Inflamed Gingiva Research Articles

Management of peri-implant soft tissue defect with connective tissue graft - A case report

The use of dental implants to replace missing and nonrestorable teeth has increased significantly in the past two decades. The health of peri-implant tissues plays an important role in the long-term success of dental implants. Soft tissue dehiscence around dental implants has frequently been reported which may lead to poor oral hygiene, plaque accumulation, inflamed gingiva, compromised esthetics, and further progression to peri-implantitis. The coverage of exposed implants is aimed to provide adequate keratinized tissue around dental implant. Complete coverage of mucosal defect around the dental implants is still a challenging prospect. This case report describes correction of soft tissue defect and building up of keratinized tissue in relation to implant in the maxillary right first premolar region with subepithelial connective tissue graft along with coronally advanced flap.

Expression of matrix metalloproteinases-2, -9 and reversion-inducing cysteine-rich protein with Kazal motifs in gingiva in periodontal health and disease

Background and objectivePeriodontitis is characterized by pathological destruction of extracellular matrix (ECM) of periodontal tissues. Matrix metalloproteinases (MMPs) promote the occurrence and development of periodontitis by degrading almost all proteins of ECM. RECK (reversion-inducing-cysteine-rich protein with kazal motifs), a novel membrane-anchored inhibitor of MMPs, could regulate the expression of MMP-2 and MMP-9 at post-transcriptional level. The study was to investigate the expression of RECK in healthy and diseased human gingival tissues and to correlate it with the production of MMP-2 and MMP-9. Material and methodsGingival biopsies were collected from chronic periodontitis patients and periodontally healthy control individuals. The protein and mRNA of RECK, MMP-2 and MMP-9 was determined by immunohistochemistry and semi-quantitative polymerase chain reaction analysis. ResultsThe expression of RECK protein was mainly confined to the gingival epithelium in inflamed and non-inflamed gingival tissues. Expression of RECK was significantly lower in tissues from chronic periodontitis patients, while the positive expression levels of MMP-2 and MMP-9 in periodontitis specimens were significantly higher. RECK protein expression was negatively correlated to the expressions of MMP-2 and MMP-9 in periodontitis. Moreover, RECK mRNA was significanly lower in diseased gingiva than in healthy samples(P<0.05), while MMP-2 and MMP-9 mRNAs were observed overexpressed in periodontal lesions, with no significant correlation between RECK and MMP-2/MMP-9 mRNA shown in periodontally diseased group. ConclusionThe expression of RECK in human healthy and diseased gingiva may contribute to periodontal physiological and pathological processes; low RECK expression may be associated with the enhanced MMP-2 and MMP-9 production in inflamed gingiva.

DIFFERENTIAL HISTOMORPHOMETRIC CHANGES IN NORMAL AND INFLAMED GINGIVAL EPITHELIUM

Introduction and aim: In recent decades, many factors such as smoking, unhealthy diet as well as high alcohol intake were marked as risk factors that can lead to increased incidence of malignant alterations, gingivitis, periodontal disease and other oral epithelium pathological changes. Having in mind that in the group of non-malignant and non-dental oral pathology gingivitis and periodontal disease are the most common oral mucosa alterations aim of our research was to investigate histomorphometric characteristics of healthy and altered oral and gingival epithelium.Material and methods: Tissue samples of 24 oral and gingival mucosa specimens were collected. Samples were fixed in 10% buffered paraformaldehyde, routinely processed and embedded in paraffin blocks. From each block sections 5 micrometer thin were made and standard H/E staining as well as immunocytochemical detection of Ki-67 proliferation marker and CD79a lymphocyte marker were performed. Measurements and image analysis was performed with Image Pro Plus software (Media Cybernetics, USA) and Axiovision (Ziess, USA).Results: We showed that inflamed gingival epithelium is increasing its thickness in proportion to the severity of adjacent inflammation. Furthermore, mitotic index is rising (up to 132%) in the same manner as well as basal lamina length (up to 70%) when normal and inflamed gingiva is compared. Architecture of epithelial ridges is changed from straightforward to mesh-like.Conclusion: Assessment of the free gingival epithelium thickness is directly related to the severity of the inflammation process in gingiva.

