Phenotypic characterisation of stellate and giant cells in giant cell fibroma by immunocytochemistry

The origin of the stromal, stellate and multinucleate cells in oral giant cell fibroma is unclear. Sixteen giant cell fibromas were stained immunocytochemically for keratin (MNF 116), vimentin, S-100 protein, neurofilaments, glial fibrillary acidic protein, α-smooth muscle actin, desmin, CD31 (PECAM-1). CD68, Factor XIIIa and prolyl 4-hydroxylase (5B5). In all cases positive staining was found with vimentin and prolyl 4-hydroxylase, indicating a functional fibroblast phenotype. Reactivity for Factor XII la was seen in two cases and in only one was a small number of giant cells stained, suggesting that the majority of oral giant cell fibromas are unrelated to the histologically similar fibrous papule of the nose or facial angiofibroma.     

Cytokeratins in epithelia of odontogenic neoplasms

Neoplasms and tumours related to the odontogenic apparatus may be composed only of epithelial tissue or epithelial tissue associated with odontogenic ectomesenchyme. The immunohistochemical detection of different cytokeratins (CKs) polypeptides and vimentin has made it easier to explain the histogenesis of many epithelial diseases. The present study aimed to describe the immunohistochemical expression of cytokeratins 7, 8, 10, 13, 14, 18, 19 and vimentin in the epithelial components of the dental germ and of five types of odontogenic tumours. The results were compared and histogenesis discussed. All cells of the dental germ were positive for CK14, except for the preameloblasts and secreting ameloblasts, in which CK14 was gradually replaced by CK19. CK7 was especially expressed in the cells of the Hertwig root sheath and the stellate reticulum. The dental lamina was the only structure to express CK13. The reduced epithelium of the enamel organ contained CK14 and occasionally CK13. Cells similar to the stellate reticulum, present in the ameloblastoma and in the ameloblastic fibroma, were positive for CK13, which indicates a nature other than that of the stellate reticulum of the normal dental germ. The expression of CK14 and the ultrastructural aspects of the adenomatoid odontogenic tumour probably indicated its origin in the reduced dental epithelium. Calcifying odontogenic epithelial tumour is thought to be composed of primordial cells due to the expression of vimentin. Odontomas exhibited an immunohistochemical profile similar to that of the dental germ. In conclusion, the typical IF of odontogenic epithelium was CK14, while CK8, 10 and 18 were absent. Cytokeratins 13 and 19 labelled squamous differentiation or epithelial cells near the surface epithelium, and CK7 had variable expression.     

Enhanced expression of podoplanin in ameloblastomas

J Oral Pathol Med (2010) 39 : 103–109 Objective: Podoplanin, a mucin‐type transmembrane glycoprotein, is specifically expressed by lymphatic but not blood vascular endothelial cells, and is also widely expressed in various specialized cell types throughout the body. Recent studies have demonstrated that it mediates a pathway leading to collective cell migration and invasion in vivo and in vitro. In the present study, we carried out an immunohistochemical investigation of podoplanin to clarify whether it is expressed in human ameloblastomas (AMs), which are characterized by locally aggressive behavior with a high rate of recurrence. In addition, we examined the localization of the epithelial marker E‐cadherin and the mesenchymal marker vimentin to clarify whether AMs show epithelial–mesenchymal transition (EMT). Methods: Paraffin‐embedded tissue specimens of 38 AMs were examined immunohistochemically using antibodies against podoplanin, E‐cadherin, and vimentin. Results: Immunohistochemical reactivity for podoplanin was detected in the cell membrane and cytoplasm of most odontogenic tumor epithelial cells in AMs. Podoplanin was expressed strongly in peripheral columnar cells and slightly in central stellate reticulum‐like cells. E‐cadherin was expressed in central stellate reticulum‐like cells and showed decreased expression in peripheral columnar cells. Immunoreactivity for E‐cadherin was weak or negative in keratinizing cells of acanthomatous AMs, suggesting terminal differentiation of the tumor cells. Immunohistochemical reactivity for vimentin was found in stromal cells, but partial or no reaction was observed in neoplastic cells. Conclusion: Expression of podoplanin in AMs is considered to be associated with neoplastic odontogenic tissues; this molecule might play a role in the collective cell migration of tumor nests in AMs. The pattern of expression of E‐cadherin and vimentin suggests that invasion in AMs occurs in the absence of EMT. The migration and invasion mediated by podoplanin in AMs could be related to cytoskeletal reorganization.     

