Department of Cariology, Restorative Sciences, Endodontics, University of Michigan School of Dentistry, Ann Arbor, MI, USA.
J Endod. 2010 Nov;36(11):1805-11. doi: 10.1016/j.joen.2010.08.031. Epub 2010 Sep 19.
Dental pulp tissue engineering is an emerging field that can potentially have a major impact on oral health. However, the source of morphogens required for stem cell differentiation into odontoblasts and the scaffold characteristics that are more conducive to odontoblastic differentiation are still unclear. This study investigated the effect of dentin and scaffold porogen on the differentiation of human dental pulp stem cells (DPSCs) into odontoblasts.
Poly-L-lactic acid (PLLA) scaffolds were prepared in pulp chambers of extracted human third molars using salt crystals or gelatin spheres as porogen. DPSCs seeded in tooth slice/scaffolds or control scaffolds (without tooth slice) were either cultured in vitro or implanted subcutaneously in immunodefficient mice.
DPSCs seeded in tooth slice/scaffolds but not in control scaffolds expressed putative odontoblastic markers (DMP-1, DSPP, and MEPE) in vitro and in vivo. DPSCs seeded in tooth/slice scaffolds presented lower proliferation rates than in control scaffolds between 7 and 21 days (p < 0.05). DPSCs seeded in tooth slice/scaffolds and transplanted into mice generated a tissue with morphological characteristics similar to those of human dental pulps. Scaffolds generated with gelatin or salt porogen resulted in similar DPSC proliferation. The porogen type had a relatively modest impact on the expression of the markers of odontoblastic differentiation.
Collectively, this work shows that dentin-related morphogens are important for the differentiation of DPSC into odontoblasts and for the engineering of dental pulp-like tissues and suggest that environmental cues influence DPSC behavior and differentiation potential.
牙髓组织工程是一个新兴领域,可能对口腔健康产生重大影响。然而,用于干细胞分化为成牙本质细胞的形态发生因子的来源以及更有利于成牙本质细胞分化的支架特性仍不清楚。本研究探讨了牙本质和支架成孔剂对人牙髓干细胞(DPSCs)分化为成牙本质细胞的影响。
用人牙第三磨牙的牙髓室中的盐晶体或明胶球作为成孔剂制备聚-L-乳酸(PLLA)支架。将牙髓切片/支架或对照支架(无牙髓切片)中的 DPSCs 接种在体外培养或免疫缺陷小鼠皮下植入。
在体外和体内,接种在牙切片/支架中的 DPSCs 表达了假定的成牙本质细胞标志物(DMP-1、DSPP 和 MEPE),而不是在对照支架中。与对照支架相比,接种在牙切片/支架中的 DPSCs 在第 7 至 21 天之间的增殖率较低(p < 0.05)。接种在牙切片/支架中的 DPSCs 并移植到小鼠中产生的组织具有类似于人牙髓的形态特征。用明胶或盐成孔剂生成的支架导致 DPSCs 增殖相似。成孔剂类型对成牙本质细胞分化标志物的表达影响相对较小。
总的来说,这项工作表明牙本质相关的形态发生因子对于 DPSC 分化为成牙本质细胞以及牙髓样组织的工程化非常重要,并表明环境线索会影响 DPSC 的行为和分化潜能。