NETs Are Double-Edged Swords with the Potential to Aggravate or Resolve Periodontal Inflammation.

Periodontitis is a general term for diseases characterised by inflammatory destruction of tooth-supporting tissues, gradual destruction of the marginal periodontal ligament and resorption of alveolar bone. Early-onset periodontitis is due to disturbed neutrophil extracellular trap (NET) formation and clearance. Indeed, mutations that inactivate the cysteine proteases cathepsin C result in the massive periodontal damage seen in patients with deficient NET formation. In contrast, exaggerated NET formation due to polymorphonuclear neutrophil (PMN) hyper-responsiveness drives the pathology of late-onset periodontitis by damaging and ulcerating the gingival epithelium and retarding epithelial healing. Despite the gingival regeneration, periodontitis progression ends with almost complete loss of the periodontal ligament and subsequent tooth loss. Thus, NETs help to maintain periodontal health, and their dysregulation, either insufficiency or surplus, causes heavy periodontal pathology and edentulism.

Impaired polymorphonuclear neutrophils in the oral cavity of edentulous individuals.

Oral health is characterized by functional oral polymorphonuclear neutrophils (oPMNs). Edentulism might be associated with a loss of oPMNs because these cells enter the oral cavity primarily through the gingival crevices. The main aim of this study was to investigate the numbers of oPMNs in rinse samples obtained from edentulous (n = 21) and dentate (n = 20) subjects. A second study aim was to investigate possible differences between oPMNs and peripheral blood polymorphonuclear neutrophils (cPMNs). Apoptosis/necrosis and cell-activation markers (CD11b, CD63 and CD66b) were analyzed using flow cytometry. Reactive oxygen species (ROS) production was determined either without stimulation (constitutive) or in response to 10 µM phorbol myristate acetate or Fusobacterium nucleatum. The edentulous subjects presented with lower oPMN counts and higher percentages of apoptotic/necrotic oPMNs compared with dentate subjects. Furthermore, oPMNs from edentulous donors expressed low levels of all three activation markers and low constitutive ROS. In contrast, oPMNs from dentate subjects expressed high levels of all three activation markers and a higher level of constitutive ROS than cPMNs. When challenged, oPMNs from edentulous subjects showed no upregulation in ROS production, whereas oPMNs from dentate subjects retained their ability to respond to stimulation. The functional characteristics of cPMNs were comparable between edentulous and dentate subjects. This study demonstrates that despite having functional cPMNs, edentulous subjects have low oPMN numbers that are functionally impaired.

Assessment of Myeloperoxidase and Nitric Levels around Dental Implants and Natural Teeth as a Marker of Inflammation: A Comparative Study.

INTRODUCTION: Dental implants form the mainstay of dental treatment involving rehabilitation of missing teeth. One of the major concerns for the clinicians doing dental implants is the postsurgical failure of dental implants. Success of dental implants is dependent upon the skills of the surgeon and the amount and quality of the bone remaining at the edentulous area where dental implant has to be placed. Myeloperoxidase (MPO) and nitrites are few of the enzymes and molecules which are said to be altered in inflammation. However, their exact role in the inflammatory processes around natural tooth and dental implant is still unclear. Hence we comparatively evaluated the levels of MPO and nitrites in the areas around the dental implants and natural teeth. MATERIALS AND METHODS: The present study comprises 42 patients who underwent prosthetic rehabilitation by dental implants from 2011 to 2014. Depth of probing value (DP), score of plaque index (SPI), gingival index (GI), and index of gingival bleeding time (GBT) were evaluated for the assessment of the periimplant soft tissue changes. Assessment of inflammation around the dental implant surface and around natural tooth was done based on the readings of these parameters. For the measurement of the MPO levels, spectrophotometric MPO assay was used. All the results were analyzed by Statistical Package for the Social Sciences (SPSS) software. RESULTS: The mean plaque index values were 1.56 and 0.97 in periodontitis cases of natural teeth and inflamed cases of dental implants respectively. While comparing mean plaque index, mean probing depth, and mean gingival bleeding index in between the two groups, significant difference was obtained. Mean MPO concentration in periodontitis and gingivitis cases in natural teeth were 0.683 and 0.875 U/µL, while in inflamed dental implant cases, the mean value was 0.622 U/µL. While comparing the total MPO levels, total nitrite levels, and total nitrite concentration in between two study groups, significant difference was obtained. On comparing the healthy and periodontitis cases in natural teeth, significant difference was obtained. CONCLUSION: In the inflammatory processes occurring around dental implant and natural teeth, MPO and NO make some amount of significant contribution. CLINICAL SIGNIFICANCE: The present study enforces on the role of MPO and nitrite as diagnostic and prognostic marker.