Immunohistochemical investigation in odontogenic myxoma

Three odontogenic myxomas are described immunohistochemically by a panel of poly- and monoclonal antibodies to characterize this tumor type. Three types of odontogenic myxoma cells were discriminated: spindle cells, stellate cells and hyaline cells. Neoplastic cells of myxomas were positively stained for transferrin, ferritin, alpha-1-antichymotrypsin (alpha 1-ACT), alpha-1-antitrypsin (alpha 1-AT), S-100 protein and vimentin; however, neuron specific enolase (NSE), S-100 alpha subunit, S-100 beta subunit, Factor VIII-related antigen (FVIII-AG) and cytokeratin (CK1) were negative. Spindle cells were positive for transferrin, ferritin, alpha 1-ACT, alpha 1-AT, S-100 protein and vimentin. Stellate cells were strongly positive for transferrin, alpha 1-AT, S-100 protein and vimentin. Hyaline cells reacted with alpha 1-ACT and alpha 1-AT. Myxomatous matrix showed negative reaction for all the antibodies used. These results have confirmed that odontogenic myxoma is a tumor of a dual fibroblastic-histiocytic origin.     

Patterns of expression of intermediate filaments in ameloblastoma and human fetal tooth germ

Monoclonal antibodies (Mab) were used to study the expression of cytokeratins and vimentin in various histological types of ameloblastoma and in human fetal tooth germ. The ameloblastoma and the tooth germ epithelia showed characteristics of both simple glandular and stratified squamous epithelial cells. Cytokeratin No. 18 was detected focally in most ameloblastomas studied but not in fetal odontogenic epithelia. Cytokeratins Nos. 8 and 19 were expressed in all epithelial elements of ameloblastomas and tooth germs. Only two tumors showed focally characteristics of keratinizing epithelia also seen in dental lamina but not in the enamel organ. All tumors except the granular cell ameloblastoma showed a variable coexpression of vimentin and cytokeratins in their neoplastic epithelia. A similar coexpression was detected in the stellate reticulum cells of the developing tooth. Ameloblastoma and human tooth germ epithelia share complex pattern of cytokeratin polypeptides together with coexpression of vimentin. The results strongly support the theory that ameloblastomas are of odontogenic origin and not direct derivatives of basal cells of oral epithelium or epidermis.     

Immunohistochemical expression of mast cell tryptase in giant cell fibroma and inflammatory fibrous hyperplasia of the oral mucosa

This study analysed the immunohistochemical expression of mast cell tryptase in giant cell fibromas (GCFs). In addition, the possible interaction of mast cells with stellate giant cells, as well as their role in fibrosis and tumour progression, was investigated. For this purpose, the results were compared with cases of inflammatory fibrous hyperplasia (IFH) and normal oral mucosa. Thirty cases of GCF, 30 cases of IFH and 10 normal mucosa specimens used as control were selected. Immunoreactivity of mast cells to the anti-tryptase antibody was analysed quantitatively in the lining epithelium and in connective tissue. In the epithelial component (p = 0.250) and connective tissue (p = 0.001), the largest mean number of mast cells was observed in IFHs and the smallest mean number in GCFs. In connective tissue, the mean percentage of degranulated mast cells was higher in GCFs than in IFHs and normal mucosa specimens (p < 0.001). Analysis of the percentage of degranulated mast cells in areas of fibrosis and at the periphery of blood vessels also showed a larger mean number in GCFs compared to IFHs and normal mucosa specimens (p < 0.001). The percent interaction between mast cells and stellate giant cells in GCFs was 59.62%. In conclusion, although mast cells were less numerous in GCFs, the cells exhibited a significant interaction with stellate giant cells present in these tumours. In addition, the results suggest the involvement of mast cells in the induction of fibrosis and modulation of endothelial cell function in GCFs.     

