RESUMO
AIM: To evaluate the mRNA expression levels of the cytokines interferon-γ, tumour necrosis factor-α, interleukin (IL)-1ß, IL-10, IL-6, VEGF, and AGT and the chemokine CCL2/MCP-1 in periapical interstitial fluid associated with root canal infections before and after the reduction of the bacterial load using a cleaning procedure. METHODOLOGY: The case group included 11 patients with chronic liver disease, and the control group included 11 healthy patients. Clinical samples were taken from teeth with pulp necrosis. After cleaning and drying the canal, three paper points were introduced into the root canal and passed through the root apex (2 mm) into the periapical tissues for 1 min. The samples were collected immediately after root canal cleaning and 7 days later to characterize those gene expression levels using real-time PCR. The data were subjected to the Shapiro-Wilk and the Wilcoxon tests. RESULTS: In the control group, significantly increased expression of the pro-inflammatory cytokines IFN-γ and TNF-α was observed in teeth with restrained bacterial loads (day 7) (P < 0.05). Similarly, increased TNF-α expression was found on day 7 in the liver group (P < 0.05). No differences were observed in the expression levels of the IL-1ß, IL-10 and, IL-6, MCP-1/CCL-2 and VEGF between the first collection (day 0) and second collection (day 7), over time in either group. CONCLUSION: Chronic liver disease patients exhibited sufficient immunologic ability showing relatively similar expression levels of cytokines, chemokines and angiogenic factors in periapical samples compared with the responses from no-chronic liver disease patients. The outcomes of this study suggest that liver impairment did not compromise the periapical immune response.
Assuntos
Quimiocinas/metabolismo , Citocinas/metabolismo , Hepatopatias/complicações , Hepatopatias/imunologia , Doenças Periapicais/imunologia , Tratamento do Canal Radicular , Dente/imunologia , Adulto , Idoso , Carga Bacteriana , Quimiocina CCL2/metabolismo , Cavidade Pulpar/microbiologia , Necrose da Polpa Dentária/imunologia , Feminino , Humanos , Interferon gama/metabolismo , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Masculino , Pessoa de Meia-Idade , Tecido Periapical/imunologia , Tecido Periapical/microbiologia , RNA Mensageiro/metabolismo , Ápice Dentário , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
The objective of this study was to observe the distribution of macrophages (MPs) expressing transforming growth factor beta-1 (TGF-ß1) in tissue samples from patients with different human chronic periapical diseases. In this study, samples were collected from 75 volunteers, who were divided into three groups according to classified standards, namely, healthy control (N = 25), periapical granuloma (N = 25), and periapical cyst (N = 25). The samples were fixed in 10% buffered formalin for more than 48 h, dehydrated, embedded, and stained with hematoxylin and eosin for histopathology. Double immunofluorescence was conducted to analyze the expression of TGF-ß-CD14 double-positive MPs in periapical tissues. The number of double-positive cells (cells/mm2) were significantly higher in the chronic periapical disease tissues (P < 0.01) compared to that in the control tissue; in addition, the density of TGF-ß1-CD14 double positive cells was significantly higher in the periapical cyst group than in the periapical granuloma group (P < 0.01). The number of TGF-ß1 expressing macrophages varied with human chronic periapical diseases. The TGF-ß1-CD14 double-positive cells might play an important role in the pathology of human chronic periapical diseases.
Assuntos
Macrófagos/metabolismo , Doenças Periapicais/metabolismo , Fator de Crescimento Transformador beta1/biossíntese , Adulto , Estudos de Casos e Controles , Doença Crônica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Doenças Periapicais/genética , Doenças Periapicais/imunologia , Granuloma Periapical/genética , Granuloma Periapical/imunologia , Granuloma Periapical/metabolismo , Cisto Radicular/genética , Cisto Radicular/metabolismo , Fator de Crescimento Transformador beta1/genéticaRESUMO
The aim of this study was to evaluate the three-dimensional (3D) parameters given by the micro-computed tomography (µCT) analysis of experimentally induced periapical lesions in wild type (WT) and knockout mice for the interleukin 22 (IL-22 KO). Periapical lesions were induced in the mandibular first molars of wild type and IL-22 KO mice (n = 12 teeth/group). The animals were euthanized after the experimental periods of 7, 21 and 42 days. The mandibles were removed and exposed to µCT scanning. The analyses were performed by the CTAn software for the tree-dimensional parameters: Tissue Volume (TV), Lesion Volume (LV), Tissue Surface (TS), Lesion Surface (LS), Intersection Surface (IS), and Trabecular Pattern factor (Tb.Pf). After that, the tissue was subjected to routine histologic procedures and to immunohistochemistry analysis. Statistical analysis was performed in the GraphPad software. A t-test was used to compare the differences between the groups with significance level of 5%. The evaluation of the 3D parameters showed statistical significant difference between the groups only at the latest period of periapical lesion development (42 days), for the TV, LV, TS, LS and IS parameters. The immunohistochemistry evaluation confirmed the immunostaining for IL-22 only in the WT mice, surrounding the periapical lesion. There were no differences regarding the trabecular alveolar bone (Tb.Pf) that could influence the lesion development. In conclusion, the 3D parameters showed that the absence of IL-22 leads to detectable differences at 42 days of lesion progression, resulting in smaller periapical lesions.
