Subgingival temperature: relation to gingival crevicular fluid enzymes, cytokines, and subgingival plaque micro-organisms.

There have been no reports on the relationship of subgingival temperature to specific gingival crevicular fluid (GCF) components. Therefore, the purpose of this cross-sectional study was to determine whether there was any relationship between subgingival temperature and GCF levels of neutrophil elastase (NE), myeloperoxidase (MPO), beta-glucuronidase (BG), interleukin-1 alpha (IL-1), and interferon alpha (IFN). Furthermore, another objective was to confirm an association of subgingival temperature with clinical parameters and specific subgingival plaque micro-organisms as has been reported earlier. 27 human subjects each having healthy (n = 50), gingivitis (n = 59) and periodontitis (n = 53) sites were evaluated. The plaque index (PI), subgingival temperature, probing depth, attachment loss, bleeding index and gingival index were measured. GCF was sampled following the measurement of the PI and removal of the supragingival plaque. GCF samples were assayed for the enzymes NE, BG, MPO and the cytokines IFN-alpha and IL-1 alpha. A sterile Gracey curette was utilized at each sampled site to collect subgingival plaque. The plaque samples were evaluated using an immunoassay. Subgingival temperature was found to directly correlate with all clinical parameters (p < 0.001). Significant, albeit not large, correlations were found between subgingival temperature and NE (r = 0.35, p < 0.001), MPO (r = 0.26, p < 0.001) and BG (r = 0.23, p < 0.01). Temperature was found to correlate positively with E. corrodens (r = 0.33, p < 0.02) and F. nucleatum (r = 0.25, p < 0.05) but not with P. intermedia (r = 0.02, p = 0.9), P. gingivalis (r = 0.20, p = 0.1) and A. actinomycetemcomitans (r = 0.01, p > 0.9). In conclusion, subgingival temperature is correlated with the GCF enzymes, NE, MPO and BG as well as the clinical parameters and specific plaque micro-organisms associated with periodontal disease.

Risk indicators for periodontal disease in a racially diverse urban population.

A cross-sectional study of 117 subjects from a dental clinic serving a diverse population (i.e., Whites, African-Americans, Native-Americans, and Asians) was performed to evaluate risk indicators of periodontal disease. Gingival crevicular fluid (GCF) and subgingival plaque were taken at the same visit from 4 posterior sites of the most diseased sextant in each subject. Age, smoking packyears, beta-glucuronidase (beta G), neutrophil elastase (NE), myeloperoxidase (MPO), Fusobacterium nucleatum (F. nucleatum), and Porphyromonas gingivalis (P. gingivalis) were significantly (p < 0.05-0.005) correlated with attachment loss. Probing depth was significantly correlated with smoking packyears, beta G, NE, MPO, F. nucleatum and Prevotella intermedia (P. intermedia) (p < 0.05-0.005). Mean NE value of Whites was lower than the mean NE values of African-Americans, Native-Americans and Asians (p < 0.05). Whites had a lower mean beta G value compared to African Americans, and a lower mean MPO value compared to African Americans and Native Americans. The %s of patients positive for F. nucleatum, P. intermedia and Eikenella corrodens (E. corrodens) were higher in Native Americans compared to Whites. Step-wise multiple regression analysis was performed to construct models for the estimation of probing depth and attachment loss. The most parsimonious regression models which had the best R2 values included the following variables and accounted for the indicated % of variability: models 1 and 2: beta G, race, and F. nucleatum accounted for 50% of the variability in mean probing depth and 39% of the variability in a single site (first molar) for probing depth, respectively; model 3: age, beta G, and F. nucleatum accounted for 53% of the variability in mean attachment loss; model 4: age, NE, and F. nucleatum explained 35% of the variability in a single site (first molar) for attachment loss. The results suggest that age, race, smoking packyears, beta G, NE, MPO, F. nucleatum, P. gingivalis and P. intermedia are risk indicators for periodontal disease in this racially diverse urban population. Regression models which include multiple variables (i.e., demographic factors, GCF enzymes and periodontopathic bacteria) can be used to estimate periodontal disease status.

Crevicular fluid enzymes from endosseous dental implants and natural teeth.

