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Background: Periodontal diagnosis is based on recording clinical parameters including bleeding on probing (BOP), probing pocket depth (PPD), and clinical attachment loss (CAL). These techniques may be prone to errors due to different factors. Available biomarkers in the oral biofluid such as interleukin (IL)-1β could provide solutions for these issues. The study aimed to determine the potential of salivary IL-1β to differentiate periodontal health from disease and between gingivitis and periodontitis., Methods: Patients with gingivitis (N.=25), periodontitis (N.=50), and healthy periodontium (N.=25) were recruited for this study. For each patient, whole unstimulated saliva was collected followed by recording periodontal parameters namely; Plaque Index (PI), BOP, PPD, CAL. Level of salivary IL-1β was assayed by using enzyme-linked immunosorbent assays. Sensitivity and specificity of IL-1β, to differentiate any given condition, was determined by Receiver operating characteristic curve and area under the curve (AUC)., Results: Both BOP and PI were significantly higher in association with gingivitis and periodontitis groups as compared to controls. Concentration of salivary IL-1β in periodontal health was significantly lower than gingivitis and periodontitis groups. The biochemical analyses showed that salivary IL-1β differentiated periodontal health from gingivitis (AUC 0.949) and periodontitis (AUC 0.852) but could not discriminate gingivitis from periodontitis (AUC 0.532). The proposed cut-off points to differentiate periodontal health from gingivitis was 103.8 pg/mL, while the value of the biomarker to differentiate periodontal health from periodontitis was 102.0 pg/mL., Conclusions: Salivary IL-1β could be a reliable biomarker with a good level of accuracy to differentiate periodontal health from disease but not to discriminate gingivitis from periodontitis.

Fluorosis is one of the global problems that have persisted for a very long time now without any specific treatment and cure. Fluoride is known for its influence on the tooth structures like enamel and dentin, but little information exists on its effect on the surrounding periodontal structures and bone. With this background, we conducted multiple studies assessing etiological, diagnostic, therapeutic and clinical criteria in subjects with and without dental fluorosis in a single group of the population residing in high fluoride water areas (1.5-3.0 ppm) of Davangere District, Karnataka, India. These preliminary studies conducted are hereby elaborately presented and discussed in this comprehensive review article along with their important conclusions., (© 2021. Springer Science+Business Media, LLC, part of Springer Nature.)

Objective: To analyse the citrulline level in the periodontium in association with the presence of or antibody levels against Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis., Design: Gingival crevicular fluid (GCF), subgingival biofilm and blood serum were sampled from 98 subjects (26 with RA, 72 without RA (NoRA)). GCF was analyzed for the level of citrulline, for interleukin (IL)-1β, IL-17, IL-10 and monocyte-chemoattractant protein (MCP)-1. Microorganisms were identified in subgingival biofilms. Antibodies againstP. gingivalis, and Aggregatibacter actinomycetemcomitans were quantified in serum., Results: GCF citrulline level was the lowest (by trend) in NoRA group without periodontitis. In NoRA, but not in RA an association between GCF citrulline level and P. gingivalis antibody levels was found and the GCF citrulline levels were higher in P. gingivalis positive samples. Any association of A. actinomycetemcomitans with GCF citrulline level did not exist. A model of univariate variance analysis (p = 0.001) showed a dependence of GCF citrulline level from the number of sites with PD (probing depth) ≥5 mm (p = 0.003) and the GCF MCP-1/CCL2 level (p = 0.019). Compared with NoRA in RA the number of teeth was lower, the number of sites with PD ≥ 5 mm was less, GCF levels of interleukin-17 and MCP-1/CCL2 were higher and those of IL-10 lower. Yeasts were only cultured in 15 RA patients (p < 0.001)., Conclusion: Citrullination in periodontium might be associated with P. gingivalis supporting the potential role as a trigger in the development of RA. Pathogenesis of periodontal disease in RA patients seems to differ from that in NoRA and should be investigated further., Competing Interests: Declaration of Competing Interest Authors declare no conflict of interest., (Copyright © 2020. Published by Elsevier Ltd.)

Interleukin-34 (IL-34), an alternative ligand for macrophage colony-stimulating factor receptor, plays an important role in osteoclastogenesis. The aim of this study was to analyze the effect of smoking on IL-34 levels in gingival crevicular fluid (GCF) and plasma in individuals with healthy periodontium and chronic generalized periodontitis (CP). A total of 60 individuals ranging in age from 25 to 55 years were enrolled in the study. The participants were divided into 4 groups: Group A, 30 samples (15 GCF and 15 plasma) obtained from 15 non-smokers with healthy periodontium; Group B, 30 samples (15 GCF and 15 plasma) from 15 smokers with healthy periodontium; Group C, 30 samples (15 GCF and 15 plasma) from 15 non-smokers with CP; and Group D, 30 samples (15 GCF and 15 plasma) from 15 smokers with CP. The Gingival Index and probing depth scores, together with the Clinical Attachment Level, were assessed in each group as clinical periodontal parameters. Levels of IL-34 in GCF and plasma were quantified using enzyme linked immunosorbent assay. The results showed that the mean IL-34 concentrations in GCF and plasma were highest in Group D, followed by Group C, Group B, and Group A, and the difference among them was statistically significant (p<0.05). The relatively elevated IL-34 levels observed here in smokers with CP suggest that this cytokine offers a potential inflammatory marker of periodontal disease in smokers.

Objectives: The levels of interleukin-1β (IL-1β), nitric oxide (NO), total antioxidant capacity (TAC), and total oxidant status (TOS) in gingival crevicular fluid (GCF) were determined during rapid maxillary expansion (RME) treatment., Materials and Methods: Fourteen patients (10-13 years old) were included. A modified hyrax appliance was used for the treatment. After periodontal parameters were recorded, GCF was collected from the first molars at each observation [T1:baseline:14 days after periodontal prophylaxis and instructions; T2:1 day later hyrax inserted, at passive position; T3:1 week later; after the first activation; T4:after 2 × 1/4 activation; T5:after 7 × 1/4 activation; T6:after 14 × 1/4 activation; T7:retention period on the 1 st month; and T8:retention period on the 3rd month]., Results: Although the levels of IL1-β, NO, and PD increased significantly from T1 to T2, the GI, BOP%, and PI remained unchanged throughout treatment. GCF volume at buccal and palatal surfaces increased significantly from T1 to T4, T6, T7, and T8. The parameters in GCF and TAC levels were not only higher at palatal side in comparison with buccal, but also TOS levels increased at both buccal and palatal sides., Conclusions: In this study, the differences of oxidative status and IL-1β levels during RME treatment could be attributable to orthopedic effect of the heavy forces on maxilla and minimal orthodontic forces on teeth applied by the RME apparatus., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: The aim of this study was to determine whether saliva interleukin-6 (IL-6) levels are elevated in patients with rheumatoid arthritis versus a control group and examine the possible relationship between the oral condition and the risk of RA., Material and Methods: In 30 patients with RA and 30 healthy controls, different periodontal indices were recorded; sialometric measurements were taken to determine resting whole saliva, stimulated whole saliva and stimulated parotid saliva flow; and the saliva IL-6 levels were measured. Logistic regression analysis was performed, with the presence or absence of RA as dependent variable., Results: The patients with RA had a greater presence of bacterial plaque, a greater periodontal pocket depth, a larger percentage of medium-sized pockets, and greater periodontal attachment loss compared with the controls. Likewise, a decrease in resting and stimulated saliva flow was observed, together with an increase in saliva IL-6 levels. Logistic regression analysis reported that the plaque index is the principal differentiating factor of patients with RA. Stimulated parotid saliva flow was also significantly correlated to the presence of RA., Conclusions: The patients with RA showed a greater tendency to develop periodontal disease than the controls, with lower salivary flow and higher levels of IL-6 in saliva.