Comparison of proliferation, apoptosis and expression of syndecan-1 and α-SMA in edentulous ridge oral mucosa of successful and early failed submerged dental implants--An immunohistochemical study.

AIMS: Pathogenic mechanisms involved in early submerged implant failure are poorly understood. In this study we immunohistochemically analyse differences in proliferation, apoptosis and inflammation in edentulous ridge oral mucosa (ERM) of successful and early failed submerged implants. MATERIALS AND METHODS: 30 samples of ERM covering successful and early failed submerged implants were obtained at the end of osseointegration period along with control samples of healthy ERM. Sections were stained with Ki-67 (proliferation), caspase-3 (apoptosis) and syndecan-1 (epithelial marker). Percentage of positive cells was analysed by Kruskal-Wallis test and Dunn's post hoc test. Co-localization of Ki-67 and caspase-3 with α-SMA, CD68 and TGF-ß was done by double immunofluorescence. RESULTS: There was no significant difference in number of Ki-67 positive cells within surface epithelium (SE) in all groups. Proliferation was significantly higher in underlying connective tissue (UCT) of ERM of early failed submerged implants (26%) compared to ERM of successful submerged implants (3%) and controls (4%). More apoptotic cells appeared in UCT of early failed submerged implants (8%) compared to UCT of successful submerged implants (4%) and UCT of control ERM (3%). Co-localization of Ki-67 and α-SMA in ERM of early failed submerged implants disclosed proliferating fibroblasts and pericytes of blood vessels. Macrophages and cells expressing TGF-ß appeared in UCT of failed implants. Expression of syndecan-1 was significantly weaker in SE of early failed submerged implants. CONCLUSIONS: Imbalance between proliferation and apoptosis, changes in syndecan-1 expression and inflammation are histopathological features of ERM of early failed submerged implants.

Salivary changes related to systemic diseases in the edentulous patients.

INTRODUCTION: The relatively frequent systemic comorbidities of geriatric patients can be linked to salivary changes, which may induce oral alteration and discomfort with the removable prosthesis. The aim of the study was to evaluate the salivary parameters in completely edentulous patients treated by removable prosthesis, in relation to their general health status. MATERIAL AND METHOD: A cross-sectional study was performed on 30 completely edentulous patients, 53% male and 47% female, aged between 53 and 84. The evaluation of the salivary parameters (oral hydration index, pH and salivary flow, viscosity and saliva buffer capacity) was performed with the Saliva Check Buffer kit (GC Corporation). RESULTS: The salivary changes encountered were the following: low hydration level (63%), high saliva viscosity (57%), below-average pH (27%), reduced salivary flow (77%) and low saliva buffer capacity (80%). A reduced salivary flow and saliva buffer capacity was found in women. A lower buffer capacity of the saliva was found in patients with respiratory and gastro-intestinal disease. CONCLUSIONS: The alterations of the salivary flow are relatively frequent in geriatric patients, removable denture wearers, with compromised systemic status. These changes may be a risk factor for denture stomatitis and oral candidiasis, with a negative effect on the patient's comfort and quality of life.

Salivary concentration of free LL-37 in edentulism, chronic periodontitis and healthy periodontium.

OBJECTIVE: The antimicrobial peptide LL-37, a component of innate immunity, has an important role in maintaining oral health. This study aimed to investigate the concentration of free LL-37 in whole saliva of periodontally healthy, edentulous and chronic periodontitis subjects. DESIGN: Unstimulated whole saliva was sampled from 154 subjects (76 periodontally healthy, 20 edentulous, and 58 subjects with chronic periodontitis). All participants were in good general health. The salivary LL-37 was determined by ELISA. RESULTS: The median salivary concentrations of free LL-37 were 30.5, 22.5, and 1.8ng/ml for the healthy, the chronic periodontitis and the edentulous group, respectively. The differences in concentration between the edentulous and the others were statistically significant (Mann-Whitney test, p<0.001). In the healthy subjects, women displayed significantly higher peptide concentrations compared to men (Mann-Whitney test, p<0.05). The intra-subject variation in LL-37 concentration was wider for the healthy (range 0.75-285ng/ml) and chronic periodontitis patients (range 1-207ng/ml) than for the edentulous subjects (range 0.15-4.4ng/ml). CONCLUSIONS: The present findings show that edentulism correlates with a substantial decrease in salivary levels of free LL-37, thus indicating the considerable contribution of the gingival tissues in the secretion of the peptide in the oral environment.

