RESUMO
Although dental implants and bone regenerative procedures are important approaches for the reestablishment of esthetics and function in young patients with a history of generalized aggressive periodontitis (GAP), no predictable outcomes have been reported, and the host osteo-immunoinflammatory response may play a relevant role in this context. In view of the lack of molecular investigations into the bone tissue condition of young patients with periodontitis, the aim of this study was to evaluate the gene expression of bone-related factors in this population. Bone biopsies were obtained from the posterior mandible in 16 individuals previously diagnosed with GAP and on periodontal support therapy and from 17 periodontally healthy (PH) patients. The gene expression of tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-ß, receptor activator of the NF-κB ligand (RANKL), osteoprotegerin (OPG), osteocalcin (OC), bone sialoprotein (BSP), and type I collagen (COL-I), important biomarkers of bone turnover, was evaluated by qRT-PCR. Lower TGF-ß and OPG mRNA levels were observed in GAP patients compared to PH individuals (p ≤ 0.05). There were no between-group differences in levels of TNF-α, BSP, RANKL, OC, or COL-I mRNA (p>0.05). In young adults, a history of periodontal disease can negatively modulate the gene expression of important bone-related factors in alveolar bone tissue. These molecular outcomes may contribute to the future development of therapeutic approaches to benefit bone healing in young patients with history of periodontitis via modulation of osteo-immuno-inflammatory biomarkers.
Assuntos
Periodontite Agressiva/genética , Expressão Gênica , Adulto , Periodontite Agressiva/metabolismo , Processo Alveolar/química , Biomarcadores , Colágeno Tipo I/análise , Colágeno Tipo I/genética , Estudos Transversais , Feminino , Humanos , Sialoproteína de Ligação à Integrina/análise , Sialoproteína de Ligação à Integrina/genética , Masculino , Osteocalcina/análise , Osteocalcina/genética , Osteoprotegerina/análise , Osteoprotegerina/genética , Ligante RANK/análise , Ligante RANK/genética , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Método Simples-Cego , Estatísticas não Paramétricas , Fator de Crescimento Transformador beta/análise , Fator de Crescimento Transformador beta/genética , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/genética , Adulto JovemRESUMO
Abstract Although dental implants and bone regenerative procedures are important approaches for the reestablishment of esthetics and function in young patients with a history of generalized aggressive periodontitis (GAP), no predictable outcomes have been reported, and the host osteo-immunoinflammatory response may play a relevant role in this context. In view of the lack of molecular investigations into the bone tissue condition of young patients with periodontitis, the aim of this study was to evaluate the gene expression of bone-related factors in this population. Bone biopsies were obtained from the posterior mandible in 16 individuals previously diagnosed with GAP and on periodontal support therapy and from 17 periodontally healthy (PH) patients. The gene expression of tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, receptor activator of the NF-κB ligand (RANKL), osteoprotegerin (OPG), osteocalcin (OC), bone sialoprotein (BSP), and type I collagen (COL-I), important biomarkers of bone turnover, was evaluated by qRT-PCR. Lower TGF-β and OPG mRNA levels were observed in GAP patients compared to PH individuals (p ≤ 0.05). There were no between-group differences in levels of TNF-α, BSP, RANKL, OC, or COL-I mRNA (p>0.05). In young adults, a history of periodontal disease can negatively modulate the gene expression of important bone-related factors in alveolar bone tissue. These molecular outcomes may contribute to the future development of therapeutic approaches to benefit bone healing in young patients with history of periodontitis via modulation of osteo-immuno-inflammatory biomarkers.
