Human dental pulp stem cells (hDPSCs) are the primary cells responsible for dentin regeneration. Typically, in order to allow for odontoblastic differentiation, hDPSCs are cultured over weeks with differentiation-inducing factors in a typical monolayered culture. However, monolayered cultures have significant drawbacks including inconsistent differentiation efficiency, require a higher BMP concentration than should be necessary, and require periodic treatment with BMPs for weeks to see results. To solve these problems, we developed a 3D-cell spheroid culture system for odontoblastic differentiation using microparticles with leaf-stacked structure (LSS), which allow for the sustained release of BMPs and adequate supply of oxygen in cell spheroids. BMPs were continuously released and maintained an effective concentration over 37 days. hDPSCs in the spheroid maintained their viability for 5 weeks, and the odontoblastic differentiation efficiency was increased significantly compared to monolayered cells. Finally, dentin-related features were detected in the spheroids containing BMPs-loaded microparticles after 5 weeks, suggesting that these hDPSC-LSS spheroids might be useful for dentin tissue regeneration.

中文翻译：

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使用负载 BMP 的微粒形成 3D 球体可增强人牙髓干细胞的成牙本质细胞分化


人牙髓干细胞（hDPSC）是负责牙本质再生的主要细胞。通常，为了允许成牙本质细胞分化，hDPSC 在典型的单层培养物中与分化诱导因子一起培养数周。然而，单层培养物具有显着的缺点，包括分化效率不一致、需要比所需浓度更高的 BMP 浓度，并且需要定期用 BMP 治疗数周才能看到结果。为了解决这些问题，我们开发了一种使用叶堆叠结构（LSS）微粒进行成牙本质细胞分化的3D细胞球体培养系统，该系统可以持续释放BMP并为细胞球体提供充足的氧气。 BMP持续释放并保持有效浓度超过37天。球体中的hDPSCs可维持5周的活力，且与单层细胞相比，成牙本质细胞分化效率显着提高。最后，5周后在含有负载BMP的微粒的球体中检测到牙本质相关特征，这表明这些hDPSC-LSS球体可能对牙本质组织再生有用。