T2 relaxation mapping MRI of healthy and inflamed gingival tissue.

To investigate the use and reproducibility of MRI transverse relaxation time (T2) mapping in healthy and inflamed gingivae. 21 subjects were recruited into 2 groups: those without evidence of gingivitis ("healthy"; n = 11, age 24.0 ± 3.66 years) by visual assessment and those with moderate to severe gingivitis ("gingivitis"; n = 10, age 28.9 ± 6.03 years) exhibited across the second mandibular premolar and first mandibular molar buccal gingivae. Subjects were imaged by MRI twice in a single day. Three T2 weighted turbo spin-echo volumes with 0.25 × 0.25 × 0.8-mm3 resolution were acquired at echo times of 16, 32 and 48 ms for T2 decay fitting. Image analysis was fully blinded; the two imaging sessions were not identifiable as coming from the same subject. Each imaging session had independent regions of interest drawn on the first echo image and applied to the calculated T2 decay maps. The coefficient of variation was low and similar in healthy and gingivitis populations: 6.10 and 5.25% populations, respectively, with 5.65% populations across both groups. Bland-Altman analysis revealed no bias (mean -2.93%; 95% confidence intervals -22.20 to 16.34%) between sessions. The intersession agreement was good (r = 0.744, ρ = 0.568, intraclass correlation coefficient = 0.68). T2 mapping did not differentiate healthy from gingivitis groups. The mean T2 value in the healthy group (63.7 ms) was similar to that of the gingivitis group (65.23 ms) (p = 0.30). Mapping of the T2 decay in the gingivae was a repeatable process; however, T2 value alone did not differentiate those with clinical examination-determined gingivitis from those without signs of gingivitis.

Localization and expression pattern of amelotin, odontogenic ameloblast-associated protein and follicular dendritic cell-secreted protein in the junctional epithelium of inflamed gingiva.

The purpose of this study is to elucidate the localization of amelotin (AMTN), odontogenic ameloblast-associated protein (ODAM) and follicular dendritic cell-secreted protein (FDC-SP) at the junctional epithelium (JE) in Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans infected mice and inflamed and non-inflamed human gingiva. We performed immunostaining to determine the localization and expression pattern of AMTN, ODAM and FDC-SP. AMTN, ODAM and FDC-SP in A. actinomycetemcomitans infected mice did not change dramatically compared with non-infected mice. AMTN and FDC-SP expressions were observed stronger in P. gingivalis infected mice at early stage. However, at the following stage, the coronal part of the AMTN expression disappeared from the JE, and FDC-SP expression decreased due to severe inflammation by P. gingivalis. ODAM expressed internal and external basal lamina, and the expression increased not only at early stage but also at the following stage in the inflammatory JE induced by P. gingivalis. In the human gingival tissues, AMTN was detected at the surface of the sulcular epithelium and JE in the non-inflamed and inflamed gingiva, and the localization did not change the process of inflammation. ODAM and FDC-SP were more widely detected at the sulcular epithelium and JE in the non-inflamed gingiva. In the inflamed gingiva, localization of ODAM and FDC-SP was spread into the gingival epithelium, compared to AMTN. These studies demonstrated that the expression pattern of AMTN, ODAM and FDC-SP at the JE were changed during inflammation process and these three proteins might play an important role in the resistance to inflammation.

Increased expression of triggering receptor expressed on myeloid cells 1 and 2 in inflamed human gingiva.

