lactoferrin and Periodontitis

Lactoferrin (LF) is a component of saliva and is suspected to be a defense factor against oral pathogens including Streptococcus mutans and Candida albicans. Periodontitis is a very common oral disease caused by periodontopathic bacteria. Antimicrobial activities and other biological effects of LF against representative periodontopathic bacteria, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia, have been widely studied. Association of polymorphisms in LF with incidence of aggressive periodontitis and the role of LF in the gingival crevicular fluid as a marker of periodontitis severity have also been reported. Periodontopathic bacteria reside as a biofilm in supragingival and subgingival plaque. Our recent study indicated that LF exhibits antibacterial activity against planktonic forms of P. gingivalis and P. intermedia at higher concentrations, and furthermore, LF effectively inhibits biofilm formation and reduces the established biofilm of these bacteria at physiological concentrations. A small-scale clinical study indicated that oral administration of bovine LF reduces P. gingivalis and P. intermedia in the subgingival plaque of chronic periodontitis patients. LF seems to be a biofilm inhibitor of periodontopathic bacteria in vitro and in vivo.

Topics: Aggregatibacter actinomycetemcomitans; Animals; Anti-Bacterial Agents; Clinical Trials as Topic; Humans; Lactoferrin; Periodontitis; Porphyromonas gingivalis; Prevotella intermedia

Topics: Adhesins, Bacterial; Animals; Bacterial Proteins; Cloning, Molecular; Cysteine Endopeptidases; Erythrocyte Aggregation; Fimbriae Proteins; Gingipain Cysteine Endopeptidases; Hemagglutinins; Hemoglobins; Humans; Iron; Lactoferrin; Mutation; Oxidative Stress; Periodontitis; Porphyromonas gingivalis; Superoxide Dismutase

Topics: Hemeproteins; Hemin; Hemoglobins; Humans; Iron; Lactoferrin; Periodontitis; Porphyromonas gingivalis; Transferrin

Biomarkers of periodontal disease activity may be obtained from potential proteolytic and hydrolytic enzymes of inflammatory cell origin. Studies that have sought to correlate these enzymes with periodontal disease activity are reviewed with special consideration given to collagenases, cysteine, aspartate and serine proteinases, beta-glucuronidase, arylsulphate, alkaline and acid phosphatases, myeloperoxidase, lysozyme and lactoferrin.

Topics: Acid Phosphatase; Alkaline Phosphatase; Arylsulfatases; Aspartic Acid Endopeptidases; Biomarkers; Collagenases; Cysteine Endopeptidases; Glucuronidase; Humans; Hydrolases; Lactoferrin; Muramidase; Peptide Hydrolases; Periodontal Diseases; Periodontitis; Peroxidase; Serine Endopeptidases

Periodontitis (PDT) has gained attention in the literature with the increase in life expectancy of people living with HIV on combined antiretroviral therapy (cART). Thus, the search for inflammatory biomarkers could be useful to understand the pathophysiology of chronic oral diseases in the cART era.. The aim of this study was to evaluate the impact of non-surgical periodontal therapy (NSPT) on clinical parameters of PDT, Candida spp. count and expression of lactoferrin (LF) and histatin (HST) in saliva and gingival crevicular fluid (GCF) of HIV-infected patients.. Bleeding index (BI), probing depth (PD), clinical attachment level (CAL), colonyforming units (CFUs) of Candida spp, and LF and HST levels were measured in saliva and GCF of both groups at three different times: baseline (before treatment), and 30 and 90 days after the NSPT. Clinical, mycological and immunoenzymatic analyses were also performed.. Twenty-two HIV-infected patients and 25 non-HIV-infected patients with PDT participated in the study. NSPT was effective in improving periodontal clinical parameters, including ≤ 4 sites with PD ≤ 5mm and BI ≤ 10%. Significant change in oral Candida spp. count occurred neither between the two groups nor after NSPT. And the salivary and GCF levels of LF and HST were not influenced by the NSPT; by contrast, except for salivary LF, HST and LF were shown to exhibit significantly higher levels in HIV-infected than in non-HIV-infected patients.. NSPT was effective in improving periodontal disease parameters in HIV-infected patients, but did not affect LF and HST expression in saliva and GCF of HIV-infected patients.

