Department of Endodontics, Henry M. Goldman School of Dental Medicine, Boston University, Boston, Massachusetts 02118, USA.
J Endod. 2011 Sep;37(9):1217-24. doi: 10.1016/j.joen.2011.05.022. Epub 2011 Jul 16.
We previously reported the presence of mesenchymal stem/progenitor cells (MSCs) in inflamed pulp tissue. Here we asked whether MSCs also exist in inflamed periapical tissues resulting from endodontic infection. The objectives of this study were to detect the expression of MSC markers in periapical inflammatory tissues and to characterize isolated cells from these tissues.
Human periapical inflammatory tissues were collected and processed to detect MSC marker expression by immunohistochemistry. Cells were isolated and tested for cell surface marker expression by using flow cytometry and examined for multiple differentiation potential into osteogenic and adipogenic pathways. In vivo formation of mineralized tissues was assessed in a mouse model.
Immunohistochemistry showed positive staining for MSC markers STRO-1, CD90, and CD146. Isolated cells at passage 0 appeared as typical fibroblastic cells, and a few cells formed colony-forming unit-fibroblasts (CFU-Fs). After passaging, the CFU-F forming ability diminished dramatically, and the population doubling was up to 26. Flow cytometry data showed that these cells at passage 2 expressed low levels of STRO-1 and CD146 and moderate to high levels of CD90, CD73, and CD105. At passage 6, the levels of these markers decreased. When incubated in specific differentiation medium, cells demonstrated a strong osteogenic but weak adipogenic capacity. After in vivo cell transplantation, mineralized tissues formed in immunocompromised mice.
Human periapical inflammatory tissues expressed MSC markers, suggesting the presence of MSCs. Isolated cells exhibited typical mesenchymal cell immunophenotype with a capacity to form mineralized matrix in vitro and in vivo.
我们之前曾报道过炎症牙髓组织中存在间充质干细胞/祖细胞(MSCs)。在此,我们探讨了来源于牙髓感染的根尖周炎症组织中是否也存在 MSCs。本研究的目的是检测根尖周炎症组织中 MSC 标志物的表达,并对这些组织中分离的细胞进行特征分析。
收集人根尖周炎症组织,通过免疫组织化学法检测 MSC 标志物的表达。通过流式细胞术检测细胞表面标志物的表达,并对其向成骨和成脂分化潜能进行检测。在小鼠模型中评估体内矿化组织的形成。
免疫组织化学染色显示 MSC 标志物 STRO-1、CD90 和 CD146 呈阳性表达。第 0 代分离的细胞呈典型的成纤维细胞样,少数细胞形成集落形成单位-成纤维细胞(CFU-F)。传代后,CFU-F 形成能力显著下降,群体倍增次数达 26 次。流式细胞术数据显示,这些细胞在第 2 代时表达低水平的 STRO-1 和 CD146,以及中高水平的 CD90、CD73 和 CD105。在第 6 代时,这些标志物的水平降低。在特定的分化培养基中孵育时,细胞表现出较强的成骨能力,但成脂能力较弱。体内细胞移植后,免疫缺陷小鼠体内形成矿化组织。
人根尖周炎症组织表达 MSC 标志物,提示存在 MSCs。分离的细胞具有典型的间充质细胞免疫表型,具有体外和体内形成矿化基质的能力。