Zhang Hongmei, Wang Jinhua, Deng Fang, Huang Enyi, Yan Zhengjian, Wang Zhongliang, Deng Youlin, Zhang Qian, Zhang Zhonglin, Ye Jixing, Qiao Min, Li Ruifang, Wang Jing, Wei Qiang, Zhou Guolin, Luu Hue H, Haydon Rex C, He Tong-Chuan, Deng Feng
Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, and The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.
Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, and The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China; Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, IL 60637, USA.
Biomaterials. 2015 Jan;39:145-54. doi: 10.1016/j.biomaterials.2014.11.007. Epub 2014 Nov 22.
Dental pulp/dentin regeneration using dental stem cells combined with odontogenic factors may offer great promise to treat and/or prevent premature tooth loss. Here, we investigate if BMP9 and Wnt/β-catenin act synergistically on odontogenic differentiation. Using the immortalized SCAPs (iSCAPs) isolated from mouse apical papilla tissue, we demonstrate that Wnt3A effectively induces early osteogenic marker alkaline phosphatase (ALP) in iSCAPs, which is reduced by β-catenin knockdown. While Wnt3A and BMP9 enhance each other's ability to induce ALP activity in iSCAPs, silencing β-catenin significantly diminishes BMP9-induced osteo/odontogenic differentiation. Furthermore, silencing β-catenin reduces BMP9-induced expression of osteocalcin and osteopontin and in vitro matrix mineralization of iSCAPs. In vivo stem cell implantation assay reveals that while BMP9-transduced iSCAPs induce robust ectopic bone formation, iSCAPs stimulated with both BMP9 and Wnt3A exhibit more mature and highly mineralized trabecular bone formation. However, knockdown of β-catenin in iSCAPs significantly diminishes BMP9 or BMP9/Wnt3A-induced ectopic bone formation in vivo. Thus, our results strongly suggest that β-catenin may play an important role in BMP9-induced osteo/ondontogenic signaling and that BMP9 and Wnt3A may act synergistically to induce osteo/odontoblastic differentiation of iSCAPs. It's conceivable that BMP9 and/or Wnt3A may be explored as efficacious biofactors for odontogenic regeneration and tooth engineering.
使用牙干细胞与牙源性因子相结合来进行牙髓/牙本质再生，可能为治疗和/或预防过早牙齿脱落带来巨大希望。在此，我们研究BMP9和Wnt/β-连环蛋白是否协同作用于牙源性分化。利用从小鼠根尖乳头组织分离出的永生化根尖乳头干细胞（iSCAPs），我们证明Wnt3A能有效诱导iSCAPs中早期成骨标志物碱性磷酸酶（ALP），而β-连环蛋白敲低可使其降低。虽然Wnt3A和BMP9相互增强在iSCAPs中诱导ALP活性的能力，但沉默β-连环蛋白会显著减弱BMP9诱导的骨/牙源性分化。此外，沉默β-连环蛋白会降低BMP9诱导的骨钙素和骨桥蛋白表达以及iSCAPs的体外基质矿化。体内干细胞植入试验表明，虽然转导BMP9的iSCAPs能诱导强大的异位骨形成，但同时用BMP9和Wnt3A刺激的iSCAPs表现出更成熟且矿化程度更高的小梁骨形成。然而，iSCAPs中β-连环蛋白的敲低会显著减弱体内BMP9或BMP9/Wnt3A诱导的异位骨形成。因此，我们的结果强烈表明β-连环蛋白可能在BMP9诱导的骨/牙源性信号传导中起重要作用，并且BMP9和Wnt3A可能协同作用以诱导iSCAPs向成骨/成牙本质细胞分化。可以想象，BMP9和/或Wnt3A可能被探索作为牙源性再生和牙齿工程的有效生物因子。
