The composition of gingival inflammatory cell infiltrates in children studied by enzyme histochemistry

Gingival biopsies were obtained from 23 children, aged 5-11 years (8.6 +/- 1.8 years). Specimens were taken from areas of the gingiva adjacent to the teeth which were to be extracted because of caries or its sequelae and which clinically had a gingival index score of at least 1. Staining for alpha-naphthyl acetate esterase with unspecific esterase at pH 5.8 (ANAE) permitted identification of T lymphocytes, monocytes/macrophages, plasma cells and non-reactive (ANAE-negative) cells. Cells which tentatively were identified as "natural killer" (NK) cells were also observed. Differential cell counting was performed for 10 specimens, selected on the basis of the presence of a well-defined inflammatory infiltrate, clear morphology throughout and good ANAE staining. Cell counts confirmed earlier studies showing that lymphocytes predominate in the inflammatory infiltrates in childrens' gingivitis. T lymphocytes dominated particularly in the periphery of the most densely infiltrated areas. Relatively few plasma cells were seen. It was concluded that T lymphocytes dominate in the inflammatory infiltrate in childrens' gingivitis.     

Effect of short chain fatty acids on human gingival epithelial cell keratins in vitro

Hemidesmosomal attachment of the junctional epithelial cells to the tooth and the ability of the attached cells to divide are essential features of the healthy dentogingival junction. Short chain fatty acids are bacterial metabolites associated with gingival inflammation and periodontal pockets. In vitro, short chain fatty acids have been shown to inhibit epithelial cell division and increase the density of their keratin filaments. This study examined these keratin changes by making use of human gingival keratinocyte cultures, gel electrophoresis and Western blot. Short chain fatty acids, butyrate and propionate, increased the relative amount of keratin proteins in the cells, most strikingly keratin K17. The distribution of K17 was further studied in a culture model for human junctional epithelium and in gingival biopsies. In butyrate-treated cultures of junctional epithelium, K17 expression was markedly increased and extended to the basal cells and to the cells mediating the attachment of the explant to the substratum. In clinically healthy gingiva, K17 was expressed predominantly in sulcular epithelium. The dividing basal cells and the cells attached to the tooth were negative. In advanced periodontitis, a strong reaction for K17 was localised to the pocket epithelium. The inhibition of epithelial cell division and the simultaneous upregulation of K17 in vitro, and the strong expression of this protein in detached pocket epithelium suggest a role for the short chain fatty acids in the degenerative process that leads to subgingival advancement of pathogens and, eventually, to periodontal pocket formation.     

Characterisation of the inflammatory cell infiltrate in chronic hyperplastic candidosis of the oral mucosa

The inflammatory cell infiltrate in biopsy material of chronic hyperplastic candidosis (CHC) from the oral mucosa was characterised using immunocytochemical techniques. Nine specimens were stained for human kappa and lambda immunoglobulin light chains. CD68 antigen (macrophages), lysozyme (macrophages, granulocytes), CD3 antigen (T-lymphocytes), CD20 antigen (B-lymphocytes) and leucocyte common antigen (LCA). In addition, these and a further 13 specimens were also examined for immunoglobulin (Ig)-containing cells (IgA, IgG and IgM). The density of the infiltrate varied considerably between cases: T-lymphocytes were the dominant cell type (153.9%), with fewer B-lymphocytes (8.2%) and macrophages (14.2%). Many Ig-containing cells were seen, and although IgG-containing cells predominated, (60.8%, SD ±9.0) there was a high proportion of IgA-containing cells (36.7%, SD ± 9.1) with few IgM-containing cells (2.5%, SD ±3.0). Many neutrophils, together with smaller numbers of T-lymphocytes and macrophages were seen in the epithelium. It is suggested that mucosal defence to Candida infection involves a cell-mediated reaction in which there is recruitment of macrophages and local production of immunoglobulin with a prominent IgA component.     

Physical activity, inflammatory biomarkers in gingival crevicular fluid and periodontitis

Aims: To examine the associations of physical activity with interleukin 1- β (IL-1 β ), C-reactive protein (CRP) and periodontitis and to investigate whether any relationship between physical activity and inflammatory mediators differs between periodontitis cases and non-cases.
Material and Methods: In this population-based case control study of Australians aged 18+ years, dentists conducted oral epidemiologic examinations identifying cases with moderate or severe periodontitis and periodontally healthy controls. Gingival crevicular fluid samples collected during examinations were analysed for inflammatory biomarkers. Subject-completed questionnaires assessed leisure-time physical activity. Exposure odds ratios (ORs) were estimated in multivariable logistic regression models adjusting for periodontitis risk indicators.
Results: Of 751 subjects (359 cases, 392 controls), those meeting a prescribed threshold for leisure-time physical activity had lower adjusted odds of elevated IL-1 β : OR=0.69, (95% CI=0.50–0.94) and detectable CRP: OR=0.70 (0.50–0.98) than less active adults. Physical activity was not associated with periodontitis: OR=1.14 (0.80–1.62). Periodontitis modified the association between levels of physical activity and detectable CRP. Increasing quartiles of physically activity were associated with decreasing probability of detectable CRP, but the effect was limited to periodontitis cases and was not apparent among non-cases.
Conclusion: Leisure-time physical activity may protect against an excessive inflammatory response in periodontitis.     

