Nitric oxide synthase type-II is synthesized by human gingival tissue and cultured human gingival fibroblasts

Nitric oxide is known to be an important inflammatory mediator, and is implicated in the pathophysiology of a range of inflammatory disorders. The aim of this study was to determine the localization and distribution of endothelial NOS (NOS-II) in human gingival tissue, and to ascertain if human gingival fibroblasts express NOS-II when stimulated with interferon gamma (IFN-gamma) and bacterial lipopolysaccharide (LPS). The distribution of NOS-II in inflamed and non-inflamed specimens of human gingivae was studied using a monoclonal antibody against nitric oxide synthase II. Cultures of fibroblasts derived from healthy human gingivae were used for the cell culture experiments. The results from immunohistochemical staining of the tissues indicated an upregulation of NOS-II expression in inflamed compared to non-inflamed gingival tissue. Fibroblasts and inflammatory cells within the inflamed connective tissue were positively stained for NOS-II. In addition, basal keratinocytes also stained strongly for NOS-II, in both healthy and inflamed tissue sections. When cultured human gingival fibroblasts were stimulated by INF-gamma and Porphyromonas gingivalis LPS, NOS-II was more strongly expressed than when the cells were exposed to LPS or IFN-gamma alone. These data suggest that, as for other inflammatory diseases, NO plays a role in the pathophysiology of periodontitis.

Immunocytochemical Localization of Substance P Receptors (NK-1 Receptors) in Human Gingival Tissue

Substance P (SP) is a member of the structurally related family of neuropeptides known as the tachykinins. In addition to neurotransmitter roles, the tachykinins are also known to modulate local inflammation which depends on signalling between the neuropeptide molecules and target cells and tissues. SP mediates its effects through a specific receptor, known as the substance P receptor or the neurokinin 1 (NK-1) receptor. The NK-1 receptor is a G-protein associated integral membrane protein and although it has been studied in a wide range of tissues, to date there has been no published data on the localisation of the NK-1 receptor in human gingival tissue. Objective: The aim of this study was to examine the distribution of the NK-1 receptor in human gingival tissue using immunocytochemistry. Method: Gingival tissue was obtained from patients undergoing periodontal surgery. Tissue was fixed in paraformaldehyde and embedded in wax for sectioning. Sections were dewaxed in xylene and then rehydrated in alcohols and phosphate buffered saline. Rehydrated sections were probed with rabbit polyclonal antibody to human NK-1 receptor which was subsequently detected using anti-rabbit horseradish peroxidase conjugate and diaminobenzidine as substrate. Results: Immunocytochemistry revealed that the NK-1 receptor was distributed along nerve fibres and blood vessel endothelial cells, suggesting these areas are main targets for the actions of SP via the NK-1 receptor. Conclusion: This is the first immunocytochemical report of NK-1 receptors in human gingival tissue and provides evidence for possible NK-1 mediated biological effects of SP in human gingival tissue from periodontitis patients.

Characterization of a Local Renin-Angiotensin System in Rat Gingival Tissue

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nicotine-Induced Damage Affects Gingival Fibroblasts in the Gingival Tissue of Rats

Background: Very limited information is available from in vivo studies about whether smoking and/or nicotine affect gingival tissues in the absence of plaque. The purpose of this study is to evaluate the effect of the systemic administration of nicotine in the proliferation and counting of fibroblast-like cells in the gingival tissue of rats.Methods: Thirty adult male Wistar rats were randomly assigned into two groups to receive subcutaneous injections of a saline solution (control group = group C) or nicotine solution (group N; 3 mg/kg) twice a day. The animals were euthanized 37, 44, or 51 days after the first subcutaneous injection. Specimens were routinely processed for serial histologic sections. Five fields of view in the connective tissue adjacent to the gingival epithelium and above the alveolar bone crest of the maxillary first molar were selected for the counting of fibroblast-like cells. Data were statistically analyzed (P<0.05).Results: The intergroup analysis detected a lower number of fibroblast-like cells in group N compared to group C on days 37 (2.65 +/- 1.41 and 6.67 +/- 3.25, respectively), 44 (2.70 +/- 1.84 and 8.57 +/- 2.37, respectively), and 51(2.09 +/- 1.41 and 7.49 +/- 2.60, respectively) (P<0.05). The quantification of fibroblast-like cells showed no significant difference (P >0.05) in the intragroup analysis of control and nicotine throughout experimental periods. In the intergroup analysis, group N had reduced proliferating cell nuclear antigen positive fibroblasts compared to group C in all periods (P<0.05).Conclusion: The daily systemic administration of nicotine negatively affected, in vivo, the number and proliferation of fibroblast-like cells in the gingival tissue of rats. J Periodontol 2011;82:1206-1211.