Topics: Candida; Gingival Crevicular Fluid; Histatins; HIV Infections; Humans; Lactoferrin; Periodontitis; Saliva

This study was conducted to evaluate the antimicrobial potential of the antimicrobial peptides (AMP) LL-37 and human Lactoferricin (LfcinH) on the planktonic growth and biofilm formation of oral pathogenic anaerobes related to caries and periodontitis. Multi-species bacterial suspensions of either facultative anaerobic bacteria (FAB: Streptococcus mutans, Streptococcus sanguinis, Actinomyces naeslundii) or obligate anaerobic bacteria (OAB: Veillonella parvula, Parvimonas micra, Fusobacterium nucleatum) were incubated with different concentrations of AMP solutions for 8 h. Planktonic growth was registered with an ATP-based cell viability assay for FAB and via plate counting for OAB. Biofilms were grown on ZrO

Topics: Antimicrobial Cationic Peptides; Bacteria, Anaerobic; Biofilms; Cathelicidins; Dental Caries; Humans; Lactoferrin; Microbial Sensitivity Tests; Microbial Viability; Oxygen; Periodontal Diseases; Periodontitis

We aimed to evaluate the effect of bovine lactoferrin (bLF) on alveolar bone destruction and remodelling under orthodontic force (OF) in periodontitis-affected rats.. After establishing the periodontitis-affected rat model with lipopolysaccharides (LPS), the left maxillary first molars were moved orthodontically under a force of 0.2N. Based on saline or bLF gavage, 54 Sprague-Dawley (SD) rats were randomized into 5 groups: A (blank), P1 (LPS+OF+bLF), P2 (LPS+OF+saline), C1 (OF+bLF), and C2 (OF+saline). Animals were evaluated using micro-computed tomography (micro-CT) followed by haematoxylin and eosin (H&E) and tartrate-resistant acid phosphatase (TRAP) staining, and the LF level was determined using ELISA in the gingival crevicular fluid (GCF) of the experimental teeth. Immunohistochemistry helped to detect expression changes in RANKL, OPG and COX-2.. Micro-CT results indicated that compared with group P2, trabecular number (Tb.N) and trabecular thickness (Tb.Th) in group P1 were higher and bone surface/bone volume (BS/BV) was lower on day 14, while trabecular separation (Tb.Sp) decreased significantly on Day 5 and Day 14 after bLF gavage (P<0.05). This was supported by changes in H&E and TRAP staining. bLF down-regulated RANKL level at both timepoints and up-regulated OPG level on Day 14 in periodontitis rats (P<0.05). The significant changes mentioned above were not observed between group C1 and C2 (P>0.05). No significant change in COX-2 levels were observed in any group (P>0.05). The lactoferrin level in GCF increased significantly after bLF gavage (P<0.05).. Bovine lactoferrin inhibited LPS-induced bone destruction, but the bone healing effect was independent of orthodontic aseptic inflammatory bone remodelling.

Topics: Alveolar Bone Loss; Animals; Lactoferrin; Lipopolysaccharides; Periodontitis; Rats; Rats, Sprague-Dawley; X-Ray Microtomography

Adjunctive use of antibiotics in periodontal treatment have limitations and disadvantages including bacterial resistance. Antimicrobial peptides (AMPs) are potential new agents that can combat bacterial infection. In this study, antimicrobial activity of different concentrations of conventional antibiotics minocycline (MH), doxycycline (DOX), and antimicrobial peptides LL-37, LL-31, Lactoferrin chimera (LFchimera) and Innate Defense Regulator Peptide 1018 (IDR-1018) against Aggregatibacter actinomycetemcomitans ATCC 43718 were determined using colony culturing assay. Subsequently, in vitro activity of the most effective drug and peptide combination was evaluated by checkerboard technique. Impact of the drug and peptide co-administration on biofilm at different stages, i.e., during adhesion and 1-day old biofilm was compared to each of the agents used alone. Results revealed that the killing effects of all AMPs range from 13-100%. In contrast, MH and DOX at 1 and 5 μM showed no killing activity and instead stimulated growth of bacteria. DOX has better killing activity than MH. LFchimera displayed the strongest killing amongst the peptides. Checkerboard technique revealed that combining DOX and LFchimera yielded synergism. Confocal laser scanning microscopy further showed that the combination of DOX and LFchimera caused significant reduction of bacterial adhesion and reduction of biomass, average biofilm thickness and substratum biofilm coverage of 1-day old biofilm compared to DOX and LFchimera alone. In conclusion, LFchimera alone and in combination with DOX exhibited strong antibacterial and anti-biofilm property against A. actinomycetemcomitans. The findings suggest that LFchimera should be considered for development as a new potential therapeutic agent that may be used as an adjunctive treatment for periodontitis.

