RESUMO
BACKGROUND: Periodontitis is a dental disease characterized by inflammation of periodontal tissues and loss of the periodontal ligaments and alveolar bone. Exosomes are a class of extracellular vesicles that are involved in a variety of diseases by releasing active substances. In this study, we aimed to investigate the effect and mechanism of exosomes from M2 polarized macrophages (M2-exos) on osteogenic differentiation in human periodontal ligament stem cells (hPDLSCs). METHODS: M2-exos were isolated from IL-4-induced RAW264.7 cells (M2 macrophages) and then treated on hPDLSCs. Osteogenic differentiation was assessed by alkaline phosphatase (ALP) staining, alizarin red S (ARS) staining, measurement of osteogenic differentiation-related genes and proteins, and inflammation was evaluated by measuring the levels of inflammatory factors. The mechanism of M2-exo was confirmed through qPCR, western blot, ALP and ARS staining. RESULTS: Results suggested that M2-exo improved osteogenic differentiation and inhibited inflammation in LPS-induced hPDLSCs. CXCL12 expression was elevated in M2 macrophages, but decreased in LPS-induced hPDLSCs. Moreover, the effect of M2-exo on osteogenic differentiation and inflammation in LPS-induced hPDLSCs was reversed by CXCL12 knockdown. CONCLUSION: We demonstrated that M2-exo facilitated osteogenic differentiation and suppressed inflammation in LPS-induced hPDLSCs through promotion of CXCL12 expression. These results suggested the potential of M2-exo in the treatment of periodontitis, which may provide a new theoretical basis for M2-exo treatment of periodontitis.
Assuntos
Diferenciação Celular , Quimiocina CXCL12 , Exossomos , Inflamação , Macrófagos , Osteogênese , Ligamento Periodontal , Células-Tronco , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , Humanos , Exossomos/metabolismo , Macrófagos/metabolismo , Células-Tronco/metabolismo , Quimiocina CXCL12/metabolismo , Inflamação/metabolismo , Camundongos , Animais , Células Cultivadas , Periodontite/metabolismo , Células RAW 264.7RESUMO
Periodontitis, a common oral disease characterized by the progressive infiltration of bacteria, is a leading cause of adult tooth loss. Periodontal stem cells (PDLSCs) possess good selfrenewal and multipotential differentiation abilities to maintain the integrity of periodontal support structure and repair defects. The present study aimed to analyze the roles of Wnt7B and frizzled4 (FZD4) in the osteogenic differentiation and macrophage polarization during periodontitis using an in vitro cell model. First, Wnt7B expression in the periodontitisaffected gingival tissue of patients and lipopolysaccharide (LPS)stimulated PDLSCs was assessed using the GSE23586 dataset and western blot analysis, respectively. In Wnt7Boverexpressing PDLSCs exposed to LPS, the capacity of osteogenic differentiation was evaluated by detecting alkaline phosphatase activity, the level of Alizarin Red S staining and the expression of genes related to osteogenic differentiation. Subsequently, conditioned medium from PDLSCs overexpressing Wnt7B was used for M0 macrophage culture. The expression of CD86 and INOS was examined using immunofluorescence staining and western blot analysis. In addition, reverse transcriptionquantitative PCR was employed to examine the expression of TNFα, IL6 and IL1ß in macrophages. The binding between Wnt7B and FZD4 was estimated using coimmunoprecipitation. In addition, FZD4 was silenced to perform the rescue experiments to elucidate the regulatory mechanism between Wnt7B and FZD4. The results demonstrated a decreased expression of Wnt7B in periodontitisaffected gingival tissue and in LPSexposed PDLSCs. Wnt7B overexpression promoted the osteogenic differentiation of LPSexposed PDLSCs and suppressed the M1 polarization of macrophages. Additionally, Wnt7B bound to FZD4 and upregulated FZD4 expression. FZD4 silencing reversed the effects of Wnt7B overexpression on the osteogenic differentiation in LPSexposed PDLSCs and the M1 polarization of macrophages. In summary, Wnt7B plays an antiperiodontitis role by binding FZD4 to strengthen the osteogenic differentiation of LPSstimulated PDLSCs and suppress the M1 polarization of macrophages.
Assuntos
Diferenciação Celular , Receptores Frizzled , Lipopolissacarídeos , Macrófagos , Osteogênese , Ligamento Periodontal , Células-Tronco , Proteínas Wnt , Humanos , Receptores Frizzled/metabolismo , Receptores Frizzled/genética , Osteogênese/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , Proteínas Wnt/metabolismo , Proteínas Wnt/genética , Células-Tronco/metabolismo , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Periodontite/metabolismo , Periodontite/patologia , Células Cultivadas , Adulto , Ligação ProteicaRESUMO
Given the posited role of oxidative stress in the pathogenesis of both periodontitis and type 2 diabetes mellitus (T2DM), it may also serve as a link between these highly prevalent chronic inflammatory diseases. This view is supported by an ample body of evidence indicating that the severity and progression of periodontitis is in part driven by diabetes, while periodontal infection may hinder the attainment of adequate glycemic control in diabetic patients. Thus, this review focuses on the potential synergistic interactions along the oxidative stress-inflammation pathway characterizing both conditions. Because periodontitis and T2DM share the same risk factors and compromise patients' quality of life, to develop effective strategies for combatting both conditions, their mutual influence needs to be explored.
