Iwasaki Akihiro, Hatakeyama Mayumi, Liu Qimei, Orimoto Ai, Fukuda Tomokazu, Kitaoka Takuya
Department of Agro-Environmental Sciences, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Fukuoka 819-0395, Japan.
Division of Endodontics and Restorative Dentistry, Kyushu Dental University, Kitakyushu 803-8580, Japan.
Carbohydr Polym. 2025 Jul 1;359:123593. doi: 10.1016/j.carbpol.2025.123593. Epub 2025 Apr 9.
Human dental pulp stem cells (hDPSCs) are a promising cell source for tooth regeneration therapies. However, conventional culture scaffold materials are often animal-derived, leading to immunogenicity concerns and limited availability. In this study, we explored phosphorylated cellulose nanofibers (P-CNFs), which have a fine fiber morphology and phosphate groups, as a novel scaffold material for cell culture. Immortalized hDPSCs were cultured on P-CNF scaffolds with different phosphate contents (0-1.42 mmol g) prepared by varying the molar ratio of urea and diammonium hydrogen phosphate and the reaction time. Cells cultured on unmodified CNFs exhibited poor adhesion and formed spheroids, indicating low bioadaptability. In contrast, P-CNF scaffolds with moderate phosphate content (0.54-0.78 mmol g) significantly improved cell adhesion; further increases in phosphate content decreased cell adhesion, indicating a strong dependence on phosphate content. Intriguingly, even in the absence of differentiation inducers, hDPSCs on P-CNF scaffolds with an optimal phosphate content of 0.78 mmol g showed equal or higher expression of hard tissue marker genes compared to collagen scaffolds with differentiation inducers, suggesting that P-CNFs can directly promote hard tissue differentiation. These findings highlight plant-derived, animal-free P-CNFs as a promising biomaterial for advanced dental tissue engineering.
人牙髓干细胞（hDPSCs）是牙齿再生治疗中一种很有前景的细胞来源。然而，传统的培养支架材料通常来自动物，这引发了对免疫原性的担忧以及可用性有限的问题。在本研究中，我们探索了具有良好纤维形态和磷酸基团的磷酸化纤维素纳米纤维（P-CNFs）作为一种新型的细胞培养支架材料。通过改变尿素和磷酸氢二铵的摩尔比以及反应时间，制备了具有不同磷酸盐含量（0 - 1.42 mmol/g）的P-CNF支架，并在其上培养永生化的hDPSCs。在未改性的CNFs上培养的细胞表现出较差的粘附性并形成球体，表明生物适应性较低。相比之下，磷酸盐含量适中（0.54 - 0.78 mmol/g）的P-CNF支架显著改善了细胞粘附；磷酸盐含量的进一步增加则降低了细胞粘附，这表明细胞粘附对磷酸盐含量有很强的依赖性。有趣的是，即使在没有分化诱导剂的情况下，磷酸盐含量为0.78 mmol/g的最佳P-CNF支架上的hDPSCs与添加了分化诱导剂的胶原支架相比，硬组织标记基因的表达相同或更高，这表明P-CNFs可以直接促进硬组织分化。这些发现突出了植物来源、无动物成分的P-CNFs作为先进牙科组织工程中有前景的生物材料的地位。
