Synergistic effects of stromal cell-derived factor-1 alpha and bone morphogenetic protein-2 treatment on odontogenic differentiation of human stem cells from apical papilla cultured in the VitroGel 3D system
Synergistic effects of stromal cell-derived factor-1 alpha and bone morphogenetic protein-2 treatment on odontogenic differentiation of human stem cells from apical papilla cultured in the VitroGel 3D system
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基质细胞衍生因子 1 α 和骨形态发生蛋白 2 治疗对 VitroGel 3D 系统中培养的根尖乳头干细胞的牙源性分化的协同作用
DOI：
10.1007/s00441-019-03045-3
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发表时间：
2019
影响因子：
3.6
通讯作者：
Yu Qing
中科院分区：
文献类型：
--
作者：
Xiao Min;Qiu Jun;Kuang Rong;Zhang Beidi;Wang Wei;Yu Qing
未成熟恒牙根尖区的牙髓牙本质再生是一个重大的临床挑战。使用生物活性分子和支架的组织工程方法可能具有再生这些牙齿的天然根尖结构的潜力，代表现有治疗方案的上级替代方案。本研究的目的是（i）评价VitroGel 3D系统（一种无动物源多糖水凝胶），探讨基质细胞衍生因子-1 α（SDF-1α）和骨形态发生蛋白-2（BMP-2）联合应用对人根尖乳头干细胞（SCAP）牙向分化的影响在VitroGel 3D系统中培养。通过扫描电镜（SEM）、活细胞和死细胞染色和CCK-8测定法测定在VitroGel 3D系统中培养的SCAP的形态、活力和增殖。采用碱性磷酸酶（ALP）活性测定、实时荧光定量RT-PCR和Western blot分析等方法，观察体外培养的SCAP向牙源性分化的情况。最后，通过异位皮下注射，在体内评估牙向分化。结果表明，在三维水凝胶中培养的SCAP具有良好的存活和增殖能力。SDF-1α和BMP-2联合处理可增强SCAP牙向分化相关基因和蛋白的表达，促进SCAP牙向分化。总之，本研究表明VitroGel 3D系统促进SCAP增殖和分化。SDF-1α对BMP-2诱导的人SCAP牙向分化具有协同作用。
Pulp-dentin regeneration in the apical region of immature permanent teeth represents a significant clinical challenge. Tissue engineering approaches using bioactive molecules and scaffolds may have the potential to regenerate the natural apical structure of these teeth, representing a superior alternative to existing treatment regimens. The aims of this study are (i) to evaluate the VitroGel 3D system, an animal origin-free polysaccharide hydrogel, as a possible injectable scaffold for pulp-dentin regeneration and (ii) to investigate the effects of stromal cell-derived factor-1α (SDF-1α) and bone morphogenetic protein-2(BMP-2) cotreatment on odontogenic differentiation of human stem cells from apical papilla (SCAP) cultured in the VitroGel 3D system. The morphology, viability and proliferation of SCAP cultured in the VitroGel 3D system were measured via scanning electron microscopy (SEM), live and dead cell staining and CCK-8 assays. Alkaline phosphatase (ALP) activity, real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR) and Western blot analysis were further used to evaluate the odontogenic differentiation of SCAP cultured in the VitroGel 3D system in vitro. Finally, the odontogenic differentiation was assessed in vivo through ectopic subcutaneous injection. The results showed that SCAP cultured in 3D hydrogel demonstrated favorable viability and proliferation. SDF-1α and BMP-2 cotreatment enhanced odontogenic differentiation-related gene and protein expression in vitro and promoted odontogenic differentiation of SCAP in vivo. In conclusion, the present study demonstrated that the VitroGel 3D system promoted SCAP proliferation and differentiation. Moreover, SDF-1α cotreatment had synergistic effects on BMP-2-induced odontogenic differentiation of human SCAP cultured in the VitroGel 3D system both in vitro and in vivo.