RESUMO
OBJECTIVE: The purpose of this study is to investigate regenerative process by immunohistochemical analysis and evaluate periodontal tissue regeneration following a topical application of BDNF to inflamed 3-wall intra-bony defects. BACKGROUND: Brain-derived neurotrophic factor (BDNF) plays a role in the survival and differentiation of central and peripheral neurons. BDNF can regulate the functions of non-neural cells, osteoblasts, periodontal ligament cells, endothelial cells, as well as neural cells. Our previous study showed that a topical application of BDNF enhances periodontal tissue regeneration in experimental periodontal defects of dog and that BDNF stimulates the expression of bone (cementum)-related proteins and proliferation of human periodontal ligament cells. METHODS: Six weeks after extraction of mandibular first and third premolars, 3-wall intra-bony defects were created in mandibular second and fourth premolars of beagle dogs. Impression material was placed in all of the artificial defects to induce inflammation. Two weeks after the first operation, BDNF (25 and 50 µg/mL) immersed into atelocollagen sponge was applied to the defects. As a control, only atelocollagen sponge immersed in saline was applied. Two and four weeks after the BDNF application, morphometric analysis was performed. Localizations of osteopontin (OPN) and proliferating cell nuclear antigen (PCNA)-positive cells were evaluated by immunohistochemical analysis. RESULTS: Two weeks after application of BDNF, periodontal tissue was partially regenerated. Immunohistochemical analyses revealed that cells on the denuded root surface were positive with OPN and PCNA. PCNA-positive cells were also detected in the soft connective tissue of regenerating periodontal tissue. Four weeks after application of BDNF, the periodontal defects were regenerated with cementum, periodontal ligament, and alveolar bone. Along the root surface, abundant OPN-positive cells were observed. Morphometric analyses revealed that percentage of new cementum length and percentage of new bone area of experimental groups were higher than control group and dose-dependently increased. CONCLUSION: These findings suggest that BDNF could induce cementum regeneration in early regenerative phase by stimulating proliferation of periodontal ligament cells and differentiation into periodontal tissue cells, resulting in enhancement of periodontal tissue regeneration in inflamed 3-wall intra-bony defects.
Assuntos
Perda do Osso Alveolar , Fator Neurotrófico Derivado do Encéfalo , Cementogênese , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Fator Neurotrófico Derivado do Encéfalo/uso terapêutico , Cães , Cementogênese/efeitos dos fármacos , Antígeno Nuclear de Célula em Proliferação/metabolismo , Osteopontina , Ligamento Periodontal/patologia , Ligamento Periodontal/efeitos dos fármacos , Masculino , Regeneração Tecidual Guiada Periodontal/métodos , Regeneração Óssea/efeitos dos fármacos , Cemento Dentário/patologia , Cemento Dentário/efeitos dos fármacos , Periodonto/patologia , Periodonto/metabolismo , Mandíbula , Proliferação de Células/efeitos dos fármacosRESUMO
The extracellular matrix (ECM) is a complex non-cellular three-dimensional macromolecular network present within all tissues and organs, forming the foundation on which cells sit, and composed of proteins (such as collagen), glycosaminoglycans, proteoglycans, minerals, and water. The ECM provides a fundamental framework for the cellular constituents of tissue and biochemical support to surrounding cells. The ECM is a highly dynamic structure that is constantly being remodeled. Matrix metalloproteinases (MMPs) are among the most important proteolytic enzymes of the ECM and are capable of degrading all ECM molecules. MMPs play a relevant role in physiological as well as pathological processes; MMPs participate in embryogenesis, morphogenesis, wound healing, and tissue remodeling, and therefore, their impaired activity may result in several problems. MMP activity is also associated with chronic inflammation, tissue breakdown, fibrosis, and cancer invasion and metastasis. The periodontium is a unique anatomical site, composed of a variety of connective tissues, created by the ECM. During periodontitis, a chronic inflammation affecting the periodontium, increased presence and activity of MMPs is observed, resulting in irreversible losses of periodontal tissues. MMP expression and activity may be controlled in various ways, one of which is the inhibition of their activity by an endogenous group of tissue inhibitors of metalloproteinases (TIMPs), as well as reversion-inducing cysteine-rich protein with Kazal motifs (RECK).