Reaction of the human gingival tissue to different suture materials used in periodontal surgery.

The gingival reaction to 4 different suture materials used in periodontal surgery was studied in 36 patients. The gingiva was sutured prior to surgery and biopsies were taken at 3, 7 and 14 days to observe the tissue reaction. The histological examination showed that silk caused the most intense and longest inflammatory response. Polyester and perlon provoked shorter, less intense tissue reactions than silk, and nylon caused the least inflammatory response, with earlier tissue repair.

Drug induced gingival overgrowth: A role for Protease Activated Receptors?

Background: Protease activated receptors (PAR) belong to a subfamily of G protein coupled receptors. They consist of seven transmembrane domains but are not classical receptors as their agonist is a circulating serine proteinase. This proteinase cleaves an N-terminal extracellular domain of the receptor to reveal a new N-terminal tethered ligand which binds intramolecularly, thus converting an extracellular proteolytic event into a transmembrane signal. Therefore, the cleavage and activation of PARs provide a mechanism whereby proteinases can directly influence the inflammatory response. Gingival hyperplasia or gingival enlargement is a side effect of some drugs such as cyclosporine, a potent immunosuppressant. To date, the potential role of PAR in the inflammation associated with the pathogenesis of gingival overgrowth has not been studied. Objectives: The present study was designed to determine whether proteinases derived from extracts of cyclosporine induced hyperplasia were capable of activating PAR in vitro. Methods: Cell lysates were derived from tissue obtained from gingival overgrowth of patients requiring surgical excision. Cell lines over-expressing PARs were maintained in Dulbecco's modified Eagle's medium (DMEM), containing 10% foetal calf serum (FCS) in 5% CO2. The cells were treated with gingival overgrowth lysates and agonist stimulated calcium release from the cells was recorded using the Fluo-4-Direct™ Calcium Assay Kit from Invitrogen, according to manufacturer's instructions. Results: Calcium release by activated PAR on tumour cells was detected in those treated with gingival hyperplasia lysates. Samples from healthy gingival fibroblasts did not elicit this response. Conclusions: The identification of mediators of the molecular events central to the inflammatory phenotype elicited by gingival hyperplasia is important. To this end, our experiments show that in vitro, enzymes derived from overgrown gingival tissue are capable of activating PAR and thereby provide evidence for the potential role of PAR in sustaining gingival hyperplasia.

Acellular Dermal Matrix Graft for Gingival Augmentation: A Preliminary Clinical, Histologic, and Ultrastructural Evaluation

Background: The aim of this study was to evaluate clinically, histologically, and ultrastructurally the integration process of the acellular dermal matrix used to increase the band of keratinized tissue while achieving gingival inflammation control. Methods: Ten patients exhibiting a mucogingival problem with bands of keratinized tissue <= 1 mm and gingival inflammation of the related teeth were included in the study. The surgical procedures were performed to augment the gingival tissue using acellular dermal matrix. Clinical measurements were assessed at baseline and after 3 months. A specimen of the allograft and surrounding tissues was obtained immediately before the surgery and 4 minutes and 1, 2, 3, 4, 6, and 10 weeks after grafting. Results: Clinically, a gain of keratinized tissue of 2.92 +/- 0.65 mm was observed after 3 months. Histologically and ultrastructurally, many macrophages were observed phagocytosing preexisting collagen fibers in the first weeks. From week 2 on, fibroblasts synthesizing new collagen, epithelial cells colonizing the graft surface, and revascularization were noticed. After 6 weeks it was difficult to find the acellular dermal matrix preexisting collagen fibers. This process of substitution was completed after 10 weeks, when the reepithelialization of the entire graft throughout a well-structured basement membrane was achieved. Conclusion: The acellular dermal matrix graft seemed to be an easily handled material for use in keratinized tissue augmentation that, in humans, was substituted and completely reepithelialized in 10 weeks according to histologic and ultrastructural results. J Periodontol 2009;80:253-259.