Assuntos
Diabetes Mellitus Tipo 2 , Estresse Oxidativo , Doenças Periodontais , Humanos , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/complicações , Doenças Periodontais/metabolismo , Doenças Periodontais/etiologia , Inflamação/metabolismo , Animais , Periodontite/metabolismo , Periodontite/complicaçõesRESUMO
OBJECTIVE: Bidirectional influences between senescence and inflammation are newly discovered. This study aimed to clarify the roles and mechanism of Porphyromonas gingivalis (P. gingivalis) in exacerbating senescence in human gingival fibroblasts (HGFs). DESIGN: Subgingival plaque and gingivae were collected from twenty-four periodontitis patients and eighteen periodontally healthy subjects. Quantities of P. gingivalis in subgingival plaque were explored using real-time PCR and the expressions of p53, p21 and SIRT6 in gingivae were detected by IHC. Moreover, senescence in HGFs was induced by P. gingivalis lipopolysaccharide (LPS) and the expressions of senescence-related ß-galactosidase (SA-ß-gal), p53, p21 and senescence-associated secretory phenotype (IL-6 and IL-8) with or without treatment by SIRT6 activator UBCS039 were explored by IHC, western blot and ELISA, respectively. In addition, the levels of SIRT6, Nrf2, HO-1 and reactive oxygen species (ROS) were examined by western blot and flow cytometry. RESULTS: Quantities of P. gingivalis in subgingival plaque and semi-quantitative scores of p53 and p21 in gingivae of periodontitis patients were increased compared with healthy controls (p < 0.05), while SIRT6 score in periodontitis patients was decreased (p < 0.001). Quantities of P. gingivalis were positively correlated with p53 and p21 scores (0.6 < r < 0.9, p < 0.01), and negatively correlated with SIRT6 score (-0.9 < r<-0.6, p < 0.01). Moreover, P. gingivalis LPS increased the levels of SA-ß-gal, p53, p21, IL-6, IL-8 and ROS and decreased the levels of SIRT6, Nrf2 and HO-1 in HGFs, which was rescued by UBCS039 (p < 0.05). CONCLUSIONS: P. gingivalis LPS could induce senescence of HGFs, which could be reversed by SIRT6 via Nrf2-HO-1 signaling pathway.
Assuntos
Senescência Celular , Fibroblastos , Gengiva , Fator 2 Relacionado a NF-E2 , Porphyromonas gingivalis , Espécies Reativas de Oxigênio , Sirtuínas , Humanos , Porphyromonas gingivalis/patogenicidade , Gengiva/microbiologia , Gengiva/metabolismo , Fibroblastos/metabolismo , Sirtuínas/metabolismo , Sirtuínas/genética , Masculino , Feminino , Adulto , Fator 2 Relacionado a NF-E2/metabolismo , Fator 2 Relacionado a NF-E2/genética , Espécies Reativas de Oxigênio/metabolismo , Lipopolissacarídeos/farmacologia , Periodontite/microbiologia , Periodontite/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Pessoa de Meia-Idade , Heme Oxigenase-1/metabolismo , Heme Oxigenase-1/genética , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Inibidor de Quinase Dependente de Ciclina p21/genéticaRESUMO
G protein-coupled receptor (GPR)40 and GPR120 are receptors for medium- and long-chain free fatty acids. It has been well documented that GPR40 and GPR120 activation improves metabolic syndrome (MetS) and exerts anti-inflammatory effects. Since chronic periodontitis is a common oral inflammatory disease initiated by periodontal pathogens and exacerbated by MetS, we determined if GPR40 and GPR120 activation with agonists improves MetS-associated periodontitis in animal models in this study. We induced MetS and periodontitis by high-fat diet feeding and periodontal injection of lipopolysaccharide, respectively, and treated mice with GW9508, a synthetic GPR40 and GPR120 dual agonist. We determined alveolar bone loss, osteoclast formation, and periodontal inflammation using micro-computed tomography, osteoclast staining, and histology. To understand the underlying mechanisms, we further performed studies to determine the effects of GW9508 on osteoclastogenesis and proinflammatory gene expression in vitro. Results showed that GW9508 improved metabolic parameters, including glucose, lipids, and insulin resistance. Results also showed that GW9508 improves periodontitis by reducing alveolar bone loss, osteoclastogenesis, and periodontal inflammation. Finally, in vitro studies showed that GW9508 inhibited osteoclast formation and proinflammatory gene secretion from macrophages. In conclusion, this study demonstrated for the first time that GPR40/GPR120 agonist GW9508 reduced alveolar bone loss and alleviated periodontal inflammation in mice with MetS-exacerbated periodontitis, suggesting that activating GPR40/GPR120 with agonist GW9508 is a potential anti-inflammatory approach for the treatment of MetS-associated periodontitis.
