Local and systemic antibody response to putative periodontopathogens in patients with chronic periodontitis: correlation with clinical indices

Specific immunoglobulin G (IgG), IgA and IgM antibody titres to Porphyromonas gingivali s and Actinobacillus actinomycetemcomitans were measured by enzyme-linked immunosorbent assay in serum and gingival crevicular fluid at 5 sites in each of 20 chronic periodontitis patients. Specific serum antibody litres correlated with mean gingival crevicular fluid litres. The 3 immunoglobulin subclass responses (IgA, IgG and IgM) to P. gingivalis correlated. A comparison of sites with probing depth < 4 mm and ≥4 mm showed that the latter group had significantly lower gingival crevicular fluid IgG titres lo P. gingivalis. Sites with a gingival index of 3 had significantly lower gingival crevicular fluid IgG litres to this organism than those with a gingival index of less than 3. These findings supporl the concepl that the humoral immune response is protective, as chronic periodontitis patients with greater pockel depths and more gingival inflammation had paradoxically lower antibody titres to suspected periodontopalhogens.     

Isotypic antibody response to plaque anaerobes in periodontal disease

BACKGROUND: It has been suggested that locally produced immunoglobulin (Ig)A could be more protective than IgG and that there could be a relationship between crevicular fluid-specific IgA levels and the onset of periodontal disease. This study was designed to investigate this hypothesis regarding specific immune responses towards 4 plaque anaerobes in gingival crevicular fluid and saliva from patients with periodontopathies and controls. METHODS: Gingival crevicular fluid (GCF) and whole saliva were collected from 35 adults with periodontitis and 24 periodontally healthy adults (controls). Antigens were extracted from Actinomyces actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum and used to set up specific enzyme-linked immunosorbent assay (ELISA) tests to assess IgA and IgG levels to these microorganisms in the fluids collected. RESULTS: The crevicular fluid of periodontitis patients contained significantly higher levels of IgG to the 4 microorganisms tested than that of controls (P < 10(-6) for all comparisons). IgA levels to the 4 bacteria were statistically significantly much higher in control crevicular fluid (P < 10(-7) for all comparisons). Controls also had statistically significantly higher levels of specific salivary IgA than patients (P < 0.02 for all comparisons). CONCLUSIONS: These data support the potentially protective role of specific IgA directed to oral microorganisms involved in the onset and development of periodontal disease.     

Levels of specific immunoglobulin G to Porphyromonas gingivalis in gingival crevicular fluid are related to site disease status

Titers of immunoglobulin G (IgG) directed against Porphyromonas gingivalis in gingival crevicular fluid of 40 periodontitis patients were measured at three sites in each patient (healthy, gingivitis and periodontitis) by enzyme-linked immunosorbent assay. When paired analyses were performed using Wilcoxon signed-rank tests, periodontitis sites were found to have lower median titers than gingivitis sites. Both systemic and locally-produced antibodies contribute to the overall gingival crevicular fluid antibody profile. Albumin, in contrast, is derived only from serum, and thus this protein serves as an indicator of serum contribution to gingival crevicular fluid. Correction was therefore made for systemic input to the gingival crevicular fluid IgG profile by expressing the results per unit of albumin. When this was done, periodontitis sites were also found to have significantly lower antibody levels than gingivitis sites. These findings suggest that a failure of local antibody production or reduction in quantities, by, for example, degradation by bacterial proteases, may contribute to the change from a gingivitis to a periodontitis lesion.     

Rapid evaluation of serum and gingival crevicular fluid immunoglobulin G subclass antibody levels in patients with early-onset periodontitis using checkerboard immunoblotting

A method was developed to evaluate the presence of immunoglobulin G (IgG) subclass (1-4) antibody to Actinobacillus actinomycetemcomitans, serotype b (strain Y4) in patients with early-onset periodontitis on a single nitrocellulose membrane. Sera from 30 early-onset periodontitis patients and gingival crevicular fluid samples from 2 patients were collected and tested with four different preparations of A. actinomycetemcomitans (Y4). The principle steps of the assay are: a) binding of the bacterial antigen (Y4) and the anti-human IgG antibody (capture antibody) in parallel lanes on nitrocellulose membranes; b) incubation of known concentrations of the IgG subclasses 1, 2, 3 and 4, as well as a dilution of serum and/or gingival crevicular fluid from patients in lanes perpendicular to the antigen lanes; c) incubation of the membranes with the corresponding peroxidase conjugated anti-human IgG subclass secondary antibody; d) detection of positive signals by enhanced chemiluminescence. The blots were evaluated by visual comparison to a series of blots containing known concentrations of IgG subclasses. The method was used to rapidly screen a relatively large number of patient sera and gingival crevicular fluid samples for IgG subclasses in a cost-effective assay. The predominant IgG subclass found in early-onset periodontitis was IgG2.     