Assuntos
Humanos , Masculino , Feminino , Adulto , Adulto Jovem , Periodontite Agressiva/genética , Expressão Gênica , Periodontite Agressiva/metabolismo , Valores de Referência , Biomarcadores , Osteocalcina/análise , Osteocalcina/genética , Método Simples-Cego , Estudos Transversais , Fator de Crescimento Transformador beta/análise , Fator de Crescimento Transformador beta/genética , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/genética , Estatísticas não Paramétricas , Colágeno Tipo I/análise , Colágeno Tipo I/genética , Ligante RANK/análise , Ligante RANK/genética , Osteoprotegerina/análise , Osteoprotegerina/genética , Sialoproteína de Ligação à Integrina/análise , Sialoproteína de Ligação à Integrina/genética , Processo Alveolar/química , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Generalized aggressive periodontitis (GAgP) presents a reduced response to non-surgical therapy. However, it is not clear if the initial clinical, microbiological or immunological characteristics are impacting the worse response to treatment. This study aimed to identify the predictive value of clinical, microbiological and immunological patterns on the clinical response to therapy in GAgP patients. Twenty-four GAgP patients were selected, and gingival crevicular fluid (GCF) and subgingival biofilm were collected. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia levels were evaluated by qPCR, and IL-1ß and IL-10 concentration by ELISA. Twelve patients were treated with SRP (scaling and root planning), and twelve with SRP plus 375 mg amoxicillin and 250 mg metronidazole (8/8 hours, 7 days) (SRP + AM). The clinical changes (Probing Pocket Depth [PPD] reduction and Clinical Attachment Level [CAL] gain) 6 months post-treatment were correlated to the initial clinical, inflammatory and microbiological variables using stepwise logistic regression (α = 5%). CAL gain at 6 months was 1.16 ± 0.77 for SRP and 1.74 ± 0.57 mm for SRP + AM (P > .05). PPD reduction was 1.96 ± 0.82 for SRP and 2.45 ± 0.77 mm for SRP + AM (P < .05). In the SRP group, IL-10 showed a predictive value for clinical response. The higher the IL-10 concentration at baseline, the higher the reduction in PPD at 6 months (P = .01, r = .68). However, when antimicrobials were administered, no significant influence was detected (P > .05). It can be concluded that the IL-10 levels in GFC act as a predictor of clinical response to GAgP. Moreover, the intake of antimicrobials appears to overlap the influence of the inflammatory response on clinical response to treatment. Clinical trial registration number: NCT03933501.
Assuntos
Periodontite Agressiva/diagnóstico , Periodontite Agressiva/metabolismo , Interleucina-10/metabolismo , Adulto , Periodontite Agressiva/etiologia , Periodontite Agressiva/terapia , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Biofilmes , Biomarcadores , Feminino , Líquido do Sulco Gengival/metabolismo , Líquido do Sulco Gengival/microbiologia , Humanos , Masculino , Prognóstico , Aplainamento Radicular/métodos , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: Emerging evidence suggests that activation of inflammasomes plays a central mechanism in pathogenesis of periodontitis. This study aims to compare salivary levels of nod-like receptor family pyrin domain containing protein (NLRP) 3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), cysteine aspartase (caspase)-1, and interleukin (IL)-1ß from individuals with aggressive (AgP) or chronic periodontitis (CP) and healthy controls (HC), as well as elucidate its association with periodontal clinical status. METHODS: Saliva samples from individuals with CP (n = 75), AgP (n = 20), and HC (n = 69) were collected. Periodontal status was assessed by measurement of probing depth, clinical attachment level, and extent and severity of disease. Salivary levels of analytes were analyzed by enzyme-linked immunosorbent assay. Association between biomarkers with CP or AgP was analyzed using multivariate binary logistic regression models. RESULTS: Significantly higher levels of NLRP3, ASC, and IL-1ß were detected in periodontitis groups in comparison to the periodontally HC group. However, no significant differences were observed for caspase-1 levels between clinical groups, and only NLRP3 salivary concentration was significantly higher in AgP compared with CP patients. Also, positive significant correlations among NLRP3, ASC, and IL-1ß salivary concentrations and clinical parameters were observed. Logistic regression analyses revealed a strong/independent association of NLRP3, ASC, and IL-1ß salivary levels with CP and AgP. CONCLUSION: Although the concentration of caspase-1 in saliva samples makes its determination useless for detection of periodontal disease and/or its severity, salivary levels of NLRP3, ASC, and IL-1ß may act as strong/independent indicators of amount and extent of periodontal breakdown in both CP and AgP and could potentially be used for prevention and therapy of this group of diseases.