Assuntos
Síndrome Metabólica , Metilaminas , Periodontite , Propionatos , Receptores Acoplados a Proteínas G , Animais , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/metabolismo , Camundongos , Síndrome Metabólica/tratamento farmacológico , Síndrome Metabólica/metabolismo , Síndrome Metabólica/complicações , Propionatos/farmacologia , Propionatos/uso terapêutico , Periodontite/tratamento farmacológico , Periodontite/metabolismo , Metilaminas/farmacologia , Masculino , Camundongos Endogâmicos C57BL , Perda do Osso Alveolar/tratamento farmacológico , Perda do Osso Alveolar/etiologia , Dieta Hiperlipídica/efeitos adversos , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Modelos Animais de Doenças , Osteogênese/efeitos dos fármacosRESUMO
Purpose: Periodontitis is a chronic infectious disease characterized by progressive inflammation and alveolar bone loss. Forkhead box O1 (FoxO1), an important regulator, plays a crucial role in maintaining bone homeostasis and regulating macrophage energy metabolism and osteogenic differentiation of mesenchymal stem cells (MSCs). In this study, FoxO1 was overexpressed into small extracellular vesicles (sEV) using engineering technology, and effects of FoxO1-overexpressed sEV on periodontal tissue regeneration as well as the underlying mechanisms were investigated. Methods: Human periodontal ligament stem cell (hPDLSCs)-derived sEV (hPDLSCs-sEV) were isolated using ultracentrifugation. They were then characterized using transmission electron microscopy, Nanosight, and Western blotting analyses. hPDLSCs were treated with hPDLSCs-sEV in vitro after stimulation with lipopolysaccharide, and osteogenesis was evaluated. The effect of hPDLSCs-sEV on the polarization phenotype of THP-1 macrophages was also evaluated. In addition, we measured the reactive oxygen species (ROS) levels, adenosine triphosphate (ATP) production, mitochondrial characteristics, and metabolism of hPDLSCs and THP-1 cells. Experimental periodontitis was established in vivo in mice. HPDLSCs-sEV or phosphate-buffered saline (PBS) were injected into periodontal tissues for four weeks, and the maxillae were collected and assessed by micro-computed tomography, histological staining, and small animal in vivo imaging. Results: In vitro, FoxO1-overexpressed sEV promoted osteogenic differentiation of hPDLSCs in the inflammatory environment and polarized THP-1 cells from the M1 phenotype to the M2 phenotype. Furthermore, FoxO1-overexpressed sEV regulated the ROS level, ATP production, mitochondrial characteristics, and metabolism of hPDLSCs and THP-1 cells in the inflammatory environment. In the in vivo analyses, FoxO1-overexpressed sEV effectively promoted bone formation and inhibited inflammation. Conclusion: FoxO1-overexpressed sEV can regulate osteogenesis and immunomodulation. The ability of FoxO1-overexpressed sEV to regulate inflammation and osteogenesis can pave the way for the establishment of a therapeutic approach for periodontitis.
Assuntos
Vesículas Extracelulares , Proteína Forkhead Box O1 , Mitocôndrias , Osteogênese , Ligamento Periodontal , Periodontite , Vesículas Extracelulares/química , Vesículas Extracelulares/metabolismo , Osteogênese/efeitos dos fármacos , Animais , Humanos , Proteína Forkhead Box O1/metabolismo , Proteína Forkhead Box O1/genética , Mitocôndrias/metabolismo , Periodontite/terapia , Periodontite/metabolismo , Camundongos , Ligamento Periodontal/citologia , Células THP-1 , Espécies Reativas de Oxigênio/metabolismo , Inflamação/metabolismo , Masculino , Diferenciação Celular/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Macrófagos/metabolismo , Regeneração , Células CultivadasRESUMO
BACKGROUND: Depression is a chronic psychiatric disease of multifactorial etiology, and its pathophysiology is not fully understood. Stress and other chronic inflammatory pathologies are shared risk factors for psychiatric diseases, and comorbidities are features of major depression. Epidemiological evidence suggests that periodontitis, as a source of low-grade chronic systemic inflammation, may be associated with depression, but the underlying mechanisms are not well understood. METHODS: Periodontitis (P) was induced in Wistar: Han rats through oral gavage with the pathogenic bacteria Porphyromonas gingivalis and Fusobacterium nucleatum for 12 weeks, followed by 3 weeks of chronic mild stress (CMS) to induce depressive-like behavior. The following four groups were established (n = 12 rats/group): periodontitis and CMS (P + CMS+), periodontitis without CMS, CMS without periodontitis, and control. The morphology and inflammatory phenotype of microglia in the frontal cortex (FC) were studied using immunofluorescence and bioinformatics tools. The endocannabinoid (EC) signaling and proteins related to synaptic plasticity were analyzed in FC samples using biochemical and immunohistochemical techniques. RESULTS: Ultrastructural and fractal analyses of FC revealed a significant increase in the complexity and heterogeneity of Iba1 + parenchymal microglia in the combined experimental model (P + CMS+) and increased expression of the proinflammatory marker inducible nitric oxide synthase (iNOS), while there were no changes in the expression of cannabinoid receptor 2 (CB2). In the FC protein extracts of the P + CMS + animals, there was a decrease in the levels of the EC metabolic enzymes N-acyl phosphatidylethanolamine-specific phospholipase D (NAPE-PLD), diacylglycerol lipase (DAGL), and monoacylglycerol lipase (MAGL) compared to those in the controls, which extended to protein expression in neurons and in FC extracts of cannabinoid receptor 1 (CB1) and to the intracellular signaling molecules phosphatidylinositol-3-kinase (PI3K), protein kinase B (Akt) and extracellular signal-regulated kinase 1/2 (ERK1/2). The protein levels of brain-derived neurotrophic factor (BDNF) and synaptophysin were also lower in P + CMS + animals than in controls. CONCLUSIONS: The combined effects on microglial morphology and inflammatory phenotype, the EC signaling, and proteins related to synaptic plasticity in P + CMS + animals may represent relevant mechanisms explaining the association between periodontitis and depression. These findings highlight potential therapeutic targets that warrant further investigation.