Topics: Aggregatibacter actinomycetemcomitans; Anti-Bacterial Agents; Antimicrobial Cationic Peptides; Biofilms; Drug Synergism; Humans; Lactoferrin; Periodontitis; Plankton; Recombinant Fusion Proteins

To examine the role of lactoferrin (LF) on Toll like receptor 4 (TLR4) stimulated by lipopolysaccharide (LPS) in human periodontal ligament cells (hPDLCs).. Primary hPDLCs were cultured by tissue block enzymolytic method. Cells obtained from four passages were identified and used in this experiment. Cells without stimulation served as the controls and cells treated with LPS (0.1 microg x mL(-1)) comprised the LPS group. The LPS + LF group was pretreated with LPS (0.1 microg x mL(-1)) for 2 h, and then treated with LF (10 microg x mL(-1)). Four hours after LF stimulation, the mRNA expression levels of TLR4 were examined by real-time quantitative polymerase chain reaction (RT-PCR). The protein expression of TLR4 was observed by cell immunofluorescence staining after LF stimulation of 24 hours.. TLR4 mRNA expression in the LPS + LF group was significantly more decreased than that in the LPS group (P < 0.05), but exhibited no difference with that in the control group (P > 0.05). Cell immunofluorescence staining showed that the protein expression of TLR4 in the LPS + LF group was significantly more decreased than that in the LPS group (P < 0.05), but exhibited no difference with that in the control group (P > 0.05).. LF can decrease the expression of TLR4 stimulated by LPS in hPDLCs, thus presenting potential application for controlling the TLR4 immune pathway of periodontitis.

Topics: Down-Regulation; Humans; Lactoferrin; Lipopolysaccharides; Periodontal Ligament; Periodontitis; Toll-Like Receptor 4

The aim of this study was to evaluate lactoferrin quantification as a sensitive and objective method of detecting the degree of periodontal inflammation, oxidative stress and to monitor the effects of periodontal therapy.. Fifty subjects were divided into two groups based on gingival index, probing pocket depth, clinical attachment loss and alveolar bone loss: healthy group and periodontitis group with generalized chronic periodontitis. Non-surgical periodontal therapy was rendered and crevicular fluid samples collected at baseline and four weeks after therapy for lactoferrin quantification using enzyme linked immunosorbent assay. The correlation between clinical parameters and lactoferrin levels was drawn and analysed for both groups.. The mean level of crevicular lactoferrin in the periodontitis group was 1857.21 ng/ml. The mean level decreased to 1415.03 ng/ml after treatment. The lowest lactoferrin concentration was seen in the healthy group (75.34 ng/ml). All clinical parameters correlated positively with lactoferrin levels.. The lactoferrin level was higher in the periodontitis group compared to the healthy group, and reduced with periodontal therapy. Higher levels were associated with higher values of clinical parameters, both before and after therapy. The data indicates that Lactoferrin plays an important role in periodontal disease and crevicular lactoferrin quantification can be a marker for detecting periodontal inflammation, oxidative stress and monitoring periodontal therapy.