Assuntos
Periodontite Agressiva/diagnóstico , Periodontite Crônica/diagnóstico , Inflamassomos/análise , Proteína 3 que Contém Domínio de Pirina da Família NLR/análise , Saliva/química , Adolescente , Adulto , Idoso , Periodontite Agressiva/metabolismo , Biomarcadores/análise , Proteínas Adaptadoras de Sinalização CARD/análise , Estudos de Casos e Controles , Caspase 1/metabolismo , Domínio de Ativação e Recrutamento de Caspases , Periodontite Crônica/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interleucina-1beta/análise , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
BACKGROUND: Inadequate antioxidant balance may play a role in the excessive tissue breakdown in periodontitis. Because aggressive periodontitis (AgP) not only differs from chronic periodontitis (CP) in terms of clinical manifestations, this study investigates whether the salivary levels of glutathione reductase (GR) may be linked with periodontal status. METHODS: Saliva samples from patients with CP (n = 121), patients with AgP (n = 18), and healthy controls (n = 69) were collected. Periodontal status was assessed by criteria based on probing depth, clinical attachment level, and extent and severity of periodontal breakdown. GR salivary levels were analyzed by spectrophotometry. The association among GR concentration and CP or AgP was analyzed individually and adjusted for confounding using multivariate binary logistic regression models. RESULTS: GR levels not only differed significantly between the two periodontitis groups, being significantly greater in patients with AgP, but also were significantly greater than those observed in healthy controls. Synchronously, positive significant correlations between salivary GR concentration and clinical parameters were observed. After binary logistic regression analysis, both GR salivary levels ≥15.38 and ≥24.20 mU/mL were associated independently with CP and AgP, respectively. A significant interaction effect was also detected between increased GR salivary concentration and aging in the CP group. CONCLUSIONS: Increased GR salivary concentration may be a strong/independent prognostic indicator of the amount and extent of oxidative stress-induced periodontal damage in both CP and AgP. Likewise, saliva samples might reflect an interactive effect of GR levels associated with the aging-related cumulative characteristics of periodontal damage in CP.
Assuntos
Periodontite Agressiva/metabolismo , Glutationa Redutase/metabolismo , Perda da Inserção Periodontal/metabolismo , Estudos de Casos e Controles , Periodontite Crônica , Humanos , Índice Periodontal , SalivaRESUMO
OBJECTIVE: Salivary mucin and amylase levels are increased in patients with chronic periodontitis (CP). Due to the fact that aggressive periodontitis (AgP) not only differs from chronic periodontitis in terms of its clinical manifestation, the aim of this study was to compare salivary mucin and amylase levels and their relation to the clinical parameters of patients with aggressive periodontitis with that of patients with chronic periodontitis. MATERIAL AND METHODS: Eighty subjects were divided into two groups: 20 patients with AgP and their 20 matched controls and 20 patients with CP and their 20 matched controls, based on clinical attachment loss (CAL), probing pocket depth (PPD) and bleeding on probing (BOP). Whole unstimulated saliva was obtained and mucin, amylase and protein were determined by colorimetric methods. Pearson's correlation analysis was used to determine the relationship between salivary mucin, amylase and protein levels and the clinical parameters. RESULTS: Salivary mucin, amylase and protein levels were increased in patients with AgP and CP but there were no differences between them or between control groups. Pearson's correlation analysis, determined in the entire subjects studied, showed a positive and significant correlation of mucin, amylase and proteins with CAL and PPD and a negative correlation with the flow rate. When Pearson's correlation analysis was carried out in each group separately, Fisher's z transformation showed no significant difference between both groups. CONCLUSION: Comparison of the salivary levels of mucin, amylase and protein and their relationship with clinical parameters of AgP patients with that of CP patients revealed no differences between both groups.