Wearing of complete dentures reduces slow fibre and enhances hybrid fibre fraction in masseter muscle.

Edentulous conditions and use of complete dentures alter the function of jaw muscles, which is presumably reflected in the myosin heavy chain (MyHC) isoform composition. This study is the first dealing with MyHC isoforms expression in edentulous persons with the aim to clarify to which extent the decreased functional load following teeth loss contributes to the changed muscle phenotype during ageing. We analysed MyHC expression in old masseter muscle at decreased and full functional load by comparing age-matched edentulous and dentate subjects. Edentulous subjects had upper and lower complete dentures. Dentate subjects had at least 24 natural teeth in continuous dental arches with two molars present in each quadrant and normal intermaxillary relationship. The adaptive response to the reduced masticatory load was lower numerical and area proportion of MyHC-1 expressing fibres and higher numerical proportion of hybrid fibres in edentulous compared with dentate subjects with no significant difference in the proportion of MyHC-neo-expressing fibres between both groups. We conclude that the observed differences in the proportion of fibre types between denture wearers and dentate subjects cannot be ascribed to degenerative changes intrinsic to the ageing muscle, but to functional differences in muscle activity and to morphological alterations of stomatognathic system accompanying the complete teeth loss.

Exploring salivary proteomes in edentulous patients with type 2 diabetes.

Type 2 diabetes and tooth loss are linked both epidemiologically and pathophysiologically. We applied label-free differential protein expression analysis using multidimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS) to explore the proteomic profile of saliva samples collected from selected type 2 diabetic edentulous patients and non-diabetic controls. Ninety-six peptides corresponding to 52 proteins were differentially expressed between the diabetic edentulous patients and controls (p < 0.05). Some diabetes-related inflammatory biomarkers including glyceraldehyde-3-phosphate dehydrogenase and serum amyloid A were detected with levels increased in diabetic samples. Other biomarkers including amylase, palate, lung and nasal epithelium associated protein (PLUNC), and serotransferrin levels were decreased in diabetic samples. In contrast with previous findings, salivary carbonic anhydrase 6 and alpha-2 macroglobulin levels, however, were decreased in this diabetic patient population. Cluster analysis and principle component analysis demonstrated a differential pattern of protein biomarker expression between diabetic and control subjects. Western blot analysis was completed to confirm the relatively lower expression level of two biomarkers, including PLUNC and amylase in the diabetic group compared to control subjects. The presence of salivary biomarkers specific for diabetes in edentulous subjects mimics those in serum, especially those related to inflammatory/lipid metabolism. While this exploratory study requires further validation with a larger population, it provides proof-of-principle for salivary proteomics for edentulous subjects with diabetes.

alpha-Defensin levels in whole saliva of totally edentulous subjects.

Salivary levels of alpha-defensins 1-4 and histatins 1, 3 and 5 were determined in 11 totally edentulous patients, 11 younger healthy adults with normal gingival mucosa (Control group I) and 8 subjects, age-matched with edentulous patients, having a minimum of 25 teeth (Control group II). Whole saliva was treated with trifluoroacetic acid and the acidic soluble fraction analyzed by High Performance Liquid Chromatography-Mass Spectrometry. The area of the extracted ion current peaks was used for peptide quantification. Levels of alpha-defensins1-4, but not of histatins, were significantly lower in totally edentulous patients with respect to both Control group I and Control group II. The two control groups did not show significant differences. The reduced level of oral alpha-defensins, which are mainly of crevicular origin, is most likely due to the absence of the gingival sulcus in the edentulous subjects. The near absence of alpha- defensins might be in part responsible for the higher vulnerability of the oral cavity to oral pathogen infections observed in totally edentulous patients.

Number of teeth and serum lipid peroxide in 85-year-olds.