Topics: Adult; Biomarkers; Case-Control Studies; Female; Gingival Crevicular Fluid; Humans; Lactoferrin; Male; Middle Aged; Oxidative Stress; Periodontal Diseases; Periodontal Index; Periodontitis

Bovine lactoferrin (bLF) modulates the production of tumor necrosis factor-alpha (TNF-α) and inhibits alveolar bone breakdown associated with periodontitis. This study is designed to examine the effects of orally administered liposomal bLF (LbLF) on orthodontic force (OF)-induced alveolar bone remodeling during experimental tooth movement.. Two groups of male Wistar rats were treated with either LbLF or control solution in drinking water 7 days before OF application. Lipopolysaccharide (LPS) was injected into the gingival sulcus in half the rats in each group. Thus, four groups: OF, OF+LbLF, OF+LPS, and OF+LPS+LbLF were established.. Orally administered LbLF significantly reduced apical migration of junctional epithelium in the OF and OF+LPS groups. In OF+LPS, osteoclast number in the alveolar crestal area was increased by LPS treatment, whereas osteoclast number was significantly reduced in OF+LPS+LbLF through suppression of TNF-α production. Osteoclastic induction in the middle part, mainly from OF application, was not affected by LbLF administration. Inhibition of tooth movement was not induced by LbLF.. Orally administered LbLF significantly inhibits LPS-induced alveolar bone resorption but not OF-induced bone remodeling. LbLF could be a potent therapeutic and preventive agent to control periodontal inflammation in patients undergoing orthodontic treatment.

Topics: Acid Phosphatase; Administration, Oral; Alveolar Bone Loss; Alveolar Process; Animals; Anti-Infective Agents; Bone Remodeling; Cattle; Cell Count; Epithelial Attachment; Escherichia coli; Isoenzymes; Lactoferrin; Lipopolysaccharides; Liposomes; Male; Osteoclasts; Periodontitis; Pilot Projects; Random Allocation; RANK Ligand; Rats; Rats, Wistar; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Tooth Cervix; Tooth Movement Techniques; Tumor Necrosis Factor-alpha

Bovine lactoferrin (bLF) modulates the production of proinflammatory cytokines including tumor necrosis factor (TNF)-alpha, and may thus control alveolar bone destruction associated with periodontitis. In this study, the effects of bLF on mRNA expression in lipopolysaccharide (LPS)-stimulated osteoblasts (OBs) and on LPS-induced osteoclastogenesis were examined. The inhibitory effects of oral administration of liposomal-bLF (L-bLF), which improved the robustness of bLF to digestive enzymes, on alveolar bone resorption using LPS-induced periodontitis rat model are also reported. Three groups of 7-week-old male Wistar rats were treated with L-bLF (L-bLF group), bLF (bLF group), or the vehicle (control group) in drinking water (n=6 in each group). On day 7, LPS was topically applied into the gingival sulcus. Number of osteoclasts and immunoexpression of TNF-alpha were analyzed. The bLF inhibited the upregulation of TNF-alpha-mRNA- and upregulation of receptor activator of NF kappaB (RANKL)-mRNA expression and eliminated downregulation of osteoprotegerin (OPG)-mRNA expression in LPS-stimulated OBs and reduced LPS-induced osteoclastogenesis in co-culture with primary OBs and bone marrow cells. In the control group, the number of osteoclasts increased after LPS treatment. The number of osteoclasts that appeared along the alveolar bone margin was significantly reduced (P<0.01) in the L-bLF but not in the bLF group. Furthermore, L-bLF suppressed upregulation of TNF-alpha immunoexpression in periodontal tissue and TNF-alpha and interleukin (IL)-1 beta-mRNA level in gingival tissue. The results of this study indicate that oral administration of L-bLF significantly reduces alveolar bone resorption induced by LPS stimulation through inhibition of TNF-alpha production and modulation of RANKL/OPG balance in OBs. It is suggested that L-bLF could be a potent therapeutic and preventive agent for attenuating alveolar bone destruction in periodontitis patients.

Topics: Alveolar Bone Loss; Animals; Bone Marrow Cells; Bone Resorption; Cattle; Coculture Techniques; Cytokines; Gingiva; Interleukin-1; Lactoferrin; Lipopolysaccharides; Male; Osteoblasts; Osteoclasts; Osteoprotegerin; Periodontitis; Rats; Rats, Wistar; Receptor Activator of Nuclear Factor-kappa B; Tumor Necrosis Factor-alpha