Assuntos
Periodontite Agressiva/metabolismo , Amilases/análise , Periodontite Crônica/metabolismo , Mucinas/análise , Saliva/química , Adolescente , Adulto , Análise de Variância , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Perda da Inserção Periodontal , Índice Periodontal , Valores de Referência , Salivação , Taxa Secretória , Estatísticas não Paramétricas , Adulto JovemRESUMO
Objective Salivary mucin and amylase levels are increased in patients with chronic periodontitis (CP). Due to the fact that aggressive periodontitis (AgP) not only differs from chronic periodontitis in terms of its clinical manifestation, the aim of this study was to compare salivary mucin and amylase levels and their relation to the clinical parameters of patients with aggressive periodontitis with that of patients with chronic periodontitis. Material and Methods Eighty subjects were divided into two groups: 20 patients with AgP and their 20 matched controls and 20 patients with CP and their 20 matched controls, based on clinical attachment loss (CAL), probing pocket depth (PPD) and bleeding on probing (BOP). Whole unstimulated saliva was obtained and mucin, amylase and protein were determined by colorimetric methods. Pearson's correlation analysis was used to determine the relationship between salivary mucin, amylase and protein levels and the clinical parameters. Results Salivary mucin, amylase and protein levels were increased in patients with AgP and CP but there were no differences between them or between control groups. Pearson's correlation analysis, determined in the entire subjects studied, showed a positive and significant correlation of mucin, amylase and proteins with CAL and PPD and a negative correlation with the flow rate. When Pearson's correlation analysis was carried out in each group separately, Fisher's z transformation showed no significant difference between both groups. Conclusion Comparison of the salivary levels of mucin, amylase and protein and their relationship with clinical parameters of AgP patients with that of CP patients revealed no differences between both groups. .
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Adulto Jovem , Periodontite Agressiva/metabolismo , Amilases/análise , Periodontite Crônica/metabolismo , Mucinas/análise , Saliva/química , Análise de Variância , Estudos de Casos e Controles , Perda da Inserção Periodontal , Índice Periodontal , Valores de Referência , Salivação , Taxa Secretória , Estatísticas não ParamétricasRESUMO
OBJECTIVES: The aim of this study was to compare the pattern of secretion and the expression of mucin glycoprotein-2 (MG2) and lactoferrin in individuals with or without periodontitis. MATERIAL AND METHODS: Five individuals with aggressive periodontitis (APG), 5 with generalized chronic periodontitis (CPG) and 5 without periodontitis (CG) were enrolled after informed consent. Non-stimulated and stimulated submandibular and sublingual saliva was collected and samples analyzed by Western blot probed with specific antibodies. RESULTS: Stimulated and non-stimulated salivary flow rates did not differ among groups. Western blot analysis revealed that stimulation led to: an increase in MG2 expression in all groups, and to lactoferrin expression in APG and CPG. In non-stimulated saliva, CG exhibited the highest expression of both glycoproteins. In stimulated saliva, CG exhibited the highest expression of MG2, whereas APG the highest of lactoferrin. CONCLUSIONS: The pattern of secretion of MG2 and lactoferrin in health and disease is complex. Although the present study analyzed samples from a limited number of participants, the reduced expression of MG2 and lactoferrin in APG and CPG under non-stimulated condition, the predominant circumstance of salivary secretion during the day, suggests that these salivary constituents may play a role in the etiopathogenesis of these diseases.