PCNA and Ki-67 immunoreactivity in multinucleated cells of giant cell fibroma and peripheral giant cell granuloma

Immunohistochemical investigation of PCNA and Ki-67, two diverse nuclear proteins essential to the cell cycle, was undertaken in archival, formalin-fixed and paraffin-embedded specimens of giant cell fibroma (GCF) and peripheral giant cell granuloma (PGCG). GCF multinucleated cell nuclei were mostly PCNA⁺, although there was variability in staining intensity. This indicates heterogeneity in nuclear PCNA metabolism of GCF multinucleated cells, and it is possible that the most intensely stained nuclei have passed through the cell cycle more recently compared to the less immunoreactive nuclei. However, the absence of Ki-67 immunoreactivity in GCF multinucleated cells, and absence of mitoses in GCF multinucleated cells, suggests that cell cycling in the absence of cytokinesis is not involved in GCF multinucleated cell formation. Alternatively, GCF multinucleated cells possibly form by fusion of mononuclear cells previously identified as fibroblasts, although this theory cannot be confirmed by the data presented in this study, and the histogenesis of GCF multinucleated cells remains unclear. In contrast, absence of either PCNA or Ki-67 immunoreactivity in PGCG multinucleated cells is consistent with an osteoclast lineage and formation from differentiated mononuclear cells.     

Collagenous fibroma (desmoplastic fibroblastoma) of the palate: a case report

Collagenous fibroma is an uncommon benign soft tissue lesion that has a wide anatomic distribution. We describe a case of a collagenous fibroma that appeared in the left soft and hard palate of a 37-year-old woman as a 5.0-cm solitary, firm nodule. Microscopically, it was composed of stellate or spindle-shaped cells embedded in hypovascular fibrous stroma. Entrapment of fat was focally identified at the edges. Mitotic figures and tumor necrosis were absent. Tumor cells were immunopositive for vimentin, and a few cells were positive for alpha-smooth muscle actin. Tumor extracellular matrix was immunopositive for type I and type III collagen, as well as for fibronectin. These findings satisfied the diagnostic criteria for collagenous fibroma (desmoplastic fibroblastoma). This case, to our knowledge, represents the first report of this tumor in the mouth. The differential diagnosis of fibrous lesions of the mouth is discussed.     

Expression of epithelial and nervous-system-related antigens on pleomorphic adenoma of the salivary glands--an analysis of 20 cases

The expression of cytokeratin (40-52 kD), carcinoembryonic antigen (CEA), vimentin, neuron-specific enolase (NSE), S-100 protein, glial fibrillary acidic protein (GFAP) were investigated in 20 cases of pleomorphic adenoma of the salivary glands and 10 cases of normal salivary glands, in order to analyze and correlate the antigens' expressions with the probably histogenetic mechanisms of the various histopathological differentiations in pleomorphic adenoma of the salivary glands and their probably original cells in normal salivary glands. Immunohistochemistry has provided some evidence for the relationship of the tumor cells to normal salivary glands: In the normal glands, the acinic cells exhibited cytokeratin, CEA and focal, predominantly nuclear S-100 protein staining. In both normal glands and pleomorphic adenomas, the duct-lining cells were immunoreactive for cytokeratin, CEA and had both cytoplasmic and nuclear S-100 positivity; The myoepithelial cells of the normal glands as well as the periduct cells, epithelial nests/cords, squamous metaplasia and the stellate/spindle/cartilaginous cells in the myxomatous-chondroid areas of the pleomorphic adenoma contain immunoreactive vimentin, NSE, S-100 proteins and GFAP, and lesser amounts cytokeratin (40-52 kD)/CEA. The varicosities of the terminal axon may lie directly on the basal membrane, or penetrate the basal membrane and lie in direct contact with the effector cells (duct-acinar-myoepithelial cells) of the salivary glands. The peripheral neurons and axons of the autonomic nervous system were identified by vimentin, NSE, S-100 proteins and GFAP. The combination of epithelial cytokeratin and nervous system-related vimentin, NSE, S-100 and GFAP immunostaining in myoepithelium of the normal glands and in all component elements (particularly the periduct cells) of pleomorphic adenoma reflects pleomorphic adenoma of the salivary glands is an epithelial tumor, the probably original cells or the probably histogenetic mechanisms of the various histopathological differentiations is correlated not only with "duct-acinar-myoepithelial cells" but also with the neuroectoderm in the normal salivary glands."     

