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PubMed1998

Interferon-gamma-producing cells and inducible nitric oxide synthase-producing cells in periapical granulomas.

Periapical granulomas contain a large number of T lymphocytes and monocytes/macrophages and a small number of B lymphocytes and polymorphonuclear leukocytes. Sections from eight periapical granulomas were stained by a variety of immunohistochemical methods. The vascular endothelial cells stained positively for intercellular adhesion molecule-1 and vascular cell adhesion molecule-1. Helper T cells were identified by immunostaining for CD4 and stained positively for interferon-gamma (IFN-gamma). However, CD4-positive T cells did not stain for interleukin-4 (IL-4). Monocytes/macrophages were identified by immunostaining for CD68 and stained positively for IL-1alpha or inducible nitric oxide synthase (iNOS). IL-1beta could not be detected in the same samples. No cytokine expression was observed in B cells identified by immunostaining for CD20. IFN-gamma- and iNOS-positive cells could not be detected in clinically healthy periodontal ligament being used as a negative control. These results suggest that the IFN-gamma-producing T cells and iNOS-positive cells may modulate the progress of disease in local inflammation sites such as in periapical granulomas.

Adult↗

PubMed2006

RANKL in human periapical granuloma: possible involvement in periapical bone destruction.

OBJECTIVES: The cytokine receptor activator of nuclear factor kappaB-ligand (RANKL) has been involved in both the physiological and pathological regulation of osteoclast life span and bone metabolism. Periapical granuloma is a periradicular lesion characterized by periapical bone destruction. The aims of this study were to associate the RANKL mRNA levels to periapical granulomas using the real-time reverse transcriptase-polymerase chain reaction (RT-PCR) technique and to determine the specific cell involved in RANKL synthesis. METHODS: In eight periapical granuloma and eight periodontal ligament samples from periodontally healthy volunteers, RANKL mRNA was detected by real-time RT-PCR. Expression of RANKL on infiltrate leukocytes was further investigated by flow cytometry in six periapical granulomas. RESULTS: Receptor activator of nuclear factor kappaB-ligand mRNA levels were higher in periapical granulomas than in healthy periodontal ligament as its RANKL mRNA cycle threshold (Ct) and DeltaCt were significantly lower than that of controls (33.07 +/- 1.24 vs 36.96 +/- 1.69 and 11.58 +/- 3.02 vs 15.60 +/- 3.31, respectively). A 16.2-fold (2.0-131.6) higher RANKL gene expression was detected in the granulomas compared with the control tissues. We determined by flow cytometry that lymphocytes were the predominant leukocyte cells (53.31%), and monocytes and dendritic cells were the main RANKL synthesizers in granuloma lesions. CONCLUSIONS: These data indicate that monocytes synthesized RANKL in periapical granulomas and suggest that RANKL is involved in bone loss associated with periapical lesions.

Adolescent↗

PubMed1996

Histological findings of human leprosy periapical granulomas.

Histological analysis of human leprosy periapical granulomas was conducted to study the histological responses to Mycobacterium leprae. Many Langhans-type giant cells and epithelioid cell tubercles were observed, although M. leprae were not detected in leprosy periapical granulomas. Although numerous T- and B-cells were infiltrated in leprosy periapical granuloma, the T-/B-cell ratio of leprosy is not changed to that of periapical granulomas isolated from normal patients. These findings suggested that human leprosy periapical granulomas develop as a result of immunological responses to M. leprae.

B-Lymphocytes↗

PubMed1982

Isolation and characterization of inflammatory cells from the human periapical granuloma.