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??Objective    To investigate the differences in detection results of periodontal pathogens in saliva and gingival crevicular fluid between patients with chronic periodontitis??CP?? and those complicated with diabetes mellitus??DM??. Methods    Of the patients treated in the First Affiliated Hospital of Henan University between Aug. 2012 and Sep. 2015??40 cases of patients with CP complicated with T2DM were selected as CP+T2DM group??65 cases of patients with simple CP were selected as CP group??and 35 cases of healthy subjects were selected as the control group. The kinds and relative contents of suspected pathogens??such as Porphyromonas gingivalis??Pg????Actinobacillus actinomycetemcomitans??Aa????Fusobacterium nucleatum??Fn????Tannerella forsythia??Tf????intermediate type Prevotella intermedia??Pi??and Treponema denticola??Td????in saliva and gingival crevicular fluid were detected by polymerase chain reaction??PCR??. Results    The detection rates of Pg??Aa and Tf in the saliva and gingival crevicular fluid showed statistically significant differences among the three groups??P??0.05????but there were no significant differences in the detection rates of Fn??Pi or Td??P??0.05??. The detection rate of Fn in saliva and gingival crevicular fluid of the three groups was the highest??and the detection rates of Pg??Aa and Tf in saliva and gingival crevicular fluid of CP +T2DM group were significantly higher than those in control group and CP group??P??0.05??. The detection rate of Pg in CP group was significantly higher than that in the control group??P??0.05??. The relative contents of Pg??Aa and Tf in the saliva and gingival crevicular fluid of CP+T2DM group and CP group were significantly higher than those in the control group??P??0.05??. The relative contents of Aa and Tf in saliva and gingival crevicular fluid of CP+T2DM group were significantly higher than those in CP group??P??0.05??. Conclusion    There are significant differences in the types and number of suspected pathogens between patients with simple CP and those with CP and DM. Pg??Aa and Tf have more advantages in patients with DM??which may be closely related to DM.     

慢性牙周炎合并糖尿病患者唾液及龈下菌斑中牙周致病菌检测临床研究

目的    探讨慢性牙周炎（CP）合并2型糖尿病（T2DM）患者与单纯CP患者唾液及龈下菌斑中牙周致病菌的差异性。方法    选择河南大学第一附属医院口腔科2012年8月至2015年9月伴T2DM的CP患者40例为CP+T2DM组,同时选取背景相似的慢性CP患者65例为CP组,并选取健康体检者35名作为对照组,采用聚合酶链反应（PCR）检测各组受试者唾液及龈下菌斑中牙龈卟啉单胞菌（Pg）、放线共生放线杆菌（Aa）、具核梭杆菌（Fn）、福塞坦菌（Tf）、中间型普里沃菌（Pi）、齿垢密螺旋体（Td）等可疑致病菌种类和相对含量。结果    3组受试者唾液及龈下菌斑中Pg、Aa和Tf检出率比较差异有统计学意义（P＜0.05）,而Fn、Pi和Td检出率差异无统计学意义（P＞0.05）,3组受试者唾液及龈下菌斑中Fn检出率均最高,CP+T2DM组唾液及龈下菌斑中Pg、Aa、Tf检出率均显著高于对照组和CP组（P＜0.05）,而CP组仅Pg检出率显著高于对照组（P＜0.05）,CP+T2DM组和CP组唾液及龈下菌斑中Pg、Aa和Tf相对含量均显著高于对照组（P＜0.05）,且CP+T2DM组唾液及龈下菌斑中Aa和Tf相对含量显著高于CP组,差异有统计学意义（P＜0.05）。结论    CP患者和伴糖尿病的CP患者牙周可疑致病菌的种类和数量存在明显差异,CP+T2DM组患者Pg、Aa和Tf检出率更高,可能与糖尿病关系更为密切。     