Assuntos
Metaloproteinases da Matriz , Periodontite , Humanos , Metaloproteinases da Matriz/metabolismo , Periodontite/metabolismo , Periodonto/metabolismo , Matriz Extracelular/metabolismo , Colágeno/metabolismo , Inflamação/metabolismo , Inibidores Teciduais de Metaloproteinases/metabolismo , Proteínas Ligadas por GPI/metabolismoRESUMO
OBJECTIVE: Increasing the effectiveness of treatment of chronic generalized periodontitis using PDT based on clinical and functional substantiation of the effects of a photosensitizer. MATERIALS AND METHODS: A clinical and functional study and treatment of moderate chronic generalized periodontitis was carried out in 62 people (26 men and 36 women) aged from 35 to 55 years without a somatic model with an orthognathic occlusion diagnosed according to ICD-10 - K05.3. Of these, 2 groups were divided depending on the type of treatment: Group 1 (main) - patients with moderate chronic generalized periodontitis - 32 people. (17 men and 15 women, average age of the group - 43.2±2.2 years); Group 2 (control) - patients with moderate chronic generalized periodontitis - 30 people. (14 men and 16 women, average age of the group - 44.0±3.3 years). Complex treatment consisted of sanitation of the mouth, removal of dental plaque and curettage of periodontal pockets in group 1, followed by PDT with Revixan gel using a special wired aligner REVIXAN DENTAL LED (16 r). The clinical condition of the periodontium was assessed using the Greene Vermillion Hygienic Index (OHI-S), the Mühlleman Bleeding Index (SBI) modified by Cowell, and the periodontal index PI. To study the state of microcirculation in the gum tissue, the laser Doppler flowmetry (LDF) method was used using the LAKK-M device (NPP «Lazma¼, Russia). The state of microcirculation was assessed by the microcirculation index (M), which characterizes the level of tissue blood flow; parameter - «σ¼, which determines the fluctuation of the erythrocyte flow. According to Wavelet analysis of LDF-grams, the shunt index (SH) of blood flow was determined. In the «LDF + spectrometry¼ mode, oxygenation in periodontal tissues was studied using optical tissue oximetry (OTO), based on the results of which the perfusion saturation index (Sm) and the specific oxygen consumption index (U, %) were determined. RESULTS: According to LDF data, after PDT (group 1), normalization of clinical indices and the level of microcirculation in periodontal tissues was established, which was accompanied by an increase in the level of blood flow (M) and its activity (σ), which persisted after 3 and 6 months. after PDT. The perfusion saturation index (Sm) and specific oxygen consumption (U) increased more significantly after PDT, which persisted after 3 and 6 months. In the control group, the dynamics of indicators was less pronounced. CONCLUSION: The use of PDT with Revixan gel normalizes the clinical condition of the periodontium, indicators of microhemodynamics and oxygen metabolism.
Assuntos
Periodontite Crônica , Microcirculação , Fotoquimioterapia , Humanos , Feminino , Masculino , Adulto , Microcirculação/efeitos dos fármacos , Pessoa de Meia-Idade , Periodontite Crônica/tratamento farmacológico , Periodontite Crônica/terapia , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Periodonto/irrigação sanguínea , Periodonto/efeitos dos fármacos , Periodonto/metabolismo , Oxigênio/metabolismoRESUMO
OBJECTIVES: To determine the abilities of salivary E-cadherin to differentiate between periodontal health and periodontitis and to discriminate grades of periodontitis. BACKGROUND: E-cadherin is the main protein responsible for maintaining the integrity of epithelial-barrier function. Disintegration of this protein is one of the events associated with the destructive forms of periodontal disease leading to increase concentration of E-cadherin in the oral biofluids. MATERIALS AND METHODS: A total of 63 patients with periodontitis (case) and 35 periodontally healthy subjects (control) were included. For each patient, periodontal parameters including bleeding on probing (BOP), probing pocket depth (PPD), and clinical attachment level (CAL) were recorded. Concentration of salivary E-cadherin was determined by ELISA. Receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to determine the diagnostic potentials of E-cadherin. RESULTS: Level of salivary E-cadherin was significantly higher in periodontitis cases than controls. The ROC analysis showed that salivary E-cadherin exhibits excellent sensitivity and specificity (AUC 1.000) to differentiate periodontal health from periodontitis with a cutoff concentration equal to 1.325 ng/mL. The AUCs of E-cadherin to differentiate grade A from grade B and C periodontitis were 0.731 (cutoff point = 1.754 ng/mL) and 0.746 (cutoff point = 1.722 ng/mL), respectively. However, the AUC of salivary E-cadherin to differentiate grade B from grade C periodontitis was lower (0.541). Additionally, BOP and PPD were significantly and positively correlated with the concentration of salivary E-cadherin. CONCLUSION: Salivary E-cadherin exhibited excellent sensitivity and specificity to differentiate periodontitis from a healthy periodontium. The level of accuracy of E-cadherin was also sufficient to recognize grade A periodontitis from grade B and C periodontitis.