Assuntos
Interleucinas/imunologia , Doenças Periapicais/diagnóstico por imagem , Doenças Periapicais/imunologia , Animais , Imageamento Tridimensional , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microtomografia por Raio-X , Interleucina 22RESUMO
Immunoregulatory mechanisms within periapical lesions (PLs) are as of yet unexplored. Considering the crucial role of DCs in controlling the immune response within PLs, the immunomodulatory properties of mesenchymal stem cells (MSCs), and the colocalization of MSCs and DCs in situ, we wondered whether MSCs from PLs modulate the development and functions of DCs. Using a model of monocyte-derived DCs, we showed that PL-MSCs inhibited differentiation of DCs via soluble factors, of which IL-6 had a minor effect, but did not impair their subsequent maturation induced by pro-inflammatory cytokines. However, upon maturation such DCs favored the production of Th2/Th17 cytokines by allogenic CD4(+) lymphocytes in coculture, compared with mature DCs differentiated without PL-MSCs. PL-MSC-differentiated DCs, cultivated with pro-inflammatory cytokines and PL-MSCs, although phenotypically mature, exhibited poor allostimulatory activity, induced anergy, Th2 polarization, differentiation of suppressive CD4(+) CD25(high) CD39(+) Treg-cell subsets via IDO-1-, ILT-3-, and ILT-4-dependent mechanisms, and increased production of TGF-ß in the coculture. In contrast, DCs cultivated with PL-MSCs only during maturation stimulated proliferation and Th1 polarization of CD4(+) T cells in an IL-12-independent manner. In conclusion, PL-MSCs significantly modulate the development and functions of DCs, depending on the phase of DCs development during which the interaction occurs.
Assuntos
Diferenciação Celular/imunologia , Células Dendríticas/citologia , Células Dendríticas/imunologia , Células-Tronco Mesenquimais/imunologia , Tecido Periapical/citologia , Tecido Periapical/imunologia , Separação Celular , Técnicas de Cocultura , Citometria de Fluxo , Humanos , Imuno-Histoquímica , Teste de Cultura Mista de Linfócitos , Monócitos/citologia , Monócitos/imunologia , Doenças Periapicais/imunologiaRESUMO
AIM: To examine cytokine expression profiles during periapical lesion development in response to synergetic human pathogens in a gnotobiotic mouse model. METHODOLOGY: Human strains of Fusobacterium nucleatum and Peptostreptococcus prevotii were inoculated into the root canals of germ-free mice in either mono- or bi-association. Animals were killed 7 and 14 days after infection, and periapical tissues were collected. mRNA expression of the cytokines IFN-γ, TNF-α, Receptor activator of nuclear factor kappa-B ligand (RANKL), IL-10, IL-4 and transforming growth factor ß (TGF-ß) was assessed using real-time PCR. Levene's test was used to assess the equality of variance of the data, whereas a t-test for independent samples was used to evaluate the significance of the differences between groups (P < 0.05). RESULTS: The mRNA expression of IFN-γ and TNF-α was up-regulated by F. nucleatum during the acute (day 7) and chronic phase (day 14) of periapical lesion development. However, in bi-infection the expression of IFN-γ and TNF-α were effectively absent at both time-points. RANKL mRNA expression was down-regulated during dual infection at the chronic phase. As IL-4 expression was similar at both time-points, IL-4 does not appear to be involved in the periapical response to these bacterial strains. IL-10 was up-regulated during the chronic phase by mono-infection with either F. nucleatum or P. prevotii. Dual infection increased TGF-ß mRNA expression on day 7, which paralleled the decrease in IFN-γ and TNF-α mRNA levels at the same time-point. F. nucleatum increased TGF-ß mRNA expression during the chronic phase. CONCLUSION: Cytokine profiles depend on the nature of the bacterial challenge. Both TGF-ß and IL-10 appeared to be regulating the proinflammatory cytokine responses at both time-points of the periapical immune response.