An uncommon presentation of an inflammatory gingival enlargement--responding to non-surgical periodontal therapy

To cite this article:
Int J Dent Hygiene 9 , 2011; 303–307 DOI: 10.1111/j.1601‐5037.2010.00497.x
Agrawal N, Agrawal K, Mhaske S. An uncommon presentation of an inflammatory gingival enlargement – responding to non‐surgical periodontal therapy. Abstract: Background: The various clinical manifestations of inflammatory gingival enlargement reported are more or less similar regardless of the underlying aetiological factors. Unusual presentation and unknown aetiology pose a diagnostic challenge for a periodontist. Methods: A 34‐year‐old Indian woman presented with the complaint of gum swelling that was sessile, lobulated, soft in consistency and bluish red in colour with ulcerated surface in some region, which was covered by the necrotic slough. This type of enlargement was unusual and some underlying systemic pathology was suspected. But a written consultation from her physician confirmed her systemic health, which was based on clinical, radiological and haematological investigations. Histopathological examination confirmed the diagnosis of inflammatory gingival enlargement. Patient was treated with oral hygiene instructions, scaling and root planning. Result: Within a month of conventional periodontal therapy, gum enlargement reduced markedly and patient was put on oral hygiene maintenance programme. Conclusion: Periodontal therapy is diagnosis‐driven and, to the extent possible, should address all the possible factors that impact development and progression of diseases that may affect periodontal tissue. In plaque‐induced periodontal diseases, non‐surgical periodontal therapy is still a gold standard among all the therapies available.     

Scleraxis在牙周韧带细胞、牙龈成纤维细胞中的表达

目的研究碱性螺旋-环-螺旋转录因子 Scleraxis 能否在人牙周韧带细胞、牙龈成纤维细胞、骨髓基质细胞中表达,及其与牙周韧带细胞分化能力的关系。方法体外培养人牙周韧带细胞、牙龈成纤维细胞和骨髓基质细胞,RT-PCR 法检测牙龈成纤维细胞、骨髓基质细胞及不同代次牙周韧带细胞中 Scleraxis 的表达。结果 Scleraxis 在牙周韧带细胞、骨髓基质细胞及牙龈成纤维细胞中的表达差异有统计学意义(P<0.05)。其 Scleraxis 与β-肌动蛋白吸光度比值分别为0.877±0.024,0.438±0.031,0.313±0.083。Scleraxis 在牙周韧带细胞中的表达最强,在牙龈成纤维细胞中表达最弱。牙周韧带细胞中 Scleraxis 的表达随培养代次的增加而减弱。结论 Scleraxis 可表达于体外培养的牙周韧带细胞、牙龈成纤维细胞和骨髓基质细胞,Scleraxis 可能在牙周韧带细胞的分化过程中起重要作用。     

Elastase in gingival crevicular fluid from smokers and non-smokers with chronic inflammatory periodontal disease

OBJECTIVE: To compare elastase concentrations in gingival crevicular fluid (GCF) from individual sites of smokers and non-smokers.
MATERIALS AND METHODS: Twelve pairs of smokers and non-smokers with untreated, moderate to advanced chronic inflammatory periodontal disease were matched for gender, age, ethnicity and the clinical and radio-graphic extent of disease. Durapore filter strip samples were collected over 30 s from two mesiopalatal sites on upper left posterior teeth. Samples were analysed for: I) polymorphonuclear neutrophil leucocyte (PMNL) cell counts; 2) PMNL elastase-αI-antitrypsin complex in the GCF supernatant by ELISA; and 3) functional elastase, free or bound to α2-macroglobulin, estimated from activity against N-tert-butoxycarbonyI-alanyl-prolyl-nor-valylg-chlorothiobenzyl ester in supernatant and lysates of GCF PMNLs.
RESULTS: There were no differences in disease parameters between groups except that bleeding on probing was less extensive in smokers (P< 0.001). Cell counts and elastase content of crevicular PMNLs showed no differences between groups. Lower concentrations of elastase were found in GCF supernatants from smokers than non-smokers. This difference was observed for functional elastase (mean [s.d.] = 30.21 [17.60] against 73.77 [75.26] ng μI^-1, P <0.05) and elastase complexed with αl-antitrypsin (8.97 [6.54] ng μl^-1 against 25.71 [22.07] ng μI^-1, P < 0.001).
CONCLUSIONS: Smokers have lower elastase concentrations in GCF than non-smokers. Further investigation is required to elucidate the underlying cause and its relationship with periodontal disease.     