Assuntos
Doenças Periodontais , Periodontite , Humanos , Periodontite/diagnóstico , Periodontite/metabolismo , Doenças Periodontais/metabolismo , Periodonto/metabolismo , Biomarcadores/metabolismo , Caderinas/metabolismo , Saliva/químicaRESUMO
The principles of periodontal therapy are based on the control of microbial pathogens and host factors that contribute to biofilm dysbiosis, with the aim of modulating the progression of periodontitis and periodontal tissue destruction. It is currently known how differently each individual responds to periodontal treatment, depending on both the bacterial subtypes that make up the dysbiotic biofilm and interindividual variations in the host inflammatory response. This has allowed the current variety of approaches for the management of periodontitis to be updated by defining the goals of target strategies, which consist of reducing the periodontopathogenic microbial flora and/or modulating the host-mediated response. Therefore, this review aims to update the current variety of approaches for the management of periodontitis based on recent target therapies. Recently, encouraging results have been obtained from several studies exploring the effects of some targeted therapies in the medium- and long-term. Among the most promising target therapies analyzed and explored in this review include: cell-based periodontal regeneration, mediators against bone resorption, emdogain (EMD), platelet-rich plasma, and growth factors. The reviewed evidence supports the hypothesis that the therapeutic combination of epigenetic modifications of periodontal tissues, interacting with the dysbiotic biofilm, is a key step in significantly reducing the development and progression of disease in periodontal patients and improving the therapeutic response of periodontal patients. However, although studies indicate promising results, these need to be further expanded and studied to truly realize the benefits that targeted therapies could bring in the treatment of periodontitis.
Assuntos
Microbiota , Periodontite , Humanos , Periodontite/microbiologia , Metagenoma , Metagenômica , Periodonto/metabolismo , Disbiose/terapiaRESUMO
OBJECTIVES: This review article aims to describe some of the roles of Matrix metalloproteinases (MMPs) in enamel, dentine, dental caries, hybrid layer degradation, pulp and periodontal tissues, throwing light on their current inhibitors. The article addresses the potential of MMPs to serve as biomarkers with diagnostic and therapeutic value. DESIGN: The sections of this review discuss MMPs' involvement in developmental, remodeling, degradational and turnover aspects of dental and periodontal tissues as well as their signals in the pathogenesis, progress of different lesions and wound healing of these tissues. The literature was searched for original research articles, review articles and theses. The literature search was conducted in PubMed and MEDLINE for articles published in the last 20 years. RESULTS: 119 published papers, two textbooks and two doctoral theses were selected for preparing the current review. CONCLUSIONS: MMPs are significant proteases, of evident contribution in dental and periapical tissue development, health and disease processes, with promising potential for use as diagnostic and prognostic disease biomarkers. Continuing understanding of their role in pathogenesis and progress of different dental, periapical and periodontal lesions, as well as in dentine-pulp wound healing could be a keystone to future diagnostic and therapeutic regimens.
Assuntos
Cárie Dentária , Biomarcadores , Humanos , Metaloproteinases da Matriz/metabolismo , Periodonto/metabolismoRESUMO
Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) are transcriptional coactivators encoded by paratactic homologous genes, shuttle-crossing between cytoplasm and nucleus to regulate the gene expression and cell behavior and standing at the center place of the sophisticated regulatory networking of mechanotransduction. Orthodontic tooth movement (OTM) is a process in which extracellular mechanical stimuli are transformed into intracellular biochemical signals to regulate cellular responses and tissue remodeling. Literature studies have confirmed that YAP/TAZ plays an important role not only in embryonic development, homeostasis and tumorigenesis, but also in mechanical-biochemical signal transduction of periodontal tissues under the mediation of various signal molecules in its upstream and downstream. Herein, we review the advances in the roles and mechanisms of YAP/TAZ in OTM to provide insights for better understanding and further study of the OTM and possible targeted clinical intervention in orthodontic treatment.
Assuntos
Processo Alveolar/metabolismo , Remodelação Óssea , Mecanotransdução Celular , Periodonto/metabolismo , Técnicas de Movimentação Dentária , Proteínas com Motivo de Ligação a PDZ com Coativador Transcricional/metabolismo , Proteínas de Sinalização YAP/metabolismo , Animais , Humanos , Estresse MecânicoRESUMO
Increase of sympathetic activity has been known to exacerbate osteoporosis through promotion of bone resorption. However, it is largely unknown about involvement of sympathetic activity in exacerbation of periodontitis. In this study, we investigated whether α2-adrenergic receptor (α2-AR) agonist guanabenz which decreases sympathetic activity, attenuates alveolar bone resorption in rats having high sympathetic activity with periodontitis. Volumes of residual alveolar bone and attachment levels in periodontium were examined using micro-computed tomography and hematoxylin-eosin staining, respectively. Furthermore, osteoclast numbers per bone surface and osteoclast surface per bone surface were measured using tartrate-resistant acid phosphatase staining. To examine the suppressive effects of guanabenz on pro-inflammatory cytokines, expression levels of tyrosine hydroxylase (TH), TNF-α, IL1-ß, and IL-6 in periodontium were measured using immunohistostaining. Administration of guanabenz attenuated loss of alveolar bone and attachment levels in rats having high sympathetic activity. Furthermore, its administration suppressed osteoclast numbers in rats having high sympathetic activity. TH, TNF-α, IL-1ß, and IL-6 positive cells in periodontium in rats treated with guanabenz for 12 weeks, were lower than those in control rats having high sympathetic activity. This study demonstrated administration of α2-AR agonist guanabenz attenuates alveolar bone resorption through decrease of sympathetic activity in rats.