Periodontitis is an infectious disease in which the host immune and inflammatory responses play essential roles in resistance to bacterial infection, as well as the induction of tissue destruction if the immune response is dysregulated. The triggering receptor expressed on myeloid cells (TREMs) modulates inflammatory and innate immune signaling. TREM-1 is considered as an amplifier of the immune response, while TREM-2 is a negative regulator that has yet to be explored in periodontal disease before. We hypothesized that TREMs participated in the innate immune responses during the pathogenesis of periodontitis. Therefore, the aim of this study was to evaluate TREM-1 and TREM-2 expression in the gingival tissues from patients with chronic periodontitis and healthy subjects as well as their correlation with clinical periodontal parameters. This study is the first to identify TREM-2 in periodontal tissue, as well as the protein expression changes of TREM-1 and TREM-2 in periodontal tissues. Gingival tissue sections were collected from 31 healthy subjects and 53 patients with chronic periodontitis. Immunohistochemistry and quantitative real-time polymerase chain reaction were employed to evaluate the protein and mRNA expression of these receptors in gingival tissues. The recorded clinical parameters were probing depth, clinical attachment loss, plaque index and bleeding on probing. In addition to myeloid cells in gingival connective tissues, TREM-1 and TREM-2 were also found expressed in gingival epithelial cells. In particular, TREM-1 was detected in almost all gingival epithelium from both healthy and inflamed biopsies. The expression levels of TREM-1 and TREM-2 were significantly increased in the periodontitis group compared to the healthy group. Increased levels of these receptors are to be positively correlated with site-specific periodontal parameters. The increased expression of TREM-1 and TREM-2 levels in periodontitis may confer diagnostic and potential therapeutic targets as well as indicating their association with the clinical severity of the disease.

Platelet activation and platelet–leukocyte interaction in generalized aggressive periodontitis

Generalized aggressive periodontitis (GAgP) is an inflammatory disease of host response to bacterial challenge. To explore the role of platelets in host-microbial interactions in patients with periodontitis, 124 patients with GAgP and 57 healthy subjects were enrolled. Reliable indicators of subclinical platelet functional status, platelet count (PLT), platelet large cell ratio (PLCR), and mean platelet volume (MPV), were significantly lower in the GAgP group than in the control group and were negatively correlated with clinical periodontal parameters. The levels of important cytosolic protein in neutrophils, calprotectin (S100A8/A9) in plasma, and gingival crevicular fluid (GCF) were significantly higher in patients with GAgP compared with healthy subjects. Moreover, the GCF calprotectin level was negatively correlated with PLCR and MPV values. To explore the possible mechanisms of changes in platelet indices in periodontitis, flow cytometry analysis was performed, and patients with GAgP were found to have a higher status of platelet activation compared with healthy controls. Porphyromonas gingivalis (P. gingivalis) and recombinant human S100A8/A9 (rhS100A8/A9) induced platelet activation and facilitated platelet-leukocyte aggregate formation in whole blood of healthy subjects. In response to P. gingivalis and rhS100A8/A9, platelets from patients with GAgP increased activation and increased formation of platelet-leukocyte aggregates compared with those from healthy subjects. Platelet aggregates and platelets attached to leukocytes were found on gingival tissues from patients with GAgP, suggesting that decreased platelet size and count in the circulation might be related to consumption of large, activated platelets at inflamed gingiva. Platelets may have a previously unrecognized role in host response to periodontal infection.

The expression of human β-defensins (hBD-1, hBD-2, hBD-3, hBD-4) in gingival epithelia

ObjectiveThe aim of this study was to analyze and compare the expression patterns of human β-defensin-4 (hBD-4) with those of hBD-1, hBD-2, and hBD-3 in healthy and chronically inflamed gingival tissues. Materials and methodsA total of 96 gingival samples were collected, comprising 46 biopsies from chronic periodontitis (CP) patients and 50 from individuals with healthy tissue. Of these, 21CP and 21 healthy samples were used to examine the protein expression of the hBDs by immunohistochemistry. The remaining 25CP and 29 healthy tissue samples were subjected to real-time reverse transcription polymerase chain reaction to quantify the mRNA expression of the hBDs. ResultsAll four types of β-defensin peptides were confined to the gingival epithelia. The percentage of samples positive for hBD-4 mRNA was significantly lower than the percentages for the other three β-defensins, both in healthy (p=0.003) and in inflamed (p=0.030) gingiva. However, in the case of the relative mRNA expression levels, that of hBD-4 was significantly higher than the levels for hBD-2 (healthy group: p<0.01; CP group: p<0.01) and hBD-3 (healthy group: p=0.004; CP group: p<0.01). ConclusionsOur study demonstrates differential expression of hBD-1, hBD-2, hBD-3, and hBD-4 in healthy and CP gingiva.