Assuntos
Citocinas/análise , Doenças da Polpa Dentária/microbiologia , Infecções por Fusobacterium/imunologia , Fusobacterium nucleatum/imunologia , Infecções por Bactérias Gram-Positivas/imunologia , Peptostreptococcus/imunologia , Doenças Periapicais/microbiologia , Animais , Coinfecção/imunologia , Doenças da Polpa Dentária/imunologia , Vida Livre de Germes , Humanos , Mediadores da Inflamação/análise , Interferon gama/análise , Interleucina-10/análise , Interleucina-4/análise , Camundongos , Doenças Periapicais/imunologia , Ligante RANK/análise , RNA Mensageiro/análise , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Tempo , Fator de Crescimento Transformador beta/análise , Regulação para Cima/imunologiaRESUMO
Bacteria are the prime cause of periapical diseases and root canal microbiology is a well-researched area of endodontics. Antigen-presenting cells (APCs) are present in periapical lesions of endodontic origin and play a substantial role in recognizing, processing and presenting pathogenic antigens to the adaptive immune system such as an effective and long-lasting immune response is generated against the specific pathogens. Toll-like receptors (TLRs) are germ-line encoded pathogen recognition receptors (PRR) expressed by various APCs which induce their maturation, lead to gene transcription in the nucleus and the production of several pro- and anti-inflammatory cytokines. Thirteen TLRs have been discovered, 10 of which have been identified in humans so far. Preliminary studies of dental pulp tissue have demonstrated various cell types expressing different TLRs in response to commonly encountered microorganisms. However, there is little information available regarding the expression and function of the various TLRs in human periapical lesions. This review discusses the interactions of various APCs in periapical lesions and the possible roles of different TLRs and APCs in pulp/periapical pathogen recognition and presentation to the adaptive immune system in the initiation and sustaining of periapical diseases.
Assuntos
Imunidade Adaptativa/fisiologia , Células Apresentadoras de Antígenos/imunologia , Polpa Dentária/citologia , Doenças Periapicais/imunologia , Receptores Toll-Like/fisiologia , Imunidade Adaptativa/imunologia , Apresentação de Antígeno/imunologia , Células Apresentadoras de Antígenos/citologia , Polpa Dentária/imunologia , HumanosRESUMO
OBJECTIVE: The aim of this study was to qualitatively and semi-quantitatively analyze mast cells in periapical lesions. MATERIALS AND METHODS: Biopsy specimens of 96 periapical lesions were stained with hematoxylin-eosin, histochemical Giemsa and immunohistochemical CD 117 (C kit) antibody. Mast cell count below 100 mast cells on 1000 fields of high power magnification was noted as 'negative', 101-400 as 'mild', 401-800 cells as 'moderate', and over 800 cells as 'severe'. RESULTS: Mast cells are found in 68 (70.8%) lesions. The presence of mast cells was greater in cysts than in granulomas (P < 0.0028). There was no difference in semi-quantitative expression of CD 117 in granulomas and cysts (P > 0.05). Mast cells were placed in both: inflammatory infiltrate and in fibroblastic areas of periapical lesions, and their presence was most frequently mild to moderate. CONCLUSIONS: The findings of present study could suggest a role of mast cells in regulation of cellular immune mechanisms in periapical lesions, balancing between alterative and reparatory processes in inflamed periapical tissue.