Assuntos
Depressão , Endocanabinoides , Microglia , Periodontite , Ratos Wistar , Transdução de Sinais , Animais , Ratos , Endocanabinoides/metabolismo , Microglia/metabolismo , Microglia/patologia , Periodontite/patologia , Periodontite/metabolismo , Transdução de Sinais/fisiologia , Depressão/metabolismo , Depressão/patologia , Masculino , Modelos Animais de Doenças , Fenótipo , Inflamação/metabolismo , Inflamação/patologiaRESUMO
This study aimed to compare several potential mouthrinse biomarkers for periodontitis including active matrix-metalloproteinase-8 (aMMP-8), total MMP-8, and other inflammatory biomarkers in diagnosing and monitoring the effects of nonsurgical periodontal therapy. Thirteen patients with stage III/IV periodontitis were recruited, along with thirteen periodontally and systemically healthy controls. These 13 patients were representative of the number of outpatients visiting any dentist in a single day. Full-mouth clinical periodontal parameters and biomarkers (the aMMP-8 point-of-care-test [POCT], total MMP-8, tissue inhibitor of MMPs (TIMP)-1, the aMMP-8 RFU activity assay, Myeloperoxidase, PMN elastase, calprotectin, and interleukin-6) were recorded at baseline and after nonsurgical therapy at 6 weeks. The aMMP-8 POCT was the most efficient and precise discriminator, with a cut-off of 20 ng/mL found to be optimal. Myeloperoxidase, MMP-8's oxidative activator, was also efficient. Following closely in precision was the aMMP-8 RFU activity assay and PMN elastase. In contrast, the total MMP-8 assay and the other biomarkers were less efficient and precise in distinguishing patients with periodontitis from healthy controls. aMMP-8, MPO, and PMN elastase may form a proteolytic and pro-oxidative tissue destruction cascade in periodontitis, potentially representing a therapeutic target. The aMMP-8 chair-side test with a cut-off of 20 ng/mL was the most efficient and precise discriminator between periodontal health and disease. The aMMP-8 POC test can be effectively used by dental professionals in their dental practices in online and real-time diagnoses as well as in monitoring periodontal disease and educating and encouraging good oral practices among patients.
Assuntos
Biomarcadores , Metaloproteinase 8 da Matriz , Periodontite , Humanos , Metaloproteinase 8 da Matriz/metabolismo , Periodontite/diagnóstico , Periodontite/terapia , Periodontite/metabolismo , Feminino , Masculino , Pessoa de Meia-Idade , Adulto , Peroxidase/metabolismo , Sistemas Automatizados de Assistência Junto ao Leito , Elastase de Leucócito/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismoRESUMO
Signal regulatory protein alpha (SIRPα) is mainly expressed by cells of myeloid origin. This membrane glycoprotein is shown to be involved in regulation of different inflammatory conditions, such as colitis and arthritis. However, SIRPα has not been investigated in relationship to periodontitis, an inflammatory condition affecting the tooth supporting tissues. We aim to investigate if resident cells in the periodontium express SIRPα and whether a possible expression is affected by inflammatory conditions. Primary human keratinocytes, fibroblasts, periodontal ligament cells, and osteoblasts were cultured with or without the pro-inflammatory cytokines tumor necrosis factor alpha (TNF-α) or interleukin-1-beta (IL-1ß). All different periodontal cell types showed a basal mRNA expression of SIRPα. Pro-inflammatory cytokines induced a 2-3-fold significant increase in SIRPα expression in both cultured human gingival fibroblasts and osteoblasts but neither in keratinocytes nor in periodontal ligament cells. Tissue sections from human gingival tissue biopsies were histochemically stained for SIRPα. Epithelial keratinocytes and gingival fibroblasts stained positive in sections from periodontally healthy as well as in sections from periodontitis. In periodontitis sections, infiltrating leukocytes stained positive for SIRPα. We highlight our finding that oral keratinocytes, gingival fibroblasts, and periodontal ligament cells do express SIRPα, as this has not been presented before. The fact that inflammatory stimulation of gingival fibroblasts increased the expression of SIRPα, while an increased expression by gingival fibroblasts in periodontitis tissue in situ could not be detected, is indeed contradictory.
Assuntos
Receptores Imunológicos , Humanos , Receptores Imunológicos/metabolismo , Células Cultivadas , Periodonto/metabolismo , Periodonto/patologia , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , Antígenos de Diferenciação/metabolismo , Queratinócitos/metabolismo , Periodontite/metabolismo , Fibroblastos/metabolismoRESUMO
Tooth-related inflammatory disorders, including caries, pulpitis, apical periodontitis (AP), and periodontitis (PD), are primarily caused by resident oral microorganisms. Although these dental inflammatory conditions are typically not life-threatening, neglecting them can result in significant complications and greatly reduce an individual's quality of life. Nuclear factor κB (NF-κB), a family formed by various combinations of Rel proteins, is extensively involved in inflammatory diseases and even cancer. This study reviews recent data on NF-κB signaling and its role in dental pulp stem cells (DPSCs), dental pulp fibroblasts (DPFs), odontoblasts, human periodontal ligament cells (hPDLCs), and various experimental animal models. The findings indicate that NF-κB signaling is abnormally activated in caries, pulpitis, AP, and PD, leading to changes in related cellular differentiation. Under specific conditions, NF-κB signaling occasionally interacts with other signaling pathways, affecting inflammation, bone metabolism, and tissue regeneration processes. In summary, data collected over recent years confirm the central role of NF-κB in dental inflammatory diseases, potentially providing new insights for drug development targeting NF-κB signaling pathways in the treatment of these conditions. Keywords: NF-κB, dental caries, pulpitis, apical periodontitis, periodontitis.