慢性牙周炎患者唾液中miR-146a的表达及其与龈沟炎症、MMP-8/TIMP-1水平的关系

目的： 探讨慢性牙周炎患者唾液中miR-146a的表达及其与龈沟炎症、基质金属蛋白酶8（MMP-8）、基质金属蛋白酶抑制剂1（TIMP-1）水平的关系。方法： 选择2015年3月—2017年1月间收治的慢性牙周炎患者68例作为慢性牙周炎组,同期在本院进行体检的健康志愿者50例作为正常对照组。检测2组研究对象唾液中miR-146a的表达量,龈沟液中炎症因子、MMP-8/TIMP-1的水平及牙周临床症状指标。采用SPSS24.软件中的Pearson检验评估慢性牙周炎患者唾液中miR-146a的表达量与病情严重程度的相关关系。结果： 慢性牙周炎患者唾液中miR-146a的表达量显著高于正常对照组（P<0.05）,龈沟液中炎症因子（IL-1β、IL-6、IL-35、TNF-α）水平、牙周临床症状指标（PD、AL、PLI、BI）以及龈沟液中MMP-8、TIMP-1的水平显著高于正常对照组（P<0.05）。Pearson检验发现,慢性牙周炎患者唾液中miR-146a表达量与龈沟炎症程度、牙周临床症状严重程度及MMP-8/TIMP-1水平呈正相关。结论： 慢性牙周炎患者唾液中miR-146a表达量异常增高,且与龈沟炎症程度、牙周损伤程度一致。     

Cystatin A in gingival crevicular fluid of periodontal patients

Cystatins are physiological inhibitors of cysteine proteinases which are widely distributed in human tissues and fluids. In the present study we analysed both the cystatin activity and the different cystatin isoforms in gingival crevicular fluid and saliva samples of nine periodontitis patients. All crevicular fluid samples, which were collected with filter paper points, showed cystatin activity ranging from 7–67 units/ing protein. The mean cystatin activity (24 units/mg protein) was significantly lower (p < 0.05) than that of the saliva samples (mean 93 units/mg protein). The cystatin isoforms in the crevicular fluid were further characterized by immunoblotting with specific antibodies against cystatin C, S, SN and A. While they were clearly present in saliva, cystatin C, cystatin S and cystatin SN could not be detected in any of the crevicular fluid samples. Remarkably, cystatin A was found in all the crevicular fluids as well as in the saliva samples. It is concluded that the cystatin activity found in crevicular fluid is caused, at least partially, by cystatin A. Furthermore, the gingival crevicular fluid is not a major contributor of cystatin C, S and SN activity in saliva.     

The gingival crevicular fluid ciprofloxacin level in subjects with gingivitis and periodontitis, and its effects on clinical parameters

OBJECTIVES: The purpose of this study, conducted on patients with gingivitis and periodontitis, was twofold: to find out the serum and gingival crevicular fluid concentration of ciprofloxacin, which is a common drug used effectively against Actinobacillus actinomycetemcomitans and to determine the effects of ciprofloxacin administration on clinical parameters. METHOD: A total of 32 adult patients, consisting of 16 subjects with gingivitis and 16 subjects with untreated chronic periodontitis, were included in the study. The subjects were divided into four groups: group I included eight subjects with chronic gingivitis who had not previously received any ciprofloxacin; group II included eight subjects with chronic gingivitis to whom three doses of ciprofloxacin were administered (Siprosan 500 mg) to establish adequate gingival crevicular fluid and serum concentrations of the agent; group III consisted of eight subjects with chronic periodontitis who had not received any ciprofloxacin; group IV included eight subjects with chronic periodontitis to whom three doses of ciprofloxacin were administered to establish adequate gingival crevicular fluid and serum concentrations of the agent. All patients were systemically healthy, free of pain and reported no current medication usage. Each patient was treated with scaling and/or root planing using specific hand instruments under local anesthesia. Gingival index, plaque index and clinical attachment levels of the teeth were used to determine the clinical condition of the subjects and findings were recorded at the beginning, seventh day, 21st day and third month of the study. Serum ciprofloxacin level was measured in venous blood. Approximately 5 ml of venous blood was drawn from subjects in groups II and IV using a standard venipuncture technique. Gingival crevicular fluid samples were sampled from six interproximal sites with six paper strips in the posterior region of upper jaw (excluding third molar) and all gingival crevicular fluid and serum samples were evaluated by high-performance liquid chromatography. RESULTS: The serum concentrations of ciprofloxacin at the first and 72nd hour were not significantly different in subjects with periodontitis compared to subjects with gingivitis. But the gingival crevicular fluid concentrations of ciprofloxacin at the same hours were significantly high in subjects with periodontitis compared to subjects with gingivitis. Both subjects with gingivitis and periodontitis had significantly higher ciprofloxacin levels in the gingival crevicular fluid than in serum. The application of ciprofloxacin did not have any positive or statistically significant effect upon the clinical parameters of the subjects with gingivitis. On the other hand, a significant decrease in the clinical attachment level scores of the subjects with periodontitis (group IV) was observed compared to group III in the 21st day and third month. CONCLUSION: According to these results, the use of ciprofloxacin as an alternative drug in subjects with periodontitis but not gingivitis can be recommended. However, long-term studies are also needed to assess the effects of ciprofloxacin on clinical parameters.     