Assuntos
Mastócitos/fisiologia , Doenças Periapicais/etiologia , Biópsia , Contagem de Células , Degranulação Celular/fisiologia , Corantes , Fibroblastos/patologia , Humanos , Imuno-Histoquímica , Linfócitos/patologia , Mastócitos/imunologia , Neutrófilos/fisiologia , Doenças Periapicais/imunologia , Doenças Periapicais/patologia , Granuloma Periapical/imunologia , Granuloma Periapical/patologia , Periodontite Periapical/imunologia , Periodontite Periapical/patologia , Plasmócitos/patologia , Proteínas Proto-Oncogênicas c-kit/análise , Cisto Radicular/imunologia , Cisto Radicular/patologiaRESUMO
AIM: The aim of this study is to determine the presence and distribution of Langerhans cells in periapical lesions, and correlate this with inflammatory cell infiltration and epithelial cell proliferation. MATERIAL AND METHODS: Seventy chronic dental periradicular lesions, obtained during periapical surgery from 70 patients, were included in this study, including: 46 granulomas, 18 scar tissue and 6 periradicular cysts. Immunohistochemical staining was performed using the following markers: CD3 to analyze the inflammatory infiltrate, CD1a to determine the presence of Langerhans cells and Ki67 to analyze the epithelial cell proliferation. The CD1a immunostaining density was established following Cincura (2007) criteria, being classified ranging from intense (3), moderate (2), discrete (1) or no (0) immunostaining. CD3 and Ki67 staining was evaluated following the Liapatas et al. scale, as: 0) no cells stained; 1) weak stain or few cells stained (11-25%); 2) moderate staining or some cells stained (26-75%); 3) intense staining or many cells stained (more than 76%). RESULTS: Langerhans cells were found in 32.8% of the periapical lesions being more intense in the epithelialized lesions. CD3 immunohistochemical staining was found in all lesions, but with different values in relation to histological subtypes. Ki67 was positive in all epithelialized lesions, although with a moderate staining. CONCLUSIONS: Langerhans cells appeared to be associated with T-lymphocyte infiltration and the proliferative potential of the epithelial tissue in periapical lesions.
Assuntos
Células Epiteliais , Células de Langerhans , Doenças Periapicais/imunologia , Doenças Periapicais/patologia , Adolescente , Adulto , Proliferação de Células , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
INTRODUCTION: Mesenchymal stem cells (MSCs) are characterized by the potential to differentiate into multiple cell lineages, high proliferation rates, and self-renewal capacity, in addition to the ability to maintain their undifferentiated state. These cells have been identified in physiological oral tissues such as pulp tissue, dental follicle, apical papilla and periodontal ligament, as well as in pathological situations such as chronic periapical lesions (CPLs). The criteria used for the identification of MSCs include the positive expression of specific surface antigens, with CD73, CD90, CD105, CD44, CD146, STRO-1, CD166, NANOG and OCT4 being the most specific for these cells. AIM: The aim of this review was to explore the literature on markers able to identify MSCs as well as the presence of these cells in the healthy periodontal ligament and CPLs, highlighting their role in regenerative medicine and implications in the progression of these lesions. METHODS: Narrative literature review searching the PubMed and Medline databases. Articles published in English between 1974 and 2020 were retrieved. CONCLUSION: The included studies confirmed the presence of MSCs in the healthy periodontal ligament and in CPLs. Several surface markers are used for the characterization of these cells which, although not specific, are effective in cell recognition. Mesenchymal stem cells participate in tissue repair, exerting anti- inflammatory, immunosuppressive and proangiogenic effects, and are therefore involved in the progression and attenuation of CPLs or even in the persistence of these lesions.
Assuntos
Células-Tronco Mesenquimais/citologia , Doenças Periapicais/patologia , Ligamento Periodontal/citologia , Endodontia Regenerativa/métodos , Adipócitos/citologia , Adipócitos/imunologia , Antígenos CD/genética , Antígenos CD/imunologia , Antígenos de Superfície/genética , Antígenos de Superfície/imunologia , Biomarcadores/metabolismo , Diferenciação Celular , Linhagem da Célula/genética , Linhagem da Célula/imunologia , Condrócitos/citologia , Condrócitos/imunologia , Polpa Dentária/citologia , Polpa Dentária/imunologia , Expressão Gênica , Humanos , Células-Tronco Mesenquimais/imunologia , Proteína Homeobox Nanog/genética , Proteína Homeobox Nanog/imunologia , Fator 3 de Transcrição de Octâmero/genética , Fator 3 de Transcrição de Octâmero/imunologia , Osteoblastos/citologia , Osteoblastos/imunologia , Osteogênese/genética , Osteogênese/imunologia , Doenças Periapicais/genética , Doenças Periapicais/imunologia , Doenças Periapicais/terapia , Ligamento Periodontal/imunologiaRESUMO
The present study aims to evaluate the longitudinal effects of induced experimental infections in gnotoxenic animals on the expression of inflammatory chemokines and their receptors in periradicular tissues. The null hypothesis tested was that Enterococcus faecalis and Fusobacterium nucleatum had no effect on CCR5, CCL5, CXCL10, CCL2/MCP-1, CXCR2 and CCR1 expression. Two groups of five animals (n = 5) aged between 8 and 12 weeks were used in this study. The animals were anaesthetized, and coronary access was performed in the first molar on the right and left sides. Microorganisms were inoculated into the left molar, and the right molar was sealed without contamination to function as a control. Animals were sacrificed 7 and 14 days after infection, and periapical tissues were collected. The cytokine mRNA expression levels were assessed using real-time PCR. The chemokine mRNA expression levels demonstrated that the experimental infection was capable of inducing increased chemokine expression on day 7 compared to that on day 14, except for CCR5 and CCL5, which showed no changes. The gnotoxenic animal model proved to be effective and allowed evaluation of the immune response against a known infection. Additionally, this study demonstrates that gene expression of chemokines and their receptors against the experimental infection preferentially prevailed during the initial phase of induction of the periradicular alteration (i.e., on day 7 post-infection).