Assuntos
Cárie Dentária , NF-kappa B , Periodontite Periapical , Periodontite , Transdução de Sinais , Humanos , NF-kappa B/metabolismo , Cárie Dentária/metabolismo , Cárie Dentária/patologia , Cárie Dentária/imunologia , Periodontite/metabolismo , Periodontite/imunologia , Periodontite/patologia , Animais , Periodontite Periapical/metabolismo , Periodontite Periapical/patologia , Periodontite Periapical/imunologia , Pulpite/metabolismo , Pulpite/patologia , Pulpite/imunologia , Polpa Dentária/imunologia , Polpa Dentária/metabolismo , Polpa Dentária/patologia , Inflamação/metabolismo , Inflamação/imunologiaRESUMO
BACKGROUND: Periodontitis results from host-microbe dysbiosis and the resultant dysregulated immunoinflammatory response. Importantly, it closely links to numerous systemic comorbidities, and perplexingly contributes to adverse pregnancy outcomes (APOs). Currently, there are limited studies on the distal consequences of periodontitis via oral-gut axis in pregnant women. This study investigated the integrative microbiome-metabolome profiles through multi-omics approaches in first-trimester pregnant women and explored the translational potentials. METHODS: We collected samples of subgingival plaques, saliva, sera and stool from 54 Chinese pregnant women at the first trimester, including 31 maternal periodontitis (Perio) subjects and 23 Non-Perio controls. By integrating 16S rRNA sequencing, untargeted metabolomics and clinical traits, we explored the oral-gut microbial and metabolic connection resulting from periodontitis among early pregnant women. RESULTS: We demonstrated a novel bacterial distinguisher Coprococcus from feces of periodontitis subjects in association with subgingival periodontopathogens, being different from other fecal genera in Lachnospiraceae family. The ratio of fecal Coprococcus to Lachnoclostridium could discriminate between Perio and Non-Perio groups as the ratio of subgingival Porphyromonas to Rothia did. Furthermore, there were differentially abundant fecal metabolic features pivotally enriched in periodontitis subjects like L-urobilin and kynurenic acid. We revealed a periodontitis-oriented integrative network cluster, which was centered with fecal Coprococcus and L-urobilin as well as serum triglyceride. CONCLUSIONS: The current findings about the notable influence of periodontitis on fecal microbiota and metabolites in first-trimester pregnant women via oral-gut axis signify the importance and translational implications of preconceptional oral/periodontal healthcare for enhancing maternal wellbeing.
Assuntos
Fezes , Metaboloma , Periodontite , Primeiro Trimestre da Gravidez , Humanos , Feminino , Gravidez , Periodontite/microbiologia , Periodontite/metabolismo , Adulto , Fezes/microbiologia , Boca/microbiologia , Microbiota , Microbioma Gastrointestinal , RNA Ribossômico 16S/genéticaRESUMO
To explore severity and progression biomarkers, we examined the clinical relevance of multiple cytokines and mediators involved in the inflammatory response in periodontitis. A cohort of 68 patients was enrolled in the study and periodontal status assessed by the current classification of periodontal diseases. Immune mediators present in saliva, of both patients and healthy controls, were quantified using a Legendplex-13 panel. Clinic parameters were significantly higher in PD patients compared with HC, with a strong significant association with the disease severity (stage) (p < 0.001), but not with progression (grade). The panel of immune mediators evidenced elevated levels of pro-inflammatory cytokines IL-6 and IL-1ß as disease established (p < 0.01). IL-1ß/IL-1RA ratio was increased in PD patients, being associated with disease stage. An anti-inflammatory response was spotted by higher IL-10. Lower levels of IL-23 and IP-10 were associated with disease severity. No significant statistical differences were found by grade classification. Moreover, salivary IL-1ß and IL-6 exhibited significant positive correlations with several clinical measurements (PI, BOP, PPD, CAL), while IP-10 showed a statistical negative correlation with BOP, PPD, and CAL. These insights highlight the complexity of the periodontitis inflammatory network and the potential of cytokines as biomarkers for refined diagnostic and therapeutic strategies.
Assuntos
Biomarcadores , Interleucina-10 , Interleucina-1beta , Interleucina-6 , Periodontite , Saliva , Índice de Gravidade de Doença , Humanos , Biomarcadores/metabolismo , Feminino , Masculino , Saliva/metabolismo , Interleucina-1beta/metabolismo , Interleucina-1beta/análise , Interleucina-6/metabolismo , Pessoa de Meia-Idade , Interleucina-10/metabolismo , Periodontite/metabolismo , Periodontite/patologia , Periodontite/imunologia , Adulto , Estudos de Casos e ControlesRESUMO
BACKGROUND: This study compared the concentrations of interleukin (IL)-6, IL-17, and IL-35 in the gingival crevicular fluid of periodontally healthy participants with individuals who had stage III and IV periodontitis. METHODS: In total, 60 participants with stage III grade B-C (n = 12)-stage IV grade C (n = 18) periodontitis and 30 healthy controls were included in this cross-sectional study. Full-mouth clinical periodontal measurements were performed. Concentrations of IL-6, IL-17, and IL-35 were determined using enzyme-linked immunosorbent assays. Parametric/nonparametric methods, Pearson's/Spearman's correlation, and logistic regression methods were used for data analyses. RESULTS: The periodontitis group exhibited significantly higher levels of IL-6, IL-17, and IL-35 compared with the healthy group (p < 0.001). IL-17 levels had a positive correlation with pocket depth (PD) (r = 0.395; p = 0.031) in the periodontitis group. IL-6, IL-17, and IL-35 levels were associated with periodontitis (odds ratio [OR] = 1.344, 95% confidence interval [CI] = 1.159-1.56; OR = 1.063, 95% CI = 1.025-1.102; OR = 1.261, 95% CI = 1.110-1.434, respectively) (p < 0.001, p = 0.001, p < 0.001, respectively). Full-mouth and sampling sites PD and clinical attachment loss (CAL) values were significantly higher in the periodontitis group than in the healthy group (p < 0.001). CONCLUSIONS: This study revealed upregulated levels of IL-6, IL-17, and IL-35 in periodontitis patients compared to healthy individuals. IL-17 shows a correlation with increased PD. These findings suggest a potential association between these cytokines and severe and advanced periodontitis. TRIAL REGISTRATION: The trial was registered in ClinicalTrials.gov with this identifier NCT05306860 on 24/01/2022.