Assuntos
Agonistas de Receptores Adrenérgicos alfa 2/administração & dosagem , Agonistas de Receptores Adrenérgicos alfa 2/farmacologia , Reabsorção Óssea/etiologia , Reabsorção Óssea/prevenção & controle , Guanabenzo/administração & dosagem , Guanabenzo/farmacologia , Periodontite/complicações , Periodontite/fisiopatologia , Animais , Reabsorção Óssea/metabolismo , Reabsorção Óssea/fisiopatologia , Citocinas/metabolismo , Modelos Animais de Doenças , Mediadores da Inflamação/metabolismo , Masculino , Periodontite/metabolismo , Periodonto/metabolismo , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Sistema Nervoso Simpático/efeitos dos fármacos , Sistema Nervoso Simpático/fisiopatologiaRESUMO
Periodontitis is a chronic complex inflammatory disease associated with a destructive host immune response to microbial dysbiosis, leading to irreversible loss of tooth-supporting tissues. Regeneration of functional periodontal soft (periodontal ligament and gingiva) and hard tissue components (cementum and alveolar bone) to replace lost tissues is the ultimate goal of periodontal treatment, but clinically predictable treatments are lacking. Similarly, the identification of biomarkers that can be used to accurately diagnose periodontitis activity is lacking. A relatively novel category of molecules found in oral tissue, circular RNAs (circRNAs) are single-stranded endogenous, long, non-coding RNA molecules, with covalently circular-closed structures without a 5' cap and a 3' tail via non-classic backsplicing. Emerging research indicates that circRNAs are tissue and disease-specific expressed and have crucial regulatory functions in various diseases. CircRNAs can function as microRNA or RNA binding sites or can regulate mRNA. In this review, we explore the biogenesis and function of circRNAs in the context of the emerging role of circRNAs in periodontitis pathogenesis and the differentiation of periodontal cells. CircMAP3K11, circCDK8, circCDR1as, circ_0062491, and circ_0095812 are associated with pathological periodontitis tissues. Furthermore, circRNAs are expressed in periodontal cells in a cell-specific manner. They can function as microRNA sponges and can form circRNA-miRNA-mRNA networks during osteogenic differentiation for periodontal-tissue (or dental pulp)-derived progenitor cells.
Assuntos
Periodontite/metabolismo , Periodonto/metabolismo , RNA Circular/metabolismo , Regeneração , Animais , HumanosRESUMO
Periodontitis is a chronic inflammatory immune disease associated with a dysbiotic state, influenced by keystone bacterial species responsible for disrupting the periodontal tissue homeostasis. Furthermore, the severity of periodontitis is determined by the interaction between the immune cell response in front of periodontitis-associated species, which leads to the destruction of supporting periodontal tissues and tooth loss in a susceptible host. The persistent bacterial challenge induces modifications in the permeability and ulceration of the sulcular epithelium, which facilitates the systemic translocation of periodontitis-associated bacteria into distant tissues and organs. This stimulates the secretion of pro-inflammatory molecules and a chronic activation of immune cells, contributing to a systemic pro-inflammatory status that has been linked with a higher risk of several systemic diseases, such as type 2 diabetes mellitus (T2DM) and gestational diabetes mellitus (GDM). Although periodontitis and GDM share the common feature of systemic inflammation, the molecular mechanistic link of this association has not been completely clarified. This review aims to examine the potential biological mechanisms involved in the association between periodontitis and GDM, highlighting the contribution of both diseases to systemic inflammation and the role of new molecular participants, such as extracellular vesicles and non-coding RNAs, which could act as novel molecular intercellular linkers between periodontal and placental tissues.