Twelve histologically-confirmed periapical granulomas were evaluated by conventional immunologic rosette assays for the presence of T-lymphocytes and complement receptor-bearing lymphocytes. A technique for dispersing the granuloma cells into suspensions was adopted to facilitate performance of the assays which were not applicable to tissue sections. Differential cell counts by an acridine orange vital dye method disclosed that the cell suspensions contained 30% macrophages, 44% lymphocytes, 15% plasma cells, and 12% neutrophils. Complement receptor-bearing cells comprised 17.9%, and T cells comprised 34.5% of the unseparated inflammatory cells. This study provides the first direct evidence of a predominance of thymic-derived lymphocytes in the lymphocyte compartment of the periapical granuloma. Analysis of the data shows that cell-mediated immunity most likely plays a role in the pathogenesis of the periapical granuloma.

Cell Count↗

PubMed1989

[Histologic study of periapical granulomas].

Sixty-three periapical granulomas were histologically examined. They were divided into four types: exudative, granulomatous, granulofibrous and fibrous types. The most frequent forms were the granulomatous types. The granulomatous type consisted of four zones: necrotic, exudative, granulomatous and fibrous. In the granulomatous zone lymphocytes, plasma cells, macrophages, Russel bodies, foam cells, neutrophil leukocytes were found. Mast cells were found in the fibrous zone among the collagen fibres.

Humans↗

PubMed1990

The microbiology of periapical granulomas.

Of the 16 periapical granulomas studied, 14 (88%) yielded a positive growth when homogenized and cultured. The concentration of colony-forming units per milliliter of the suspension ranged from 10(1.3) to 10(4.0) (mean 10(2.2)). A total of 47 isolates comprising 26 (55%) facultative anaerobes and 21 (45%) strict anaerobes were obtained. The organisms most commonly cultured were Veillonella species (15%), Streptococcus milleri (11%), Streptococcus sanguis (11%), Actinomyces naeslundii (11%), Propionibacterium acnes (11%), and Bacteroides species (10%). Most of the organisms (96%) were sensitive to either amoxicillin, clindamycin, or tetracycline, whereas only 45% were sensitive to metronidazole.

Adolescent↗

PubMed1983

Immunoglobulin producing cells in human periapical granulomas.

Ten cases of periapical granuloma were stained for igG, IgA, IgM, IgD and IgE by the unlabelled antibody peroxidase-antiperoxidase complex method to investigate the class distribution of plasma cells within the lesions. The results indicated that IgG containing plasma cells predominated in all lesions and that IgG, IgA, IgM, IgE and IgD cells represented 81.9, 11.4, 5.4, 1.1 and 0.2 per cent respectively, of the total positive cell population. All specimens contained IgE plasma cells in addition to many non-lymphoid cells showing membrane-like staining for IgE. The results suggest that in addition to tissue damage via local deposition of immune complexes IgE mediated immediate hypersensitivity reactions may have a role in pathogenesis.

Humans↗

PubMedDate not supplied

Ultrastructural analysis of periapical granulomas.

A large portion of periapical granulomas is composed by cells of the mononuclear-phagocyte and lymphoid system. The ultrastructural features of the monocytes/macrophages differ in relationship to the state of activity of the cells (phagocytosis or secretion). A close correlation between cells of these two systems can be surmised by their tight contacts. An important role in the bone resorption is probably played by cytokines like IL-1 and TNF.

Humans↗

PubMed1989

Cellular composition of periapical granulomas and its function. Histological, immunohistochemical and electronmicroscopic study.

