IADR Abstract Archives
Single-Cell Atlas Identifies New Inflammation-Promoting Cell Subsets in Diabetic Gingiva
Objectives: The oral gingival barrier is a constantly stimulated and dynamic environment where homeostasis is often disrupted, resulting in inflammatory periodontal diseases. Type 2 diabetes (T2D) is a recognized risk factor for periodontal disease. It has been reported to be associated with gingival barrier dysfunction, but the effects and underlying mechanisms are inconclusive.
Methods: Single-cell RNA sequencing of gingiva from diabetic mice (control/db/db, n=5/group) was performed to examine the heterogeneity of the gingival barrier. We further analyzed the differential genes and potential functional variation between control and T2D gingival barrier. The alveolar bone resorption was detected by Micro-CT scanning. Immunohistochemical staining and flow cytometry for the mice/human samples were used to verify the observations.
Results: Periodontal health is characterized by populations of Krt14+-epithelial cells and Col1a1+-fibroblasts mediating immune homeostasis primarily through the enrichment of innate lymphoid cells. The untreated diabetic mice exhibited spontaneous attachment loss. Immunohistochemical staining of the diabetic gingiva showed a profound leukocytes’ infiltration, diffuse fibrosis, and reduced expression of claudin-1 (an epithelial cell-to-cell adhesion molecule), reflecting a dysfunctional gingival barrier in T2D status. Immunofluorescent assays of mice and human gingiva confirmed the decreased epithelial/stromal ratio and increased transcriptional activity of fibroblasts in T2D. We further observed stromal, particularly fibroblast immune hyperresponsiveness, linked to the recruitment of myeloid-derived cells at the diabetic gingiva. Notably, the “immune-like” stromal cells also contributed to the hyperresponsive γδT cells. Both sequencing and histological analysis suggest the inflammatory signaling, between the “immune-like” fibroblasts and innate immune cells (specifically neutrophils and γδT cells), as the potential driver of diabetes-induced periodontal lesions.
Conclusions: Our work characterizes the cellular transcriptional landscape of gingiva in the setting of T2D. We define a fibroblast subset with a “stromal-immune” dual feature in diabetic gingiva and highlight the diversity of this subset and its interaction with innate immune cells.
Methods: Single-cell RNA sequencing of gingiva from diabetic mice (control/db/db, n=5/group) was performed to examine the heterogeneity of the gingival barrier. We further analyzed the differential genes and potential functional variation between control and T2D gingival barrier. The alveolar bone resorption was detected by Micro-CT scanning. Immunohistochemical staining and flow cytometry for the mice/human samples were used to verify the observations.
Results: Periodontal health is characterized by populations of Krt14+-epithelial cells and Col1a1+-fibroblasts mediating immune homeostasis primarily through the enrichment of innate lymphoid cells. The untreated diabetic mice exhibited spontaneous attachment loss. Immunohistochemical staining of the diabetic gingiva showed a profound leukocytes’ infiltration, diffuse fibrosis, and reduced expression of claudin-1 (an epithelial cell-to-cell adhesion molecule), reflecting a dysfunctional gingival barrier in T2D status. Immunofluorescent assays of mice and human gingiva confirmed the decreased epithelial/stromal ratio and increased transcriptional activity of fibroblasts in T2D. We further observed stromal, particularly fibroblast immune hyperresponsiveness, linked to the recruitment of myeloid-derived cells at the diabetic gingiva. Notably, the “immune-like” stromal cells also contributed to the hyperresponsive γδT cells. Both sequencing and histological analysis suggest the inflammatory signaling, between the “immune-like” fibroblasts and innate immune cells (specifically neutrophils and γδT cells), as the potential driver of diabetes-induced periodontal lesions.
Conclusions: Our work characterizes the cellular transcriptional landscape of gingiva in the setting of T2D. We define a fibroblast subset with a “stromal-immune” dual feature in diabetic gingiva and highlight the diversity of this subset and its interaction with innate immune cells.