Assuntos
Quimiocinas/análise , Cavidade Pulpar/imunologia , Doenças da Polpa Dentária/imunologia , Infecções por Fusobacterium/imunologia , Vida Livre de Germes , Infecções por Bactérias Gram-Positivas/imunologia , Receptores de Quimiocinas/análise , Animais , Quimiocinas/genética , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/microbiologia , Expressão Gênica , Camundongos , Doenças Periapicais/imunologia , Doenças Periapicais/microbiologia , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Quimiocinas/genética , Valores de Referência , Fatores de TempoRESUMO
The chronic inflammatory immune response triggered by the infection of the tooth root canal system results in the local upregulation of RANKL, resulting in periapical bone loss. While RANKL has a well-characterized role in the control of bone homeostasis/pathology, it can play important roles in the regulation of the immune system, although its possible immunoregulatory role in infectious inflammatory osteolytic conditions remains largely unknown. Here, we used a mouse model of infectious inflammatory periapical lesions subjected to continuous or transitory anti-RANKL inhibition, followed by the analysis of lesion outcome and multiple host response parameters. Anti-RANKL administration resulted in arrest of bone loss but interfered in the natural immunoregulation of the lesions observed in the untreated group. RANKL inhibition resulted in an unremitting proinflammatory response, persistent high proinflammatory and effector CD4 response, decreased regulatory T-cell (Treg) migration, and lower levels of Treg-related cytokines IL-10 and TGFb. Anti-RANKL blockade impaired the immunoregulatory process only in early disease stages, while the late administration of anti-RANKL did not interfere with the stablished immunoregulation. The impaired immunoregulation due to RANKL inhibition is characterized by increased delayed-type hypersensitivity in vivo and T-cell proliferation in vitro to the infecting bacteria, which mimic the effects of Treg inhibition, reinforcing a possible influence of RANKL on Treg-mediated suppressive response. The adoptive transfer of CD4+FOXp3+ Tregs to mice receiving anti-RANKL therapy restored the immunoregulatory capacity, attenuating the inflammatory response in the lesions, reestablishing normal T-cell response in vivo and in vitro, and preventing lesion relapse upon anti-RANKL therapy cessation. Therefore, while RANKL inhibition efficiently limited the periapical bone loss, it promoted an unremitting host inflammatory response by interfering with Treg activity, suggesting that this classic osteoclastogenic mediator plays a role in immunoregulation.