Levels of Aggregatibacter actinomycetemcomitans,Porphyromonas gingivalis,inflammatory cytokines and species‐specific immunoglobulin G in generalized aggressive and chronic periodontitis

Casarin RCV, Del Peloso Ribeiro É, Mariano FS, Nociti FH Jr, Casati MZ, Gonçalves RB. Levels of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, inflammatory cytokines and species‐specific immunoglobulin G in generalized aggressive and chronic periodontitis. J Periodont Res 2010; 45: 635–642. © 2010 John Wiley & Sons A/S Background and Objective: Aggressive periodontitis pathogenesis still is not completely understood in the literature regarding the relationship between microbial and inflammatory aspects. So this study aimed to compare microbial and inflammatory patterns in the gingival crevicular fluid of generalized aggressive and chronic periodontitis patients. Material and Methods:  Forty aggressive and 28 chronic periodontitis patients were selected. Biofilm and gingival crevicular fluid were collected from a deep pocket (periodontal probing depth >7 mm) and a moderate pocket (periodontal probing depth = 5 mm) of each patient, and microbiological and immunoenzymatic assays were performed. Real‐time PCR was used to determine quantities of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Enzyme‐linked immunosorbent assay (ELISA) was employed to determine gingival crevicular fluid levels of interleukin‐1β, interferon‐γ, prostaglandin E₂ and interleukin‐10. In addition, immunoglobulin G (IgG) levels against A. actinomycetemcomitans and P. gingivalis lipopolysaccharide were also determined by ELISA. Analysis of variance/Tukey test, Mann–Whitney U‐test and the Pearson correlation test were used to determine differences and correlations between variables analysed (α = 5%). Results:  Patients suffering from generalized aggressive periodontitis had their mouth colonized by higher amounts of A. actinomycetemcomitans and P. gingivalis than chronic periodontitis patients. Conversely, the gingival crevicular fluid levels of IgG against both pathogens were statistically inferior in aggressive periodontitis patients (p < 0.05). With regard to gingival crevicular fluid levels of cytokines, aggressive periodontitis patients presented reduced levels of interleukin‐10 (p < 0.05). Conclusion:  In comparison to chronic periodontitis, generalized aggressive periodontitis patients have an imbalance in the host response, with reduced levels of interleukin‐10 and IgG, and increased periodontal pathogens.     

Distribution of systemic clarithromycin to gingiva

Background: Aggressive and recurrent forms of periodontitis are associated with infections by Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) and Porphyromonas gingivalis. Because these pathogens invade tissue, they are difficult to eliminate by root planing alone. The use of systemic antibiotics in conjunction with root planing significantly enhances clinical and microbiologic treatment outcomes. Although it is not widely prescribed by periodontists, clarithromycin is potentially useful because it is taken up by host cells and has favorable antimicrobial activity. Methods: Experimental gingivitis was induced in eight healthy subjects at one randomly selected maxillary posterior site. The contralateral maxillary site served as the healthy control. Thereafter, subjects were administered six doses of clarithromycin, 500 mg, every 12 hours. Blood was then drawn, and samples of gingiva were harvested from both sites. The samples were extracted, and clarithromycin content was analyzed by liquid chromatography. Results: Mean clarithromycin concentrations in healthy control and inflamed gingiva (2.4 and 3.0 mug/g, respectively) were significantly higher than in serum (0.5 mug/ml; P <0.05). Clarithromycin levels at control and gingivitis sites were higher than serum by 5.7- and 7.0-fold, respectively (difference between sites was significant; P = 0.02). At control sites, a significant decrease in gingival crevicular fluid flow rate was evident at the conclusion of the clarithromycin regimen (P = 0.018). Conclusions: Clarithromycin can attain higher levels in gingiva than serum and reach higher levels in inflamed gingiva than in healthy gingiva. Its distribution profile seems to be suitable for the treatment of periodontitis. The reduction in crevicular fluid flow at control sites suggested that clarithromycin may produce anti-inflammatory effects.     