Assuntos
Diabetes Gestacional , Periodontite , Periodonto , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/microbiologia , Diabetes Gestacional/metabolismo , Diabetes Gestacional/microbiologia , Feminino , Humanos , Periodontite/etiologia , Periodontite/metabolismo , Periodontite/microbiologia , Periodonto/metabolismo , Periodonto/microbiologia , GravidezRESUMO
The aim of the study was to clarify whether orthodontic forces and periodontitis interact with respect to the anti-apoptotic molecules superoxide dismutase 2 (SOD2) and baculoviral IAP repeat-containing protein 3 (BIRC3). SOD2, BIRC3, and the apoptotic markers caspases 3 (CASP3) and 9 (CASP9) were analyzed in gingiva from periodontally healthy and periodontitis subjects by real-time PCR and immunohistochemistry. SOD2 and BIRC3 were also studied in gingiva from rats with experimental periodontitis and/or orthodontic tooth movement. Additionally, SOD2 and BIRC3 levels were examined in human periodontal fibroblasts incubated with Fusobacterium nucleatum and/or subjected to mechanical forces. Gingiva from periodontitis patients showed significantly higher SOD2, BIRC3, CASP3, and CASP9 levels than periodontally healthy gingiva. SOD2 and BIRC3 expressions were also significantly increased in the gingiva from rats with experimental periodontitis, but the upregulation of both molecules was significantly diminished in the concomitant presence of orthodontic tooth movement. In vitro, SOD2 and BIRC3 levels were significantly increased by F. nucleatum, but this stimulatory effect was also significantly inhibited by mechanical forces. Our study suggests that SOD2 and BIRC3 are produced in periodontal infection as a protective mechanism against exaggerated apoptosis. In the concomitant presence of orthodontic forces, this protective anti-apoptotic mechanism may get lost.
Assuntos
Proteína 3 com Repetições IAP de Baculovírus/genética , Regulação da Expressão Gênica , Ligamento Periodontal/metabolismo , Periodonto/metabolismo , Superóxido Dismutase/genética , Animais , Apoptose/genética , Proteína 3 com Repetições IAP de Baculovírus/metabolismo , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Células Cultivadas , Fibroblastos/citologia , Fibroblastos/metabolismo , Fusobacterium nucleatum/fisiologia , Gengiva/citologia , Gengiva/metabolismo , Interações Hospedeiro-Patógeno , Humanos , Ligamento Periodontal/citologia , Ligamento Periodontal/microbiologia , Periodonto/citologia , Periodonto/microbiologia , Ratos , Superóxido Dismutase/metabolismoRESUMO
Nitric oxide (NO) binds to soluble guanylyl cyclase (sGC), activates it in a reduced oxidized heme iron state, and generates cyclic Guanosine Monophosphate (cGMP), which results in vasodilatation and inhibition of osteoclast activity. In inflammation, sGC is oxidized and becomes insensitive to NO. NO- and heme-independent activation of sGC requires protein expression of the α1- and ß1-subunits. Inflammation of the periodontium induces the resorption of cementum by cementoclasts and the resorption of the alveolar bone by osteoclasts, which can lead to tooth loss. As the presence of sGC in cementoclasts is unknown, we investigated the α1- and ß1-subunits of sGC in cementoclasts of healthy and inflamed human periodontium using double immunostaining for CD68 and cathepsin K and compared the findings with those of osteoclasts from the same sections. In comparison to cementoclasts in the healthy periodontium, cementoclasts under inflammatory conditions showed a decreased staining intensity for both α1- and ß1-subunits of sGC, indicating reduced protein expression of these subunits. Therefore, pharmacological activation of sGC in inflamed periodontal tissues in an NO- and heme-independent manner could be considered as a new treatment strategy to inhibit cementum resorption.
