Institute of Animal Physiology and Genetics, Brno, Czech Republic; Masaryk University, Brno, Czech Republic.
Institute of Animal Physiology and Genetics, Brno, Czech Republic; Veterinary University, Brno, Czech Republic.
Ann Anat. 2023 Oct;250:152149. doi: 10.1016/j.aanat.2023.152149. Epub 2023 Aug 12.
Teeth and their associated tissues contain several populations of mesenchymal stem cells, one of which is represented by dental pulp stem cells (DPSCs). These cells have mainly been characterised in vitro and numerous positive and negati ve markers for these cells have been suggested. To investigate the presence and localization of these molecules during development, forming dental pulp was examined using the mouse first mandibular molar as a model. The stages corresponding to postnatal (P) days 0, 7, 14, and 21 were investigated. The expression was monitored using customised PCR Arrays. Additionally, in situ localization of the key trio of markers (Cd73, Cd90, Cd105 coded by genes Nt5e, Thy1, Eng) was performed at prenatal and postnatal stages using immunohistochemistry. The expression panel of 24 genes assigned as in vitro markers of DPSCs or mesenchymal stem cells (MSCs) revealed their developmental dynamics during formation of dental pulp mesenchyme. Among the positive markers, Vcam1, Fgf2, Nes were identified as increasing and Cd44, Cd59b, Mcam, Alcam as decreasing between perinatal vs. postnatal stages towards adulthood. Within the panel of negative DPSC markers, Cd14, Itgb2, Ptprc displayed increased and Cd24a decreased levels at later stages of pulp formation. Within the key trio of markers, Nt5e did not show any significant expression difference within the investigated period. Thy1 displayed a strong decrease between P0 and P7 while Eng increased between these stages. In situ localization of Cd73, Cd90 and Cd105 showed them overlap in differentiated odontoblasts and in the sub-odontoblastic layer that is speculated to host odontoblast progenitors. The highly prevalent expression of particularly Cd73 and Cd90 opens the question of potential multiple functions of these molecules. The results from this study add to the in vitro based knowledge by showing dynamics in the expression of DPSC/MSC markers during dental pulp formation in an in vivo context and thus with respect to the natural environment important for commitment of stem cells.
牙齿及其相关组织包含几种间充质干细胞群体，其中一种由牙髓干细胞 (DPSCs) 代表。这些细胞主要在体外进行了表征，并提出了许多针对这些细胞的阳性和阴性标记物。为了研究这些分子在发育过程中的存在和定位，使用小鼠第一下颌磨牙作为模型检查形成的牙髓。研究了对应于出生后 (P) 第 0、7、14 和 21 天的阶段。使用定制的 PCR 阵列监测表达。此外，在产前和出生后阶段使用免疫组织化学对关键三标志物（基因 Nt5e、Thy1、Eng 编码的 Cd73、Cd90、Cd105）的原位定位进行了研究。24 个基因的表达谱被指定为 DPSCs 或间充质干细胞 (MSCs) 的体外标记物，揭示了它们在牙髓间充质形成过程中的发育动力学。在阳性标记物中，Vcam1、Fgf2、Nes 被鉴定为在围产期与出生后阶段向成年期增加，而 Cd44、Cd59b、Mcam、Alcam 则减少。在阴性 DPSC 标记物的表达谱中，Cd14、Itgb2、Ptprc 在牙髓形成的后期显示出增加的水平，而 Cd24a 则减少。在关键的三标志物中，Nt5e 在研究期间没有显示出任何显著的表达差异。Thy1 在 P0 和 P7 之间表达强烈下降，而 Eng 在这些阶段之间增加。Cd73、Cd90 和 Cd105 的原位定位显示它们在分化的成牙本质细胞和推测容纳成牙本质细胞祖细胞的亚成牙本质层中重叠。这些分子的高普遍表达尤其引发了对这些分子的潜在多种功能的疑问。这项研究的结果通过在体内背景下显示 DPSC/MSC 标记物在牙髓形成过程中的表达动态，从而与对干细胞具有重要意义的自然环境相关，补充了基于体外的知识。