Crevicular fluid and serum IgG subclasses and corresponding mRNA expressing plasma cells in periodontitis lesions

Recent reports suggest that specific serum IgG subclasses are a feature of several forms of periodontitis. GCF antibodies are both serum-derived and locally produced by the abundant plasma cells of the diseased periodontal tissue. Previous work has shown that crevicular fluid (GCF) levels of IgG may be reduced in active and deep periodontal pockets when compared to other sites in chronic periodontitis patients (7). These findings, and more recent findings for IgA levels in GCF (5), suggest that GCF immunoglobulins may indicate "high risk" sites for periodontitis. In these studies, the relative distribution of IgG isotypes was not investigated, nor was the relative contribution of local and serum antibodies to the GCF immunoglobulin profile. Therefore, more precise investigation of the tissue distribution of local gingival IgG subclass producing plasma cells and their protein levels in the GCF from the same sites and in serum, was undertaken.     

High levels of soluble intercellular adhesion molecule-1 (ICAM-1) in crevicular fluid of periodontitis patients with plaque

Abstract. The intercellular adhesion tnolecule-1 (ICAM-1) is a membrane-bound molecule involved in cell-cell adhesive interactions which is upregulated on inflammatory epithelial cells. The levels of soluble ICAM-1 (sICAM-1) shed into the gingival crevicular fluid (GCF) were studied in healthy patients and patients with gingivitis, adult periodontitis or rapidly progressive periodontitis, using an ELISA technique. Clinical parameters including plaque index, gingival index, probing depth, and bleeding on probing were recorded following careful sampling of GCF with standardised filter strips. In GCF, sICAM-1 levels were higher for patients with plaque ( p =0.04) and for patients with inflammation ( p =0.02), but did not correlate with disease classifications. These results suggest that elevated GCF sICAM-1 levels may represent increased shedding of this molecule in the interstitial fluid as a result of membrane-bound ICAM-1 upregulation on ICAM-1 gingival-bearing cells in relation with plaque accumulation and inflammation.     

Osteogenic activity of cells from dental pulp, periodontal ligament, bone marrow and muscle in vitro: an ultrastructural study and alkaline-phosphatase activity.

Dental pulp, periodontal ligament, bone marrow and muscle tissue from the same rat were cultured in vitro in order to investigate their osteogenic activity by transmission electron microscope. Immunohistochemical methods with various antibodies were utilized and alkaline-phosphatase (ALPase) activities of these cells were also measured biochemically. Dental pulp cells were stellate in shape, showed an intense ALPase reaction, and had lipid-like droplets. Periodontal ligament cells were composed of spindle fibroblasts and epithelial cells. The former revealed a positive reaction for ALPase and possessed microfilaments. Bone marrow cells were spindle shaped, resembling fibroblasts, but some of them were similar to osteoblasts. Muscle cells were long, slender in shape, and showed no positive reaction for ALPase. The cells from pulp tissue showed the highest activity of ALPase, followed by periodontal ligament and bone marrow; there was no activity in muscle tissue. All the cells except the epithelial-like cells of the periodontal ligament and muscle cells were positive in reaction with ALPase which is a marker for osteogenic cells, and vimentin which is a marker of fibroblastic characteristics. Osteogenic activity and cellular differentiation of these cells were discussed.     