Increase in receptor activator of nuclear factor κB ligand/osteoprotegerin ratio in peri-implant gingiva exposed to Porphyromonas gingivalis lipopolysaccharide

Background/purposeThe prevalence of peri-implant diseases, including peri-implant mucositis and peri-implantitis, is increasing. The aim of this study was to elucidate the pathological mechanisms of inflammation and alveolar bone resorption in peri-implant tissues. To do this, we fabricated inflamed gingiva around mini-implants in the palatine processes of rats using lipopolysaccharide derived from Porphyromonas gingivalis (P.g-LPS). Materials and methodsPure titanium mini-implants were implanted into the palatine processes of rats, and then intermittent injections of P.g-LPS were made into the gingival tissues surrounding the mini-implants. The expression patterns of tumor necrosis factor-α, interleukin-1β, chemokine (C–C motif) ligand 2, receptor activator of nuclear factor κB ligand (RANKL), and osteoprotegerin (OPG) in the tissues were examined using real-time reverse transcriptase polymerase chain reaction or enzyme-linked immunosorbent assays. Immunohistochemical analysis was also performed to compare the T and B cells expressing RANKL. ResultsP.g-LPS increased the expressions of tumor necrosis factor-α, interleukin-1β, chemokine (C–C motif) ligand 2, and RANKL in the gingival tissues surrounding the mini-implants. In contrast, the expression of OPG in the P.g-LPS samples was decreased. Consequently, the RANKL/OPG ratio was significantly increased. Moreover, cells stained positively for both anti-CD3 and anti-RANKL antibodies were only found in the samples treated with P.g-LPS. ConclusionThese data revealed that P.g-LPS injections increased the RANKL/OPG ratio in the gingival tissues surrounding mini-implants in the rat model. In addition, the CD3-positive cells in the gingival tissues injected with P.g-LPS expressed RANKL. This suggests that the activated T cells capable of infiltrating gingival tissues affected by P.g-LPS may be one of the sources of RANKL and may also be involved in the disease progression from peri-implant mucositis to peri-implantitis.

Gingivitis and Associated Factors Among Children in Boxes Toddlers in Senegal

The aim of this study was to determine the prevalence of gingivitis and the factors associated in boxes toddlers among 3-6 year-old preschool children in 3 regions in Senegal. This was a descriptive and cross sectional study covering 984 preschool children aged 3 to 6 years and attending boxes toddlers in the regions of Dakar (urban area), Saint-Louis (semi-urban and urban areas) and Kaolack (semi-urban and rural areas). They were selected through both a cluster sampling and a stratified sampling and the clusters were drawn at random. These children obtained parental permission to participate in the survey. The socio-demographic data, the gingivitis status and the parental knowledge on oral preventive methods applied to children, were collected. The data was analysed using SPSS Version 19 software with a significance level of 5%. In total 901 children were examined: 299 in Dakar, 302 in Saint-Louis and 300 in Kaolack with respective completeness rate of 93%, 91% and 90%. Girls constituted 56.6% of the sample; children aged six were less represented (10.7%), while children of non-salaried parents constituted 77.3% of the sample. Fourteen percent of the children had an inflamed gingiva. The distribution of gingivitis according to socio-demographic data does not show a significant association. Depending on the region, gingivitis differently distributed (p 0.90). Gingivitis is significantly associated with the frequency of brushing (p=0.03), it is not significantly associated with visiting the dentist (p>0.20). These results suggest the importance of hygiene and perhaps the parent oral health literacy in the prevention of this infection often overlooked because painless most of the time. To avoid damage as noma more prevalent in rural area, health officials may develop integrated community programs against gingivitis in boxes toddlers in Senegal.