Several neutrophil-derived enzymes that are present in the gingival crevicular fluid have been evaluated for use as risk markers for periodontal disease progression. However, very little information is available about the presence of these enzymes in peri-implant tissues. The purpose of this cross-sectional study was to compare levels of enzymes in gingival crevicular fluid between natural teeth and endosseous dental implants and between well-integrated and failing implants. Scores of plaque and gingivitis were recorded for 68 integrated implants, five failing implants, and 34 natural teeth in 12 completely edentulous and 18 partially edentulous subjects. Samples of gingival crevicular fluid were obtained from these sites using filter paper strips and were assayed for levels of neutral protease, neutrophil elastase, myeloperoxidase, and beta-glucuronidase. Neutral protease levels were higher (P = .066) at moderately to severely inflamed implant sites (Gingival Index of 2, 3) compared to mildly or noninflamed sites (Gingival Index of = 0, 1). Despite the small number (n = 5) of failing implants evaluated in this study, levels of neutrophil elastase, myeloperoxidase, and beta-glucuronidase were significantly higher (P < or = .001) around failing implants compared to successful implants. Neutral protease levels were also elevated around failing implants, but the difference was not statistically significant. Results of this study indicate that neutrophil elastase, myeloperoxidase, and beta-glucuronidase levels in GCF appear to be good candidates for study as risk markers of implant failure.

Neutrophil elastase in crevicular fluid: comparison of a middle-aged general population with healthy and periodontitis groups.

Neutrophil elastase (NE) was measured in crevicular fluid (GCF) collected from 3 subject groups. GCF was harvested at a single visit of subjects with periodontal health (n = 21) and with periodontitis (n = 28). Samples were obtained from 132 middle-aged, middle-class health conscious patients of a health maintenance organization (HMO) at baseline and 1 year later. GCF NE was higher in periodontitis than in health. Mean GCF NE of HMO subjects was much closer to health than to periodontitis. Few members of the HMO population had enzyme levels typical of periodontitis. Subjects and sites of the HMO population were segregated into 3 categories based on enzyme levels of the healthy and periodontitis subjects. Most HMO subjects and sites were in the activity category corresponding to healthy subjects. Only a small portion were in the activity category common in periodontitis. Enzyme levels in the highest activity category at both samplings were infrequent. High enzyme levels in the HMO population were not associated with attachment loss. Thus, assay of GCF NE provided little evidence of disease in a middle-aged, middle-class health conscious general population. This finding confirms an analysis of epidemiological surveys which concluded that a population such as studied here would not benefit from periodontal diagnostic testing.

Development of a risk profile for periodontal disease: microbial and host response factors.

Advances in our understanding of the relationship between the microbial challenge and the host response in periodontal disease have led to the search for pathogenesis-based risk indicators or risk factors for disease progression. This evaluation is based on analysis of non-invasive or minimally invasive samples that allow measurement of the subgingival plaque microflora or the host response in gingival crevicular fluid (GCF), serum, or saliva. Studies conducted by us have indicated that in GCF, persistently elevated levels of beta-glucuronidase (beta G, a marker for primary granule release from polymorphonuclear leukocytes) are associated with clinical attachment loss in patients with periodontitis. This finding has been confirmed in a multicenter trial. We have also observed that a statistically significant positive correlation exists between beta G in GCF and measures of the subgingival microbial challenge, but the correlation was less than 0.5, suggesting variations in the host response to the challenge. Furthermore, beta G levels in GCF were inversely correlated with the IgG serum antibody titer to a panel of periodontal pathogens, suggesting the essentially protective function of the systemic humoral response in periodontal disease. Data in the literature support this concept. In addition, recent studies of the relationship of antibody isotypes in GCF to progression of clinical attachment loss have suggested that IgA in GCF has a protective function. This may relate to the lack of complement activation by IgA. Alternately, the development of IgA antigen-specific responses are T-cell dependent, and reductions in local levels of IgA may indicate a decrease in T-helper cell function. These data have allowed development of strategies for identifying individual risk profiles for patients with periodontal disease based on the host response to the microbial challenge. With identification of these risk indicators/risk factors for active periodontal disease, the next challenge is to provide clinicians with access to the tests and analyses that are required for this approach to periodontal diagnosis. Improved patient management should result from the incorporation of these tests into clinical practice.

Inhibition of crevicular fluid neutrophil elastase by alpha 1-antitrypsin in periodontal health and disease.