OBJECTIVES: To investigate influence of dental status on systemic oxidative stress, we evaluated the association between number of teeth and serum lipid peroxide, an oxidative stress index, in 85-years old residents of Japan. METHODS: In October 2003, 207 subjects 85-years old agreed to participate in the present follow-up study after five years from the 8020 Data Bank Survey of Fukuoka prefecture in 1998. Dental health condition including number of teeth was examined by dentists. Data from 204 subjects (88 male, 116 female) who completed nonfasting venous blood examination including lipid peroxide and blood chemistry were analyzed. The examination included a medical questionnaire regarding smoking history, physical activity, alcohol consumption, educational duration, and regular dental care, anthropometric and manometric measurements. RESULTS: Albumin, lipids, and lipid peroxide in serum all were within the normal range. Number of teeth correlated positively with height and white blood cell count, and correlated negatively with lipid peroxide. In a multiple regression analysis to adjust for confounding factors, tooth number retained this correlation with lipid peroxide. By analysis of variance with a Bonferroni-Dunn correction, edentulous subjects showed significantly higher lipid peroxide than those retaining 20 teeth or more. CONCLUSION: The negative association between number of teeth and lipid peroxide links more teeth remaining with less oxidative stress in an 85-year-old population; this may decrease risk of atherosclerotic complications.

Cyclosporin-induced downregulation of the expression of E-cadherin during proliferation of edentulous gingival epithelium in rats.

BACKGROUND: To examine the role of E-cadherin in epithelial hyperplasia of cyclosporin A (CsA)-induced gingival enlargement, mRNA and protein levels of E-cadherin, beta-catenin, proliferating cell nuclear antigen (PCNA), and Cyclin D1 were examined in the edentulous gingiva of rats following CsA treatment. METHODS: Three weeks after the extraction of all maxillary molars, 20 male Sprague-Dawley rats were assigned to a CsA-fed group (30 mg/kg daily) or a control group. Five rats per group were sacrificed at weeks 1 and 4. Edentulous ridge specimens were taken, and the expression levels of E-cadherin, beta-catenin, Cyclin D1, and PCNA mRNAs were estimated by reverse transcription-polymerase chain reaction (RT-PCR). Tissue specimens of the week 4 groups were examined using immunohistochemical (IHC) staining for proteins. RESULTS: The mRNA expression of E-cadherin was significantly weaker in the CsA-treated group than the control group at both times. Using IHC staining, a weaker level of membrane-bonded E-cadherin was also observed in the gingival epithelial cells in the CsA group than in controls. By contrast, significantly stronger beta-catenin and Cyclin D1 mRNA expressions and protein levels were found in CsA-treated rats than controls by RT-PCR and immunohistochemistry at week 4, whereas PCNA production was stronger at both times. CONCLUSIONS: CsA treatment reduced the production of E-cadherin but increased the production of beta-catenin, Cyclin D1, and PCNA. Thus, CsA may downregulate E-cadherin gene expression, leading to the epithelial cell proliferation of gingival overgrowth.

Quantification of oxidative metabolism in masseter muscle of denture wearers.

This study aimed to quantify oxidative metabolism in masseter muscle using near-infrared spectroscopy, in particular for denture wearers. Fourteen normal dentate subjects without malocclusion (ND group, 25-50 years) participated in the quantification of oxidative metabolism. Eleven partially edentulous patients without occlusal stops (PD group, 64-80 years) and ten edentulous patients (CD group, 57-84 years) also participated after prosthodontic treatment. Oxidative metabolism was recorded during gum chewing, maximum clenching and regulated clenching at 5 kgf. The oxygenated hemoglobin at 5 kgf clenching level was normalized to the oxygenated hemoglobin at the lowest blood flow and expressed as oxygen consumption rate (OCR). The relationship of the OCR to the maximum clenching force was analyzed using Pearson's correlation coefficient, and differences between the PD and CD groups were tested by unpaired Student's t-test. The OCR showed a significant negative correlation with maximum clenching force in the ND group. The OCR of the PD group was significantly greater than that of the CD group, although the difference in maximum clenching force was not significant between both groups. These results suggest that the aerobic ability of masseter muscle in complete denture wearers is relatively greater than in partial denture wearers with same age level.

Detection of platelet-activating factor in gingival tissue surrounding failed dental implants.

BACKGROUND: Dental implant therapy has entered routine clinical practice. However, the failure rate of implants at 5 years, due to biological factors, is still around 7%. The pathogenesis of implant loss involves a complex network of cells and inflammatory mediators. This study evaluated platelet-activating factor (PAF), a potent phospholipid mediator of inflammation, in soft tissue surrounding failed dental implants versus healthy implants. METHODS: PAF was estimated on extracted lipids by bioassay on washed rabbit platelets; inflammatory cell populations were assessed semiquantitatively after staining, and microvessel density was evaluated after immunohistochemical staining. RESULTS: Biologically active PAF was detected in the lipid extracts of samples excised from gingival tissue of patients with failed implants, but not in samples from patients with osseointegrated implants or from healthy edentulous subjects. The amount of PAF detected in failed implants was significantly higher than in healthy implants, suggesting a local production of this mediator. CONCLUSIONS: The presence of PAF was associated with histopathological findings of local inflammation and increased blood vessel density.