Immunohistochemical study in pleomorphic adenomas of human parotid gland--with special reference to cytoskeletal filamentous proteins

The purpose of this immunohistochemical study was to know the degree and direction of differentiation, the mechanism of mesenchymal appearance, and histogenesis in pleomorphic adenomas which were obtained from surgical removed specimens. All specimens were fixed in 10% neutral phosphoate buffered formalin, and embedded in paraffin. Sections were steined with the avidin-biotin peroxidase complex method for keratin, vimentin, desmin and S-100 protein, the peroxidase-antiperoxidase method for lysozyme, and the direct immunoperoxidase method for IgA and secretory component. Normal salivary glands were employed as controls. 1. Solid portion of pleomorphic adenomas were consisted of the focal hypline cells and scattered the duct cell-like cells. The hyaline cells were positive for vimentin and S-100 protein, and the duct cell-like cells were positive for keratin. But one case was found the bundles of spindle-shaped cells which was similar to myoepithelial cells were positive for actin and another case was consisted of the focal duct cell-like cells were observed. 2. In the tubular portion, the duct-like structures were classified into three types. One type was mono-layered structure, another type was double-layered structure and the other type is duct-like structure in the solid portion. Any inner layer of the duct-like structure were positive for keratin. 3. Double-layered duct-like structures were most common type. Among the double-layered structures, hyaline cells mostly made up the outer layered structures. At times, the spindleshaped cells of the outer layer were found. This structures were similar to intercalated portion of normal salivary glands. In one case, inner and outer layer was consisted of keratin positive cells, but outer layer was steined for keratin stronger than inner. This structure was similar to excretory duct of normal salivary glands. 4. IgA and sc were seen at inner cells and in luminal accumulation of the duct-like structures. 5. In the myxoid regions, stellate cells or spindle cells were positive for vimentin, S-100 protein and partial positive for desmin. In the chondroid regions, chondrocyte-like cells were positive for vimentin and S-100 protein. These results seemed to suggest that pleomorphic adenomas were composed of duct cell-like cells and hyaline cells. Duct cell-like cells consisted of the duct-like structure in order to transport secretion which was similar to the duct of normal salivary glands.(ABSTRACT TRUNCATED AT 250 WORDS)     

Expression of interleukin-2 receptor and HLA-DR on lymphocyte subsets of gingival crevicular fluid in patients with periodontitis

Expression of interleukin-2 receptor (IL2R) and HLA-DR on lymphocytes of gingival crevicular fluid (GCF) was examined by two-color flow cytometric analysis. GCF from 15 patients with periodontitis was collected by crevicular washing. Mononuclear cells were isolated by Ficoll-paque gradient centrifugation from inflamed gingival tissue (GT) and peripheral blood (PB) sampled from each of the 15 patients. Lymphocyte subsets were detected by using monoclonal antibodies (mAb) of Leu 12 (CD19), Leu 4 (CD3), Leu 3a (CD4) and Leu 2a (CD8) directed to B cells, T cells, helper/inducer T cells (Th) and suppressor/cytotoxic T cells (Ts), respectively. Anti-IL2R (CD25) and anti-HLA-DR were used as lymphocyte activation markers. IL2R- or HLA-DR-positive fractions in Th, Ts and B cells were calculated. Percentage of IL2R-positive fraction in Th (IL2R+ Th) of GCF (34.0%) was significantly higher than those of GT (18.4%) and PB (13.7%). IL2R-positive fraction in B cells (IL2R+ B) of GCF was the highest among the three groups (23.9% in GCF, 12.5% in GT, 6.3% in PB). Ts did not express IL2R regardless of the origin of the samples. Compared with PB and GT, GCF showed significantly higher HLA-DR expression on Th and Ts in GCF (PB: 8.7% and 27.1%; GT: 27.9% and 50.3%; GCF: 44.7% and 65.3%). These results suggest that lymphocytes in GCF were highly activated and are related to the local host immune response in periodontitis.     

Production of prostaglandin E2 by polymorphonuclear neutrophils isolated from gingival crevicular fluid and peripheral blood of dogs in periodontal health and disease

We examined PGE2 synthesis using inflamed and non-inflamed GCF PMNs and PB PMNs in the presence and absence of certain stimulators. The basal levels of PGE2 release from GCF PMNs isolated from ligature-induced gingival sulcus with a GI value over 2.2 were significantly lower than those from GCF PMNs isolated from sham operated sites with a GI value below 2.1. Levels were comparable to those from PB PMNs isolated at each experimental period, indicating that the amount of PGE2 synthesized by GCF PMNs is not correlated exactly with the severity of periodontitis. Calcium ionophore A23187 stimulated PGE2 synthesis by all PMN preparations. When compared to those with inflamed and non-inflamed GCF PMNs, stimulation was higher with PB PMNs. However, the chemotactic factor fMLP inhibited the synthesis by inflamed and non-inflamed GCF PMNs. PGE2 synthesis by PB PMNs isolated after periodontal operation was stimulated by the drug, but those cells isolated before the operation did not respond.     