Gingival crevicular fluid (GCF) was collected from two healthy, two gingivitis and two periodontitis sites of two groups of individuals presenting for treatment of chronic adult periodontitis (group 1, 25 subjects; group 2, seven subjects) and from distal approximal sites of two incisors and one molar of 10 subjects with periodontal health. GCF eluates of periodontitis group 1 and controls, prepared by a technique that lysed polymorphonuclear leucocytes (PMN) in the samples, were assayed for functional neutrophil elastase (NE) and immunoreactive alpha 1-antitrypsin (alpha 1-AT) and alpha 1-antitrypsin-neutrophil elastase complex (alpha 1-AT-NE). Periodontitis group 2 GCF eluates, generated by a method that did not disrupt PMNs, were assayed for functional NE in the presence and absence of a specific NE inhibitor. A greater amount of NE (ng/5-s sample) was found in eluates of GCF from diseased sites irrespective of whether or not the eluates contained products of lysed PMNs. However, the GCF eluates prepared without disrupting PMNs contained only about one-tenth as much NE as eluates of corresponding sites that included constituents of lysed PMNs. The amount of alpha-AT in GCF was insufficient to inactivate most of the NE available for release into the gingival sulcus at either healthy or diseased sites. In addition, much of the alpha 1-AT in GCF was not complexed with NE under conditions of excess NE. More than 90% of the NE in GCF from each site category was inactivated by the NE specific inhibitor. It is concluded, because of the large quantity of NE available in PMNs compared to the amount of NE inhibitors in GCF, that at least locally transient free NE occurs, which contributes to tissue destruction in chronic adult periodontitis.

Salivary amylase in crevicular fluid.

Extrasulcular substances such as saliva, supragingival plaque and salivary sediment may be contaminants in gingival crevicular fluid (GCF) collected with Periopaper. This report provides data obtained with salivary amylase as a marker for these substances in GCF. Amylase was a common constituent of GCF collected from sites with clinical health and with clinical signs of periodontitis. Rinsing the mouth with water reduced, but did not eliminate amylase in GCF. More frequent (p < 0.01) and greater (p < 0.001) contamination of GCF with amylase occurred in samples from periodontitis than from healthy subjects. The volume of saliva required to give the amylase in the GCF was calculated. This volume exceeded the GCF volume in 21% of samples collected without a water rinse. Thus, oral constituents other than saliva likely contribute to GCF amylase. Small quantities of plaque and salivary sediment (9.6 +/- 5.9, 3.4 +/- 2.0 micrograms protein) provided amylase from a saliva volume equal to the GCF volume in health (0.23 microliters). The above and other data presented here show that extrasulcular substances likely are frequent constituents of GCF collected with Periopaper. Reporting GCF constituents as quantities/sample appears least subject to error from the contamination by extrasulcular substances.

Five parameters of gingival crevicular fluid from eight surfaces in periodontal health and disease.

Volume and amounts of myeloperoxidase (MPO), lactoferrin (LF), aryl sulfatase (AS) and lactate dehydrogenase (LDH) were measured in gingival crevicular fluid (GCF) collected from the mesial and distal proximal surfaces of the premolars and first and second molars of 3 subject groups. Group assignment was based on subject mean gingival index (GI) and probing depth (PD) of sampled sites as follows: healthy, GI less than or equal to 0.5, PD less than or equal to 3.0; disease 1, GI greater than or equal to 1.0, PD greater than or equal to 3.0 mm; disease 2, PD greater than or equal to 4.0 mm. Attachment loss (ATL) of most sites in the 3 groups was: healthy, 0-1 mm; disease 1, 1-2 mm; and disease 2, 4-9 mm. GCF volume differed among surfaces and teeth in each of the 3 groups. The greater amount of GCF collected from posterior locations was not related to the GI and PD. Differences with sampling location in amounts of GCF constituents were restricted to MPO and LF. Most of these differences (greater amounts at posterior sites) were associated with more severe disease. Variability in amount and composition of GCF collected from different sites, therefore, should be considered in experiments which include quantitation of GCF parameters. The ratio of MPO in disease group 2 to disease group 1 was greater than similar ratios for GCF volume and LF, AS and LDH. The quantity of MPO was the only measure which differed between the 2 disease groups at all surfaces. MPO thus appears to have the greatest potential, among the measured parameters, to serve as a marker for advanced periodontal disease.

Repeated measurement of crevicular fluid parameters at different sites.

Few systematic studies have been made of amounts or of the composition of gingival crevicular fluid (GCF) from different sites or of the stability of GCF parameters over time. These data are needed to better understand the relation of GCF composition to periodontal health status. This report gives the volume and the amounts of lactate dehydrogenase (LDH), aryl sulfatase (AS) and neutrophil elastase (NE) for GCF collected from 6 samplings of 6 standard gingival sites in 11 young adult subjects over a 6-week period. Attachment loss (3 mm) was noted at only 1 of the 66 sites. The mean gingival index of the 11 subjects ranged from 0.33 to 1.67. The GCF volume and activity/sample of LDH and AS but not NE differed among subjects. However, differences among subjects were not found when the GCF enzyme activities were expressed as activity/microliter GCF. GCF volume and LDH, AS and NE activity all differed among the 6 sites when the activities were expressed as either quantity/sample or microliter GCF. These data show that differences among sites must be carefully considered in evaluation of GCF data. Fluid volume and LDH, AS and NE activity all varied from sampling to sampling. However, differences among sites were retained throughout the experimental period.