Assuntos
Osteólise/imunologia , Doenças Periapicais/imunologia , Ligante RANK/imunologia , Linfócitos T Reguladores/imunologia , Transferência Adotiva , Perda do Osso Alveolar/imunologia , Perda do Osso Alveolar/microbiologia , Animais , Anticorpos Monoclonais/farmacologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Expressão Gênica , Imunidade nas Mucosas , Inflamação/imunologia , Inflamação/microbiologia , Infliximab/farmacologia , Interleucina-10/imunologia , Ativação Linfocitária/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Osteólise/microbiologia , Doenças Periapicais/microbiologia , Ligante RANK/antagonistas & inibidores , Reação em Cadeia da Polimerase em Tempo Real , Fator de Crescimento Transformador beta/imunologiaRESUMO
Dentoalveolar bacterial infections cause localized tissue and bone destruction, but usually remain well-localized within teeth in immunocompetent hosts. However, in certain cases these infections may invade head and neck tissues, resulting in orofacial abscesses, cellulitis and sepsis, with resultant high morbidity and even mortality. In the present studies, we developed a novel model of spreading dentoalveolar infections in mice by treatment with neutralizing antibodies against both interleukin-1α (IL-1α) and IL-1ß. Surprisingly male but not female mice given anti-IL-1 antibodies developed orofacial abscesses, weight loss, splenomegaly and sepsis. Female mice developed abscesses and sepsis comparable to males following ovariectomy (OVX), which was reversed by estrogen supplementation. Anti-IL-1 blockade inhibited IL-12, interferon γ (IFNγ) and IL-6 but not IL-10 expression in infrabony lesions, suggestive of a local anti-inflammatory response. There was greater infiltration of neutrophils and other inflammatory cells into lesions in anti-IL-1-treated animals; however, blood leukocytes had reduced bacterial phagocytic and killing activity ex vivo. Estrogen directly stimulated IL-1 production by macrophages, suggesting that the resistance of females to disseminating dentoalveolar infections may be due to their heightened pro-inflammatory responses following bacterial challenge, leading to enhanced localization of these infections.
Assuntos
Infecções Bacterianas/tratamento farmacológico , Estradiol/farmacologia , Interleucina-1alfa/farmacologia , Interleucina-1beta/farmacologia , Doenças Periapicais/tratamento farmacológico , Animais , Infecções Bacterianas/imunologia , Infecções Bacterianas/microbiologia , Ensaio de Imunoadsorção Enzimática , Estradiol/imunologia , Interleucina-1alfa/imunologia , Interleucina-1beta/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Doenças Periapicais/imunologia , Doenças Periapicais/microbiologia , Fatores SexuaisRESUMO
INTRODUCTION: The pathogenesis of periapical lesions is determined by the balance between host proinflammatory immune response and counteracting anti-inflammatory and reparative responses, which include regulatory T cells (Tregs) as potential immunoregulatory agents. In this study, we investigated (in a cause-and-effect manner) the involvement of CCL22-CCR4 axis in Treg migration to the periapical area and the role of Tregs in the determination of outcomes in periapical lesions. METHODS: Periapical lesions were induced in C57Bl/6 (wild-type) and CCR4KO mice (pulp exposure and bacterial inoculation) and treated with anti-glucocorticoid-induced TNF receptor family regulated gene to inhibit Treg function or alternatively with CCL22-releasing, polylactic-glycolic acid particles to induce site-specific migration of Tregs. After treatment, lesions were analyzed for Treg influx and phenotype, overall periapical bone loss, and inflammatory/immunologic and wound healing marker expression (analyzed by real-time polymerase chain reaction array). RESULTS: Treg inhibition by anti-glucocorticoid-induced TNF receptor family regulated gene or CCR4 depletion results in a significant increase in periapical lesion severity, associated with upregulation of proinflammatory, T-helper 1, T-helper 17, and tissue destruction markers in parallel with decreased Treg and healing marker expression. The local release of CCL22 in the root canal system resulted in the promotion of Treg migration in a CCR4-dependent manner, leading to the arrest of periapical lesion progression, associated with downregulation of proinflammatory, T-helper 1, T-helper 17, and tissue destruction markers in parallel with increased Treg and healing marker expression. CONCLUSIONS: Because the natural and CCL22-induced Treg migration switches active lesion into inactivity phenotype, Treg chemoattractant may be a promising strategy for the clinical management of periapical lesions.
Assuntos
Quimiotaxia de Leucócito , Doenças Periapicais/imunologia , Doenças Periapicais/terapia , Linfócitos T Reguladores/imunologia , Animais , Quimiocina CCL22/imunologia , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Receptores CCR4/imunologia , Linfócitos T Reguladores/efeitos dos fármacosRESUMO
T helper (TH) and T suppressor (TS) cells were enumerated in developing rat periapical lesions in order to identify the cell type associated with the active phase of lesion formation. Experimental pulp exposures were made in all molar teeth of 29 Sprague-Dawley rats, and teeth were left open to the oral environment. On days 15, 20, 30, and 90 after induction, inflammatory cells were isolated from periapical lesions, and T helper (W3/25+), T suppressor (OX8+), and total T cells (W3/13+) were enumerated by reactivity with monoclonal antibodies. During the early, most active phase of lesion development (day 15), TH cells outnumbered TS cells (TH/TS = 1.7). However, by day 20, when lesion expansion had slowed, the TH/TS ratio was reversed (0.9) and remained so for up to 90 days after induction (TH/TS = 0.7). Total T cell numbers did not change over the period of observation. TH cells thus predominated during the active phase of lesion development, whereas TS cells were associated with the chronic phase. This dynamic TH/TS reversal indicates that immune responses which occur within periapical lesions are highly regulated. In addition, the predominance of TH during the active phase of lesion development suggests that TH-mediated activities may be involved in the bone destruction and immune cell activation within periapical lesions.