Osteopontin in gingival crevicular fluid

Osteopontin (OPN) is a major glycosylated phosphoprotein in bone matrix and is produced by several cells including osteoblasts, osteoclasts and macrophages. OPN levels increase in active sites of bone metabolism. Recently, several bone-related proteins were identified in gingival crevicular fluid (GCF) to seek markers of alveolar bone resorption in periodontal disease. In this study, we investigated the existence of OPN in GCF and the correlation between OPN level in GCF and probing depth (PD) of sampling sites in 98 periodontitis patients and 35 healthy subjects. An immunoblotting analysis using 10% polyacrylamide gel showed that two forms of OPN with molecular masses of 54 and 66 kDa and several degraded fragments were detected in most GCF samples from diseased sites (PD > 4 mm). In GCF samples from healthy sites (PD < or = 3 mm), only one form (54 kDa) was observed, but any degraded fragments were not detected. When OPN amounts in GCF samples were determined by ELISA, a weak. but significant correlation was observed between OPN amount in GCF and PD (r=0.32, p=0.0013). These results demonstrate that OPN exists in GCF and that OPN level in GCF increases with the progression of periodontal disease.     

Presence of activated eosinophils, high IgE and sCD23 titers in gingival crevicular fluid of patients with adult periodontitis

Our previous studies showed that the expression of CD23 on polymorphonuclear leukocytes (PMNLs) in gingival crevicular fluid (GCF) from adult periodontitis (AP) patients was higher than in autologous peripheral blood (PB). Percentages of eosinophils in GCF PMNLs ranged between 6 and 14%. The purpose of the present studies was to increase understanding of the potential role of eosinophils and their products, including CD23, in periodontal disease. We analysed the eosinophil fraction in GCF and PB by flow cytometry using monoclonal antibodies to CD23b (BB10), eosinophil cationic protein (ECP) in stored and secretory forms (EG1 and EG2), and CD67 (80H3). Simultaneously, we measured IgE and soluble CD23 titer and GCF and serum by ELISA. Flow cytometric analysis of BB10, EG2 and 80H3 binding showed that GCF eosinophils from AP were activated. A large BB10⁺ EG2⁺ cellular fraction was detected in GCF from AP whereas it was very low in autologous serum (9.30±2.460 vs 0.16±0.10, p>0.001). GCF from gingivitis patients exhibited no flow cytometric evidence for the presence of BB10+ EG2+ cells. BB10+ EG1+ cells, or inactivated eosinophils rated lower in GCF than in PB both in gingivitis and periodontitis patients (0.45±0.63 vs 1.83±0.96 and 0.15±0.30 vs 1.30±0.20, p>0.05. respectively). IgE titer in AP patients reached 1208.1 ±421.2 IU/ml in GCF while only 49.1 ±50.4 in sera. Soluble CD23 in GCF reached 236.1 ±81.3 ng/ml in GCF and 5.6±1.8 ng/ml in sera. GCF of gingivitis patients, however, contained no detectable sCD23. Thus. GCF from AP patients displayed a high rate of activated eosinophils secreting ECP. while GCF of gingivitis patients did not. These results suggest that ECP-secreting activated eosinophils are relevant to the pathology of adult periodontitis.     