Crevicular fluid myeloperoxidase at healthy, gingivitis and periodontitis sites.

The volume and myeloperoxidase (MPO) activity of gingival crevicular fluid (GCF) collected with filter paper strips for 30 s from the sulcus of healthy, gingivitis and periodontitis sites of Chinese subjects were measured. MPO/site and MPO/microliter GCF were both greater at gingivitis and periodontitis sites than at healthy sites. Enzyme activity was similar at the 2 categories of diseased sites. Mean GCF volume and MPO activity were calculated for all samples from healthy, gingivitis and periodontitis sites with GI 0, 1, 2 and 0 + 1. GCF volume, MPO/site and MPO/microliter GCF all were greater at GI 2 than GI 0 or 0 + 1. These data indicate that increased GCF MPO previously observed at periodontitis sites is not specific to such sites. Rather increased GCF MPO likely occurs when additional polymorphonuclear leukocytes enter the sulcus as a result of gingival inflammation. A second sample was obtained from 22 sites 4 weeks after the initial collection. These samples were collected for 5 s rather than 30 s. The GCF volume, MPO/site and MPO/microliters GCF were each greater in samples collected for 30 s rather than 5 s. Correlation coefficients showed that the amount of GCF and MPO activity of the fluid collected for 5 s and 30 s was dependent upon the site even though the 5-s and 30-s samples were collected 4 weeks apart.

Relationship between lactate dehydrogenase and myeloperoxidase levels in human gingival crevicular fluid and clinical and microbial measurements.

The present study was designed to determine, in a cross-sectional study, whether there was any relationship between levels of lactate dehydrogenase (LDH) and myeloperoxidase (MPO) in gingival crevicular fluid (GCF) and clinical periodontal status or microbial parameters. Another objective was to determine, in a longitudinal study, the effect of a single session of root planning on GCF levels of LDH and MPO and the relation to changes in clinical and microbial measurements. 15 and 12 test subjects with moderate to severe periodontal disease were seen in the cross-sectional and longitudinal study, respectively. 1 healthy and 2 diseased sites were evaluated in each subject. Higher LDH and MPO levels in GCF were closely associated with higher clinical and microbial signs of periodontal disease. Root planing was effective in reducing these enzymes in GCF, with an accompanying decrease in clinical and microbial signs associated with disease. The return of LDH to baseline levels at 3 months after instrumentation, without a corresponding return of clinical signs of disease, may serve as a marker for subclinical periodontal pathology.

Phenytoin and 5-(p-hydroxyphenyl)-5-phenylhydantoin do not alter the effects of bacterial and amplified plaque extracts on cultures of fibroblasts from normal and overgrown gingivae.

Local irritation of gingival tissue by plaque is among the factors which affect development of gingival overgrowth in patients undergoing chronic phenytoin (PHT) therapy. Variability in the cytotoxicity of plaque components or of plaque substances plus PHT and/or its metabolites toward gingival fibroblasts may relate to whether gingival overgrowth forms in a particular patient. Fibroblasts from healthy and overgrown gingivae were incubated with (a) PHT and its major human metabolite, 5-(p-hydroxyphenyl)-5-phenylhydantoin (HPPH), (b) microbial and "amplified" plaque extracts, and (c) microbial and "amplified" plaque extracts plus PHT and HPPH. Cell numbers and cell-associated protein were determined for each incubation preparation. A wide range in cytotoxic response to a particular microbial or plaque extract occurred among cell strains. Plaque extracts from different subjects had variable cytotoxicity toward a cell strain. The differences among fibroblast strains in response to an extract and the variability in cytotoxicity of different plaque extracts toward a cell strain were not related to their source from normal or overgrown gingivae. Cell numbers and cell-associated protein were similar for incubation mixtures containing extracts with and without PHT and HPPH. These data do not show differences among cytotoxicity levels of plaque extracts, the response of particular gingival fibroblast strains to plaque components, or interaction between drugs and certain plaque samples which explain development of gingival overgrowth in some subjects receiving chronic PHT therapy.

Psychiatric issues of adolescent chemical dependence.

