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Interkingdom Endodontic Biofilm Supernatant Induces Biphasic Inflammatory and Metabolic Reprogramming in Dental Pulp Stem Cells

ABSTRACT: This study investigate how soluble by-products derived from a four-species endodontic biofilm model impacts upon the viability, transcriptomic profile, and inflammatory response of human dental pulp stem cells (DPSCs). The sterile-filtered supernatant was extracted from an established interkingdom endodontic biofilm model comprising Streptococcus gordonii, Fusobacterium nucleatum, Porphyromonas gingivalis, and Candida albicans. DPSCs were exposed to the microbial biofilm supernatant (BSN) for 4 and 24 hours. Cellular responses were evaluated via MTT, CCK-8, LDH assays, and Annexin V/PI staining. Whole transcriptomic sequencing was performed to assess gene expression dynamics, with GO and KEGG pathway enrichment analyses. IL6 and IL8 expression was validated by qPCR and ELISA.It was found that BSN significantly suppressed DPSC metabolic activity without inducing apoptosis or necrosis. RNA-seq revealed 723 significantly differentially expressed genes at 4 h and 1667 at 24 h. Early responses were dominated by upregulation of inflammatory mediators (e.g., CXCL8, IL6, TNFAIP2), with enrichment of TNF, NF-κB, and JAK-STAT signalling pathways. At 24 h, the expression profile shifted toward redox regulation and metabolic suppression, including downregulation of glycolytic and purine metabolism pathways. IL6 and IL8 expression was markedly increased at both transcript and protein levels. Indeed, soluble factors produced by a biofilm model representative of deep caries and carious pulp exposures are capable of reprogramming DPSC function in a time-dependent manner. The soluble factors initiate a biphasic response characterized by early immune activation and later metabolic adaptation. The findings underscore the potential role of biofilm-derived products associated with deep caries and carious pulp exposures in compromising DPSC function. In addition, they emphasise the need for the development of vital pulp therapy strategies that not only neutralise microorganisms but also their secreted products.

ORGANISM(S): Homo sapiens

PROVIDER: GSE308847 | GEO | 2026/07/22

REPOSITORIES: GEO

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Project description:Tooth decay, also known as dental caries, is a prevalent oral health problem that significantly reduces an individual's quality of life; however, it can be effectively managed through restorative treatments. The condition primarily affects the enamel and dentin structures and can be treated before it progresses to the pulp chamber. Even in cases where the caries does not reach the pulp, microbial products from the lesion can still penetrate the pulp chamber, potentially inducing stress on pulp cells, including dental pulp stem cells (DPSCs). Even in the absence of direct pulpal exposure, microbial by-products originating from carious lesions may diffuse through dentinal tubules, potentially exerting stress on pulpal cells, including dental pulp stem cells (DPSCs). In this study, we performed a comparative analysis of the biological and proteomic profiles of DPSCs isolated from carious teeth that had not reached the pulp and those isolated from healthy teeth. In this study, we conducted a comparative analysis of the biological and proteomic profiles of dental pulp stem cells (DPSCs) isolated from clinically asymptomatic teeth with deep dentinal caries that had not reached the pulp and those isolated from healthy teeth. After evaluation of biological mechanisms such as senescence and apoptosis, we examined both the whole-cell proteome and secretome of these DPSCs by conducting a shotgun proteomics approach. Our findings indicate that the DPSC population isolated from decayed teeth exhibited a significantly higher proportion of senescent cells compared to the control group. Proteomic analysis further revealed that DPSCs from decayed teeth activated specific signaling pathways in response to microbial products, manifesting an inflammatory profile. Additionally, we observed an upregulation in the expression of proteins associated with extracellular matrix (ECM) remodeling and components of the senescence-associated secretory phenotype (SASP), both of which are hallmarks of the senescence process. This study highlights that even in the absence of direct pulp involvement, microbial stress resulting from caries can impact DPSCs within the pulp. The study reveals that DPSCs can be affected by microbial signals from carious lesions, even when the pulp appears clinically intact. The activation of senescence and inflammatory mechanisms in these affected cells may have deleterious effects on other critical tissues within the organism. Consequently, restorative treatments should consider targeting not only the decayed tissue but also the senescent cells within the pulp that may have been affected by the microbial stress induced by caries.

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Interkingdom Endodontic Biofilm Supernatant Induces Biphasic Inflammatory and Metabolic Reprogramming in Dental Pulp Stem Cells

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