Interaction of oral bacteria with gingival epithelial cell multilayers

Primary gingival epithelial cells were cultured in multilayers as a model for the study of interactions with oral bacteria associated with health and periodontal disease. Multilayers maintained at an air–liquid interface in low‐calcium medium displayed differentiation and cytokeratin properties characteristic of junctional epithelium. Multilayers were infected with fluorescently labeled Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum or Streptococcus gordonii, and bacterial association was determined by confocal microscopy and quantitative image analysis. Porphyromonas gingivalis invaded intracellularly and spread from cell to cell; A. actinomycetemcomitans and F. nucleatum remained extracellular and showed intercellular movement through the multilayer; whereas S. gordonii remained extracellular and predominantly associated with the superficial cell layer. None of the bacterial species disrupted barrier function as measured by transepithelial electrical resistance. P. gingivalis did not elicit secretion of proinflammatory cytokines. However, A. actinomycetemcomitans and S. gordonii induced interleukin‐1β (IL‐1β), tumor necrosis factor‐α (TNF‐α), IL‐6 and IL‐8 secretion; and F. nucleatum stimulated production of IL‐1β and TNF‐α. Aggregatibacter actinomycetemcomitans, F. nucleatum and S. gordonii, but not P. gingivalis, increased levels of apoptosis after 24 h infection. The results indicate that the organisms with pathogenic potential were able to traverse the epithelium, whereas the commensal bacteria did not. In addition, distinct host responses characterized the interaction between the junctional epithelium and oral bacteria.     

In situ characterization of the inflammatory cell infiltrates of hyperplastic denture stomatitis

Cryostat sections from 14 surgical specimens were examined to determine whether selected factors of the immune response related to histopathological reactions are present in the palatal mucosa affected by hyperplastic denture stomatitis. By means of various immunological techniques the presence of IgG, IgA, IgM, complement factor C3c, receptors for the Fc region of IgG (FcR) and for complement factor C3b (C3bR), T lymphocytes, and macrophages were studied. The inflammatory infiltrate was mainly located in the papillary part of the lamina propria. IgG, IgA, and IgM appeared both in plasma cells and intercellularly. FcR, C3bR, and T lymphocytes were present in the areas with inflammatory cell infiltrate. Macrophages were found in the papillary part of the lamina propria and within the epithelium. The immunological response in the mucosa affected by denture stomatitis was in many respects similar to that of marginal and apical periodontitis. We conclude that hyperplastic denture stomatitis is a complex inflammatory lesion showing elements of both humoral and cellular immune responses.     

同一个体来源正常与炎性牙周膜干细胞生物学性能的比较

目的：比较同一个体来源正常与炎性牙周膜干细胞的生物学性能。方法：分别分离培养同一个体来源正常牙周膜干细胞（hPDLSCs）及炎性牙周膜干细胞（i－hPDLSCs）。流式细胞仪鉴定两种干细胞表面抗原标记;MTT法、克隆形成率检测并比较两者的增殖能力;茜素红染色及油红O染色检测并比较其分化能力;RT－PCR检测成骨相关基因COL－1、Runx－2、BSP－mRNA及成脂相关基因LPL与PPAR γmRNA的表达水平。结果：i－hPDLSCs及hPDLSCs均具有间充质干细胞特性,i－hPDLSCs增殖能力强于hPDLSCs,但分化能力明显弱于hPDLSCs（P＜0．05）;两者成脂能力比较无统计学差异（P＞0．05）。结论：炎症微环境对牙周膜干细胞的生物学性能有一定的影响。     

Expression of CDIa on monocytes cultured with supernatants from periodontally diseased gingival epithelial cells

OBJECTIVE: Langerhans cells are believed to originate from the monocyte lineage and have been reported to increase in number with plaque accumulation and gingival inflammation. The aim of this study was to investigate the effects of local gingival epithelial factors on the induction of CDla, a Langerhans cell phenotype, on monocyte rich populations.
MATERIALS AND METHODS: Peripheral blood monocyte rich populations from healthy subjects were cultured for 24 h with either healthy gingival or periodontally diseased gingival epithelial supernatants. Additionally, the monocyte rich populations were cultured with cytokines IL-Iα, IL-Iβ, IL-6 and TNF-α which are known to be produced by epithelial cells or co-cultured with autologous epithelial cells. The percent CDIa positive cells was determined using FACS analysis.
RESULTS: Healthy gingival supernatants did not induce CDIa expression in monocyte rich populations, however, a significant increase in per cent CDla⁺ cells for monocyte rich populations cultured with five (P < 0.01) of six periodontal gingival epithelial supernatants was found. IL-lα or TNF-α (10ng/well) resulted in a significant increase in the per cent CDla⁺ cells (P < 0.01). Depletion of CDla⁺ Langerhans cells from healthy gingival epithelium did not enhance induction of CDIa expression in monocyte rich populations. Monocyte rich populations cultured together with non-depleted epithelial cultures resulted in a decreased percent of CDla⁺ cells.
CONCLUSION: These findings indicated that epithelial factor/s associated with periodontally involved epithelia, may be involved in inducing a Langerhans cell phenotype in monocyte rich populations. The data also provide indirect evidence for a role of Langerhans cells in inhibiting induction of CDIa in healthy epithelium.