Background and Objective: The aim of this 12 mo prospective study was to assess the effect of smoking cessation on periodontal tissue without periodontal intervention, using matrix metalloproteinase (MMP)-8, MMP-9 and interleukin (IL)-1ß in gingival crevicular fluid, and nicotine and cotinine in saliva., Material and Methods: Of the 122 male smokers enrolled in a smoking cessation clinic, 11 quitters, nine non-quitters, six oscillators and 13 non-smokers participated in all experiments done at follow-up week 2, and follow-up months 2, 4, 6 and 12. The following were measured: gingival index; dental plaque index and sites of 3.5 mm < probing depth < 5.5 mm using a WHO probe for the full mouth; amounts of MMP-8, MMP-9 and IL-1β in gingival crevicular fluid of the upper anterior teeth area using enzyme-linked immunosorbent assay; and concentrations of nicotine, cotinine and hydroxycotinine in saliva using high-performance liquid chromatography-tandem mass spectrometry., Results: No significant differences in MMP-8 and MMP-9 in gingival crevicular fluid were detected between smokers, quit-smokers, oscillators and non-smokers for 1 year. Only the amount of IL-1β showed that smokers (90.14 ± 65.32 pg/mL) had a significantly higher value compared with non-smokers (37.70 ± 40.90 pg/mL), quit-smokers (32.11 ± 40.50 pg/mL) and oscillators (11.90 ± 12.46 pg/mL) at 2 mo follow-up (p = 0.007). IL-1β had a positive correlation with nicotine (r = 0.351) and the cotinine (r = 0.376), nicotine (r = 0.492) and hydroxycotinine (r = 0.358), and hydroxycotinine (r = 0.413) levels at 2 wk and 4 and 6 mo follow-up, respectively., Conclusions: This 1-year prospective smoking cessation study without nonsurgical periodontal therapy shows IL-1β in gingival crevicular fluid could have a positive relationship with the nicotine and cotinine levels in saliva., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Defensins are antimicrobial peptides that exhibit direct microbicidal activity as well as mediator-like functions by, for example, activating immature dendritic cells. This review focuses on defensins and other antimicrobial peptides that are present in periodontal tissues. Their antimicrobial capacity against periodontal microorganisms, their regulation and their expression profiles during periodontal diseases is discussed. As antimicrobial peptides may possess great potential for new diagnostic, preventive and therapeutic strategies, a better understanding of how antimicrobial peptides are regulated as part of the innate host immune response is crucial., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

There is scarce evidence on suitable approaches for the treatment of unresponsive or residual periodontal sites in diabetic patients. This study assessed the effects of surgical debridement (SD) and nonsurgical debridement (NSD), associated with amoxicillin and metronidazole, on clinical and immunological outcomes of residual pockets and adjacent healthy sites in patients with type 2 diabetes. A split-mouth, randomized controlled trial was conducted in 21 patients presenting at least 2 residual pockets in contralateral quadrants 12 months after basic nonsurgical periodontal therapy. Patients received systemic antibiotics, and contralateral quadrants were assigned to receive SD or NSD. The changes in clinical parameters were evaluated from baseline to 12 months. Local levels of 14 cytokines and chemokines were measured with multiplex bead immunoassays at baseline and 3 and 12 months after therapy. There were no statistically significant differences between SD and NSD for changes in clinical parameters from baseline to 12 months (P > 0.05). There was a significantly greater increase in the levels of granulocyte-macrophage colony-stimulating factor and interleukin 6 from baseline to 3 months in the healthy sites adjacent to residual pockets receiving SD (P < 0.05). A significant decrease in the levels of monocyte chemoattractant protein-1 and macrophage inflammatory protein 1α occurred from baseline to 12 months in the residual pockets treated by SD (P < 0.05). In conclusion, SD and NSD resulted in similar clinical benefits at 12 months. The short-term increase in the levels of proinflammatory biomarkers in SD sites probably can be attributed to tissue trauma and healing, and the long-term decrease in the levels of chemotactic factors in residual pockets treated by surgery may reflect remission of infection and stable wound healing in these sites at 12 months.

Purpose: Implant placement entails disruption of the epithelial continuity, which can lead to various complications. Therefore, the area of mucosal penetration is of particular interest clinically. The goal of the present study was to compare gene expression in peri-implant soft tissue (PIST) with that in oral mucosal tissue (OMT) using microarray analysis, and to investigate which genes were specifically expressed in PIST., Materials and Methods: The bilateral upper first molars were extracted from 4-week-old rats and titanium alloy implants placed only in the left-side extraction sockets. Four weeks after surgery, samples were harvested from the left-side PIST and right-side OMT and total RNA samples isolated. Microarray analysis was used to compare gene expression in PIST and OMT, which was then confirmed using quantitative real-time polymerase chain reaction. Immunohistochemical staining was also performed to confirm protein level expression., Results: The number of genes expressed with more than a twofold change in PIST compared with OMT was 1,102, of which 750 genes were upregulated and 352 genes were downregulated. The messenger RNA (mRNA) expression of three selected genes-Ceacam1, Ifitm1, and MUC4-were more significantly expressed in PIST than in OMT(P < .01). Immunohistochemical localization of CEACAM1, IFITM1, and MUC4 was observed in PIST, but no immunoreaction was recognized in OMT., Conclusion: The result of microarray analysis showed that, because of implant placement, 750 genes were upregulated in PIST compared with OMT. CEACAM1, IFITM1, and MUC4 were specifically upregulated in PIST.

