Single-cell transcriptomics reveals cell atlas and identifies cycling tumor cells responsible for recurrence in ameloblastoma.

Ameloblastoma is a benign tumor characterized by locally invasive phenotypes, leading to facial bone destruction and a high recurrence rate. However, the mechanisms governing tumor initiation and recurrence are poorly understood. Here, we uncovered cellular landscapes and mechanisms that underlie tumor recurrence in ameloblastoma at single-cell resolution. Our results revealed that ameloblastoma exhibits five tumor subpopulations varying with respect to immune response (IR), bone remodeling (BR), tooth development (TD), epithelial development (ED), and cell cycle (CC) signatures. Of note, we found that CC ameloblastoma cells were endowed with stemness and contributed to tumor recurrence, which was dominated by the EZH2-mediated program. Targeting EZH2 effectively eliminated CC ameloblastoma cells and inhibited tumor growth in ameloblastoma patient-derived organoids. These data described the tumor subpopulation and clarified the identity, function, and regulatory mechanism of CC ameloblastoma cells, providing a potential therapeutic target for ameloblastoma.

Sox9+ cells are required for salivary gland regeneration after radiation damage via the Wnt/ß-catenin pathway.

Radiotherapy for head and neck cancer can cause serious side effects, including severe damage to the salivary glands, resulting in symptoms such as xerostomia, dental caries, and oral infection. Because of the lack of long-term treatment for the symptoms of xerostomia, current research has focused on finding endogenous stem cells that can differentiate into various cell lineages to replace lost tissues and restore functions. Here, we report that Sox9+ cells can differentiate into various salivary epithelial cell lineages under homeostatic conditions. After ablating Sox9+ cells, the salivary glands of irradiated mice showed more severe phenotypes and the reduced proliferative capacity. Analysis of online single-cell RNA-sequencing data reveals the enrichment of the Wnt/ß-catenin pathway in the Sox9+ cell population. Furthermore, treatment with a Wnt/ß-catenin inhibitor in irradiated mice inhibits the regenerative capability of Sox9+ cells. Finally, we show that Sox9+ cells are capable of forming organoids in vitro and that transplanting these organoids into salivary glands after radiation partially restored salivary gland functions. These results suggest that regenerative therapy targeting Sox9+ cells is a promising approach to treat radiation-induced salivary gland injury.

Effect of gelatin sponge with colloid silver on bone healing in infected cranial defects.

Oral infectious diseases may lead to bone loss, which makes it difficult to achieve satisfactory restoration. The rise of multidrug resistant bacteria has put forward severe challenges to the use of antibiotics. Silver (Ag) has long been known as a strong antibacterial agent. In clinic, gelatin sponge with colloid silver is used to reduce tooth extraction complication. To investigate how this material affect infected bone defects, methicillin-resistant Staphylococcus aureus (MRSA) infected 3-mm-diameter cranial defects were created in adult female Sprague-Dawley rats. One week after infection, the defects were debrided of all nonviable tissue and then implanted with gelatin sponge with colloid silver (gelatin/Ag group) or gelatin alone (gelatin group). At 2 and 3days after debridement, significantly lower mRNA expression levels of IL-6 and TNF-α and lower plate colony count value were detected in gelatin/Ag group than control. Micro-CT analysis showed a significant increase of newly formed bone volume fraction (BV/TV) in gelatin/Ag treated defects. The HE stained cranium sections also showed a faster rate of defect closure in gelatin/Ag group than control. These findings demonstrated that gelatin sponge with colloid silver can effectively reduce the infection caused by MRSA in cranial defects and accelerate bone healing process.

Smoking May Lead to Marginal Bone Loss Around Non-Submerged Implants During Bone Healing by Altering Salivary Microbiome: A Prospective Study.

BACKGROUND: This prospective and controlled study elucidates the impact of smoking on the salivary microbiome and its further influence on marginal bone loss (MBL) around an implant during a 3-month bone-healing period. METHODS: Saliva samples were collected preoperatively from 20 periodontally healthy patients with single-tooth replacement in the posterior mandible (smokers [n = 10] and non-smokers [n = 10]). Sequencing of 16S recombinant RNA gene amplicons was used to characterize the salivary microbiome. Each patient received implant surgery after oral clinical assessment, and MBL around the implant was measured during a 3-month healing period. RESULTS: In total, 871,389 sequences were compared against the Human Oral Microbiome Database for bacterial identification. Microbial signatures of smokers exhibited lower diversity and richness, with a significant decrease in uncultured species. The phyla Gracilibacteria and Saccharibacteria showed a significant decrease in smokers. The genera Streptococcus, Lachnoanaerobaculum, Stomatobaculum, and Eubacterium were significantly increased in smokers, whereas Selenomonas, Selenomonas [G-3], and Catonella were significantly decreased. Specifically, Porphyromonas gingivalis was significantly more abundant in smokers, which was positively related to the severity of MBL during bone healing. CONCLUSIONS: Smoking shapes the salivary microbiome in states of clinical health, and further may influence MBL during bone healing by creating high at-risk-for-harm communities. Understanding of the distinctly divergent oral microbiome in smokers and non-smokers is a base for personalized therapeutics for this high-risk cohort and also a base for further study on the pathologic mechanisms.