Insufficient data exist regarding the longitudinal influence of involuntary smoking on periodontitis progression. This study examined the relationship between involuntary smoking and periodontitis progression and the effects of involuntary smoking on salivary inflammatory and microbiologic markers related to periodontitis.. Participants were recruited during annual health checkups in 2003 and 2005. In 2005, 200 of 273 (73%) Japanese employees examined at baseline underwent periodontal measurements, including clinical attachment level (CAL) and probing depth (PD). Periodontitis progression was identified when a subject displayed one or more teeth with an increase > or = 2.0 mm in CAL and PD during the 2 years. Salivary marker levels, including cotinine, were determined by enzyme assay, including enzyme-linked immunosorbent assay. The proportions of six periodontal pathogens in saliva were assessed using real-time polymerase chain reaction methodology. Based on receiver-operating characteristic analysis, non-, involuntary, and active smokers were defined as subjects exhibiting salivary cotinine levels of 0, 1 to 7, and > or = 8 ng/ml, respectively.. By simple logistic regression analysis, age, alcohol consumption, smoking, breakfast habits, and working hours were related to the risk for significant periodontitis progression. Multiple logistic regression analysis revealed significantly higher periodontitis odds ratios (OR) in involuntary (OR = 2.23; 95% confidence interval [CI]: 1.03 to 4.83) and active (OR = 2.27; 95% CI: 1.02 to 5.04) smokers relative to non-smokers following adjustment for covariates. Levels of salivary markers, including albumin, aspartate aminotransferase, and lactoferrin, were significantly elevated in involuntary smokers relative to non-smokers. In contrast, the percentages of periodontal pathogens did not differ between the smoking groups, with the exception of Prevotella nigrescens, which displayed significantly lower levels in involuntary smokers compared to non-smokers.. Involuntary smoking increased the inflammatory response and was associated with a greater risk for periodontitis progression.

Topics: Adolescent; Adult; Age Factors; Albumins; Alcohol Drinking; Aspartate Aminotransferases; Biomarkers; Cotinine; Disease Progression; Feeding Behavior; Female; Humans; Indicators and Reagents; Lactoferrin; Longitudinal Studies; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Pocket; Periodontitis; Prevotella nigrescens; Risk Factors; Saliva; Smoking; Tobacco Smoke Pollution; Work; Young Adult

Lactoferrin (Lf) is a member of the transferrin family of iron-binding anti-bacterial proteins, present in most exocrine secretions, such as saliva, and plays an important role in mucosal defense. In this study, we identified small Lf peptides with Con A low-affinity in the parotid saliva of chronic periodontitis patients by Con A two-dimensional immunoelectrophoresis, Con A affinity chromatography and Western blotting using anti-human Lf polyclonal Ab. N-terminal amino acid sequencing of the four Con A low-affinity Lf peptides confirmed them to be fragments of intact Lf. The detection ratio of the proteinase 3 (PR3)-like activity was elevated in the parotid saliva of periodontitis patients and was associated with the severity of clinical symptoms. PR3 protein was also detected in the parotid saliva of periodontitis patients, and PR3, but not human leukocyte elastase and cathepsin G, degraded intact Lf. Con A low-affinity saliva Lf peptides showed no anti-bacterial activity against Escherichia coli, and had a reduced iron-chelating capacity. Con A low-affinity saliva Lf peptides, PR3-treated Lf preparation and two of four synthetic polypeptides induced the production of interleukin IL-6, monocyte chemoattractant protein-1 and IL-8, and the activation of NF-kappaB in human oral epithelial HSC-2 cells. Furthermore, concentrations of the Lf peptides in the parotid saliva of periodontitis patients were increased with a correlation to the severity of clinical symptoms. These results suggest that Lf in the parotid saliva of periodontitis patients was degraded into small peptides by the PR3-like activity with the capability to induce inflammatory mediators.

Topics: Amino Acid Sequence; Chemokines; Concanavalin A; Cytokines; Electrophoresis, Gel, Two-Dimensional; Humans; Lactoferrin; Molecular Sequence Data; Mouth Mucosa; Myeloblastin; NF-kappa B; Parotid Gland; Peptide Fragments; Periodontitis; Saliva