Assuntos
Inflamação/genética , Óxido Nítrico/genética , Periodonto/metabolismo , Guanilil Ciclase Solúvel/genética , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , GMP Cíclico/genética , Regulação da Expressão Gênica/genética , Heme/genética , Humanos , Inflamação/patologia , Ferro/metabolismo , Osteoclastos/metabolismo , Oxirredução/efeitos dos fármacos , Ligamento Periodontal/metabolismo , Ligamento Periodontal/patologia , Periodonto/patologiaRESUMO
We wished to evaluate whether epigenetic modifiers have a beneficial effect on treating experimental periodontitis and mechanisms for regulating the cell fate of mesenchymal stem cells (MSCs) in inflammatory microenvironments. We isolated MSCs from healthy and inflamed gingival tissues to investigate whether trichostatin A (TSA) could improve osteogenic differentiation and resolve inflammation in vitro. The tissue regenerative potentials were evaluated when treated with a temperature-dependent, chitosan-scaffold-encapsulated TSA, in a rat model of periodontitis. After induction with the conditioned medium, TSA treatment increased the osteogenic differentiation potential of inflamed MSCs and healthy MSCs. In addition, interleukin-6 and interleukin-8 levels in supernatants were significantly decreased after TSA treatment. Moreover, TSA promoted osteogenic differentiation by inhibiting nuclear factor-κB (p65) DNA binding in MSCs. In rats with experimental periodontitis, 7 weeks after local injections of chitosan-scaffold-encapsulated TSA, histology and microcomputed tomography showed a significant increase in alveolar bone volume and less inflammatory infiltration compared with vehicle-treated rats. The concentrations of interferon-γ and interleukin-6 were significantly decreased in the gingival crevicular fluid after TSA treatment. This study demonstrated that TSA had anti-inflammatory properties and could promote periodontal tissue repair, which indicated that epigenetic modifiers hold promise as a potential therapeutic option for periodontal tissue repair.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Ácidos Hidroxâmicos/farmacologia , Osteogênese/genética , Periodonto/crescimento & desenvolvimento , Animais , Proliferação de Células/efeitos dos fármacos , Proteínas de Ligação a DNA/genética , Modelos Animais de Doenças , Epigênese Genética/efeitos dos fármacos , Epigênese Genética/genética , Inibidores de Histona Desacetilases/farmacologia , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , NF-kappa B/genética , Osteogênese/efeitos dos fármacos , Periodonto/diagnóstico por imagem , Periodonto/metabolismo , Periodonto/patologia , Ratos , Microtomografia por Raio-XRESUMO
This study aims to investigate the levels of SLIT3 in gingival crevicular fluid (GCF) of healthy and periodontal disease subjects, and their correlations to periodontal disease. A total of 45 periodontal patients and 45 periodontally healthy volunteers were enrolled. The clinical parameters, radiographic bone loss and the levels of SLIT3, receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) in GCF were measured. The prevalences of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in subgingival plaque were also analyzed. The expression of SLIT3 and RANKL was detected in the periodontium of experimental periodontitis in rats and lipopolysaccharide (LPS)-induced mouse macrophage. The total amounts and concentrations of SLIT3 and RANKL were significantly higher in periodontitis than those in healthy, while the level of OPG was significantly lower (p < .05). Significant positive correlations were observed between the level of GCF SLIT3 and clinical attachment level and radiographic bone loss (p < .05). There existed a significant positive correlation between SLIT3 and RANKL (p < .05). Increased expression of SLIT3 and RANKL was observed in the periodontium of periodontal rats. SLIT3 expression was induced by LPS stimulation in macrophages. These results suggest that SLIT3 may act as a diagnostic indicator of periodontal disease and should be further investigated.
Assuntos
Líquido do Sulco Gengival/química , Proteínas de Membrana/metabolismo , Periodontite/metabolismo , Adulto , Animais , Placa Dentária/microbiologia , Feminino , Humanos , Masculino , Camundongos , Osteoprotegerina/metabolismo , Periodonto/metabolismo , Porphyromonas gingivalis/isolamento & purificação , Ligante RANK/metabolismo , Células RAW 264.7 , Ratos , Ratos Sprague-Dawley , Tannerella forsythia/isolamento & purificação , Treponema denticola/isolamento & purificaçãoRESUMO
Resistin, a proinflammatory adipokine, is elevated in many inflammatory diseases. However, little is known about its performance in periodontitis. The present study is aimed at evaluating resistin expression and synthesis in periodontal cells and tissues under inflammatory/microbial stress in addition to its effects on the periodontium. In vivo, 24 male rats were randomly divided into two groups: control and ligature-induced periodontal disease. After 6 and 12 days, animals were sacrificed to analyze gene expression of adipokines, bone loss, inflammation, and resistin synthesis. In vitro, human periodontal ligament (PDL) fibroblasts were used to evaluate the expression of resistin after inflammatory stimuli. In addition, PDL fibroblasts were exposed to resistin to evaluate its role on soft and hard tissue metabolism markers. The periodontitis group demonstrated significant bone loss, an increase in the number of inflammatory cells and vascular structures, an increase in resistin expression and synthesis, and a decrease in the expression of adiponectin, leptin, and its functional receptor. PDL fibroblasts showed a significant increase in resistin expression and synthesis in response to the inflammatory stimulus by IL-1ß. Resistin induced an increase in cytokine expression and a decrease in the regulation of some hard tissue and matrix formation genes in PDL fibroblasts. These data indicate that resistin is produced by periodontal cells and tissues, and this effect is enhanced by inflammatory stimuli. Moreover, resistin seems to interfere with soft and hard tissue metabolism during periodontitis by reducing markers related to matrix formation and bone tissue.