Pediatricians, as well as other health care professionals, must always consider psychoactive drug use by adolescents as possible causes of general health or emotional problems, such as poor appetite, insomnia, problem behavior, academic underachievement, and loss of energy and motivation. Early diagnosis and intervention with youth can result in enormous benefits for the lives of the youth, families, and society. The physician must be alert to the adolescent who manifests such signs of abuse. This article details the recognition and management of these problems.

Relationships between levels of lysozyme, lactoferrin, salivary peroxidase, and secretory immunoglobulin A in stimulated parotid saliva.

Recent studies suggest that salivary lysozyme (Lz), lactoferrin (Lf), peroxidase (Spx), and secretory immunoglobulin A (sIgA) may interact in a common antimicrobial system. A multiple protein approach therefore may be needed to determine the role of this system in oral health and ecology. In the present study we investigate the relationships between levels of Lz, Lf, Spx, and sIgA (adjusted for flow rate and total protein) in stimulated parotid saliva from 44 dental students. Principal components analysis was used to determine major patterns of intercorrelation between variables; cluster analysis was used to identify groups of subjects with similar salivary profiles for Lz, Lf, Spx, and sIgA. Spx tended to vary independently of Lz and Lf, which, in turn, tended to vary together. sIgA showed a weak negative relationship with Spx and a weak positive relationship with Lz and Lf. Six major clusters of subjects with similar antimicrobial protein profiles were found. These were significantly different at P less than 0.0001. Spx was the most important determinant of cluster membership followed (in order of importance) by Lz, Lf, and sIgA. Cluster profiles were Spx-, sIgAmu, Lf-, Lz-; Spx-, sIgA+, Lfmu, Lz+; Spxmu, sIgAmu, Lfmu, Lzmu; Spx+, sIgA-, Lf-, Lz-; Spx+, sIgAmu, Lf+, Lz-; and Spx+, sIgAmu, Lf+, Lz+ (-, mu, and + refer to the position of the cluster mean each variable relative to the overall mean for that variable). Results suggest that clusters may be a product of independent variation in the secretory activity of acinar and intercalated duct cells.

Correlations between human salivary levels of lysozyme, lactoferrin, salivary peroxidase and secretory immunoglobulin A with different stimulatory states and over time.

Within-subject correlations for the levels of these salivary proteins were determined in unstimulated and stimulated parotid saliva collected from 8 subjects and for stimulated parotid saliva collected from the same subjects once a week for a 4-week period. Initial correlations between unstimulated and stimulated samples were high and statistically significant (p less than 0.05) for all four proteins. When data were adjusted for variation attributable to flow rate and total protein, some correlations remained the same and those for lysozyme, lactoferrin and salivary peroxidase increased. However, the correlation for secretory immunoglobulin A decreased to a point where it was no longer statistically significant. In the weekly comparison, within-subject correlations across weeks were significant (p less than or equal to 0.05) for lysozyme, lactoferrin and salivary peroxidase, but not for immunoglobulin A. After adjustment for flow rate and total protein, the pattern of correlation was unchanged. Thus the relative rankings of subjects for levels of lysozyme, lactoferrin or salivary peroxidase may be consistent across stimulatory states, even though absolute concentrations may change; levels of these proteins in stimulated parotid saliva may be maintained over time. Secretory immunoglobulin A appears to be more subject to short-term variation.

Salivary myeloperoxidase of young adult humans.

Leukocytes, principally polymorphonuclear leukocytes (PMNs), enter the oral cavity where they release a portion of their constituents, including myeloperoxidase, into oral fluids. A greater number of PMNs in the oral cavity are associated with oral inflammation. However, the quantitative contribution of the PMN to oral fluids, including saliva, during various conditions is poorly understood. An assay method based on the adsorbance loss at 278 nm from the reaction of the myeloperoxidase product hypochlorous acid with monochlorodimedon to yield dichlorodimedon was developed for the quantitation of salivary myeloperoxidase. Myeloperoxidase was determined in supernatants of whole saliva obtained at low and moderate flow rates and in parotid saliva collected during moderate and pronounced stimulation from young adults with minimal oral inflammation. The greatest myeloperoxidase activity was in whole saliva supernatants collected at low flow rates where PMN products have an opportunity to accumulate. Lesser quantities of myeloperoxidase were found in both the whole saliva supernatants and parotid saliva obtained at the faster flow rates. Low flow rate whole saliva supernatants contained about 25% of the myeloperoxidase in the PMNs which enter the oral cavity. Myeloperoxidase is responsible for a significant portion (15-20%) of the total peroxidase activity in supernatants of whole saliva obtained at low flow rates. Preliminary results indicate that young adults with phenytoin-associated gingival overgrowth or who smoke have more myeloperoxidase activity in low flow rate whole saliva.