Tooth pulpitis is a prevalent oral disorder. Understanding the underlying mechanisms of pulpitis and developing effective treatment strategies hold great significance. Ferroptosis has recently emerged as a new form of cell death, but the role of ferroptosis in pulpitis remains largely unknown. In our study, single-cell RNA sequencing (scRNA-seq) was used to identify cellular heterogeneity between 3 pulpitis tissue and 3 healthy pulp tissue, and explored ferroptosis occurrence in pulpitis tissue and inflamed dental pulp cells (DPCs). In scRNA-seq, 40 231 cells (Pulpitis: 17 814; Healthy pulp: 22 417) were captured, and visualized into 12 distinct cell clusters. Differentially expressed ferroptosis-related genes (DE-FRGs) were almost presented in each cluster in pulpitis vs healthy pulp. ROS and Fe²⁺ levels significantly rose, and immunohistochemistry showed low expression of GPX4 and high expression of PTGS2 in pulpitis. In LPS-stimulated DPCs, thymosin α1 increased the expression of GPX4 and FTL, and decreased expression of TNF-α, IL-1β, IL-6, and Fe²⁺ levels. In rat pulpitis models, both prothymosin α (PTMA, precursor of thymosin α1) gelatin sponge placed at the hole of pulp (LPS-P(gs)) and PTMA injection in pulp (LPS-P(i)) significantly reduced infiltration of inflammatory cells and expression of PTGS2, and increased the expression of GPX4. In RNA sequencing, the expression of DE-FRGs were reversed when thymosin α1 were added in LPS-stimulated DPCs. Collectively, single-cell atlas reveals cellular heterogeneity between pulpitis and healthy pulp, and ferroptosis occurrence in pulpitis. Thymosin α1 may reduce ferroptosis in DPCs to alleviate pulpitis and thus potentially has the ability to treat pulpitis.
Ferroptosis/drug effects* ; Dental Pulp/drug effects* ; Animals ; Pulpitis/pathology* ; Rats ; Thymalfasin/pharmacology* ; Humans ; Male ; Thymosin/pharmacology* ; Disease Models, Animal ; Rats, Sprague-Dawley

The purpose of this study was to observe histopathologically the influence of advanced periodontitis on pulp tissue, and to conclude the correlation between the results with clinical madifestations. The samples were teeth with over 7mm pocket depth and over 50% radiographic bone loss. These were diagnosed to have very poor prognosis and thus planned to be extracted. Those with any of following conditions were excluded from the samples, loss of vitality, periapical pathology, restoration or prosthesis, dental caries, and attrition or abrasion. It was because these conditions could affect pulp without any correlation with periodontal disease. For the experiment, 17 teeth from 11 patients were selected. Average age of patient was 47. Each tooth was examined for following categoris; pocket depth, gingival recession, electric pulp test, mobility, percussion test, sensitivity test. The extracted teeth were fixed buffered neutral formalin solution. It was decalcified using 4% nitric acid. Sliced histological samples observed using light microscope, for pulp status, and severeity of inflammation. 4 samples were excluded due to histologic sample discrepency. Thus 13 samples were subject to observation. 4 showed normal conditions. Focal reversable pulpitis was shown in 5 samples. Chronic pulpitis was observed 1 samples. Pulpal abscess observed in 3 samples.
Abscess ; Adult* ; Chronic Periodontitis* ; Dental Prosthesis ; Formaldehyde ; Gingival Recession ; Humans ; Inflammation ; Nitric Acid ; Pathology ; Percussion ; Periodontal Diseases ; Periodontitis ; Prognosis ; Pulpitis ; Tooth