EKSTRAK CYMBOPOGON CITRATUS DAN EUGENIA AROMATICUM EFEKTIF UNTUK PENYEMBUHAN GINGIVITIS

Introduction: Gingivitis is an inflammatory gingiva which has early sign is a little bleeding and mild swelling of the gingiva and if this gingivitis is untreated it could be the periodontitis. The process of gingivitis can be prevented with an antibacterial compound.Purpose: The aim of this research is to determine the effectiveness of topical extracts of lemongrass and clove to cure gingivitis.Methods: This experimental research used post test control group design. The samples were 28 male Wistar rats 2-3 months that were divided into 4 groups. Group I as a positive control was given amoxicillin and metronidazole, group II was given lemongrass extract, group III was given clove extracts, and the fourth group was given a combination of lemongrass and clove extracts. The addition of an extract is done in twice a day for 2 days.Results: Healing of gingivitis seen from the number of polymorphonuclear leukocytes/PMN leukocytes. The mean number of PMN in group I = 5.00, group II = 9.43, group III = 8:57, and group IV = 5.86. Based on the Kruskal Wallis test there was not a significant difference of the PMN number withinfourth experiment group.Conclusion: The conclusion of this study was combination of lemongrass and clove extract had an effectiveness level that almost similar to metronidazole dan amoxicillin in decreasing the number of polymorphonuclear leukocytes in the gingivitis healing process.

Functional Local Renin-Angiotensin System in Human and Rat Periodontal Tissue.

The initiation or progression of periodontitis might involve a local renin-angiotensin system (RAS) in periodontal tissue. The aim of this study was to further characterize the local RAS in human and rat periodontal tissues between healthy and periodontally-affected tissue. Components of the RAS were investigated using in vitro, ex vivo and in vivo experiments involving both human and Wistar rat periodontium. Although not upregulated when challenged with P. gingivalis-lipopolysaccharide, human gingival and periodontal ligament fibroblasts expressed RAS components. Likewise, healthy and inflamed human gingiva expressed RAS components, some of which were shown to be functional, yet no differences in expression were found between healthy and diseased gingiva. However, in inflamed tissue the immunoreactivity was greater for the AT1R compared to AT2R in fibroblasts. When compared to healthy tissue, ACE activity was increased in human gingiva from volunteers with gingivitis. Human-gingiva homogenates generated Ang II, Ang 1-9 and Ang 1-7 when incubated with precursors. In gingiva homogenates, Ang II formation from Ang I was nearly abolished only when captopril and chymostatin were combined. Ang 1-7 formation was significantly greater when human gingiva homogenates were incubated with chymostatin alone compared to incubation without any inhibitor, only captopril, or captopril and chymostatin. In rat gingiva, RAS components were also found; their expression was not different between healthy and experimentally induced periodontitis (EP) groups. However, renin inhibition (aliskiren) and an AT1R antagonist (losartan) significantly blocked EP-alveolar-bone loss in rats. Collectively, these data are consistent with the hypothesis that a local RAS system is not only present but is also functional in both human and rat periodontal tissue. Furthermore, blocking AT1R and renin can significantly prevent periodontal bone loss induced by EP in rats.

Involvement of smad2 and Erk/Akt cascade in TGF-β1-induced apoptosis in human gingival epithelial cells