Cytokine production and bone remodeling in patients wearing overdentures on oral implants.

The stability of titanium dental implants is determined by osseointegration. Bone is a dynamic tissue continuously remodeled through resorption and formation, processes controlled by local cytokine production. This study investigated osseotropic cytokine expression in gingival mucosa, in the intraforamina and inferior first molar zones, during rehabilitation with implant-retained overdentures. Specimens were taken from six patients prior to placement of implants in the intraforamina bone; at connection of healing abutments; and 4, 8, and 12 months after prosthetic anchorage. Through semi-quantitative reverse-transcriptase polymerase chain-reaction, the following constitutively expressed cytokines were found at first surgical stage: interleukin-1, -6, and -8; small amounts of interleukin-11; stem cell factor; and transforming growth factor-beta1, -beta2, and -beta3. From the connection of healing abutments to 12 months after prosthetic anchorage, transforming growth factor-beta1, -beta2, and -beta3 were markedly higher than initial values. Expression of interleukin-6 and -8 decreased 8 months after prosthetic anchorage, while that of interleukin-1 increased at 12 months. In cultured gingival fibroblasts, modulation of cytokine secretion was also time-dependent. Cell culture supernatants influenced osteoclast-like multinucleated cell formation in long-term human marrow culture or osteoblast function, depending on the cytokine profile produced. These results are consistent with functional contributions of cytokines to osseointegration and minimization of posterior edentulous zone bone resorption.

Human gingival crevicular fluid contains MRP8 (S100A8) and MRP14 (S100A9), two calcium-binding proteins of the S100 family.

Human gingival crevicular fluid contains unidentified proteins which might play a role as markers in periodontal diseases. Therefore, low-molecular-weight proteins found in human gingival crevicular fluid (GCF), but absent from serum, were identified in the present study by means of two-dimensional electrophoresis (2-D PAGE) analysis. GCF, serum, and whole saliva were collected from periodontitis and healthy subjects, as well as from edentulous and newborn subjects. Protein samples were separated by two-dimensional polyacrylamide gel electrophoresis, stained with silver, and compared with reference protein maps in the SWISS-2D PAGE database. In GCF and saliva from periodontitis patients and healthy subjects, four dominant low-molecular-mass (from 8 to 14 kDa) acidic spots were observed. They were not found in serum and were less visible in saliva from edentulous and newborn subjects. From N-terminal amino acid sequencing, the two 2-D protein spots of 8 kDa and isoelectric points between 6.5 and 7.0 were both identified as protein MRP8 (SI00A8), a member of the S100 family of calcium-binding proteins. Using peptide mass fingerprinting and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), we identified the other two protein spots, with mass of 14 kDa and isoelectric points between 5.5 and 6.0, as protein MRP14 (S100A9), also belonging to the S100 family. The presence of MRP8 and MRP14 in GCF was confirmed by Western blot, with monoclonal antibodies. The two polypeptides, MRP8 and MRP14, identified in GCF represent the major difference between the 2-D PAGE patterns of serum and GCF, and we hypothesize that they may play an important role in the gingival sulcus and could represent possible markers for periodontal diseases.

Salivary flow and composition in elderly patients referred to an acute care geriatric ward.

OBJECTIVE: The purpose of this investigation was to study oral health and salivary aspects of the frail elderly. The study hypothesis was that elderly patients with many concomitant diseases and drugs would have different salivary secretion rates and biochemical constituents than healthier patients. STUDY DESIGN: The stimulated flow, pH buffering capacity, and biochemical constituents were analyzed from salivas of 169 elderly subjects (51 men and 118 women, mean age 81.2 years, range 69 to 96 years) admitted to an acute geriatric ward because of sudden worsening of their health. Common statistical methods were used to analyze the differences among patient groups. The patients were grouped according to the number of concomitant diseases and daily used drugs and on the basis of salivary flow rate values. RESULTS: Reduced salivary flow (< 0.7 ml/min) was found in 48% of the men and 62.5% of the women, and a low buffering capacity was found in 31.9% of the men and 36.7% of the women. Age did not significantly affect the salivary flow rate. The factors that showed the strongest influence on salivary flow were endocrinologic diseases, ophthalmologic and respiratory drugs, and potassium chloride. Salivary immunoglobulin A and immunoglobulin M concentrations were significantly higher in older patients. Immunoglobulin A, lysozyme, and amylase concentrations were significantly higher in older patients taking many drugs. Patients with many concomitant diseases had significantly higher salivary urea concentrations than healthier patients. Edentulous patients had significantly higher salivary immunoglobulin A, immunoglobulin M, lysozyme, and amylase concentrations. CONCLUSIONS: In this study, hyposalivation was a frequent observation, and the elderly who took many drugs and had several systemic diseases had higher concentrations of most of the analyzed biochemical constituents.