Background and Objective: Both gingival tissue destruction and regeneration are associated with chronic periodontitis, although the former overwhelms the latter. Studies have shown that transforming growth factor beta 1 (TGF-β1), a growth factor largely involved in tissue regeneration and remodeling, is upregulated in chronic periodontitis. However, the gingival expression of connective tissue growth factor (CTGF or CCN2), a TGF-β1-upregulated gene, in patients with periodontitis remains undetermined. Although both CTGF/CCN2 and TGF-b1 increase the production of extracellular matrix, they have many different biological functions. Therefore, it is important to delineate the impact of periodontitis on gingival CTGF/CCN2 expression., Material and Methods: Periodontal tissue specimens were collected from seven individuals without periodontitis (group 1) and from 14 with periodontitis (group 2). The expression of CTGF and TGFβ1 mRNAs were quantified using real-time PCR., Results: Analysis using the nonparametric Mann-Whitney U-test showed that the levels of expression of both CTGF/CCN2 and TGFβ1 mRNAs were significantly increased in individuals with periodontitis compared with individuals without periodontitis. Furthermore, analysis using a nonparametric correlation (Spearman r) test showed a positive correlation between TGFβ1 and CTGF/CCN2 mRNAs., Conclusion: The gingival expression levels of CTGF/CCN2 and TGFβ1 mRNAs in individuals with periodontitis are upregulated and correlated., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: The aims of this study are to estimate the profile of bone loss biomarkers in peri-implant tissues and to identify potential prognostic biomarkers of peri-implantitis., Methods: Peri-implant crevicular fluid samples collected from 164 participants (52 patients with peri-implantitis, 54 with mucositis, and 58 with healthy peri-implant tissues) were analyzed using enzyme-linked immunosorbent assays to evaluate concentrations of the receptor activator of nuclear factor-κB (RANK), soluble RANK ligand (sRANKL), osteoprotegerin (OPG), cathepsin-K, and sclerostin., Results: Concentrations of RANK, sRANKL, OPG, and sclerostin were significantly increased in patients with peri-implantitis compared with patients with healthy peri-implant tissues. Comparisons between peri-implantitis and mucositis demonstrated significantly higher values of sclerostin in peri-implantitis samples. Comparisons between mucositis and healthy peri-implant tissues showed significantly increased levels of RANK and cathepsin-K in mucositis., Conclusion: These results are suggestive of a role of sRANKL, OPG, and sclerostin as prognostic biomarkers in peri-implantitis.

Background and Objective: Connective tissue growth factor (CTGF) is a downstream mediator of transforming growth factor-beta1 (TGF-β1), and TGF-β1-induced CTGF expression is regulated through the SMAD pathway. CTGF is implicated in the development of cartilage, bone and tooth. However, its expression in the developing periodontium is unclear. Therefore, we aimed to investigate the immunolocalization of CTGF, TGF-β1 and phosphorylated SMAD2/3 (pSMAD2/3) in the developing periodontium of rats., Material and Methods: The maxillaries of Wistar rats, 2, 3, 7 and 12 wk of age, were used and the localization of CTGF, TGF-β1 and pSMAD2/3 was detected using immunoperoxidase techniques., Results: Hertwig' s epithelial root sheath (HERS) cells were strongly positive for CTGF and TGF-β1, but not for pSMAD2/3. Positive staining for CTGF, TGF-β1 and pSMAD2/3 was found in bone and periodontal ligament. In cementum, most cementoblasts associated with cellular cementum and some cementocytes stained strongly for CTGF, whereas cementoblasts associated with acellular cementum did not express CTGF. No signal for TGF-β1 was observed in cellular and acellular cementum. In addition, most cementocytes were strongly positive for pSMAD2/3., Conclusion: CTGF, TGF-β1 and pSMAD2/3 are localized in bone and periodontal ligament, but are differentially expressed in HERS and cementum. The results of our study indicate that the regulation of CTGF expression by TGF-β1 might be cell-type specific in periodontium., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Periodontal disease is an immune-inflammatory disease characterized by connective tissue breakdown, loss of attachment, and alveolar bone resorption. Under normal physiological conditions, a dynamic equilibrium is maintained between the reactive oxygen species (ROS) and antioxidant defense capacity. Oxidative stress occurs when this equilibrium shifts in favor of ROS. Oxidative stress is thought to play a causative role in the pathogenesis of periodontal diseases., Aim: The present study was designed to estimate and compare the superoxide dismutase (SOD) and glutathione (GSH) levels in the serum of periodontitis, gingivitis, and healthy individuals before and after nonsurgical periodontal therapy., Materials and Methods: The present study was conducted in the Department of Periodontics, A. B. Shetty Memorial Institute of Dental Sciences, Deralakatte, Mangalore. The study was designed as a single blinded interventional study comprising 75 subjects, inclusive of both sexes and divided into three groups of 25 patients each. Patients were categorized into chronic periodontitis, gingivitis, and healthy. The severity of inflammation was assessed using gingival index and pocket probing depth. Biochemical analysis was done to estimate the SOD and GSH levels before and after nonsurgical periodontal therapy. RESULTS obtained were then statistically analyzed using ANOVA test and paired t-test., Results: The results showed a higher level of serum SOD and GSH in the healthy group compared to the other groups. The difference was found to be statistically significant (P < 0.0001). The post-treatment levels of SOD were statistically higher than the pre-treatment levels in periodontitis and gingivitis group.

Objective: To simulate the expression of TNF-α and PGE2 of periodontal tissues in rat periodontitis model., Methods: 40 Wistar rats were randomly divided into the periodontitis group and the control group (n=20). After the successful establishment of periodontitis rat model, raising for six weeks before the animals were sacrificed. The periodontal tissues were obtained and made into slices. Observed the histopathological changes of the periodontal tissues and measured TNF-α, PGE2 levels change by immunohistochemistry, Western blot analysis and ELISA., Results: TNF-α, PGE2 expression of the periodontitis group was significantly higher than that in the control group, the difference was significant (P<0.05)., Conclusions: The TNF-α, PGE2 expression of the rat periodontal tissue in the periodontitis group was significantly higher than the control group., (Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.)

The aim of this study was to evaluate the effects of host modulation therapy on periodontal and biochemical parameters. Sixteen rheumatoid arthritis patients newly scheduled for anti-tumour necrosis factor (TNF) therapy were screened for 30 days. Periodontal parameters (clinical attachment level, probing pocket depth, bleeding on probing, plaque index and gingival index) as well as salivary and gingival crevicular fluid (GCF), interleukin (IL)-1β, IL-8 and monocyte chemoattractant protein-1 (MCP-1) levels of the patients were evaluated at baseline and on the 30th day of therapy. GCF volume, IL-1β and IL-8 levels (p = 0.007, p = 0.017 and p = 0.009, respectively) of the periodontitis patients significantly decreased. Although there was a decrease in all these parameters in healthy patients, it was below statistical significance. Salivary IL-8 and MCP-1 levels significantly decreased in periodontitis patients (p = 0.028 and p = 0.013, respectively), but IL-1β levels remained unchanged. These results suggest that TNF blockers may significantly modify host response in terms of biochemical parameters of the periodontium and may mask significant associations such as those reported between periodontitis and rheumatoid arthritis.

A functional endocannabinoid system is present in several mammalian organs and tissues. The endogenous cannabinoid anandamide (AEA) is a bioactive lipid neurotransmitter that is present in a variety of tissues and has multiple functions. Recently, this cannabinoid system was reported to play important roles in osteoblastic and osteoclastic cells. Human periodontal ligament (hPDL) cells modulate alveolar bone remodeling by producing cytokines when stimulated by many factors. The aim of this study was to investigate the AEA level in periodontal regions and the osteogenic effect of AEA on hPDL cells. The levels of AEA in the gingival crevicular fluid (GCF) of periodontitis patients were measured using liquid chromatography/mass spectrometry (LC/MS). Expressions of cannabinoid receptor mRNA were detected by RT-PCR in hPDL cells and expression of the transient receptor potential cation channel subfamily V member 1 (TRPV1) was observed by confocal laser scanning microscope (CLSM). IL-11 production from hPDL cells was measured using an ELISA, with or without AEA in the presence or absence of capsazepine, a selective TRPV1 antagonist AEA secreted into GCF was detected, but there was no correlation between the probing pocket depth (PPD) and AEA level. TRPV1 mRNA was detected in hPDL cells and the TRPV1 expression was observed by CLSM. IL-11 production from hPDL cells was significantly enhanced by AEA stimulation and this IL-11 production was suppressed by capsazepine. Our findings indicate that this endogenous cannabinoid system has a possible role in bone metabolism in periodontitis through TRPV1.

