Chang Mei-Chi, Chao Yi-Chi, Chen Yi-Chieh, Chang Hsueh-Wei, Zhong Bor-Hao, Pan Yu-Hwa, Jeng Jiiang-Huei, Chang Hsiao-Hua
Biomedical Science Team, Chang Gung University of Science and Technology, Taoyuan, Taiwan.
Department of Dentistry, Chang Gung Memorial Hospital, Taipei, Taiwan.
J Dent Sci. 2025 Jan;20(1):646-659. doi: 10.1016/j.jds.2024.11.002. Epub 2024 Nov 17.
BACKGROUND/PURPOSE: Revascularization procedures are used over apexification to treat teeth with necrotic pulp tissues and incomplete root formation. Clinically, inducing proliferation, migration, matrix deposition, and differentiation of stem cells from apical papilla (SCAPs) are critical for pulp regeneration. The study aimed to elucidate the impact of bone morphogenetic protein-4 (BMP-4) on plasminogen activation molecules and the osteogenic/odontogenic differentiation of SCAPs, as well as understand the related signaling mechanisms.
SCAPs were exposed to BMP-4 with or without signal transduction inhibitors. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. mRNA levels were quantified using real-time PCR. Protein expression in SCAPs was analyzed through immunofluorescent staining or western blotting. Cellular protein production was measured with enzyme-linked immunosorbent assay.
BMP-4 induced suppressor of mother against decapentaplegic (Smad)1/5/8 and Smad2/3 phosphorylation and activation. It also promoted higher expression of osteogenic and odontogenic markers, including Osterix, N-cadherin, and secreted protein acidic and rich in cysteine (SPARC), in SCAPs. Additionally, BMP-4 stimulated connective tissue growth factor (CTGF), plasminogen activator inhibitor-1 (PAI-1), and urokinase plasminogen activator receptor (uPAR) expression, but inhibited uPA expression and production in SCAPs, indicating its role in matrix remodeling and cell migration. Inhibition of Smad2/3 with SB431542 and Smad1/5/8 with LDN193189 attenuated the BMP-4-induced expression Osx, N-cadherin, CTGF, SPARC, uPAR and PAI-1.
These results indicate that BMP-4 stimulates the osteogenic and odontogenic differentiation of SCAPs by regulating matrix turnover and mineralization-related proteins. Furthermore, these processes are associated with the induction of Smad2/3 and Smad1/5/8 of SCAPs by BMP-4.
背景/目的：与根尖诱导成形术相比，血管再生术用于治疗牙髓组织坏死且牙根未完全形成的牙齿。临床上，诱导根尖乳头干细胞（SCAPs）增殖、迁移、基质沉积和分化对牙髓再生至关重要。本研究旨在阐明骨形态发生蛋白-4（BMP-4）对纤溶酶原激活分子以及SCAPs成骨/成牙分化的影响，并了解相关信号机制。
将SCAPs暴露于有或无信号转导抑制剂的BMP-4中。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法评估细胞活力。使用实时PCR定量mRNA水平。通过免疫荧光染色或蛋白质印迹分析SCAPs中的蛋白质表达。用酶联免疫吸附测定法测量细胞蛋白质产量。
BMP-4诱导母亲对果蝇dpp蛋白抑制因子（Smad）1/5/8和Smad2/3磷酸化及激活。它还促进SCAPs中包括osterix、N-钙黏蛋白和富含半胱氨酸的酸性分泌蛋白（SPARC）在内的成骨和成牙标记物的更高表达。此外，BMP-4刺激结缔组织生长因子（CTGF）、纤溶酶原激活物抑制剂-1（PAI-1）和尿激酶型纤溶酶原激活物受体（uPAR）表达，但抑制SCAPs中uPA表达和产生，表明其在基质重塑和细胞迁移中的作用。用SB431542抑制Smad2/3以及用LDN193189抑制Smad1/5/8减弱了BMP-4诱导的Osx、N-钙黏蛋白、CTGF、SPARC、uPAR和PAI-1表达。
这些结果表明，BMP-4通过调节基质周转和矿化相关蛋白刺激SCAPs的成骨和成牙分化。此外，这些过程与BMP-4诱导SCAPs的Smad2/3和Smad1/5/8有关。
