Department of Nanobiomedical Science & BK21 FOUR NBM Global Research Center for Regenerative Medicine, Dankook University, Cheonan 31116, Korea.
Department of Oral Biochemistry, School of Dentistry, Dankook University, Cheonan 31116, Korea.
Genes (Basel). 2022 Apr 8;13(4):659. doi: 10.3390/genes13040659.
Ligament-fibroblastic cells and cementoblasts, two types of progenitor cells that differentiate from periodontal ligament stem cells (hPDLSCs), are responsible for the formation of the adhesive tissues in the tooth root. Since one of the factors that determines the fate of stem cell differentiation is the change in the microenvironment of the stem/progenitor cells, this study attempted to compare and analyze the molecular differences in the membrane and ECM of the two progenitor cells. Single cells derived from hPDLSCs were treated with TGF-β1 and BMP7 to obtain ligament-fibroblastic and cementoblastic cells, respectively. The transcriptome profiles of three independent replicates of each progenitor were evaluated using next-generation sequencing. The representative differentially expressed genes (DEGs) were verified by qRT-PCR, Western blot analysis, and immunohistochemistry. Among a total of 2245 DEGs identified, 142 and 114 DEGs related to ECM and cell membrane molecules were upregulated in ligament-fibroblastic and cementoblast-like cells, respectively. The major types of integrin and cadherin were found to be different between the two progenitor cells. In addition, the representative core proteins for each glycosaminoglycan-specific proteoglycan class were different between the two progenitors. This study provides a detailed understanding of cell-cell and cell-ECM interactions through the specific components of the membrane and ECM for ligament-fibroblastic and cementoblastic differentiation of hPDLSCs.
成纤维细胞-韧带细胞和成牙骨质细胞是两种前体细胞，它们分别由牙周膜干细胞（hPDLSCs）分化而来，负责形成牙根的黏附组织。由于决定干细胞分化命运的因素之一是干细胞/前体细胞微环境的变化，因此本研究试图比较和分析这两种前体细胞的细胞膜和细胞外基质（ECM）的分子差异。用 TGF-β1 和 BMP7 处理从 hPDLSCs 中分离得到的单细胞，分别获得成纤维细胞-韧带细胞和成牙骨质细胞样细胞。使用下一代测序对每个前体细胞的三个独立重复的转录组谱进行评估。通过 qRT-PCR、Western blot 分析和免疫组织化学验证代表性差异表达基因（DEGs）。在鉴定的总共 2245 个 DEGs 中，142 个和 114 个与 ECM 和细胞膜分子相关的 DEGs 在成纤维细胞-韧带细胞和成牙骨质细胞样细胞中分别上调。发现两种前体细胞之间整合素和钙黏蛋白的主要类型不同。此外，两种前体细胞中每个糖胺聚糖特异性蛋白聚糖类的代表性核心蛋白也不同。本研究通过 hPDLSCs 成纤维细胞-韧带和成牙骨质分化的细胞膜和 ECM 的特定成分，提供了对细胞-细胞和细胞-ECM 相互作用的详细了解。