Assuntos
Doenças Periapicais/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Reguladores/imunologia , Animais , Feminino , Contagem de Leucócitos , Doenças Periapicais/etiologia , Ratos , Ratos EndogâmicosRESUMO
Ten cats were immunized with subcutaneous injections of keyhole limpet hemocyanin (KLH). After it was determined that the experimental animals had developed circulating antibodies to KLH, challenge doses of KLH were administered via the root canal system. The radiographic and histologic findings suggest that antigen-antibody complex reactions can occur in periapical tissues of teeth, and that they can play a role in the pathogenesis of periapical lesions. In contrast, no radiographic and histologic changes were noted in non-immunized cats.
Assuntos
Complexo Antígeno-Anticorpo , Doenças Periapicais/imunologia , Tecido Periapical/imunologia , Animais , Formação de Anticorpos , Reações Antígeno-Anticorpo , Gatos , Cavidade Pulpar/imunologia , Hemocianinas/imunologia , Tecido Periapical/anatomia & histologia , Tecido Periapical/diagnóstico por imagem , RadiografiaRESUMO
T-helper and B-lymphocytes may contribute to mechanisms that result in bone-resorptive cytokine production in periapical lesion. Mice with severe combined immunodeficiency (scid) lack functional B- and T-cell immunity. The purpose of this study was to investigate the progression of pulp necrosis and the histomorphometric features of periapical lesions in scid vs. normal mice. The expression of the bone-resorptive cytokines IL-1 alpha and TNF-alpha was also investigated. Sixteen five-week-old homozygous scid mice and 14 normal BALB/cJ mice were used. The pulps of mandibular first molars were exposed for 1, 2, 3, or 4 weeks. Blocks of tissue containing the mandibular teeth and supporting structures were processed for both light microscopic examination and immunohistochemical staining for IL-1 alpha dna TNF-alpha. Central sections were randomized, their images were blindly digitized into a computer, and the areas of the lesions surrounding the distal root apices were measured. The cells that stained positively for the cytokines in the same area of adjacent sections were counted. Pulp necrosis progressed at similar rates in teeth from both strains. A progressive and significant increase in the periapical lesion size in both strains was observed. The scid mice lesions were significantly smaller than the controls at only the three-week period. There was heavy cytokine staining in periapical lesions from both strains, especially in areas that contained a mixed inflammatory infiltrate or fibroblasts. The number of positively staining cells was proportional to the lesion size. Therefore, pulpal and periapical pathosis were independent of the presence of functional T- and B-cells in this model.
Assuntos
Regulação da Expressão Gênica , Síndromes de Imunodeficiência/imunologia , Interleucina-1/genética , Doenças Periapicais/imunologia , Fator de Necrose Tumoral alfa/genética , Perda do Osso Alveolar/imunologia , Perda do Osso Alveolar/patologia , Análise de Variância , Animais , Linfócitos B/imunologia , Reabsorção Óssea/imunologia , Reabsorção Óssea/patologia , Corantes , Exposição da Polpa Dentária/imunologia , Exposição da Polpa Dentária/patologia , Necrose da Polpa Dentária/imunologia , Necrose da Polpa Dentária/patologia , Progressão da Doença , Ensaio de Imunoadsorção Enzimática , Fibroblastos/imunologia , Fibroblastos/patologia , Processamento de Imagem Assistida por Computador , Técnicas Imunoenzimáticas , Síndromes de Imunodeficiência/patologia , Inflamação , Interleucina-1/análise , Linfócitos/imunologia , Linfócitos/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Neutrófilos/imunologia , Neutrófilos/patologia , Doenças Periapicais/patologia , Plasmócitos/imunologia , Plasmócitos/patologia , Linfócitos T Auxiliares-Indutores/imunologia , Fator de Necrose Tumoral alfa/análiseRESUMO
Twelve periapical lesions from symptomatic and asymptomatic patients (six each) were obtained and immediately frozen in liquid nitrogen. Six pulps from unerupted third molars as well as chronically inflamed gingival tissues were also obtained, frozen, and used as negative and positive controls, respectively. The concentration of leukotriene (LT) B4 was determined by reverse phase high pressure liquid chromatography. Representative samples from each group were fixed in formalin, sectioned, and stained with hematoxylin and eosin. Extremely low levels of LTB4 were detected in the uninflamed pulpal samples in comparison to those found in chronically inflamed gingival tissues and periradicular lesions. A significant statistical difference was noted between concentrations of LTB4 in periapical lesions of symptomatic patients and those found in asymptomatic patients and samples of chronically inflamed gingival tissues (p < 0.05). In addition, a positive correlation was found between the presence of symptoms, the concentration of LTB4, and presence of polymorphonuclear leukocytes in symptomatic periapical lesions. The results show presence of high concentrations of LTB4 in symptomatic human periapical lesions.
