by ELISA. Results CCK-8 showed that the effect of cell proli-feration was most obvious when the SP concentration was 10^-6mol·L^-1 (P<0.01). The ELISA results demonstrated that ALP expression significantly increased at day 5 compared with that in the control group (P<0.01), whereas the expression of CollaⅠand OCN significantly increased at day 7 (P<0.05). Immunofluorescence results showed that ALP activity was strongest at day 5. The expression of ALP, CollaⅠ, and OCN decreased after Noggin addition (P<0.05). Conclusion SP can promote the proliferation and osteogenic differentiation of ST2 cells, and the bone morphogenetic protein signaling pathway may be involved in this process.

Key words: neuropeptide substance P, bone mesenchy-mal stem cells, bone morphogenetic protein signaling pathway, osteogenic differentiation

CLC Number:

Q257

Ting Hui, Guangcan Zhang, Dandan Feng, Ping Ji. Role of neuropeptide substance P and the bone morphogenetic protein signaling pathway in osteogenic differentiation of ST2 cells[J]. West China Journal of Stomatology, 2018, 36(4): 378-383.

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