Assuntos
Líquido do Sulco Gengival , Interleucina-17 , Interleucina-6 , Interleucinas , Periodontite , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos de Casos e Controles , Estudos Transversais , Ensaio de Imunoadsorção Enzimática , Líquido do Sulco Gengival/química , Interleucina-17/análise , Interleucina-17/metabolismo , Interleucina-6/análise , Interleucina-6/metabolismo , Interleucinas/análise , Interleucinas/metabolismo , Índice Periodontal , Periodontite/diagnóstico , Periodontite/metabolismo , Periodontite/patologia , Estudos Prospectivos , Adulto Jovem , IdosoRESUMO
Adipose tissue dysfunction is a key feature of metabolic syndrome, which increases the risk of periodontitis, an inflammatory disease induced by bacteria that affects the gingiva and other components of periodontal tissue. Recent studies indicate that molecules from inflamed periodontal tissue contribute to adipose tissue dysfunction. However, the cellular mechanisms and interactions between adipose tissue and gingiva driving the progression of metabolic and periodontal conditions remain unclear. To address this, we developed a chimeric (mouse/human) co-culture tissue model (which identifies the origins of species-specific cytokines) to investigate these interactions. Using tissue-specific functional cells and immunocytes, we constructed equivalents of adipose tissue (ATE) and gingiva (GTE), co-cultivating them under inflammatory conditions induced by bacterial endotoxin, lipopolysaccharide (LPS). Our findings showed that exposure to LPS resulted in a notable reduction in lipid accumulation, GLUT4 expression, and adiponectin secretion in ATE, along with increased macrophage colonies forming around lipid droplets, as well as elevated levels of triglyceride, leptin, and IL-6. In GTE, LPS triggered significant inflammatory responses, characterized by increased macrophage accumulation, elevated COX-2 expression, and heightened secretion of inflammatory cytokines. LPS also reduced epithelial thickness and the expression of keratin 19 and collagen IV, indicating impaired barrier function and gingival integrity. Co-culturing ATE with GTE exacerbated these LPS-induced harmful effects in both tissues. In conclusion, our findings suggest that interplay between gingiva and adipose tissue can intensify the inflammatory and dysfunctional changes caused by LPS. This co-culture tissue model offers a valuable tool for future studies on periodontitis and metabolic syndrome.
Assuntos
Tecido Adiposo , Técnicas de Cocultura , Gengiva , Inflamação , Lipopolissacarídeos , Gengiva/metabolismo , Gengiva/patologia , Animais , Tecido Adiposo/metabolismo , Humanos , Camundongos , Inflamação/metabolismo , Inflamação/patologia , Periodontite/metabolismo , Periodontite/patologia , Citocinas/metabolismo , Camundongos Endogâmicos C57BL , Macrófagos/metabolismo , Masculino , Síndrome Metabólica/metabolismoRESUMO
Periodontal disease (PD) during pregnancy may trigger systemic inflammation, increasing the risk of developing cardiometabolic disease (CMD). As a consequence, PD may result in the activation of cellular and molecular pathways, affecting the disease course and pregnancy outcome. Although microRNAs (miRNAs) are considered ideal biomarkers for many diseases, few studies have investigated salivary miRNAs and their role in pregnancy or neonatal outcomes. In this study, we sought to investigate the associations between salivary miRNAs of pregnant women with oral diseases and their effects on neonatal outcomes. Eleven (n = 11) salivary miRNAs from a cohort of pregnant women with oral diseases (n = 32; oral health, H; gingivitis, G; and periodontitis, P) were detected using a previous profiling analysis with an FDR < 0.20 and a fold change (FC) < 0.5 or FC > 2 for the most highly expressed miRNAs. Spearman correlations were performed for 11 salivary microRNAs associated with oral-derived inflammation, which could affect neonatal outcomes during pregnancies at risk for cardiometabolic disease (CMD), defined by the presence of a high pregestational BMI. In addition, ROC curves demonstrated the diagnostic accuracy of the markers used. Upregulation of miR-423-5p expression and a decrease in miR-27b-3p expression were detected in the P-group (p < 0.05), and ROC analysis revealed the diagnostic accuracy of miR-423-5p for discriminating oral diseases, such as gingivitis versus periodontitis (P vs. G, AUC = 0.78, p < 0.05), and for discriminating it from the healthy oral cavity (P vs. H, AUC = 0.9, p < 0.01). In addition, miR-27b-3p and miR-622 were also able to discriminate the healthy group from the P-group (AUC = 0.8, p < 0.05; AUC = 0.8, p < 0.05). miR-483-5p was able to discriminate between the G-group (AUC = 0.9, p < 0.01) and the P-group (AUC = 0.8, p < 0.05). These data support the role of salivary miRNAs as early biomarkers for neonatal outcomes in pregnant women with periodontal disease at high risk for CMD and suggest that there is cross-talk between salivary miRNAs and subclinical systemic inflammation.