Progressive periodontal disease has a simultaneous incremental elevation of gingival crevicular fluid and serum CRP levels

Aim: Increased C‐reactive protein levels have been found in all active inflammations, including periodontitis. This study aims to assess the C‐reactive protein levels in periodontal disease progression. Methods: Forty‐five patients were divided into the following three groups (n = 15) based on gingival index, probing pocket depth, and clinical attachment level: healthy (group I), gingivitis (group II), and chronic periodontitis (group III). Gingival crevicular fluid and serum samples were quantified for C‐reactive protein using enzyme‐linked immunosorbent assay. Results: The mean C‐reactive protein concentration in gingival crevicular fluid and serum was found to be highest in group III (1233.33 ng/mL for gingival crevicular fluid, 5483.33 ng/mL for serum), and least in group I (60 ng/mL and 413 ng/mL for gingival crevicular fluid and serum, respectively) The mean C‐reactive protein concentration in group II (453.33 ng/mL for gingival crevicular fluid and 3565.33 ng/mL for serum) was found to be intermediate. Conclusions: C‐reactive protein levels in gingival crevicular fluid and serum increased proportionately with the severity of periodontal disease. They correlated positively with clinical parameters, including gingival index, probing pocket depth, and clinical attachment level. Thus, it can be considered as a periodontal inflammatory biomarker and deserves further consideration.     

Immunoglobulin-degrading enzymes in localized juvenile periodontitis

Previous reports have indicated the association of periodontal diseases with elevated levels of serum immunoglobulin G (IgG) antibodies to periodontally relevant bacteria. Recent results from this laboratory suggest that enzymes proteolytic for immunoglobulins are important virulence factors of several periodontal bacteria. Specifically, enzymes from Porphyromonas (Bacteroides) gingivalis culture supernatant fluid (SF) cleaved human IgG (4 subclasses), IgA1 and IgA2, IgM, IgD and IgE. Proteolytic enzymes from Actinobacillus actinomycetemcomitans culture SF cleaved IgG, IgA and IgM. An enriched Ig proteolytic preparation from Capnocytophaga ochracea culture SF was shown to extensively cleave all 4 subclasses of human IgG. Extensive degradation of IgG and IgA in crevicular fluid samples on SDS-PAGE from periodontal disease sites of localized juvenile periodontitis (LJP) patients in comparison to little degradation in healthy sites indicated the potential role the proteolytic enzymes from periodontopathogenic bacteria may play in situ. Treatment of IgG with P. gingivalis, A. actinomycetemcomitans and C. ochracea SF resulted in similar patterns of degradation. LJP patients had significantly higher levels of IgG and IgA proteolytic activity in whole saliva than age-, sex-, and race-matched periodontal disease-free controls. However, not all of the proteolytic activity could be ascribed to bacterial proteases since neutrophils are also present in large numbers at diseased sites. Using similar techniques, lysates of neutrophils from healthy controls cleaved IgG, IgA and IgM. The observation of enhanced Ig cleavage activity in crevicular fluid and saliva in LJP patients suggest a role for Ig proteolytic enzymes in LJP.     