Periodontitis is the most prevalent infectious disease caused by periodontopathic bacteria and is also a chronic inflammatory disease. Gingival crevicular fluid (GCF) is an inflammatory exudate that seeps into the gingival crevices or periodontal pockets around teeth with inflamed gingiva, and contains various materials including leukocytes and cytokines. Since gingival epithelial cells, which form a barrier against bacterial challenges, are affected by GCF, cytokines or other materials contained within GCF are engaged in the maintenance and disruption of the epithelial barrier. Accordingly, its compositional pattern has been employed as a reliable objective index of local inflammation. Transforming growth factor β1 (TGF-β1) levels in GCF were previously shown to be markedly higher in patients with periodontitis than in healthy subject. However, it currently remains unclear how TGF-β1 affects gingival epithelial cell growth or apoptosis; therefore, elucidating the mechanism responsible may lead to a deeper understanding of pathogenic periodontitis. In the present study, the human gingival epithelial cell line, OBA9 cells were stimulated with recombinant TGF-β1. Apoptosis-related protein levels were determined by Western blotting. Caspase-3/7 activity was measured with a Caspase-Glo assay kit. Surviving and apoptotic cells were detected using an MTS assay and TUNEL staining, respectively. TGF-βRI siRNA and smad2 siRNA were transfected into cells using the lipofectamine RNAiMAX reagent. TGF-β1 elevated caspase-3 activity and the number of TUNEL-positive apoptotic cells in OBA9 cells. Furthermore, while the levels of the pro-apoptotic proteins Bax, Bak, Bim, and Bad were increased in OBA9 cells stimulated with TGF-β1, the TGF-β1 treatment also decreased the levels of anti-apoptotic proteins such as Bcl-2 and Bcl-xL in a time-dependent manner. Additionally, TGF-β1 up-regulated the protein levels of cleaved caspase-9. These results indicated that TGF-β1-induced apoptosis was involved in a mitochondria-related intrinsic pathway. TGF-β1 phosphorylated smad2 in OBA9 cells and this phosphorylation was clearly reduced by SB431542 (a TGF-β type I receptor inhibitor). Consistent with this result, SB431542 or smad2 siRNA-induced reductions in smad2 protein expression levels attenuated TGF-β1-induced apoptosis. On the other hand, the ligation of TGF-β1 on its receptor also stimulated the phosphorylation of Erk and Akt, which are smad2-independent pathways. However, the inhibition of Erk/Akt signaling pathways by U0126, a MEK-Erk inhibitor and LY294002, a PI3Kinase-Akt inhibitor, augmented TGF-β1-induced apoptosis in OBA9 cells. Taken together, the results of present study demonstrated that TGF-β1 activated both the smad2 and Erk/Akt cascades via its receptor on gingival epithelial cells, even though these two pathways have opposite roles in cell death and survival, and the culmination of these signaling events induced mitochondria-dependent apoptosis in gingival epithelial cells. Based on the results of the present study, we herein proposed for the first time, that TGF-β1 is a novel target cytokine for monitoring the progression of periodontal disease.

Biomarkers of Regenerative Responses to Phenytoin in Periosteal Fibroblasts and Modulation by Histamine; Relevance to Inflammatory Repair

Objectives: Pronounced wound healing responses induced by phenytoin in inflamed gingivae have potential for matrix regeneration in this context. It enhances matrix formation in tissues and impairs their breakdown. The matrixregenerative responses of human periosteal fibroblasts to phenytoin and histamine were studied using the metabolite 5 alpha- dihydrotestosterone (DHT) as a marker of inflammatory wound repair; derived from 14C-testosterone and 14C-4- androstenedione as independent substrates in order to ascertain metabolic responses. Methods: Confluent monolayer cultures of human periosteal fibroblasts of the 5th-9th passage from eight subjects were incubated individually in 24-well multiwell plates in Eagle’s MEM. Independent incubations were performed with each of the substrates 14C-T and 14C-4- A, with optimal concentrations of histamine (H; 8μg/ml) and phenytoin at 1(Ph1) and 5μg/ml (Ph5) alone and in combination (H+Ph1; H+Ph5). Following 24h incubation the eluent was prepared for TLC in a benzene. Acetone solvent system (4:1) for the separation of metabolites. They were quantified using a radioisotope scanner. Results: The substrates a.14CT and b.14C-4A were metabolized to DHT, diol and 4-A or T respectively. With each substrate, yields of DHT were significantly increased over controls in response to H (a.74%; b.2.3-fold), Ph1, 5 (a.70%, 2-fold;b. 2.8-,1.9-fold) and the combinations (a.60%; b.2-fold and 73%), respectively (n = 8; p < 0.01/0.001). There were also significant increases in the yields of diol and T respectively in response to agents tested. Conclusion: Significant increases in yields of DHT in response to Ph and H with both substrates indicate their anabolic potential in periosteal fibroblasts. This has implications for diverse applications in tissue repair in an inflammatory environment. Patents relevant to matrix synthesis and its modulation by microRNAs during inflammatory healing responses have been discussed. Keywords: Biomarkers, histamine, inflammation, periosteal fibroblasts, phenytoin, wound repair.