Assuntos
Ligamento Periodontal/metabolismo , Periodonto/metabolismo , Resistina/metabolismo , Animais , Osso e Ossos , Fibroblastos/metabolismo , Gengiva/metabolismo , Humanos , Inflamação , Periodontite/metabolismo , Fenótipo , RatosRESUMO
The present study evaluated the quality of single-cone root canal fillings with bioceramic (BC) sealer using three different techniques by means of micro-computed tomography (micro-CT). The canals of 30 extracted single-rooted permanent teeth were shaped with R40 Reciproc blue files and filled with the single-cone technique (SCT). BioRoot RCS BC sealer was placed inside the canals with one of the following master cones: R40 cone to working length (RWL, n = 10); R40 cone trimmed 1 mm short of working length (RWL-1, n = 10); non-standardized gutta-percha cone to working length (NSWL, n = 10). A quantitative and qualitative micro-CT analysis assessed the filling quality and internal/external voids formation. Collected data underwent statistical analysis by multivariate one-way analysis of variance (α = 0.05). In all groups, the voids were minimal and prevalently external. The NSWL and RWL-1 groups had increased sealer ratios in the whole canal and the apical canal portion, respectively. The lowest amounts of voids were found in the RWL group; the void volumes were slightly greater in the RWL-1 mm and NSWL groups, especially at the apical level. Two alternative SCTs showed satisfactory filling ability, uniform distribution of the BC sealer, and a minimally increased voids formation compared to the standard SCT with dedicated cone. The two tested alternative SCTs could take advantage of the beneficial characteristics of the BC sealer, which evenly filled the endodontic space, ideally sealing both the major and the accessory communications with the periodontium.
Assuntos
Cerâmica/química , Guta-Percha/química , Materiais Restauradores do Canal Radicular , Obturação do Canal Radicular/métodos , Microtomografia por Raio-X/métodos , Cavidade Pulpar , Endodontia/instrumentação , Endodontia/métodos , Humanos , Teste de Materiais , Dente Molar , Periodonto/metabolismoRESUMO
The recent identification of senescent cells in periodontal tissues has the potential to provide new insights into the underlying mechanisms of periodontal disease etiology. DNA damage-driven senescence is perhaps one of the most underappreciated delayed consequences of persistent Gram-negative bacterial infection and inflammation. Although the host immune response rapidly protects against bacterial invasion, oxidative stress generated during inflammation can indirectly deteriorate periodontal tissues through the damage to vital cell macromolecules, including DNA. What happens to those healthy cells that reside in this harmful environment? Emerging evidence indicates that cells that survive irreparable genomic damage undergo cellular senescence, a crucial intermediate mechanism connecting DNA damage and the immune response. In this review, we hypothesize that sustained Gram-negative bacterial challenge, chronic inflammation itself, and the constant renewal of damaged tissues create a permissive environment for the abnormal accumulation of senescent cells. Based on emerging data we propose a model in which the dysfunctional presence of senescent cells may aggravate the initial immune reaction against pathogens. Further understanding of the role of senescent cells in periodontal disease pathogenesis may have clinical implications by providing more sophisticated therapeutic strategies to combat tissue destruction.
Assuntos
Senescência Celular , Suscetibilidade a Doenças , Doenças Periodontais/etiologia , Doenças Periodontais/metabolismo , Infecções Bacterianas/complicações , Infecções Bacterianas/microbiologia , Microambiente Celular , Dano ao DNA , Gerenciamento Clínico , Suscetibilidade a Doenças/imunologia , Interações Hospedeiro-Patógeno/imunologia , Humanos , Inflamação/complicações , Inflamação/etiologia , Inflamação/metabolismo , NF-kappa B/metabolismo , Saúde Bucal , Doenças Periodontais/patologia , Doenças Periodontais/terapia , Periodonto/imunologia , Periodonto/metabolismo , Periodonto/patologia , Transdução de Sinais , Estresse FisiológicoRESUMO
Cytomorphometry is used in the sampling of biological materials and diagnostic procedures. The use of cytological studies in periodontal diseases is not well described in the literature. Our study aimed to quantitatively assess the inflammation dynamics using cytomorphometric analysis of the periodontium before and after the use of fixed dental prostheses. Following ethics approval, a total of 105 subjects were divided in 3 groups as gingivitis (n = 23), periodontitis (n = 58), and healthy periodontium (control) (n = 24). The fixed dental prostheses (crowns and fixed partial dentures) were fabricated from cobalt-chrome metal-ceramic prostheses using the conventional method (C/M-CoCr), cobalt-chrome metal-ceramic prostheses by the computer-aided design and computer-aided manufacturing (CAD/CAM) technique (C/C-CoCr), and zirconia-based ceramic prostheses by the CAD/CAM technique (C/C-Zr) among subjects with gingivitis and periodontitis. The gingival crevicular fluid (GCF) was obtained from subjects before and after the use of the prostheses. The total count of epithelial cells and the connective tissue cells or polymorphonuclear neutrophils (PMNs) in GCF were studied using cytomorphometric analysis. The Statistical Package Tor the Social Sciences (SPSS), Version 20 (IBM Company, Chicago, IL, USA) was used to analyze the results and the significance level was set at p = 0.05. The data for before and after the use of the prostheses were compared using independent t-Tests. Similarly, the results after the use of prostheses in gingivitis, periodontitis, and control in each type of prostheses were compared using One-way ANOVA with post hoc using Scheffe. The total epithelial cells and the PMNs were determined along with the epithelium/leukocyte index. Regardless of the prostheses type used, no significant change in the parameters was identified among patients with a healthy periodontium, before and after prosthetic treatment. In all study groups, a statistically increase (p value < 0.05) was observed in the oral epithelial cell counts and a statistically decrease (p < 0.05) in the PMNs count following the use of the fixed prostheses. Data on cytomorphometric analysis could enable the selection of the most appropriate prostheses for use in patients with periodontal pathologies. When choosing prostheses, changes in the composition of GCF could be considered as a useful criterion for their use.