Assuntos
MicroRNAs , Doenças Periodontais , Resultado da Gravidez , Saliva , Humanos , MicroRNAs/genética , Feminino , Gravidez , Saliva/metabolismo , Adulto , Doenças Periodontais/metabolismo , Doenças Periodontais/genética , Recém-Nascido , Biomarcadores , Doenças Cardiovasculares/metabolismo , Doenças Cardiovasculares/genética , Doenças Cardiovasculares/diagnóstico , Doenças Cardiovasculares/etiologia , Complicações na Gravidez/metabolismo , Complicações na Gravidez/genética , Periodontite/metabolismo , Periodontite/genética , Gengivite/metabolismo , Gengivite/diagnóstico , Gengivite/genética , Curva ROCRESUMO
Periodontitis is featured as the periodontium's pathologic destruction caused by the host's overwhelmed inflammation. Omentin-1 has been reported to be aberrantly downregulated in patients with periodontitis, but the specific regulation of Omentin-1 during the pathogenesis of periodontitis remains unclear. In this study, human periodontal ligament stem cells (hPDLSCs) were stimulated by lipopolysaccharide (LPS) from Porphyromonas gingivalis to establish an in vitro inflammatory periodontitis model. hPDLSCs were treated with recombinant human Omentin-1 (250, 500 and 750â¯ng/mL) for 3â¯h before LPS stimulation. Results revealed that Omentin-1 significantly inhibited LPS-induced inflammation in hPDLSCs through reducing the production of proinflammatory cytokines (tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß and IL-6) and downregulating the expression of Cox2 and iNOS. Meanwhile, Omentin-1 significantly enhanced alkaline phosphatase (ALP) activity and Alizarin red-stained area, accompanied by increasing expression osteogenic markers BMP2, OCN and Runx2, confirming that Omentin-1 restores osteogenic differentiation in LPS-induced hPDLSCs. In addition, the conditioned medium (CM) from LPS-induced hPDLSCs was harvested to culture macrophages, which resulted in macrophage polarization towards M1, while CM from Omentin-1-treated hPDLSCs reduced M1 macrophages polarization and elevated M2 polarization. Furthermore, Omentin-1 also inhibited LPS-triggered endoplasmic reticulum (ER) stress in hPDLSCs, and additional treatment of the ER stress activator tunicamycin (TM) partially reversed the functions of Omentin-1 on inflammation, osteogenic differentiation and macrophages polarization. In summary, Omentin-1 exerted a protective role against periodontitis through inhibiting inflammation and enhancing osteogenic differentiation of hPDLSCs, providing a novelty treatment option for periodontitis.
Assuntos
Diferenciação Celular , Citocinas , Estresse do Retículo Endoplasmático , Proteínas Ligadas por GPI , Inflamação , Lectinas , Lipopolissacarídeos , Macrófagos , Osteogênese , Ligamento Periodontal , Células-Tronco , Ligamento Periodontal/citologia , Ligamento Periodontal/efeitos dos fármacos , Ligamento Periodontal/metabolismo , Humanos , Lipopolissacarídeos/farmacologia , Osteogênese/efeitos dos fármacos , Citocinas/metabolismo , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo , Lectinas/farmacologia , Diferenciação Celular/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Proteínas Ligadas por GPI/metabolismo , Proteínas Ligadas por GPI/farmacologia , Inflamação/patologia , Inflamação/metabolismo , Periodontite/patologia , Periodontite/metabolismo , Porphyromonas gingivalis , Células CultivadasRESUMO
OBJECTIVE: This study investigated the clinical importance of long noncoding RNA myocardial infarction-associated transcript (MIAT) in periodontitis and its impact on the functional regulation of human periodontal ligament fibroblasts (hPDLFs). METHODS: Ninety-eight periodontitis patients and 74 healthy controls were enrolled. In vitro cellular models were created using Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) to stimulate hPDLFs. Real-time quantitative polymerase chain reaction was used to measure mRNA levels of MIAT and osteogenic factors. Inflammation factor concentration was assessed using an enzyme-linked immunosorbent assay. Cell viability and apoptosis were examined by cell counting kit -8 and flow cytometry assay. The targeting relationship was verified by the dual-luciferase reporter and RNA Immunoprecipitation assay. RESULTS: Highly expressed MIAT and Dicckopf-1 (DDK1), and lowly expressed miR-204-5p were found in the gingival crevicular fluid of periodontitis patients and Pg-LPS induced hPDLFs. MIAT has a sensitivity of 76.53 % and a specificity of 86.49 % for identifying patients with periodontitis among healthy individuals. MIAT acts as a sponge for miR-204-5p and upregulates DDK1 mRNA expression. Silencing of MIAT diminished the promotion of apoptosis and inflammation in hPDLFs by Pg-LPS and enhanced osteogenic differentiation. However, a miR-204-5p inhibitor significantly reversed the effect of silenced MIAT. CONCLUSIONS: MIAT may act as a promising biomarker for periodontitis. It modulates apoptosis, inflammation, and osteogenic differentiation of PDLFs by focusing on the miR-204-5p/DKK1 axis, indicating its potential as a new therapeutic target for treating periodontitis.
Assuntos
Fibroblastos , Peptídeos e Proteínas de Sinalização Intercelular , MicroRNAs , Ligamento Periodontal , Periodontite , RNA Longo não Codificante , Humanos , RNA Longo não Codificante/metabolismo , RNA Longo não Codificante/genética , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , MicroRNAs/metabolismo , Periodontite/metabolismo , Masculino , Fibroblastos/metabolismo , Feminino , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Adulto , Apoptose , Reação em Cadeia da Polimerase em Tempo Real , Ensaio de Imunoadsorção Enzimática , Pessoa de Meia-Idade , Porphyromonas gingivalis , Líquido do Sulco Gengival/metabolismo , Células Cultivadas , Sobrevivência Celular , Estudos de Casos e Controles , Lipopolissacarídeos/farmacologia , Citometria de FluxoRESUMO
Periodontitis is a chronic inflammatory disease that affects about 45 %-50 % of adults worldwide, but the efficacy of current clinical therapies is unsatisfactory due to the complicated periodontal immune microenvironment. Thus, developing drugs that can regulate innate immune cells (e.g., macrophages) is a potent strategy to treat periodontitis. Here, we report that phloretin, a food plant-derived natural compound, is sufficient to alleviate periodontitis through immune regulation. In vivo, phloretin treatment could significantly reduce alveolar bone resorption and periodontal inflammation in mouse periodontitis models. In vitro, phloretin could suppress proinflammatory (M1-like) polarization and cytokine release in macrophages induced by LPS. Mechanistically, the immune regulatory role of phloretin in macrophages may be due to its metabolic regulation effect. Phloretin might restore the balance of M1/M2 macrophage transition in periodontitis by inhibiting HIF-1α-mediated glycolysis and PI3k/Akt pathways, thereby reducing the proinflammatory effect and immune disorder caused by over-activated M1 macrophages. Together, this study highlights that natural compound, such as phloretin, can restore periodontal immune homeostasis by metabolic regulation of macrophages, which may provide novel insight into the treatment of periodontitis.