Periapical granulomas have been investigated histologically, immunohistologically using polyclonal and monoclonal antibodies, as well as electronmicroscopically. Lesions were formed by inflammatory granulation tissue frequently with foci of purulent exudation and fibrosis. Most numerous were plasma cells usually in cellular regions of the granulation tissue where they were tightly pressed. Of other cellular types were numerous lymphocytes, fibroblasts, less frequent were macrophages, scattered granulocytes and mast cells. More than a half of the plasma cells were IgG positive, about 20% IgA positive, up to 10% IgM, rarely IgE and sporadically IgD positive cells. In the vascular walls and their surrounding as well as in the phagocytes fine granular to granular positivities of C3 and C4 components of the complement were present. The majority of lymphocytes beared markers of T lymphocytes of which the T-suppressor markedly prevailed over the T-helper lymphocytes. In electron microscopy the plasma cells were most frequent. They were usually close to each other, sometimes with a disintegrated cytoplasmic membrane and non-damaged organelles being free around the nucleus. Mast cells were numerous and did not show any signs of marked degranulation. Rich production of immunoglobulins as well as the presence of IgG and IgM positive material in phagocytes, and the presence of positivities of the C3 and C4 components of the complement in the surrounding of the vessels and in phagocytes on the other hand supported the presumption that immune complexes participate in the pathogenesis of periapical granulomas. In spite of the presence of the IgE producing cells the morphological picture of mast cells did not suggest the presence of anaphylactic reaction in periapical lesions. Diffuse distribution of T lymphocytes, moreover with the prevalence of T-suppressor/cytotoxic over T-helper lymphocytes and not numerous macrophages in the inflammatory infiltrates did not suggest the participation of a typical cell-mediated immunity reaction in the development of periapical granulomas. Numerous T-suppressor/cytotoxic lymphocytes and low numbers of macrophages can be important factors of the chronicity of periapical inflammatory diseases.

Antibodies, Monoclonal↗

PubMed2001

Hydrogen peroxide and superoxide anion production by polymorphonuclear neutrophils in patients with chronic periapical granuloma, before and after surgical treatment.

Chronic periapical granuloma represents a localized tissue injury with well established signs of systemic immunological reactions. The aim of the study was to investigate changes in superoxide anion and hydrogen peroxide production by polymorphonuclear neutrophils (PMN) in patients with chronic periapical granuloma before and after surgical treatment. The affected teeth were extracted from 20 patients with chronic periapical lesions. Blood samples were obtained at admission, before extraction and on day 14. PMNs were isolated from blood samples and superoxide anion (O2-) and hydrogen peroxide (H2O2) production were estimated without stimulation and after stimulation of the cells with opsonized zymosan. Similar procedures were performed with blood samples obtained from 20 healthy controls. Superoxide anions as well as hydrogen peroxide production by unstimulated cells obtained from patients before treatment were significantly higher in comparison with controls. Fourteen days after extraction O2- production by unstimulated cells was higher than the controls and significantly lower in comparison to PMNs obtained before treatment, while H2O2 production was not significantly higher when compared to controls and significantly lower in comparison with PMNs obtained before extraction. The results obtained strongly imply the termination of a generalized inflammatory response after elimination of local inflammation by tooth extraction.

Adult↗

PubMed2003

Examination on Candida spp. in refractory periapical granulomas.

AIM: To examine the occurrence of Candida spp. in refractory periapical granulomas. METHODOLOGY: One hundred and three surgically removed periapical granulomas were subjected to molecular analysis for the occurrence of Candida albicans. DNA was extracted from the samples using a modified phenol/chloroform/isoamyl alcohol method and was subjected to polymerase chain reaction (PCR) with OPA-03 and repetitive sequence (GACA)4 primers. The PCR products were separated in agarose gel electrophoresis, stained with ethidium bromide, visualized using UV light and the sequences were analysed. Samples indicating possible occurrence of Candida were further investigated by histological and immunohistological methods. Periodic acid-Shiff staining (PAS) was used to detect yeast cells and hyphae, and specific monoclonal antibodies to recognize high molecular mass mannoproteins present in the C. albicans cell wall. DNA extraction was controlled by running PCR using beta-actin primers (a housekeeping gene). C. albicans CCUG19915, C. tropicalis ATCC750, C. krusei ATCC6258, C. guilliermondii ATCC6260 and C. glabrata CCUG32725 served as positive controls in PCR. A tissue preparation of chronic atrophic candidosis in oral buccal mucosa served as a positive control for histological and immunohistological examinations. RESULTS: Polymerase chain reaction with beta-actin primers indicated successful DNA extraction in 68 out of 103 samples. The majority of the samples (50) were negative whereas 18 of the samples showed PCR products indicating possible occurrence of Candida spp. PAS-staining and immunohistological examination of these samples were, however, negative. Further analysis of the PCR products revealed sequences not typical for Candida spp. CONCLUSIONS: Candida spp. do not seem to occur in periapical granuloma.