Full-mouth profile of active MMP-8 in periodontitis patients

Kraft‐Neumärker M, Lorenz K, Koch R, Hoffmann T, Mäntylä P, Sorsa T, Netuschil L. Full‐mouth profile of active MMP‐8 in periodontitis patients. J Periodont Res 2012; 47: 121–128. © 2011 John Wiley & Sons A/S Background and Objective: MMP‐8 in gingival crevicular fluid is considered as a protease with high destructive potential because of its ability to degrade collagen in periodontitis‐affected patients. The aim of this study was to investigate whether there was a relationship between clinical diagnostic parameters and the concentration of active MMP‐8 (aMMP‐8) in gingival crevicular fluid in a site‐level full‐mouth analysis. Based on these data, the prognostic value of aMMP‐8 levels in relation to pocket depth may be evaluated. Material and Methods: Clinical measurements of pocket depth, bleeding on probing (BOP), plaque index (PlI) and gingival index (GI), as well as samples of gingival crevicular fluid, were obtained from four sites of each tooth of nine healthy female patients with chronic generalized periodontitis. The aMMP‐8 concentration in gingival crevicular fluid was quantified by ELISA using specific monoclonal antibodies. Multiple linear regression models for the single measures of aMMP‐8 and pocket depth were calculated with GI and BOP as additional variables. Results: Between 92 and 112 recordings were obtained for each parameter in each patient. Mean values of between 31.5 and 88.8% were calculated for pocket depths of ≥ 4 mm. Mean pocket depths ranged from 3.11 to 4.73 mm, the mean BOP values ranged from 34.0 to 96.7% and the mean full‐mouth gingival crevicular fluid aMMP‐8 concentration ranged from 3.2 to 23.7 ng/mL. Conclusion: In this sample of female periodontitis patients, a broad range of intra‐individual and interindividual aMMP‐8 values was found. Although the explained variance was rather weak, a statistically significant relationship between aMMP‐8 and pocket depth was proven.     

Gingival crevicular fluid and plasma levels of neuropeptide Substance-P in periodontal health, disease and after nonsurgical therapy

Background and Objective: The level of Substance‐P in gingival crevicular fluid has been found to correlate with clinical measures of periodontal disease. The present study was designed to assess the relationship between clinical parameters and levels of Substance‐P in the gingival crevicular fluid from inflamed gingiva, periodontitis sites and after treatment of periodontitis sites, and to correlate them to the Substance‐P levels of plasma. Material and Methods: Thirty, age‐ and gender‐matched subjects were divided into three groups (healthy, gingivitis and chronic periodontitis) based on modified gingival index scores and clinical attachment loss. A fourth group consisted of 10 subjects from the periodontitis group, 6–8 wk after initial therapy. Plasma and gingival crevicular fluid samples were collected and quantified for Substance‐P using an enzyme immunoassay. Results: The mean concentration of Substance‐P, both in gingival crevicular fluid and plasma, was observed to be highest in the periodontitis group (45.13 pg/mL in gingival crevicular fluid and 67.8 pg/mL in plasma) and lowest in the healthy group (6.07 pg/mL in gingival crevicular fluid and below the detection level in plasma). The mean Substance‐P concentration in the gingivitis group (11.42 pg/mL in gingival crevicular fluid and 38.8 pg/mL in plasma) and in the after‐treatment group (7.58 pg/mL in gingival crevicular fluid and 39.7 pg/mL in plasma) lay between the highest and lowest values. In all groups the gingival crevicular fluid levels showed a statistically significant positive correlation with that of plasma and clinical attachment loss. Conclusion: Substance‐P levels were highest in the gingival crevicular fluid from sites with periodontal destruction; however, periodontal treatment resulted in the reduction of Substance‐P levels. Gingival crevicular fluid and plasma Substance‐P levels showed a positive correlation in all of the groups.     

Crevicular Porphyromonas gingivalis‐specific immunoglobulin A levels in healthy and periodontitis‐affected Thai cohorts

Abstract Aim: Immunoglobulin A is a key humoral immune component involved in defense mechanisms against infections. Periodontitis, the chronic inflammatory disease causing periodontal destruction, adversely affects adults worldwide, including Thailand. As the development of periodontitis is partly mediated by immune components, levels of total and Porphyromonas gingivalis‐specific immunoglobulin A in gingival crevicular fluid of Thai cohorts were studied. Methods: Gingival crevicular fluid was collected from 24 patients with severe generalized chronic periodontitis and 22 healthy controls. The amount and concentration of total and Porphyromonas gingivalis‐specific immunoglobulin A in each gingival crevicular fluid sample were determined by enzyme‐linked immunosorbent assay. Results: The control group contained the highest concentrations of both types of gingival crevicular fluid–immunoglobulin A, but the lowest levels of these antibodies were found in the deep sites of the periodontitis group. Moreover, the concentrations of gingival crevicular fluid–immunoglobulin A and the degree of periodontitis severity appeared to have an inverse relationship. There was no significant difference in the amounts of gingival crevicular fluid–immunoglobulin A in the control and periodontitis groups. Conclusions: This study supports the hypothesis that high concentrations of specific gingival crevicular fluid–immunoglobulin A antibodies directed against Porphyromonas gingivalis, a potent periodontic microorganism, could retard periodontitis development.     