Objective: Interleukin-17 (IL-17) is a cytokine that induces neutrophil recruitment and the release of inflammatory mediators in several inflammatory conditions; nevertheless, the involvement of IL-17 gene polymorphisms in chronic periodontitis (CP) has not been addressed yet. Our aim was to evaluate the association between periodontal status and the polymorphisms IL-17A G197A and IL-17F C7488T in subjects with CP along with their impact on levels of inflammatory mediators., Material and Methods: Genomic DNA was obtained from 30 CP patients and 30 healthy controls (HCs). IL-17A G197A and IL-17F C7488T polymorphisms were determined using PCR-RFLP. Serum and periodontal tissues were collected and processed for ELISA, myeloperoxidase (MPO), and/or microscopic analysis., Results: The frequencies of genotypes in the CP group were significantly different from those of HC. Odds ratio indicated that increased risks for CP were associated with the -197A allele, not with the -7488T allele. In addition, the -197A allele was correlated with worse clinical parameters, higher MPO activity, and increased expression of inflammatory mediators (IL-17A and IL-8) than the other genotypes., Conclusions: These results indicate that the IL-17A -197A allele is associated with increased risk for CP, likely because this genotype relates to the enhanced inflammation in periodontal tissues.

Diabetes mellitus (a group of metabolic disorders characterized by hyperglycemia) and periodontitis (a microbially induced inflammatory disorder that affects the supporting structures of teeth) are both common, chronic conditions. Multiple studies have demonstrated that diabetes mellitus (type 1 and type 2) is an established risk factor for periodontitis. Findings from mechanistic studies indicate that diabetes mellitus leads to a hyperinflammatory response to the periodontal microbiota and also impairs resolution of inflammation and repair, which leads to accelerated periodontal destruction. The cell surface receptor for advanced glycation end products and its ligands are expressed in the periodontium of individuals with diabetes mellitus and seem to mediate these processes. The association between the two diseases is bidirectional, as periodontitis has been reported to adversely affect glycemic control in patients with diabetes mellitus and to contribute to the development of diabetic complications. In addition, meta-analyses conclude that periodontal therapy in individuals with diabetes mellitus can result in a modest improvement of glycemic control. The effect of periodontal infections on diabetes mellitus is potentially explained by the resulting increase in levels of systemic proinflammatory mediators, which exacerbates insulin resistance. As our understanding of the relationship between diabetes mellitus and periodontitis deepens, increased patient awareness of the link between diabetes mellitus and oral health and collaboration among medical and dental professionals for the management of affected individuals become increasingly important.

The comparative analysis of "lipid peroxidation-antioxidant protection" system status (LP-AOP) in mixed saliva of patients with chronic periodontitis (CP) of a mild (n=90) and moderate (n=42) stages was performed. It was established that CP in a moderate stage of severity was characterized by the content decrease of isopropanol dissolved products of LP in saliva with simultaneous worsening of periodontal and affected status in comparison with CP of a slight stage. Correlation analysis of integral cohort of patients (n=132) demonstrated the AOP enhancement due to the increase of ceruloplasmin concentration in the saliva as periodontal disorders was aggravated. It appeared that adaptive enhancement of AOP in progressing CP caused the limitation of LP and restrains the escalation of CP-associated affective disorders.

The aim of this study was to explore further the preservation of tissues and the mineral distribution in 1.6 million-year-old fossil hominin material from Koobi Fora, Kenya attributed to Paranthropus boisei (KNM-ER 1817). Bone, dentine and cementum microstructure were well preserved. Electron microprobe analysis of dentine and bone revealed an F-bearing apatite. Calcite now filled the original soft tissue spaces. The average Ca/P atomic ratio was 1.93, as compared to 1.67 in biological hydroxyapatite, indicating that the Ca-content had increased during fossilization. Analytical sums for mineral content were approximately 90 wt%. Some of the remaining 10 wt% may be preserved organic material. Demineralized dentine fragments showed irregularly distributed tubules encircled with a fibrous-like electron-dense material. A similar material was observed in demineralized dentine. Within this, structures resembling bacteria were seen. In demineralized bone an electron-dense material with a fibrous appearance and a banding pattern that repeated every 64 nm, similar to that of collagen, was noted. SEM of an enamel fragment (KNM-ER 6081) showed signs of demineralization/remineralization. Retzius lines, Hunter-Schreger bands and prism cross-striations spaced 3.7-7.1.microm apart were noted. Prisms were arranged in a pattern 3 configuration and deeper areas containing aprismatic enamel were occasionally observed. We conclude that a great deal of informative microstructure and ultrastructure remains preserved in this fossil material. We also hypothesize that the high mineral content of the tissues may 'protect' parts of the organic matrix from degradation, since our findings indicate that some organic matrix may still be present., (Copyright (c) 2009 S. Karger AG, Basel.)

In this study, a comparison between structure, chemical composition and mechanical properties of collagen fibers at three regions within a human periodontium, has enabled us to define a novel tooth attachment mechanism. The three regions include, (1) the enthesis region: insertion site of periodontal ligament (PDL) fibers (collagen fibers) into cementum at the root surface, (2) bulk cementum, and (3) the cementum-dentin junction (CDJ). Structurally, continuity in collagen fibers was observed from the enthesis, through bulk cementum and CDJ. At the CDJ the collagen fibers split into individual collagen fibrils and intermingled with the extracellular matrix of mantle dentin. Under wet conditions, the collagen fibers at the three regions exhibited significant swelling suggesting a composition rich in polyanionic molecules such as glycosaminoglycans. Additionally, site-specific indentation illustrated a comparable elastic modulus between collagen fibers at the enthesis (1-3 GPa) and the CDJ (2-4 GPa). However, the elastic modulus of collagen fibers within bulk cementum was higher (4-7 GPa) suggesting presence of extrafibrillar mineral. It is known that the tooth forms a fibrous joint with the alveolar bone, which is termed a gomphosis. Although narrower in width than the PDL space, the hygroscopic CDJ can also be termed as a gomphosis; a fibrous joint between cementum and root dentin capable of accommodating functional loads similar to that between cementum and alveolar bone. From an engineering perspective, it is proposed that a tooth contains two fibrous joints that accommodate the masticatory cyclic loads. These joints are defined by the attachment of dissimilar materials via graded stiffness interfaces, such as: (1) alveolar bone attached to cementum with the PDL; and (2) cementum to root dentin with the CDJ. Thus, through variations in concentrations of basic constituents, distinct regions with characteristic structures and graded properties allow for attachment and the load bearing characteristics of a tooth.

