Sichuan University, West China Hospital of Stomatology, State Key Laboratory of Oral Diseases and National Clinical Research Center for Oral Diseases, Chengdu, China.
Northwest Minzu University, Key Laboratory of Stomatology of State Ethnic Affairs Commission, Key Laboratory of Oral Diseases of Gansu Province, Lanzhou, Gansu, China.
J Appl Oral Sci. 2023 Jan 23;31:e20220313. doi: 10.1590/1678-7757-2022-0313. eCollection 2023.
Bone morphogenetic protein 9 (BMP9) tends to be associated with various inflammatory responses of diseases, but its relationship with pulpitis remains unknown.
This study aimed to evaluate the effects and mechanisms of BMP9 in pulpitis.
A rat model of pulpitis was used to evaluate the expression of BMP9, which was also analysed in Porphyromonas gingivalis lipopolysaccharide (Pg-LPS)-stimulated human dental pulp cells (hDPCs). The effects and mechanism of BMP9 on the regulation of inflammatory factors and matrix metalloproteinase-2 (MMP2) were evaluated using real-time quantitative PCR, western blotting, and immunocytofluorescence. Moreover, the migration ability of THP-1 monocyte-macrophages, treated with inflammatory supernate inhibited by BMP9, was previously tested by a transwell migration assay. Finally, a direct rat pulp capping model was used to evaluate in vivo the influence of the overexpression of BMP9 in pulpitis.
The expression of BMP9 decreased after 24 h and increased after 3 and 7 d in rat pulpitis and inflammatory hDPCs. The overexpression of BMP9 inhibited the gene expression of inflammatory factors (IL-6, IL-8, and CCL2) and the secretion of IL-6 and MMP2 in Pg-LPS-stimulated hDPCs. The level of phosphorylated Smad1/5 was upregulated and the levels of phosphorylated ERK and JNK were downregulated. The inflammatory supernate of hDPCs inhibited by BMP9 reduced the migration of THP-1 cells. In rat pulp capping models, overexpressed BMP9 could partially restrain the development of dental pulp inflammation.
This is the first study to confirm that BMP9 is involved in the occurrence and development of pulpitis and can partially inhibit its severity in the early stage. These findings provided a theoretical reference for future studies on the mechanism of pulpitis and application of bioactive molecules in vital pulp therapy.
骨形态发生蛋白 9(BMP9)与多种疾病的炎症反应有关,但与牙髓炎的关系尚不清楚。
本研究旨在评估 BMP9 在牙髓炎中的作用及机制。
采用大鼠牙髓炎模型评估 BMP9 的表达,并分析牙龈卟啉单胞菌脂多糖(Pg-LPS)刺激下人牙髓细胞(hDPCs)中 BMP9 的表达。采用实时定量 PCR、Western blot 和免疫细胞荧光法评估 BMP9 对炎症因子和基质金属蛋白酶-2(MMP2)调节的作用及机制。此外,通过 Transwell 迁移实验检测抑制 BMP9 的炎症上清液对 THP-1 单核-巨噬细胞迁移能力的影响。最后,通过直接大鼠牙髓盖髓模型评估 BMP9 过表达对牙髓炎的体内影响。
大鼠牙髓炎和炎症 hDPCs 中,BMP9 的表达在 24 h 后降低,3 和 7 d 后升高。BMP9 的过表达抑制了 Pg-LPS 刺激的 hDPCs 中炎症因子(IL-6、IL-8 和 CCL2)的基因表达和 IL-6 和 MMP2 的分泌。磷酸化 Smad1/5 水平上调,磷酸化 ERK 和 JNK 水平下调。BMP9 抑制的 hDPCs 炎症上清液减少了 THP-1 细胞的迁移。在大鼠牙髓盖髓模型中,过表达的 BMP9 可部分抑制牙髓炎症的发展。
本研究首次证实 BMP9 参与牙髓炎的发生发展,可在早期部分抑制其严重程度。这些发现为今后研究牙髓炎的机制和生物活性分子在活髓治疗中的应用提供了理论参考。