Department of Stomatology, Nanfang Hospital, Southern Medical University, No. 1838, Guangzhou Road North, Baiyun District, Guangzhou, 510000, Guangdong, China.
School of Stomatology, Southern Medical University, Guangzhou, Guangdong, China.
Stem Cell Res Ther. 2023 May 15;14(1):130. doi: 10.1186/s13287-023-03353-0.
Signal peptide-CUB-EGF domain-containing protein 3 (SCUBE3), a secreted multifunctional glycoprotein whose transcript expression is restricted to the tooth germ epithelium during the development of embryonic mouse teeth, has been demonstrated to play a crucial role in the regulation of tooth development. Based on this, we hypothesized that epithelium-derived SCUBE3 contributes to bio-function in dental mesenchymal cells (Mes) via epithelium-mesenchyme interactions.
Immunohistochemical staining and a co-culture system were used to reveal the temporospatial expression of the SCUBE3 protein during mouse tooth germ development. In addition, human dental pulp stem cells (hDPSCs) were used as a Mes model to study the proliferation, migration, odontoblastic differentiation capacity, and mechanism of rhSCUBE3. Novel pulp-dentin-like organoid models were constructed to further confirm the odontoblast induction function of SCUBE3. Finally, semi-orthotopic animal experiments were performed to explore the clinical application of rhSCUBE3. Data were analysed using one-way analysis of variance and t-tests.
The epithelium-derived SCUBE3 translocated to the mesenchyme via a paracrine pathway during mouse embryonic development, and the differentiating odontoblasts in postnatal tooth germ subsequently secreted the SCUBE3 protein via an autocrine mechanism. In hDPSCs, exogenous SCUBE3 promoted cell proliferation and migration via TGF-β signalling and accelerated odontoblastic differentiation via BMP2 signalling. In the semi-orthotopic animal experiments, we found that SCUBE3 pre-treatment-induced polarized odontoblast-like cells attached to the dental walls and had better angiogenesis performance.
SCUBE3 protein expression is transferred from the epithelium to mesenchyme during embryonic development. The function of epithelium-derived SCUBE3 in Mes, including proliferation, migration, and polarized odontoblastic differentiation, and their mechanisms are elaborated for the first time. These findings shed light on exogenous SCUBE3 application in clinic dental pulp regeneration.
信号肽-CUB-表皮生长因子结构域蛋白 3（SCUBE3）是一种分泌型多功能糖蛋白，其转录本表达在胚胎小鼠牙齿发育过程中仅限于牙胚上皮细胞。已经证明，它在牙齿发育的调控中起着至关重要的作用。基于这一点，我们假设上皮细胞衍生的 SCUBE3 通过上皮-间充质相互作用有助于牙间质细胞（Mes）的生物功能。
免疫组织化学染色和共培养系统用于揭示 SCUBE3 蛋白在小鼠牙胚发育过程中的时空表达。此外，人牙髓干细胞（hDPSCs）被用作 Mes 模型，以研究 rhSCUBE3 的增殖、迁移、成牙本质分化能力和机制。构建了新的牙髓牙本质样类器官模型，以进一步证实 SCUBE3 的成牙本质诱导功能。最后，进行了半原位动物实验以探索 rhSCUBE3 的临床应用。数据采用单因素方差分析和 t 检验进行分析。
在上皮细胞衍生的 SCUBE3 通过旁分泌途径在小鼠胚胎发育过程中转位到间充质中，随后在出生后的牙胚中分化的成牙本质细胞通过自分泌机制分泌 SCUBE3 蛋白。在 hDPSCs 中，外源性 SCUBE3 通过 TGF-β 信号通路促进细胞增殖和迁移，并通过 BMP2 信号通路加速成牙本质分化。在半原位动物实验中，我们发现 SCUBE3 预处理诱导的极化成牙本质样细胞附着在牙壁上，具有更好的血管生成性能。
SCUBE3 蛋白表达在胚胎发育过程中从上皮细胞转移到间充质。首次详细阐述了上皮细胞衍生的 SCUBE3 在 Mes 中的功能，包括增殖、迁移和极化的成牙本质分化及其机制。这些发现为外源性 SCUBE3 在临床牙髓再生中的应用提供了依据。
