Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), Medical Faculty and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany.
Institute of Medical Statistics and Computational Biology, Faculty of Medicine, University of Cologne, 50924 Cologne, Germany.
Int J Mol Sci. 2024 May 22;25(11):5617. doi: 10.3390/ijms25115617.
The periodontium comprising periodontal ligament (PDL), gingiva, and epithelium play crucial roles in maintaining tooth integrity and function. Understanding tissue cellular composition and gene expression is crucial for illuminating periodontal pathophysiology. This study aimed to identify tissue-specific markers via scRNA-Seq. Primary human PDL, gingiva, and epithelium tissues ( = 7) were subjected to cell hashing and sorting. scRNA-Seq library preparation using 10× Genomics protocol and Illumina sequencing was conducted. The analysis was performed using Cellranger (v3.1.0), with downstream analysis via R packages Seurat (v5.0.1) and SCORPIUS (v1.0.9). Investigations identified eight distinct cellular clusters, revealing the ubiquitous presence of epithelial and gingival cells. PDL cells evolved in two clusters with numerical superiority. The other clusters showed varied predominance regarding gingival and epithelial cells or an equitable distribution of both. The cluster harboring most cells mainly consisted of PDL cells and was present in all donors. Some of the other clusters were also tissue-inherent, while the presence of others was environmentally influenced, revealing variability across donors. Two clusters exhibited genetic profiles associated with tissue development and cellular integrity, respectively, while all other clusters were distinguished by genes characteristic of immune responses. Developmental trajectory analysis uncovered that PDL cells may develop after epithelial and gingival cells, suggesting the inherent PDL cell-dominated cluster as a final developmental stage. This single-cell RNA sequencing study delineates the hierarchical organization of periodontal tissue development, identifies tissue-specific markers, and reveals the influence of environmental factors on cellular composition, advancing our understanding of periodontal biology and offering potential insights for therapeutic interventions.
牙周组织包括牙周韧带 (PDL)、牙龈和上皮，在维持牙齿完整性和功能方面发挥着关键作用。了解组织细胞组成和基因表达对于阐明牙周病理生理学至关重要。本研究旨在通过 scRNA-Seq 鉴定组织特异性标记物。将原代人牙周韧带、牙龈和上皮组织（n = 7）进行细胞哈希和分选。使用 10× Genomics 方案和 Illumina 测序进行 scRNA-Seq 文库制备。使用 Cellranger（v3.1.0）进行分析，通过 R 包 Seurat（v5.0.1）和 SCORPIUS（v1.0.9）进行下游分析。研究确定了八个不同的细胞簇，揭示了上皮和牙龈细胞的普遍存在。PDL 细胞在两个具有数量优势的簇中进化。其他簇显示出不同程度的牙龈和上皮细胞优势或两者的均衡分布。包含最多细胞的簇主要由 PDL 细胞组成，存在于所有供体中。其他一些簇也存在于组织固有，而另一些簇则受到环境影响，显示出供体之间的变异性。两个簇表现出与组织发育和细胞完整性分别相关的遗传特征，而所有其他簇则以免疫反应特征的基因区分。发育轨迹分析表明 PDL 细胞可能在上皮和牙龈细胞之后发育，提示固有 PDL 细胞主导的簇作为最终发育阶段。这项单细胞 RNA 测序研究描绘了牙周组织发育的层次组织，鉴定了组织特异性标记物，并揭示了环境因素对细胞组成的影响，增进了我们对牙周生物学的理解，并为治疗干预提供了潜在的见解。