Tissues contacting Ti dental implants were subjected to X-ray absorption fine structure (XAFS) analysis to examine the chemical state of Ti transferred from the placed implant into the surrounding tissue. Nine tissues that contacted pure Ti cover screws for several months were excised in a second surgery whereby healing abutments were set. Six tissues that surrounded implants retrieved due to their failure were also excised. Ti distributions in the excised specimens were confirmed by X-ray scanning analytical microscopy (XSAM), and the specimens were subjected to fluorescence XAFS analysis to determine the chemical states of the low concentrations of Ti in the tissues surrounding Ti dental implants. Ti mostly existed in the metallic state and was considered to be debris derived from the abrasion of implant pieces during implant surgery. Oxidized forms of Ti, such as anatase and rutile, were also detected in a few specimens-and existed in either a pure state or mixed state with metallic Ti. It was concluded that the existence of Ti in the tissue did not cause implant failure. Moreover, the usefulness of XAFS for analysis of the chemical states of rarely contained elements in biological tissue was demonstrated.

Background: The number of transplanted solid organs and life expectancy after transplantation are steadily rising worldwide. Inflammation is widely recognized as playing a pivotal role in transplant rejection, and several studies have shown that serum interleukin-6 (IL-6) levels can identify individuals who are at greater risk for rejection. Given the known association between IL-6 and chronic periodontitis, the aim of our study was to assess the periodontal status of solid-organ transplant subjects compared to systemically healthy controls, to quantify the IL-6 levels in the serum and periodontal tissues, and to explore their association., Methods: Forty-seven heart and kidney transplant and 18 systemically healthy age-matched individuals were recruited. Subjects received a complete periodontal examination, and blood and periodontal tissue samples were collected for quantification of IL-6 protein and mRNA levels, respectively., Results: Transplant subjects had significantly higher serum IL-6 levels and slightly but statistically significantly increased mean probing depths than healthy controls. Multivariable linear regression analysis adjusting for gender, diabetes, smoking, and immunosuppressant dose showed that the mean probing depth, number of missing teeth, and mean percentage of sites with > or =4 mm attachment loss were independent predictors for elevated serum IL-6 levels. Transplant subjects with chronic periodontitis had higher mean serum IL-6 levels than those without chronic periodontitis, and there was a positive correlation between periodontal IL-6 gene expression levels and serum IL-6 protein levels., Conclusions: Periodontal tissue destruction and local IL-6 synthesis are associated with elevated serum IL-6 levels in transplant recipients. This may have serious implications in solid-organ transplant deterioration and chronic rejection.

It has been reported that not only selective alveolar-bone resorption, but also receptor activator of nuclear factor kappa B ligand (RANKL) expression is induced on the compressed side of an orthodontically moving tooth. Numerous reports have described the pharmacological acceleration of tooth movement (TM) through the activation of osteoclasts. However, because of rapid flush out by blood circulation, daily systemic administration or daily local injection is needed. Previously, we discovered that every-3-days OPG gene transfer to the periodontal-tissue inhibited RANKL-mediated osteoclastogenesis and diminished experimental TM. Therefore, we hypothesized that local RANKL gene transfer into the periodontal tissue would accelerate TM. The upper first molars of 6-week-old male Wistar rats were moved palatally using fixed orthodontic wires. The inactivated hemagglutinating-virus of Japan (HVJ) envelope vector containing the mouse RANKL expression plasmid was injected periodically into the palatal periodontal tissue of the upper first molars during TM. Local RANKL gene transfer significantly enhanced RANKL expression and osteoclastogenesis in periodontal tissue without any systemic effects. The TM rate was significantly increased in the RANKL gene transfer side. In conclusion, we demonstrated that transfer of the RANKL gene to the periodontal-tissue activated osteoclastogenesis and accelerated the amount of experimental TM. Local RANKL gene transfer might be a useful tool not only for shortening orthodontic treatment, but also for moving ankylosed teeth where teeth, fuse to the surrounding bone.

Objective: To assess the correlation between sulfide levels in periodontal and volatile sulfide compounds (VSC) in oral cavity and to assess the correlation between sulfide levels in periodontal pocket (pS) and periodontal clinical parameters., Methods: VSC in oral cavity was measured by halimeter one week after grossly supragingival cleaning. Then pS in periodontal pockets was measured by Diamond probe/Perio 2000 system, followed by probing using Williams periodontal probe to get the records of probing depth (PD). Based on the above examination results, 4 - 8 sites with PD > or = 5 mm and/or pS > 0 were selected for the following longitude observation in each patient. The plaque index (PLI), bleeding index (BI), clinical attachment level (CAL) of the target site were recorded. The same examination was repeated during the recalling time of 6 weeks, 3 and 6 months after the initial periodontal therapy., Results: VSC levels are significantly correlated with the numbers of sites with pS > 0 (r = 0.592, P < 0.001) and pS value. pS values were positively correlated with BI at low level during the maintenance time of 6 weeks, 3 and 6 months (r = 0.183, 0.411, 0.350, P < 0.05) respectively. After the initial therapy pS, PD, CAL, BI, PLI and VSC in oral cavity significantly improved at 6 weeks (P < 0.001) and further improved and kept stable at 3 and 6 months., Conclusions: pS reflects the sulfide levels in pockets with site specificity and may be considered as objective parameter to evaluate the treatment effect, and to indicate the early inflammation in periodontal tissue during the maintenance period.

It has been shown that prostaglandin E2 (PGE2) locally released adjacent to the mandible over a 20-day period increases alveolar bone area, in part, due to a reduction in the percentage of eroded surface. To determine the effect of PGE2 on alveolar bone resorption, left mandibles from 24 Lewis rats were treated over a 20-day period with a local application of PGE2 (0.1, 0.05 or 0.025 mg/day) or placebo. The right side served as the non-treated matched control. Tissue sections were stained for tartrate resistant acid phosphatase (TRAP) calcitonin receptor (CTR) and metalloproteinase-2 (MMP-2). Matched samples were analysed by Wilcoxon matched pairs test and, a non-parametric one-way analysis of variance compared groups of treatment. Those tissues treated with PGE2 at doses of 0.1 and 0.05 mg/day showed significantly reduced numbers of TRAP and CTR-positive multinucleated cells compared with matched controls (p<0.005), as well as significantly reduced numbers of TRAP- and CTR-positive multinucleated cells when compared with the placebo-treated group (p<0.001). The number of periodontal ligament cells expressing MMP-2 was also significantly reduced in tissues treated with the two higher doses of PGE2 (p<0.001) comparing with both matched controls and the placebo-treated group. Following a 20-day period, locally released PGE2 at doses of 0.1 and 0.05 mg/day appears to affect alveolar bone resorption in the periodontium of rats, as the number of multinucleated cells expressing TRAP and CTR are significantly reduced. Furthermore, the same doses of PGE2 also significantly reduced the expression of MMP-2 by the periodontal cells.

