VEGF-C–Driven Lymphatic Survival Signaling Promotes Periodontal Repair

Lymphatic vessels play a pivotal role in tissue homeostasis and repair, yet their function in periodontal healing remains poorly defined. In this study, we investigated the spatial dynamics and functional significance of lymphatics during periodontal wound repair using a mouse model of ligature-induced periodontitis. By combining Prox1-tdTomato transgenic reporter mice with advanced tissue clearing and light sheet microscopy, we generated high-resolution 3-dimensional images of gingival lymphatic networks under both physiological and repair conditions. Our observations revealed that lymphatic vessels undergo active and spatially organized reassembly during the healing phase. Notably, we found a region-specific increase in lymphatic branching and density in the marginal gingiva, suggesting localized lymphangiogenesis. Time course analysis confirmed that this lymphatic remodeling peaks during the granulation tissue formation phase. We further identified epithelial upregulation of Vegfc during the repair phase and demonstrated that local administration of vascular endothelial growth factor C (VEGF-C) C156S, a VEGFR-3-specific agonist, enhanced lymphatic vessel function and alveolar bone repair. To explore the underlying molecular mechanisms, we successfully established, for the first time, primary cultures of gingiva-derived lymphatic endothelial cells (LECs). Comparative transcriptomic analysis revealed distinct signatures of gingival LECs compared to lymph node or dermal LECs. These cells maintained typical LEC characteristics in vitro and responded robustly to VEGF-C stimulation. RNA sequencing and functional apoptosis assays revealed significant modulation of apoptosis-related genes, including downregulation of <i>Ell3</i> and upregulation of <i>Siah1b</i>, suggesting a survival-promoting role for VEGF-C signaling in LECs. Collectively, our findings highlight a previously underappreciated regenerative role of lymphatic vessels in the periodontium and establish lymphangiogenic activation as a potential therapeutic strategy for enhancing periodontal repair. The successful culture of primary gingival LECs further provides a novel platform for mechanistic studies in oral lymphatic biology.