Assuntos
Glicólise , Homeostase , Subunidade alfa do Fator 1 Induzível por Hipóxia , Macrófagos , Camundongos Endogâmicos C57BL , Periodontite , Floretina , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Animais , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Floretina/farmacologia , Floretina/uso terapêutico , Glicólise/efeitos dos fármacos , Camundongos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Periodontite/tratamento farmacológico , Periodontite/imunologia , Periodontite/metabolismo , Homeostase/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Masculino , Lipopolissacarídeos/imunologia , Humanos , Células RAW 264.7 , Modelos Animais de Doenças , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Citocinas/metabolismo , Perda do Osso Alveolar/tratamento farmacológicoRESUMO
PURPOSE: To investigate the effects of laser combined with periodontal basic treatment on periodontal indices, subgingival flora, adiponectin, matrix metalloproteinase-13 (MMP-13) and interleukin-1ß (IL-1ß) in patients with periodontitis. METHODS: A retrospective analysis was performed on 100 patients with periodontitis diagnosed and treated in Hengshui People's Hospital from December 2022 to July 2023. According to treatment methods, the patients were divided into control group (n=51) and experimental group (n=49). The control group received periodontal basic treatment, and the experimental group received laser treatment on the basis of the control group. The periodontal indexes, subgingival microflora, adiponectin, MMP-13, IL-1ß and bone metabolic factors of gingival crevicular fluid before and after treatment were compared between the two groups, as well as the clinical therapeutic effect. Statistical analysis was performed with SPSS 22.0 software package. RESULTS: After treatment, probing depth(PD), bleeding on probing(BOP), gingival index(GI) and plaque index (PLI) in the experimental group were lower than before treatment (Pï¼0.05), PD, BOP and PLI in the control group were lower than before treatment (Pï¼0.05), and PD, BOP, GI and PLI in the experimental group were significantly lower than those in control group (Pï¼0.05). After treatment, Lactobacillus, Clostridium and Bacteroides in both groups were significantly lower than before treatment (Pï¼0.05), and the experimental group was significantly lower than the control group(Pï¼0.05). After treatment, adiponectin in gingival crevicular fluid increased in both groups compared with before treatment(Pï¼0.05), and MMP-13 and IL-1ß in gingival crevicular fluid decreased in both groups compared with before treatment (Pï¼0.05), and adiponectin in gingival crevicular fluid in the experimental group was significantly higher than that in the control group (Pï¼0.05), MMP-13 and IL-1ß in the experimental group were significantly higher than that in the control group (Pï¼0.05). After treatment, procollagenâ type N-terminal peptide (PINP), cross linked C-telopeptide of type â collagen(CXT) and bone glaprotein (BGP) were significantly higher than those before treatment (Pï¼0.05), and the experimental group was significantly higher than the control group (Pï¼0.05). The total effective rate of the experimental group was significantly higher than that of the control group (Pï¼0.05). CONCLUSIONS: Laser combined with periodontal basic treatment can effectively improve periodontal indexes, reduce subgingival flora, increase the levels of adiponectin and bone metabolic factor in gingival crevicular fluid, reduce the levels of MMP-13 and IL-1ß in gingival crevicular fluid, and improve the clinical therapeutic effect in patients with periodontitis.
Assuntos
Adiponectina , Líquido do Sulco Gengival , Interleucina-1beta , Metaloproteinase 13 da Matriz , Índice Periodontal , Periodontite , Humanos , Líquido do Sulco Gengival/química , Líquido do Sulco Gengival/metabolismo , Adiponectina/metabolismo , Interleucina-1beta/metabolismo , Periodontite/terapia , Periodontite/microbiologia , Periodontite/metabolismo , Metaloproteinase 13 da Matriz/metabolismo , Estudos Retrospectivos , Gengiva/microbiologia , Gengiva/metabolismo , Terapia a Laser/métodosRESUMO
BACKGROUND: Activation of the IL-33/ST2 axis leads to the production of proinflammatory cytokines and thus to the triggering of osteoclastogenesis, which is why it plays an important role in the immunopathogenesis of periodontitis. The aim of this study was to compare IL-33 levels in serum, plasma, saliva and gingival crevicular fluid (GCF) of subjects with chronic periodontitis (CP) in comparison with the control group (CG). METHODS: This systematic review and meta-analysis followed the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) and was registered in the Open Science Framework (OSF): https://doi.org/10.17605/OSF.IO/YHUWA . Six electronic databases were used for study identification; PubMed, Google Scholar, ScienceDirect, Web of Science, Scopus and Dentistry & Oral Sciences Source from March 10, 2012 to April 30, 2024. The Joanna Briggs Institute (JBI) tool was used to assess the quality of the included cross-sectional articles and clinical trials. RESULTS: Of the 949 articles identified, 14 were included according to the inclusion and exclusion criteria. The total number of individuals studied in the included investigations was 814 of whom 445 had CP and 369 were healthy. The reported age range was from 20 to 50 years, with a mean age ± standard deviation of 40.29 ± 7.83 years. Four hundred and twenty-six (52%) patients were men and 388 (48%) were women. Meta-analysis revealed that there is an increase in IL-33 levels in plasma, saliva and GCF of subjects with CP compared to CG (p = * < 0.05). CONCLUSIONS: This study found a significant increase in IL-33 levels in different biological samples (plasma, saliva and GCF) of individuals with CP compared to CG, thus IL-33 has potential to be a biomarker in the diagnosis of periodontitis.