Investigations into the cellular contribution to host tissue proteases and inhibitors in gingival crevicular fluid

Abstract Gingival crevicular fluid (GCF) was collected from chronic periodontitis patients using plastic micropipettes and coverslip smears stained with antibodies for leukocyte markers and Toluidine Blue for mast cells. The smears consisted of 70–80% granulocytes, 10–20% monocytes/macrophages. 5% mast cells and 5% T lymphocytes: no B lymphocytes were found. Proteases and inhibitors in GCF cells were investigated by enzyme cytochemistry using 2-methoxy-4-naph-thylamine-linked peptide substrates and simultaneous coupling to Fast Blue B and immunocytochemistry using biotinylated secondary antibodies and an alkaline phosphatase/new fuchsin detecting system. Elastase was detected in granulocytes, cathepsin B in macrophages. dipeptidyl peptidases II and IV in a small proportion of macrophages, dipeptidyl peptidase IV in a few T lymphocytes, tryptase in mast cells and α-1-proteinase inhibitor and α-2-macroglobulin in some macrophages. GCF was also collected on filter paper strips and eluted into buffer for biochemical enzyme assays. Lysis of cells by addition of detergent to the elution buffer increased activities to 140–240% of control values. Removal of cells by centrifugation reduced measured activities to 1–30% of original figures; this effect was less if samples were pre-treated with detergent. Proteases from inflammatory cells therefore appear to make up most of the measured enzyme activity in GCF, and this association may explain recent correlations with periodontal disease progression.     

Gingival crevicular fluid monocyte chemoattractant protein-1 and RANTES levels in patients with generalized aggressive periodontitis

BACKGROUND: Local and systemic inflammatory and immune mechanisms may be implicated in the pathogenesis of the aggressive forms of periodontal disease. Chemokines, monocyte chemoattractant protein-1 (MCP-1) and regulated on activation, normal T cells expressed and secreted RANTES (regulated on activation, normal T cells expressed and secreted), are involved in the activation and recruitment of inflammatory and immune cells to the infected sites and thereby mediating a variety of pathophysiological conditions. The aim of the present study was to examine the gingival crevicular fluid (GCF) levels of MCP-1 and RANTES in patients with generalized agressive periodontitis (G-AgP). METHODS: MCP-1 and RANTES levels were investigated in GCF samples of 10 patients with G-AgP and 10 periodontally healthy subjects. Periodontal status was evaluated by measuring probing depth, clinical attachment loss, presence of bleeding on probing and plaque. In the G-AgP group, GCF samples were collected from the two approximal sites; from one single-rooted tooth and from one first molar tooth with > or =6 mm probing depth. In the healthy group, GCF samples were collected from one of the single-rooted teeth. GCF MCP-1 and RANTES levels were quantified by enzyme immunoassay. RESULTS: The G-AgP patients had significantly higher GCF MCP-1 and RANTES levels compared to the healthy group (p<0.05). GCF MCP-1 and RANTES levels were positively correlated with both probing depth and clinical attachment loss (p<0.05). There was no correlation between GCF MCP-1 and RANTES levels and the percentage of sites with bleeding (p>0.05). CONCLUSIONS: The results of the present study suggest that MCP-1 and RANTES could play key roles in both activation and recruitment of inflammatory and immune cells in periodontal environment of G-AgP patients. In conclusion, these CC chemokines may be considered in the biological mechanism underlying the pathogenesis and progression of G-AgP.     

Experimental tooth pain elevates substance P and matrix metalloproteinase-8 levels in human gingival crevice fluid

Objective. Tooth pain can induce a neurogenic inflammatory reaction in gingiva in association with local elevations of matrix metalloproteinase (MMP)-8, which is considered the major tissue destructive protease in gingival crevice fluid (GCF). The pro-inflammatory neuropeptides released by sensory nerves coordinate the activities of the immuno-effector cells and may influence the secretion of MMP-8. With this background, we studied whether experimental tooth pain can trigger changes in GCF levels of the neuropeptide substance P (SP) and MMP-8. Material and methods. The GCF SP levels of stimulated and non-stimulated teeth were analyzed for SP using a competitive enzyme immunoassay (EIA). The GCF MMP-8 levels were determined by quantitative immunofluorometric assay (IFMA). Results. Painful stimulation of the upper central incisor caused significant elevations in GCF SP and MMP-8 levels of the stimulated tooth. At the same time, the GCF SP and MMP-8 levels of non-stimulated control teeth were unchanged. Conclusions. These data indicate that experimental tooth pain can induce local elevations of SP and MMP-8 levels in GCF simultaneously. This supports the possibility of a local neurogenic spread of inflammatory reactions from intrapulpal to surrounding periodontal tissues.