Candida↗

PubMedDate not supplied

Evaluation of immunoglobulin G synthesizing plasma cells in periapical granuloma and cyst.

Immunoglobulin synthesizing plasma cells for IgG were quantitated in 20 periapical granulomas and 20 periapical cysts, using unlabelled antibody peroxidase-antiperoxidase complex method. Result showed that immunoglobulin G producing plasma cells were predominant in periapical cyst as compared with periapical granuloma. A statistical significant relation was observed between these two lesions.

Humans↗

PubMed2006

Co-production of vascular endothelial cadherin and inducible nitric oxide synthase by endothelial cells in periapical granuloma.

AIM: To clarify the mechanisms of inflammatory cell migration in human periapical granulomas by examining vascular endothelial (VE) cadherin and inducible nitric oxide synthase (iNOS)-producing cells. METHODOLOGY: Periapical tissues were obtained from patients during endodontic surgery and were divided into two portions. After fixing the tissues with acetone or 4% paraformaldehyde in phosphate-buffered saline, 5-microm-thick paraffin or cryostat sections were prepared, respectively. The paraffin sections of the inflamed tissues were evaluated histologically with haematoxylin-eosin stains. Cryostat sections of the tissue, diagnosed as periapical granulomas, were then examined by either immunohistochemistry using anti-human VE-cadherin or iNOS antibodies (Abs) for the characterization of infiltrating cells. In addition, co-localization of VE-cadherin and iNOS production was also analysed by two-colour immunofluorescence image analysis. RESULTS: Endothelial cells were strongly stained with iNOS Abs. Macrophages, lymphocytes, polymorphonuclear leucocytes and fibroblasts also exhibited iNOS production. These iNOS-positive cells accumulated around the blood vessels. On the other hand, VE-cadherin production was exhibited in only endothelial cells. Two-colour immunofluorescence image analysis using VE-cadherin and iNOS Abs demonstrated that iNOS-producing endothelial cells also showed VE-cadherin production. CONCLUSIONS: Vascular endothelial-cadherin produced by endothelial cells could be regulated by iNOS-producing cells in periapical granulomas and might play a pivotal role in vascular permeability.

Adult↗

PubMed1989

Characteristics of collagenase in experimental periapical granulomas from the teeth of dogs.

Periapical granulomas were induced, with a success rate of about 60% (66 granulomas produced out of 109 roots treated) in mandibular premolars. The average wet weight of the granulomas, 46.1 +/- 34.5 mg (mean +/- SD, n = 22), was sufficient to allow individual specimens to be used for most of the biochemical analyses. High collagenase activity was extracted directly from the granulomas with 4 M urea solution. The enzyme was a typical animal collagenase (EC 3.4.24.7) which clove native collagen molecules into three-quarter (alpha A) and one-quarter (alpha B) length fragments. The collagenase was activated by 1 mM p-aminophenylmercuric acetate. This activated enzyme broke down collagen I rather than collagen III preferentially, which is similar to the activity of human polymorphonuclear leucocyte collagenase. The molecular weights of the latent and activated collagenases were 67 and 49 K, respectively.

Animals↗

PubMed1991

Immunoreactivity for interleukin 1-beta and tumor necrosis factor-alpha and ultrastructural features of monocytes/macrophages in periapical granulomas.