Gingival primary extranodal non-Hodgkin's lymphoma as the first manifestation of acquired immunodeficiency syndrome

Background: Non-Hodgkin's lymphomas (NHLs) are a heterogeneous group of lymphoproliferative malignancies that may be associated with acquired immunodeficiency syndrome (AIDS). NHL can disseminate to extranodal sites; however, its dissemination to the jaws and mouth is not common. This report presents and discusses two unusual cases of gingival primary extranodal non-Hodgkin's lymphoma (PE-NHL) as the first manifestation of AIDS.Methods: Two mates presented with asymptomatic gingival swelling. They were examined clinically. Biopsies of the gingival tissue were evaluated using routine histologic techniques and immunohistochemistry. The patients were tested for human immunodeficiency virus (HIV) infection.Results: The clinicopathological evaluation and the serological HIV examination of the patients led us to the final diagnosis of gingival PE-NHL as the first manifestation of AIDS. Both patients were referred to an oncologist and to an infectious disease specialist and were given antineoplastic chemotherapy and highly active antiretroviral therapy. Only one patient presented a favorable clinical evolution.Conclusion: The present case reports have important clinical implications; the two unusual presentations of gingival PE-NHL contribute to information about the differential diagnosis of rapidly progressing gingival swelling.

A clinical trial of the effect of a mouth-rinse prepared with Lippia sidoides Cham essential oil in dogs with mild gingival disease

We evaluated the effect of a mouth-rinse prepared using Lippia sidoides essential oil (EO) in dogs with marginal gingivitis. German Shepherd dogs were distributed in two groups: control (control mouth-rinse) and EO (EO mouth-rinse). Both mouth-rinses were applied on the dogs' teeth every 2 days for 2 weeks. At day 0 and day 15, the scores for plaque-bacteria (P), calculus (C), gingivitis (G) and the inflammatory infiltrate (INF) were evaluated blindly. The results were analyzed by the Wilcoxon signed-rank and Mann-Whitney tests (P less than or equal to 0.05). P, C, G, and INF did not show any alteration in the control group, while in the EO group there were significant reductions in these scores. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Morphological evaluation of combined effects of cyclosporin and nifedipine on gingival overgrowth in rats

It has previously been shown that, while cyclosporin A (CsA) and nifedipine both cause gingival overgrowth in the rat. the combined use of these drugs increases the severity of overgrowth. The aim of this study was to describe the histometry and densities of fibroblasts, collagen fibers and vessels in the gingival tissue of rats that were treated with CsA and nifedipine, either alone or in combination. Rats were treated for 60 days with a daily subcutaneous injection of 10 mg/kg body weight of CsA and/or with 50 mg/kg body weight of nifedipine added to the chow. The results confirmed that CsA causes a more severe overgrowth than nifedipine, and that the combined use of these drugs increases the overgrowth severity. All the rat groups that were studied showed that, as the severity of overgrowth increased, there was a parallel increase in fibroblasts and collagen, and a decrease in vessel content. Therefore, independently of whether the gingival overgrowth was caused by CsA alone, nifedipine alone, or both treatments in combination, the fibroblast and collagen density increased in parallel with the severity of the overgrowth. © Blackwell Munksgaard, 2002.