Assuntos
Periodontite Agressiva/metabolismo , Periodontite Crônica/metabolismo , Lactoferrina/análise , Mucina-2/análise , Saliva/química , Glândulas Salivares/metabolismo , Adulto , Western Blotting , Estudos de Casos e Controles , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Lactoferrina/metabolismo , Masculino , Pessoa de Meia-Idade , Mucina-2/metabolismo , Taxa Secretória , Fatores de TempoRESUMO
OBJECTIVES:The aim of this study was to compare the pattern of secretion and the expression of mucin glycoprotein-2 (MG2) and lactoferrin in individuals with or without periodontitis. MATERIAL AND METHODS: Five individuals with aggressive periodontitis (APG), 5 with generalized chronic periodontitis (CPG) and 5 without periodontitis (CG) were enrolled after informed consent. Non-stimulated and stimulated submandibular and sublingual saliva was collected and samples analyzed by Western blot probed with specific antibodies. RESULTS: Stimulated and non-stimulated salivary flow rates did not differ among groups. Western blot analysis revealed that stimulation led to: an increase in MG2 expression in all groups, and to lactoferrin expression in APG and CPG. In non-stimulated saliva, CG exhibited the highest expression of both glycoproteins. In stimulated saliva, CG exhibited the highest expression of MG2, whereas APG the highest of lactoferrin. CONCLUSIONS: The pattern of secretion of MG2 and lactoferrin in health and disease is complex. Although the present study analyzed samples from a limited number of participants, the reduced expression of MG2 and lactoferrin in APG and CPG under non-stimulated condition, the predominant circumstance of salivary secretion during the day, suggests that these salivary constituents may play a role in the etiopathogenesis of these diseases.
Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Periodontite Agressiva/metabolismo , Periodontite Crônica/metabolismo , Lactoferrina/análise , /análise , Saliva/química , Glândulas Salivares , Western Blotting , Estudos de Casos e Controles , Eletroforese em Gel de Poliacrilamida , Lactoferrina/metabolismo , /metabolismo , Taxa Secretória , Fatores de TempoRESUMO
Periodontal diseases result from the interaction of bacterial pathogens with the host's gingival tissue. Gingival epithelial cells are constantly challenged by microbial cells and respond by altering their transcription profiles, inducing the production of inflammatory mediators. Different transcription profiles are induced by oral bacteria and little is known about how the gingival epithelium responds after interaction with the periodontopathogenic organism Aggregatibacter actinomycetemcomitans. In the present study, we examined the transcription of genes involved in signaling transduction pathways in gingival epithelial cells exposed to viable A. actinomycetemcomitans. Immortalized gingival epithelial cells (OBA-9) were infected with A. actinomycetemcomitans JP2 for 24 h and the transcription profile of genes encoding human signal transduction pathways was determined. Functional analysis of inflammatory mediators positively transcribed was performed by ELISA in culture supernatant and in gingival tissues. Fifteen of 84 genes on the array were over-expressed (P < 0.01) after 24 h of infection with viable A. actinomycetemcomitans. Over-expressed genes included those implicated in tissue remodeling and bone resorption, such as CSF2, genes encoding components of the LDL pathway, nuclear factor-κB-dependent genes and other cytokines. The ELISA data confirmed that granulocyte-macrophage colony-stimulating factor/colony-stimulating factor 2, tumor necrosis factor-α and intercellular adhesion molecule-1 were highly expressed by infected gingival cells when compared with control non-infected cells, and presented higher concentrations in tissues from patients with aggressive and chronic periodontitis than in tissues from healthy controls. The induction in epithelial cells of factors such as the pro-inflammatory cytokine CSF2, which is involved in osteoclastogenesis, may help to explain the outcomes of A. actinomycetemcomitans infection.