Interleukin 1 (IL-1) and tumor necrosis factor (TNF) have recently been shown to be involved in bone resorption, and because macrophages constitute a significant part of human periapical granulomas, it is reasonable to suspect that they may secrete IL-1 and TNF. The purpose of our investigation was to detect and characterize IL-1 beta- and TNF-alpha-producing cells in human periapical granulomas. Fresh tissue samples obtained during surgery from 10 patients with previously untreated teeth and histologically established periapical granulomas were studied with light and electron microscopy and with immunohistochemical analysis with monoclonal antibodies against IL-1 beta and TNF-alpha. There were very few IL-1 beta + and TNF-alpha + cells present in periapical granulomas, and the positive cells had monocyte/macrophage morphology. The IL-1 beta + cells were located mainly in areas of active exudation and were surrounded by and/or were in close contact with lymphoid cells, whereas TNF-alpha + cells were scattered and in contact with or near other inflammatory cells at the periphery of active granulation tissue. This suggests that the IL-1 beta + cells may act in a paracrine manner to activate lymphoid cells. The ultrastructural findings showed that only some macrophages are adapted to extracellular secretion rather than phagocytosis. These modified macrophages could be the major producers of interleukins in tissues. Occasionally, they have plasmacytic or plasmacytoid features resembling the so-called "plasmacytoid monocytes". Only a minor fraction of the monocytes/macrophages (representing about 40% of the inflammatory cells) is in an active cytokine-producing state.(ABSTRACT TRUNCATED AT 250 WORDS)

Bone Resorption↗

PubMed1999

Cytotoxic T lymphocytes versus streptococcal colonization in periapical granulomas.

Seventeen dental periapical lesions were investigated to study bacterial colonization. Periapical lesions, obtained after apicotomy, were also enzymatically desegregated to quantitatively analyze lymphocyte subpopulations by flow cytometry. Fourteen samples yielded a positive bacterial growth when homogenized and cultured. We isolated enough lymphocytes to make flow cytometric analysis from 12 samples. A significant increase in interleukin-2 receptor and ICAM-1 molecule expression on T cells was found, compared with peripheral blood lymphocytes. Furthermore, a decreased expression of interleukin-2 receptors and HLA DR molecules on CD8+ T cells was found in granulomas predominantly colonized by Streptococcus spp., compared with lesions predominantly colonized by strict anaerobes.

Bacteria, Anaerobic↗

PubMed2002

Involvement of substance P, mast cells, TNF-alpha and ICAM-1 in the infiltration of inflammatory cells in human periapical granulomas.

BACKGROUND: Substance P (SP), a potent proinflammatory peptide present in sensory neurons, is believed to be a major mediator of neurogenic inflammation. The aim of this study was to examine the localization and involvement of SP, mast cells and tumor necrosis factor-alpha (TNF-alpha)-positive cells in human periapical granulomas. METHODS: Sections from seven periapical granulomas were stained using a variety of immunohistochemical methods. RESULTS: SP-immunoreactive nerve fibers, mast cells and TNF-alpha-positive cells were found localized in the vicinity of blood vessels in all the samples of periapical granulomas. The vascular endothelial cells stained positively for E-selectin and intercellular adhesion molecule-1. SP, TNF-alpha-positive cells and E-selectin could not be detected in clinically healthy periodontal ligament, and served as a negative control. CONCLUSION: Our findings suggest that SP, mast cells, TNF-alpha-positive cells and E-selectin may modulate the pathogenesis of apical periodontitis and may be responsible for stimulating the formation of granuloma with the resorption of periapical bone.

Adult↗

PubMed1991

[Cellular immunity in human periapical granuloma].

Numerous authors have produced different types of immunoglobulins in analyses of the human periapical granuloma. The present study examines the cellular immunity of the human periapical granuloma, and in particular the distribution of the lymphocyte sub-populations and the macrophage population. The technique used was that of cell surface marking, using monoclonal antibodies on frozen sections. The results reveal equal proportions of inductor T lymphocytes and suppressor T lymphocytes (whereas healthy tissue shows a ratio of 2/1), which explains the chronic nature of the lesion as far as the immune reaction is concerned. The presence of numerous macrophage cells shows that there are important local immune reactions.

Adult↗