Drug-induced gingival overgrowth: A clinical and histopathological report

Gingival overgrowth is a significant problem faced by periodontists and is particularly associated with the use of certain drugs such as nifedipine, a high-specificity calcium channel blocker used for the treatment and prophylaxis of certain cardiovascular diseases. Development of gingival overgrowth is characterized by increased collagen in gingival tissue. In general is asymptomatic, at times associated with spontaneous bleeding and ulceration and can promote aesthetic changes and compromise hygiene habits and mastication of the patient. The severity of the symptoms is associated with the presence of risk factors such association with other drugs. This paper aims to present a case report of a patient with generalized gingival overgrowth, with more severe characteristics in the anterior mandible induced by chronic use of nifedipine who underwent basic non-surgical periodontal treatment including supra and subgingival scaling and root planning in both jaws associated with rigorous oral hygiene instructions and surgical therapy in the anterior mandible, the most affected area, to remove the excess of gingival tissue. Nifedipine was replaced by the patient cardiologist to propanolol hydrochloride (40 mg/kg) in an attempt to minimize unwanted side effects. After 6 month follow-up, no recurrence was observed, oral hygiene had improved and the patient had clinical periodontal health and esthetic satisfaction.

Forced orthodontic eruption for augmentation of soft and hard tissue prior to implant placement

Forced orthodontic eruption (FOE) is a non-surgical treatment option that allows modifying the osseous and gingival topography. The aim of this article is to present a clinical case of a FOE, which resulted in an improvement of the amount of available bone and soft-tissues for implant site development. Patient was referred for treatment of mobility and unesthetic appearance of their maxillary incisors. Clinical and radiographic examination revealed inflamed gingival tissue, horizontal and vertical tooth mobility and interproximal angular bone defects. It was chosen a multidisciplinary treatment approach using FOE, tooth extraction, and immediate implant placement to achieve better esthetic results. The use of FOE, in periodontally compromised teeth, promoted the formation of a new bone and soft-tissue in a coronal direction, without additional surgical procedures, enabling an esthetic, and functional implant-supported restoration.

An alternative maneuver to treat gingival recession

This article reports a clinical case in which was applied autologous bone graft associated with subepithelial connective tis- sue graft, harvested by gingivectomy procedure with technical modifications to increase gingival graft extension, also to be used as guided tissue regeneration, to treat a single gingival recession. After 1 year and 2 months of follow-up, the cover- age of the recession was 4.0 mm, which corresponded to the gain of attached keratinized gingival tissue. An increase in the gingival tissue thickness was observed, without significant probing depth. The procedures applied to treat this case may be biologically and clinically useful to treat gingival recession.

Signaling transduction analysis in gingival epithelial cells after infection with Aggregatibacter actinomycetemcomitans

Periodontal diseases result from the interaction of bacterial pathogens with the hosts gingival tissue. Gingival epithelial cells are constantly challenged by microbial cells and respond by altering their transcription profiles, inducing the production of inflammatory mediators. Different transcription profiles are induced by oral bacteria and little is known about how the gingival epithelium responds after interaction with the periodontopathogenic organism Aggregatibacter actinomycetemcomitans. In the present study, we examined the transcription of genes involved in signaling transduction pathways in gingival epithelial cells exposed to viable A.actinomycetemcomitans. Immortalized gingival epithelial cells (OBA-9) were infected with A.actinomycetemcomitans JP2 for 24 h and the transcription profile of genes encoding human signal transduction pathways was determined. Functional analysis of inflammatory mediators positively transcribed was performed by ELISA in culture supernatant and in gingival tissues. Fifteen of 84 genes on the array were over-expressed (P < 0.01) after 24 h of infection with viable A.actinomycetemcomitans. Over-expressed genes included those implicated in tissue remodeling and bone resorption, such as CSF2, genes encoding components of the LDL pathway, nuclear factor-?B-dependent genes and other cytokines. The ELISA data confirmed that granulocytemacrophage colony-stimulating factor/colony-stimulating factor 2, tumor necrosis factor-a and intercellular adhesion molecule-1 were highly expressed by infected gingival cells when compared with control non-infected cells, and presented higher concentrations in tissues from patients with aggressive and chronic periodontitis than in tissues from healthy controls. The induction in epithelial cells of factors such as the pro-inflammatory cytokine CSF2, which is involved in osteoclastogenesis, may help to explain the outcomes of A.actinomycetemcomitans infection.