Quantitative analysis of macrophages in gingivitis and periodontal disease

Periodontal disease is a chronic inflammatory disease, the pathogenesis of which involves different innate immune system as well as adaptive immune system cells, including macrophages. Macrophages are professional phagocytes whose primary role is ingestion and destruction of microorganisms. Apart from phagocytosis, these cells are engaged in production of proinflammatory cytokines and antigen presentation to T cells, and therefore are considered to be the key cells of the innate immune response. Our study focused on distribution and density of macrophages in healthy and inflamed gingiva with the purpose of evaluating their relevance to pathogenesis of periodontal disease. The material used for this purpose were gingival biopsies taken from 96 patients assorted in 4 groups: the healthy gingiva, gingivitis, moderate periodontal disease and severe periodontal disease. The immunohistochemical staining method based on the use of monoclonal antibodies anti-CD68 was used for identification of macrophages. Our study showed that the density of macrophages is higher in gingivitis and periodontal disease compared to healthy gigiva. There were no significant differences found regarding the density of macrophages in gingivitis and periodontal disease. Grouping of macrophage populations close to partially lysed collagen fibers in periodontal disease indicates their collagenic activity. The increase in the number of macrophages in inflamed gingival tissue could be linked to bone resorption due to the fact that these cells are capable of differentiating into osteoclasts.

Scaling and Root Planning decreases the Number of Melanosomes within Keratinocytes in Human Gingiva: Ultrastructural Analysis of Three Cases.

The aim of the present report was to evaluate the number of melanosomes within keratinocytes on pigmented gingiva, after and before scaling and root planning. Inflamed gingiva biopsies were taken from three patients (group 1). Forty days after scaling and root planning, biopsies were collected from the homologous contralateral areas (group 2). Samples were fixed in 2% glutaraldehyde-2.5% formaldehyde (freshly prepared from paraformaldehyde) in 0.1 M sodium cacodylate buffer, pH 7.4 for 4 hours, and then processed for transmission electron microscopy. Eighty electron micrographs were evaluated for recording the number of granules by a cross-section grid. The granules that were on intersections were recorded as well as the points that appeared on the cytoplasm for calculating the volumetric density (Vd), i.e the volume that the melanosomes occupied into the cytoplasm of keratinocytes. The presence of melanosomes in different stages of maturation and distribution into the cells were recorded with the aid of a magnifying glass. For the statistical analysis, a student t-test was applied. Results of the present report showed that melanosomes within keratinocytes were present in a higher number in inflamed gingiva A (11.08 ± 1.47), B (3.16 ± 0.38) and C (4.92 ± 0.89) and decreased after resolving of gingival inflammation A (9.46 ± 0.88), B (1.73 ± 0.25) and C (0.76 ± 0.18). There is a possibility that inflammation influences the intensity of gingival melanin pigmentation. The periodontal treatment appears to have an effect on gingival melanin pigmentation.

Effectiveness of CoQ10 Oral Supplements as an Adjunct to Scaling and Root Planing in Improving Periodontal Health.

Deficiency of CoQ 10 was found in human inflamed gingiva and has been found to be responsible for periodontal destruction. To evaluate the effectiveness of CoQ 10 supplementation as an adjunct to scaling and rootplaning in reducing gingival inflammation and periodontal pocket depth. The study was a randomized, double-blind, controlled, parallel group design clinical trial. Thirty subjects with plaque induced gingival inflammation and having atleast three nonadjacent interproximal sites with a probing pocket depth ≥ 5mm were included in the study. The subjects were randomly divided into two groups. The test group (n=15) in which patients were given oral CoQ10 supplements after scaling and root planing and the control group (n=15) in which patients were given an oral placebo after scaling and rootplaning. The plaque index, gingival index and probing depth were recorded at baseline, 1 month and 3 months. Statistical analysis done by using Student's paired t-test for intragroup comparison and unpaired t-test for inter-group comparison. Both the groups showed marked reduction of afore mentioned periodontal parameters at one month and three months when compared to baseline. Though there was no significant difference in plaque index and probing pocket depth between the two groups at any given time period, test group showed significant difference in gingival inflammation at one month and three months when compared to control group. In the present study use of Coenzyme Q10 oral supplements as an adjunct to scaling and root planing showed significant reduction in gingival inflammation when compared to scaling and rootplaning alone.