Assuntos
Prótese Dentária/efeitos adversos , Inflamação/imunologia , Inflamação/metabolismo , Periodonto/imunologia , Periodonto/metabolismo , Adulto , Idoso , Células Epiteliais , Líquido do Sulco Gengival/imunologia , Líquido do Sulco Gengival/metabolismo , Gengivite/imunologia , Gengivite/metabolismo , Humanos , Pessoa de Meia-Idade , Neutrófilos/imunologia , Neutrófilos/metabolismo , Periodontite/imunologia , Periodontite/metabolismoRESUMO
OBJECTIVES: Cementogenesis seems to be significantly compromised during tissue inflammation. In dental practice, surgical procedures are performed with the aim to regenerate periodontium including cementum. However, inflammation that occurs during the initial healing phases after surgery may impair regeneration of this tissues. The aim of the present study was to assess if surgical procedures designed to regenerate periodontium might affect levels of cementum protein-1 (CEMP-1) in periodontal wound fluid during early phase of healing. MATERIALS AND METHODS: In 36 patients, 18 intrabony periodontal defects were treated with regenerative therapy (REG group) and 18 suprabony periodontal defects were treated with open flap debridement (OFD group). In the experimental sites, gingival crevicular fluid was collected immediately before surgery, and periodontal wound fluid was collected 4, 7, 14, and 21 days after surgery. CEMP-1 levels were detected by indirect enzyme-linked immunosorbent assay technique. RESULTS: At the analysis, it resulted that there was a significant average difference in CEMP-1 values between the REG and OFD groups at baseline (p = 0.041), the CEMP-1-modeled average in the OFD group was lower by 0.45 ng/ml. There was a significant trend in CEMP-1 over time, and this trend was different among the 2 groups: the REG group showed a statistically significant rising CEMP-1 trend (0.18 ng/ml a week p = 0.012), while the OFD had a trend that was significantly lower (-0.22 ng/ml a week compared to the REG group trend p = 0.023), the OFD group lost on average 0.05 ng/ml a week. In REG sites, GCF protein levels resulted also related to clinical parameters. CONCLUSIONS: During the initial inflammatory phase of periodontal healing, CEMP-1 levels decrease regardless of the surgical protocol applied. The surgical procedures used to regenerate periodontal tissue are able to reverse this trend and to induce significant increase of CEMP-1 in periodontal wound fluid after the first week postop.
Assuntos
Líquido do Sulco Gengival/metabolismo , Periodonto/metabolismo , Proteínas/metabolismo , Cicatrização/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Inflamação/metabolismo , Masculino , Pessoa de Meia-IdadeRESUMO
Different collection methods may influence the ability to detect and quantify biomarker levels in saliva, particularly in the expression of DNA/RNA methylation regulators of several inflammations and tissue turnover markers. This pilot study recruited five participants and unstimulated saliva were collected by either spitting or drooling, and the relative preference for each method was evaluated using a visual analogue scale. Subsequently, total RNA, gDNA and proteins were isolated using the Trizol method. Thereafter, a systematic evaluation was carried out on the potential effects of different saliva collection methods on periodontium-associated genes, DNA/RNA epigenetic factors and periodontium-related DNA methylation levels. The quantity and quality of DNA and RNA were comparable from different collection methods. Periodontium-related genes, DNA/RNA methylation epigenetic factors and periodontium-associated DNA methylation could be detected in the saliva sample, with a similar expression for both methods. The methylation of tumour necrosis factor-alpha gene promoter from drooling method showed a significant positive correlation (TNF α, r = 0.9) with clinical parameter (bleeding on probing-BOP). In conclusion, the method of saliva collection has a minimal impact on detecting periodontium-related genetic and epigenetic regulators in saliva. The pilot data shows that TNF α methylation may be correlated with clinical parameters.