Salivary albumin of edentulous patients.

In order to evaluate the contribution of the periodontal sulcus or pocket to the presence of albumin in the mouth, this protein was analysed in whole saliva of 20 completely dentate adults, aged between 63 and 83 years, and in 23 edentulous patients of similar age (51-88 years). In spite of the considerable intra- and interindividual variations revealed by a preliminary trial, the concentration of salivary albumin was significantly higher (range 60-1080 mg/l) in the dentate than in edentulous individuals (range 2-690 mg/l). The low albumin content in saliva of old edentulous people was similar to that in a group of younger individuals with a healthy periodontium.

High-performance liquid chromatography analysis of chondroitin sulphate isomers in human whole saliva in a variety of clinical conditions.

OBJECTIVES: Tests have been carried out to assess the level of unsaturated disaccharide isomers obtained from chondroitin sulphate in whole saliva, which contains chondroitin sulphate derived from gingival crevicular fluid (GCF). MATERIALS AND METHODS: Whole saliva was collected from periodontally diseased subjects (PDS), clinically healthy subjects (CHS) and edentulous subjects (ES). Glycosaminoglycans (GAG) were liberated by digestion with Pronase E, and precipitated with cetylpyridinium chloride and ethanol. The unsaturated disaccharides obtained by chondroitinase ACII digestion of the liberated GAG were analysed by high-performance liquid chromatography. The unsaturated disaccharides included delta Di-0S, delta Di-6S and delta Di-4S. RESULTS AND CONCLUSIONS: Analysis of data indicated that delta Di-0S, delta Di-6S and delta Di-4S were found in all PDS samples. The amount (ng ml-1 collected whole saliva) of delta Di-0S, delta Di-6S and delta Di-4S (P < 0.01) indicated significant differences between CHS and PDS whole saliva samples. The quantities of delta Di-0S and delta Di-4S (P < 0.01) indicated significant differences between PDS and ES whole saliva. The amount of delta Di-0S (P < .05) and delta Di-6S (P < 0.01) also indicated significant differences between CHS and ES whole saliva. These results indicate that chondroitin sulphate in PDS and CHS whole saliva is representative of that previously reported in gingival crevicular fluid and so provides a useful and alternative means of assessing the role of GAG as indicators of periodontal disease.

Salivary levels of alpha 2-macroglobulin, alpha 1-antitrypsin, C-reactive protein, cathepsin G and elastase in humans with or without destructive periodontal disease.

Five host-response indicators were measured by enzyme-linked immunosorbent assays on unstimulated whole saliva samples from 45 adults (19 male, 26 female). The participants were distributed among four dentate groups representing oral health (I), gingivitis (II), moderate periodontitis (III), and severe periodontitis (IV), and one group of edentulous volunteers (V). Levels of the host-response indicators varied widely, from zero, primarily with groups I and V, to relatively high values with groups II, III and IV. The levels ranged as follows: alpha 2-macroglobulin, 0-4941 ng/ml; alpha 1-antitrypsin, 2-2271 ng/ml; C-reactive protein, 0-472 pg/ml; cathepsin G, 0-6035 ng/ml; elastase, 0-164 ng/ml (free), 0-732 ng/ml (bound to alpha 1-antitrypsin), and 0-318 ng/ml (bound to alpha 2-macroglobulin). Statistical evaluation by planned contrasts showed that levels of host-response indicators for group I were significantly lower (except for alpha 1-antitrypsin) than for groups II, III, and IV. A trend analysis of groups I-IV showed that mean scores (again, except for alpha 1-antitrypsin) increased significantly in a positive, monotonic manner. Group V showed significantly lower values for elastase than in the other groups. The findings demonstrate that these factors can be detected in whole saliva and suggest that, except for alpha 1-antitrypsin, their levels are directly related to an individual's periodontal status.