The growth of long and polarized ameloblast-like cells has long been heralded as a major prerequisite for enamel tissue engineering. In this study, we have designed three-dimensional bioreactor/scaffold microenvironments to propagate and assess the ability of cervical loop derivatives to become long and polarized ameloblast-like cells. Our studies demonstrated that cervical loop/periodontal progenitor coculture in a growth-factor-enriched medium resulted in the formation of ameloblast-like cells expressing high levels of amelogenin and ameloblastin. Coculture of cervical loop cells with dental pulp cells on tailored collagen scaffolds enriched with leucine-rich amelogenin peptide (LRAP) and early enamel matrix resulted in singular, elongated, and polarized ameloblast-like cells that expressed and secreted ameloblastin and amelogenin enamel proteins. Bioreactor microenvironments enriched with enamel matrix and LRAP also proved advantageous for the propagation of HAT-7 cells, resulting in a ∼20-fold higher expression of amelogenin and ameloblastin enamel proteins compared with controls growing on plain scaffolds. Together, studies presented here highlight the benefits of microgravity culture systems combined with ameloblast-specific microenvironments and tailored scaffolds for the growth of ameloblast-like cells.

中文翻译：

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生物反应器中颈环/牙髓共培养物中表达成釉细胞的极化、牙釉蛋白样细胞


长且极化的成釉细胞样细胞的生长长期以来一直被认为是牙釉质组织工程的主要先决条件。在这项研究中，我们设计了三维生物反应器/支架微环境来繁殖和评估宫颈环衍生物变成长且极化的成釉细胞样细胞的能力。我们的研究表明，在富含生长因子的培养基中进行宫颈环/牙周祖细胞共培养，可形成表达高水平釉原蛋白和成釉细胞的成釉细胞样细胞。将宫颈环细胞与牙髓细胞在富含亮氨酸的牙釉质肽（LRAP）和早期牙釉质基质的定制胶原支架上共培养，产生了表达和分泌成釉细胞和牙釉质蛋白的单一、细长和极化的成釉细胞样细胞。富含牙釉质基质和 LRAP 的生物反应器微环境也被证明有利于 HAT-7 细胞的增殖，与在普通支架上生长的对照相比，导致牙釉质蛋白和成釉细胞牙釉质蛋白的表达高出约 20 倍。总之，这里介绍的研究强调了微重力培养系统与成釉细胞特异性微环境和定制支架相结合用于成釉细胞样细胞生长的好处。