Human gingival crevicular fluid keratin at healthy, chronic gingivitis and chronic adult periodontitis sites

Abstract The present study was designed to determine, in a cross-sectional study, whether there was any relationship between the keratin-positive material in gingival crevicular fluid and the clinical periodontal status. Keratins were selected as putative indicators of degradation of epithelial cells cytoskeletal proteins. Keratin positive material was determined by enzyme-linked immunosorbent assay-in 42 subjects exhibiting clinical sites of health, chronic gingivitis and chronic periodontitis. The concentration of keratin in parotid saliva was also measured for each subject. Keratin concentration in gingival crevicular fluid samples was significantly greater at sites exhibiting signs of gingivitis and periodontitis compared with healthy sites. No differences were detected between sites exhibiting gingivitis and periodontitis. No differences were found between the 3 groups for the saliva keratin-positive material which was significantly less than that detected in gingival crevicular fluid. These results suggest that gingival crevicular fluid keratin concentration may serve as a marker of gingival damage.     

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.     

Specific antibodies against Actinobacillus actinomycetemcomitans in serum and saliva of patients with advanced periodontitis

The aim of the present study was to discover any possible correlation between specific antibodies against Actinobacillus actinomycetemcomitans (A.a.) in serum and saliva. The test group consisted of 38 patients aged 31–68 yr (mean 49) with advanced periodontitis. Twenty-nine subjects aged 23–67 yr, without periodontal destruction, formed a control group with a reference level of specific salivary antibodies against A.a. A subgingival plaque sample for culturing A.a. , a specimen of stimulated whole saliva, and a sample of venous blood were taken from each subject of the test group. Specific IgG and IgA antibodies against A.a. were determined from serum and stimulated whole saliva by means of the ELISA test. Fifteen of the patients (39%) had cultivable A.a. Six of the 15 A.a. culture-positive patients and one of the 29 reference subjects exhibited very high antibody titers against A.a. in saliva. Specific IgG and IgA antibodies in saliva correlated highly significantly with the corresponding antibody values in serum among the patients in the test group. It was concluded that among patients with severe adult periodontitis, the less invasive saliva sample has a diagnostic value equal to that of the serum sample concerning specific antibodies against A.a.     

Plasma and crevicular fluid osteopontin levels in periodontal health and disease

BACKGROUND AND OBJECTIVE: The level of osteopontin in gingival crevicular fluid has been found to correlate with clinical measures of periodontal disease. The present study was designed to assess the relationship between clinical parameters and osteopontin levels of the gingival crevicular fluid from inflamed gingivae, periodontitis sites and after treatment of periodontitis sites, and to correlate them to the osteopontin levels of the plasma. MATERIAL AND METHODS: Thirty, gender-matched subjects were divided into three groups--healthy, gingivitis and chronic periodontitis--based on modified gingival index scores and clinical attachment loss. The fourth group consisted of 10 subjects in the periodontitis group, 6-8 wk after initial therapy. Plasma and gingival crevicular fluid samples were collected and quantified for osteopontin using an enzyme immunoassay. RESULTS: The highest mean gingival crevicular fluid and plasma osteopontin concentrations were observed in the periodontitis group (1575.01 and 1273.21 ng/mL, respectively) and the lowest in the healthy group (1194.80 and 476.35 ng/mL, respectively). After treatment of the periodontitis group, the level of osteopontin decreased to 1416.15 in gingival crevicular fluid and to 1051.68 ng/mL in plasma. In all groups the gingival crevicular fluid osteopontin levels showed a statistically significant positive correlation with that of plasma and clinical attachment loss. CONCLUSION: Osteopontin levels were highest in the gingival crevicular fluid from sites with periodontal destruction; however, periodontal treatment resulted in the reduction of osteopontin levels. Gingival crevicular fluid and plasma osteopontin levels showed a positive correlation in all of the groups.