Objective: To study the distribution and expression of fibromodulin in normal periodontium, so as to understand its role in periodontal tissue homeostasis., Methods: Five normal male Lewis rats were killed and their bilateral mandibular first molars with surrounding alveolar bones and gingival tissues were taken out. Human gingival fibroblasts, fibroblasts of periodontal ligament, and osteoblasts were cultured. Immunohistochemistry with anti-fibromodulin, anti-decorin, anti-biglycan, anti-type I collagen, and anti-type III collagen antibodies and RT-PCR were used to detect the tissue distribution and cellular localization of fibromodulin and related proteoglycans, decorin and biglycan, and type I and type III collagens., Results: Fibromodulin was strongly expressed in the suprabasal gingival epithelium, periodontal ligament, alveolar bone, gingival and periodontal fibroblasts as well as their matrices. Strong expression was noted in the area close to oral gingival aspect and the interfaces of periodontal ligament-alveolar bone and periodontal-ligament-cementum. Decorin was strongly expressed in the area close to the gingival sulcus in gingival tissue. Biglycan was stained evidently in gingival epithelium. Fibromodulin, decorin and biglycan mRNAs were strongly expressed in osteoblasts. mRNAs of type I and type III collagens were strongly expressed in gingival fibroblasts., Conclusion: Fibromodulin may interact with other small proteoglycans to regulate the network formation of periodontal collagen fibers, and may be involved in mineralization of the alveolar bone and cementum.

Osteoprotegerin (OPG) is a novel secreted member of the tumor necrosis factor (TNF) receptor superfamily that negatively regulates osteoclastogenesis. The receptor activator of the NFKB ligand (RANKL) is one of the key regulatory molecules in osteoclast formation and binds to OPG. In this study, it was suggested that OPG and RANKL are involved in alveolar bone remodeling during orthodontic tooth movement. We examined RANKL localization and osteoclast induction in periodontal tissues during experimental movement of incisors in OPG-deficient mice. To produce orthodontic force, an elastic band was inserted between the upper right and left incisors for 2 or 5 days, and the dissected maxillae were examined for cytochemical and immunocytochemical localization of tartrate-resistant acid phosphatase (TRAP), vacuolar-type H(+)-ATPase, and RANKL. Compared to wild-type OPG (+/+) littermates, TRAP-positive multinucleated cells were markedly induced in the periodontal ligament (PDL) on the compressed side and in the adjacent alveolar bone of OPG-deficient mice. These multinucleated cells exhibited intense vacuolar-type H(+)-ATPase along the ruffled border membranes. Because of accelerated osteoclastic resorption in OPG-deficient mice, alveolar bone was severely destroyed and partially perforated at 2 and 5 days after force application. In both wild-type and OPG-deficient mice, RANKL expression became stronger at 2 and 5 days after force application than before force application. There was no apparent difference in intensity of RANKL expression between OPG (+/+) littermates and OPG-deficient mice. In both wild-type and OPG-deficient mice, expression of RANKL protein was detected in osteoblasts, fibroblasts, and osteoclasts mostly located in resorption lacunae. These results suggest that during orthodontic tooth movement, RANKL and OPG in the periodontal tissues are important determinants regulating balanced alveolar bone resorption., (Copyright 2002 Wiley-Liss, Inc.)

Collagen is one of the chief components of the extracellular matrix of gingival connective tissue, where five different types have been identified to date. The molecular mechanism of collagen loss in periodontitis still needs to be explored. In the present study total collagen content was investigated in gingival connective tissue of adult periodontitis (AP) as well as early onset periodontitis patients (EOP) and clinically healthy subjects. Furthermore, collagen type I, III, IV, V andVI content was evaluated in gingival biopsies obtained from periodontitis patients. There was a statistically significant difference between AP (25.1 +/- 8.1 microg/mg) and EOP (15.6 +/- 4.0microg/mg) groups with regard to the total collagen content (P < 0.05). In the clinically healthy control group the total collagen content was 20.7 +/- 4.6microg/mg. Moreover, the distribution of collagen types exhibited variations in pooled homogenates of each periodontitis group. The total collagen loss seemed to be greater in the EOP patients than in the AP patients. When the ratio of fibril forming collagens to nonfibrillar collagens was evaluated, it seems to be decreased in AP patients in comparison to EOP patients. The findings of the present study suggest that different collagen types present in various periodontitis categories may be related with diverse pathogenic mechanisms acting in these diseases.

E11 antigen, originally characterized in a rat osteosarcoma cell line, is known to be expressed during late stages of the osteogenic cell lineage both in vitro and in vivo. The aim of the present study was to monitor the occurrence and distribution patterns of the E11 antigen using monoclonal antibodies (mAb E11 and MEP-1) during different stages of tooth germ development of new-born rats by means of immunohistochemistry. Both antibodies strongly bound to plasma membranes of ameloblasts in presecretory and secretory stages. In addition, odontoblasts and cells of the periodontium were immunoreactive for E11 and MEP-1. During maturation, the immunoreactivity of ameloblast plasma membranes decreased significantly. Our data suggest that E11 and MEP-1 might be important as markers for cell differentiation and mineralization processes during tooth germ development.

Objective: To observe the distribution and change of transforming growth factor beta 1 (TGF-beta 1) in periodontal tissue and understand the role of TGF-beta 1 in orthodontic tooth movement., Methods: Apply force on right maxillary first molars in rats, and examine the decalcified alveolodental connected tissue in different stage of tooth movement with immunohistochemical staining technique (SP method)., Results: TGF-beta 1 exists in normal periodontal tissue. The expression of it in periodontium increases during orthodontic tooth movement. Especially from 5 to 10 days after force application, the expression at tension side is much more than those at pressure side., Conclusion: TGF-beta 1 plays an important role in remodeling of periodontal tissue during orthodontic tooth movement, and can powerfully promote bone formation.

Vascular endothelial growth factor (VEGF) is a multifunctional angiogenic cytokine of importance in inflammation and wound healing but its presence in chronic inflammatory periodontal disease has never been reported. The aims of this study were to investigate the presence of VEGF in human periodontal tissue and gingival crevicular fluid (GCF) in periodontal health and disease. VEGF in tissue was localized by immunohistochemistry. GCF and unstimulated saliva were collected from patients and clinically healthy subjects and VEGF was assessed by using an ELISA. VEGF was detected within vascular endothelial cells, neutrophils, plasma cells and junctional, pocket and gingival epithelium. In periodontitis patients, the volume of GCF and total amount of VEGF collected from diseased sites were both greater than from clinically healthy sites (Wilcoxon test p < 0.01). However, the concentration of VEGF per unit volume of GCF was higher at healthy sites compared with diseased sites (Wilcoxon test p < 0.05). Higher concentrations of VEGF were detected in healthy sites in patients compared with similar sites in clinically healthy subjects (Mann-Whitney U-test p < 0.05). A logistic regression approach indicated that there was variation in VEGF between subjects (p < 0.01), and that age (p < 0.05), plaque (p < 0.05) and pocket depth (p < 0.07) were explanatory variables. VEGF was also detected in all saliva samples and was significantly higher in patients than in healthy controls (p < 0.05). This study suggests that VEGF could be relevant to angiogenic processes in healthy as well as diseased periodontal tissue and that the periodontal status influences the salivary level of VEGF.