Assuntos
Aggregatibacter actinomycetemcomitans/fisiologia , Periodontite Agressiva/genética , Periodontite Crônica/genética , Citocinas/biossíntese , Gengiva/microbiologia , Transdução de Sinais/genética , Periodontite Agressiva/metabolismo , Apoptose , Aderência Bacteriana , Estudos de Casos e Controles , Linhagem Celular Transformada , Periodontite Crônica/metabolismo , Fatores Estimuladores de Colônias/biossíntese , Meios de Cultivo Condicionados , Células Epiteliais/microbiologia , Perfilação da Expressão Gênica , Gengiva/citologia , Gengiva/metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/biossíntese , Humanos , Molécula 1 de Adesão Intercelular/biossíntese , Transcrição Gênica , Fator de Necrose Tumoral alfa/biossínteseRESUMO
AIM: To examine changes in levels of gingival crevicular fluid (GCF) cytokines, after periodontal therapy of generalized aggressive periodontitis (GAgP). MATERIALS AND METHODS: Twenty-five periodontally healthy and 24 GAgP subjects had periodontal clinical parameters measured and gingival crevicular fluid (GCF) samples collected from up to 14 sites/subject. GCF samples were analysed using multiplex bead immunoassay for: GM-CSF, IFN-γ, IL-10, IL-1ß, IL-2, IL-6 and TNF-α. Aggressive periodontitis subjects were randomly assigned to either scaling and root planing (SRP) alone or SRP plus systemic amoxicillin (500 mg) and metronidazole (400 mg) 3 times a day for 14 days. Clinical parameters and GCF cytokines were re-measured 6 months after treatment. Differences over time were analysed using the Wilcoxon test and between groups using the Mann-Whitney test. RESULTS: Significant reductions in GCF GM-CSF, IL-1ß and the ratio IL-1ß/IL-10 and increases in GCF IL-6 were detected after therapy. The mean change in GCF cytokines did not differ significantly between groups. CONCLUSIONS: Periodontal therapy improved GCF cytokine profiles by lowering IL-1ß and increasing IL-10 levels. The reduction in GCF GM-CSF after therapy implicates this cytokine in the pathogenesis of GAgP. There was no difference between therapies in changes of GCF cytokines.
Assuntos
Periodontite Agressiva/metabolismo , Periodontite Agressiva/terapia , Antibacterianos/uso terapêutico , Citocinas/metabolismo , Raspagem Dentária , Líquido do Sulco Gengival/química , Adulto , Periodontite Agressiva/patologia , Amoxicilina/uso terapêutico , Análise de Variância , Citocinas/análise , Método Duplo-Cego , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/análise , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Humanos , Interferon gama/análise , Interferon gama/metabolismo , Interleucina-10/análise , Interleucina-10/metabolismo , Interleucina-1beta/análise , Interleucina-2/análise , Interleucina-2/metabolismo , Interleucina-6/análise , Interleucina-6/metabolismo , Masculino , Metronidazol/uso terapêutico , Estatísticas não Paramétricas , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/metabolismo , Adulto JovemRESUMO
OBJECTIVES: Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are known to be involved in the periodontal disease process. Results of in vivo MMPs and TIMPs gene expressions in the gingiva, though, are still controversial. In the present study, we compared the gene expression of MMP-1, -2, -9, -13 and TIMP-1, -2 in healthy and inflamed gingiva. METHODS: 38 gingival samples were collected from gingivitis (n = 10), advanced chronic periodontitis (n = 10), generalized aggressive periodontitis (n = 8) and periodontally healthy individuals (n = 10). Total RNA isolated from those samples was subjected to reverse transcription followed by amplification by polymerase chain reaction (RT-PCR). Products were visualized in agarose gels and quantified by optical densitometry. Samples were also processed for gelatin zymography and Western blotting for MMP-2 and MMP-9 in order to assess for post-transcriptional MMP regulation at the protein level. RESULTS: The frequencies and levels of transcripts encoding MMPs and TIMPs were found to be not significantly different among groups (p > 0.05, Fisher's Exact and Kruskall-Wallis tests). There is a trend towards higher MMP-2 and -9 gelatinase activities in the inflamed samples, although not statistically significant. In contrast, zymography and Western blotting studies show that MMP-2 is virtually absent in the chronic periodontitis group. CONCLUSION: These results could reflect a complex regulation of MMPs and TIMPs' gene expression in the course of gingival inflammation. They also reveal a great biological diversity even among individuals with similar periodontal status.