Assuntos
Leucotrieno B4/análise , Doenças Periapicais/imunologia , Análise de Variância , HumanosRESUMO
The presence of immunoglobulins has been demonstrated in periapical inflammatory lesions associated with endodontic disease. The purpose of this study was to determine if IgG is synthesized in vitro in explant cultures of untreated periapical inflammatory lesions and to determine the level of IgG in isolated samples. Periapical lesions associated with infected root canals were removed from the roots and cultured in tissue culture medium containing tritiated amino acids. Supernatant fluids from the explant tissue cultures were passed through staphylococcal protein A affinity columns to isolate IgG. When the staphylococcal protein A eluents (24-h samples) from six periapical lesions were used in double diffusion in agarose assays, the presence of IgG was demonstrated in all the samples. Radial immunodiffusion assays to quantitate the IgG in staphylococcal protein A eluents showed that the levels of IgG detected in each successive daily supernatant fluid always decreased or else fell below the lower limits of detection. The in vitro biosynthesis of IgG in explant cultures of periapical lesions was demonstrated by the incorporation of tritiated amino acids into isolated IgG.
Assuntos
Imunoglobulina G/biossíntese , Doenças Periapicais/imunologia , Análise de Variância , Técnicas de Cultura , Humanos , Imunodifusão , Proteína Estafilocócica ARESUMO
The presence of immunoreactive cells in periapical inflammatory lesions suggests that immune responses participate in the disease process. The purpose of this study was to determine the presence and concentration of immunoglobulins in the supernatant fluids of explant cultures of periapical lesions. Ninety periapical lesions that had been contiguous with the apex of a root were removed and maintained in explant cultures for 96 h. Tissue culture medium was replenished at 24, 48, 72, and 96 h. Double diffusion in agarose assays demonstrated the presence of IgG in 100% of the 24-h supernatant fluids and IgA in 65% of the 24-h supernatant fluids. However, IgM was not detected. Radial immunodiffusion assays were used to detect and quantitate IgG, IgA, and IgM in samples of 24-h supernatant fluids from 90 explant cultures. IgG was the predominant immunoglobulin followed by IgA. A radioimmunosorbent test was used to detect and quantitate IgE in samples of 24-h supernatant fluids from 90 explant cultures. Forty of the 90 supernatant fluids contained measurable IgE. All detected immunoglobulins decreased in concentration in daily supernatant fluids with time (24, 48, 72, and 96 h) in the culture.
Assuntos
Isotipos de Imunoglobulinas/análise , Doenças Periapicais/imunologia , Adolescente , Adulto , Idoso , Técnicas de Cultura , Doenças da Polpa Dentária/imunologia , Feminino , Humanos , Imunodifusão , Masculino , Pessoa de Meia-Idade , Tecido Periapical/imunologia , Teste de RadioimunoadsorçãoRESUMO
The concentration of secretory IgA in fluids present in the canals of 33 teeth was determined by the rocket immunoelectrophoresis technique. Except for the presence or absence of communication between the oral cavity and the root canals of the affected teeth, no other clinical finding showed significant statistical correlation with the presence of secretory IgA. The canals which were open to the oral flora had significantly higher concentrations of secretory IgA. Leaving canals open to the oral cavity may result in formation of periapical cysts.