Type VI collagen was immunolocalized in several soft connective tissues at the light and electron microscopic level. Positive labeling was found in all tissues examined, periodontal ligament, gingiva, mesenterium and periosteum. The labeled structures could be divided into 2 categories: microfibrils intermingling with collagen fibrils, and those that formed bundles (oxytalan fibres and elastin-associated microfibrils). Control sections incubated with antibody preabsorbed to purified type VI collagen, or with non-immune antibody, proved to be negative. Our observations indicate that the structural organization of type VI collagen varies from small microfibrillar structures associated with the collagen and elastin fibre systems to highly ordered parallel arrays of oxytalan bundles.

The aim of this study was to measure tissue levels of immunoreactive prostaglandin E2 (iPGE2), immunoreactive leukotriene B4 (iLTB4), and pain after periodontal surgery and to evaluate the effect of the non-steroidal anti-inflammatory drug (NSAID), ibuprofen, on these levels. Two contralateral quadrants in each of nine patients were selected to undergo separate surgical procedures, one with ibuprofen (800 mg 1 hour presurgery and 400 mg postsurgery) and one with a placebo. Intra-operatively, a custom-made microdialysis probe, with a 3,000 dalton molecular weight cut-off, was inserted beneath the soft tissue flap and a dialysate collected every 20 minutes for 4 hours after surgery. Pain perception was measured at the same time intervals using two pain scales. Dialysate samples were assayed using two enzyme immunoassays. Mean tissue levels of iPGE2 in the placebo group increased from 74 nM at 40 minutes to a peak of 261 nM at 200 minutes. Mean tissue levels of iLTB4 in the placebo group fluctuated between 0.2 and 0.6 nM. Pain levels in this group increased continuously with time, peaking at 4 hours. Mean tissue levels of iPGE2 in the ibuprofen group were significantly suppressed, exhibiting more than a 95% reduction. This was accompanied by a significant reduction in pain. Ibuprofen had no detectable effect on tissue levels of iLTB4. These data indicate that iPGE2 and iLTB4 are present at relatively high concentrations in the periodontal tissues after surgery. Since these concentrations exceed the Kd values for binding to their respective receptors, PGE2 and LTB4 may be associated with the development of postsurgical pain and inflammation. These data also indicate that ibuprofen can successfully inhibit iPGE2 production in the periodontal tissues and in this way may help reduce postoperative pain and inflammation.

Previous studies have shown a perivascular hyaline thickening affecting restricted regions of the microcirculation in gingivitis and moderate periodontitis and in the pulpal vessels in chronic pulpitis. In the present study of the lesion of advanced periodontitis, immunostaining for type IV collagen and laminin demonstrated widespread deposition of basement membrane material, with manifest involvement of the venous network. Some vessels were associated with an increased deposition of both basement membrane proteins, while others showed preferential deposition of either laminin or type IV collagen. Immunostaining also revealed an extensive trabecular network of type IV collagen throughout the affected gingival tissue that was related to recognizable vessels but was co-extensive with less intense staining for laminin. This network was not associated with viable endothelial cells demonstrable by staining with the endothelial marker Ulex agglutinin (UEA-1). The results indicate extensive vascular pathology in advanced periodontitis that could explain the attenuation of the inflammatory reaction and the restricted ability to develop reparative granulation tissue in this disease.

Polypeptide extracts from the pig spleen, liver, kidney, thymus, pancreas and periodont were obtained by the acid extraction in presence of Zn and MG cations. This extracts were analyzed by HPLC with determining amino acid sequence using Edman's method. According to the analysis results several fragments of haemoglobin are present in the group of parenchymal organs (epsylon-chain pig haemoglobin 56-66 and 55-66, alpha-chain 48-61), the main mass of substances is localized in more hydrophilic part of chromatogram with 20 aminoacid N-terminal ubiquitin fragments; hydrophylic polypeptides ubiquitin and 2 hydrophobic polypeptides were also found in the group, but haemoglobin peaks were absent. The main mass of polypeptide substances in periodont extract is localized in the hydrophobic region, the peaks of haemoglobin and ubiquitin are absent. We suppose that each group of similar morpho-functional tissues has similar chromatographic picture of polypeptide extracts. It can be connected with the processing of protein structures specific for each organ or tissue. This characteristic for each tissue peptide can take part in the signal transduction at the level of single population cells or between physiologically correlated organs.

Guided tissue regeneration (GTR) is a concept that evolved from the development of membrane barrier techniques which allow the repopulation of periodontal wounds by desirable cells, resulting in a so-called new attachment apparatus. To understand the biological mechanisms involved in membrane barrier-led periodontal healing, the histological localization of macromolecules phenotypical of bone and cementum formation was investigated in regenerating human periodontal tissues harvested after healing by placing barriers on teeth untreatable except by extraction. Using immunolocalization techniques, frozen sections of soft tissues and hard tissues under GTR barriers were stained with antibodies to osteonectin (LB-BON-II) and bone sialoprotein (BSP) (LF-6); alkaline phosphatase (AP) was detected histochemically. Frozen sections of regenerating periodontal tissue demonstrated the presence of spindle-shaped, fibroblast-like cells entrapped in a dense fibrillar extracellular matrix. Rounded cells aggregated to form nodules heavily stained by the Alcian blue method, indicating the presence of proteoglycans and strongly resembling those noted in hard-tissue sections. At the electron-microscopic level, the cytoplasm of the elongated cells had numerous cisternae of endoplasmic reticulum and Golgi saccules, indicating metabolic activity. Striated collagen fibres were scattered throughout the field of the sections. AP-stained soft-tissue sections demonstrated the presence cell-bound and extracellular AP. Osteonectin antibody staining confirmed the presence of this macromolecule in the extracellular matrix, particularly in the area of the cellular nodules. The dense network of connective tissue fibres was also stained.(ABSTRACT TRUNCATED AT 250 WORDS)

Biopsy of the periodontal tissues was carried out in 29 patients with chronic generalized periodontitis in order to detect sex steroid receptors. Estrogen receptors in different levels were found in 58.6% of biopsy specimens. This implies that the trophic effect of sex steroids on the periodontium should not be ruled out.

The intensity of protein biosynthesis in clinically healthy gingival tissues is the maximal at young age (20 to 30) and reduced in older subjects (aged 40 to 50). In generalized periodontitis the intensity of protein biosynthesis is reliably increased, more markedly in exacerbations and less so in a chronic course. An acute course is characterized by an approximately similar increases in the activity of protein synthesis in disease of varying severity. In an acute course the intensity of biosynthesis was the minimal in cases with the slight and the maximal in patients with grave generalized periodontitis. The content of sulfhydryl groups in the gingiva in generalized periodontitis is increased; it does not depend on the disease severity in an acute course of the disease, whereas in a chronic course this value is the maximal in cases with slight and the minimal in grave generalized periodontitis.