The Influence of Diabetes Mellitus on Expression of Stromelysins and Membrane type Matrix Metalloproteinases in Human Chronic Periodontitis

The purposes of this study were to observe influence of diabetes mellitus(DM) on the expression of MMP?3 of stromelysin type and MMP?14 of membrane type matrix metalloproteinase in the gingival tissues of patients with type 2 DM and healthy adults with chronic periodontitis. Gingival samples were devided into three groups. Group 1 is clinically healthy gingiva without bleeding, bone resorption or periodontal pockets. Group 2 is inflammed gingiva from patients with chronic periodontitis. Group 3 is inflammed gingiva from patients with chronic periodontitis associated with type 2 DM. Tissue samples were analyzed by Western blotting. The quantification of MMP?3 and MMP?14 were performed using a densitometer. MMP?3 and MMP?14 expressions were similar in group 1 and 2. While expression of MMP-14 increases in proportion to MMP-3 expression, expression of both type of MMP showed increasing tendency in chronic periodontitis associated to type 2 DM and mean amount of it was more increased in group 3 comparing to other group with statistically significant difference.BR In conclusion, MMP?3 and MMP?14 expression levels were similar in healthy and inflamed gingival tissue from systemically healthy patients. And tissue with chronic periodontitis associated to type 2 DM showed significantly increased MMP?3 and MMP?14 levels compared to healthy and non?diabetic inflamed gingiva. Also, MMP?3 may be related to MMP?14 activity in chronic periodontitis associated with type 2 DM. Therefore, the expression levels of MMP?3 and MMP?14 will be inflammatory markers of periodontal inflammed tissue with type 2 DM.

Transcriptional regulation of amelotin gene by proinflammatory cytokines in gingival fibroblasts

Amelotin (AMTN) is a secreted protein expressed during the late stages of enamel formation and in the junctional epithelium. Among many differentially expressed genes, we found significantly increased AMTN mRNA level in inflamed gingiva by DNA microarray. The inductions of AMTN mRNA expressions in inflamed gingiva and human gingival fibroblasts (HGF) were confirmed by real-time polymerase chain reaction. To determine the molecular basis of the expression of AMTN and its regulation by proinflammatory cytokines, we have isolated and characterized the promoter region of mouse AMTN gene. Transient transfection assays were performed using luciferase constructs including mouse AMTN gene promoter. Interleukin-1β, Interleukin-6 and tumor necrosis factor-α induced AMTN mRNA levels in HGF. These cytokines increased the luciferase activities of the AMTN promoter constructs in HGF. The results suggest that proinflammatory cytokines induce AMTN gene transcription and a role for AMTN in gingival inflammation.

The Expressions of Xanthine oxidase and Malondialdehyde in Chronic Periodontitis with Hypertension

The purpose of this study was to quantify the expressions of XO and MDA in the gingival tissues of the chronic periodontitis associated with HTN. Xanthine oxidase (XO) generates reactive oxygen species. Malondialdehyde (MDA) is one of many low molecular weight end-products of LPO and is the most often measured as an index of peroxidation. Depending on the patients systemic condition and clinical criteria of gingiva, each gingival sample was divided into three groups. Group 1 is clinically healthy gingiva. Group 2 is inflamed gingiva from with chronic periodontitis. Group 3 is inflamed gingiva from with chronic periodontitis associated with HTN. The expression levels of XO increased in order of group 1, group 2 and group 3, and significantly increased in group 3 as compared to group 1. The expression levels of MDA increased in order of group 1, group 2 and group 3. MDA expression was significantly higher in group 2 than in group 1, and the quantitative analysis of MDA level was significantly higher in group 3 than in group 1. In conclusion, this study demonstrated that the expression levels of XO and MDA might be inflammatory and bone resorption marker in periodontal inflammed tissue. It is assumed that hypertension may be associated with the progression of periodontal inflammation and alveolar bone resorption.