To elucidate the defective dentin formation in osteogenesis imperfecta (OI), we analyzed the expression of selected fibronectin (FN) isoforms in the dentin matrix of a patient with dentinogenesis imperfecta (DI) associated with OI, and in normal teeth. Frozen tooth sections were immunostained with three monoclonal antibodies (MAbs). The MAb recognizing the major cell-binding region (f-33), shared by plasma FN (pFN) and cellular FN (cFN), stained the pulp of normal adult permanent teeth intensely, while no reactivity was present in predentin, (demineralized) dentin, or dental cementum. The periodontal ligament stained unevenly. The dentin matrix of the patient with OI displayed reactive zones, alternating layerwise or concentrically with non-reactive ones. Staining throughout the connective tissue of adult oral mucosa, analyzed for the form of FN present, was intense, and in dermis, which was also studied, it was moderate. Reactivities in dental tissues with the MAb specific for the ED-A region (IST-9), included in cFN but not pFN, were similar to those with MAb f-33. The mucosal connective tissue stained weakly and dermis was negative, except that nerves and endothelia of some large blood vessels stained clearly. The MAb specific for the ED-B segment (BC-1), also included in cFN only, did not stain any of the tissues analyzed. The results suggest that, unlike mucosal and dermal FNs, FNs in the dental tissues are largely cellular, and also that dentin formation in OI may be completed by successive generations of pulpal fibroblasts differentiated into hard-tissue-forming cells.

Patients with malignant melanomas and breast cancer (83 men and women) were examined. On the basis of study of the DNA contents in nuclei of epitheliocytes from the mucous oral cavity, index of their nuclei heterogeneity we have revealed disturbances in maturation and differentiation of cells. They were arranged to the spinous layer of the oral mucosa. The disturbances were caused by the tumour influence on the functional state of the mucosa.

The molecular specificity of the dental papilla of a bell-stage tooth was studied by production of dental-papilla-reactive monoclonal antibodies (Mabs). One of the Mabs, designated 7C5, recognized an epitope present in glycosaminoglycan. Several lines of evidence suggested that the 7C5-epitope consists of chondroitin 6-sulfate. The Mab did not react with mouse dental epithelium, but reacted uniformly with mesenchymal tissue in the mandibular process and accumulated in the dental sac and in the papilla of bell-stage tooth germs. The 7C5-staining was lost from the differentiating odontoblasts, while the staining in the molar tooth papilla was accumulated in the subodontoblastic layer. In the developing mouse incisor, the 7C5-epitope was restricted to the lingual-posterior area. The 7C5-epitope was also present in pulpal tissue and predentin of different types of teeth of various mammalian species, including man, sheep, swine, and rat. Collagenase pre-treatment of tissue sections abolished the bulk of the 7C5-reactivity in peridental mesenchyme during embryonic stages while leaving the staining of the dental papilla intact. In newborn and adult teeth, collagenase also impaired the reactivity in the pulp except for the subodontoblastic layer. This suggests the existence of different subpopulations of the 7C5-epitope containing proteoglycans in dental papilla and pulp. A high-molecular-weight proteoglycan, sensitive to chondroitinase ABC but not to heparinase or heparitinase, was immunoprecipitated by 7C5 from extracts of bell-stage mouse tooth germs. We suggest that the evolutionary conservation of chondroitin 6-sulfate in the dental pulp reflects its properties as non-terminally differentiated tissue and perhaps the retention of a potential to differentiate to odontoblasts.

Proteoglycans (PGs) are extracellular and cell surface-associated macromolecules that regulate cell adhesion, cell growth, matrix formation, and bind growth factors. In this work we studied the distribution of core proteins of four PGs (decorin, biglycan, a large molecular weight PG, and CD44) in human gingiva and periodontal ligament by immunohistochemical staining of frozen tissue sections with specific antibodies. Decorin, a major PG of this tissue, was localized on collagen fiber bundles in the gingival and periodontal connective tissues. Staining for decorin was most intense at the subepithelial region. Biglycan was a minor PG component of the human periodontium, showing some accumulation in connective tissue under the oral epithelium. At the immunohistochemical level, biglycan appeared to form fine filament-like structures on extracellular matrix fibers. Localization of large molecular weight PG differed from that of decorin and biglycan. It was concentrated in deep connective tissue areas of the gingiva and in the periodontal ligament, and was only weakly present at the subepithelial region. CD44 was mainly concentrated in cell-cell contact areas of basal and spinous layers of oral epithelium. In the connective tissue of gingiva and periodontal ligament, CD44 was localized on fibroblast cell surfaces. Connective tissue area under the junctional epithelium contained relatively small amounts of PGs. The results indicate that different parts of human periodontium contain a typical variety of PGs, suggesting a specific function for each PG species in the location at which they accumulate.

We investigated the chondroitin sulfate in human periodontal samples (gingiva, periodontal ligament, cementum and alveolar bone) collected for orthodontic reasons. Glycosaminoglycans (GAGs) were extracted from the periodontium by enzyme digestion, and unsaturated disaccharide isomers of chondroitin sulfate were obtained by chondroitinase ACII and hyaluronidase digestion. The isomers were analyzed by high-performance liquid chromatography. Chondroitin sulfate was found in all four types of periodontal tissue; its unsaturated disaccharide isomers consisted in delta Di-0S, delta Di-6S, delta Di-4S, delta Di-diSE and delta Di-triS. These four types of periodontal tissue showed different molar ratios of the unsaturated disaccharides. The ratio of delta Di-4S to delta Di-6S was greater in the calcified than in the uncalcified tissue.

The aim of this study was to compare hydroxyproline (Hyp) and total protein levels both in the gingiva and gingival crevicular fluid (GCF) from juvenile (JP), rapidly progressive (RPP), and adult periodontitis (AP) patients and periodontally healthy controls (C). A total of 60 individuals, 15 from each group, were studied. GCF was obtained before gingival sampling. Clinical measurements were recorded. The gingival samples were harvested by full thickness flap operation from patients and immediately after the extraction of the teeth for orthodontic reasons from controls. The samples were analyzed biochemically. GCF Hyp levels were significantly higher in the AP group than those of the RPP and C groups. The strong positive correlations between gingival and GCF Hyp levels were determined in the disease groups. Total protein levels in both the gingiva and GCF were significantly higher in disease groups. However, the differences between the disease groups were not statistically significant. Correlations between the clinical parameters and Hyp levels in gingiva and GCF were determined. The findings indicate the Hyp and total protein levels both in the gingiva and GCF appeared to be increased in the disease groups. These findings suggest that both the synthesis and degradation of collagen and total protein are elevated in periodontal disease. However, our findings do not support the concept that measurement of either Hyp or total protein levels in gingiva or GCF is a suitable or reliable criterion in determination of disease activity.

A wide panel of monoclonal antibodies to various keratins together with an anti-vimentin antibody to label mesenchymal cells were used to label cells attached to expanded polytetrafluorethylene (ePTFE) membranes removed 6 weeks postoperatively from 7 patients. All membranes demonstrated variable amounts of adherent tissue, within which vimentin-positive cells were detected. In 8 of the 18 samples, no epithelial cells were detected. Seven of the 18 samples demonstrated small groups of epithelial cells on both sides of the membranes, staining predominantly as basal cells. The remaining 3 samples were more densely populated by epithelium which expressed a diverse keratin profile. Thus, the membranes were successful in establishing mesenchymal cell proliferation, as evidenced by vimentin positivity, but did not totally exclude epithelium.