Insufficient data exist regarding the long-term influence of lifestyle factors including smoking on periodontal health. The objective of this study was to examine the prospective association between smoking and periodontal disease progression and the effects of smoking on salivary biomarkers related to periodontitis.. Probing depth (PD) was measured at health checkups of workers in 1999 and 2003; additionally, lifestyle information was obtained through a questionnaire. In 2003, 219 of 256 (86%) workers examined at baseline completed PD measurements; saliva samples were also collected. Change in PD was used for assessment of periodontitis progression when three or more sites displayed an increase of >or=2 mm over 4 years. Salivary biomarker levels were determined by real-time polymerase chain reaction and enzyme assay. Statistical methods included bivariate and multivariate regression analyses.. In the multiple logistic model, in which lifestyle-related factors served as independent variables, significant variables were current smoking and hours of sleep; respective odds ratios were 2.3 and 2.1. Additionally, 38.5% of periodontal disease progression was attributable to current smoking. Moreover, pack-years of smoking showed a dose-response relationship with disease progression. Levels of salivary markers including prostaglandin E(2), lactoferrin, albumin, aspartate aminotransferase, lactate dehydrogenase, and alkaline phosphatase were significantly lower in current smokers than in non-current smokers. However, no meaningful differences in the proportions of six periodontal pathogens were observed between current and non-current smokers.. Smoking exerted the greatest influence on periodontitis risk among lifestyle factors. Smoking may suppress the host-defense system, which may promote periodontal disease progression.

Topics: Adolescent; Adult; Albumins; Alkaline Phosphatase; Aspartate Aminotransferases; Bacteroidaceae; Biomarkers; Dental Health Surveys; Dinoprostone; Disease Progression; Female; Humans; Japan; L-Lactate Dehydrogenase; Lactoferrin; Life Style; Logistic Models; Longitudinal Studies; Male; Middle Aged; Periodontitis; Population Surveillance; Regression Analysis; Risk Factors; Saliva; Sex Factors; Smoking

The mechanism of passive smoking in terms of development of periodontitis has not been investigated. This study examined the effect of passive smoking on salivary markers related to periodontitis.. Periodontal status was evaluated on the basis of probing pocket depth and clinical attachment level in 273 workers. Salivary marker levels were determined by enzyme assay including enzyme-linked immunosorbent assay. Six periodontal pathogens in saliva were assessed using real-time PCR methodology. Non-, passive and active smokers were defined as subjects exhibiting salivary cotinine levels of 0 (53 subjects), 1-7 (118) and > or = 8 ng/ml (102).. Levels of salivary markers, including IL-1beta, lactoferrin, albumin and aspartate aminotransferase (AST), were elevated significantly in passive smokers relative to non-smokers. Additionally, these marker levels, with the exception of IL-1beta, decreased significantly in active smokers in comparison with passive smokers. However, no meaningful differences in percentages of periodontal pathogens were observed between non- and passive smokers. Multiple linear regression analyses were performed for each marker utilizing age, gender, cotinine level and periodontal status as independent variables. IL-1beta, albumin and AST were independently associated with cotinine level.. Passive smoke exposure leads to elevation of IL-1beta, albumin and AST levels in saliva.

Topics: Adolescent; Adult; Albumins; Aspartate Aminotransferases; Biomarkers; Cotinine; Female; Humans; Interleukin-1beta; Lactoferrin; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Porphyromonas gingivalis; Prevotella intermedia; Prevotella nigrescens; Saliva; Smoking; Tobacco Smoke Pollution

The aim of this study was to compare the activity of neutrophilic granulocytes in patients with severe periodontitis and patients with gingivitis alone.. The study population comprised 22 patients with gingivitis and 44 with periodontitis. Samples of gingival crevicular fluid (GCF) were collected from untreated patients with gingivitis and from shallow and deep pockets in untreated patients with periodontitis. GCF samples were analyzed for lactoferrin, elastase, matrix metalloproteinase-8 and -9, and collagenolytic activity.. The free elastase activity and the neutrophil activity, estimated as the ratio between elastase and lactoferrin, were significantly higher in the samples from the periodontitis patients. These differences were also observed in shallow pockets in periodontitis patients compared to similar pockets in patients with gingivitis.. This study shows higher levels of free elastase in untreated patients with periodontitis, relative to inflammation-matched controls, which may explain the tissue destruction seen in periodontitis.

Topics: Animals; Cattle; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Lactoferrin; Male; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Middle Aged; Neutrophil Activation; Neutrophils; Pancreatic Elastase; Periodontal Pocket; Periodontitis; Statistics, Nonparametric

Lactoferrin is an antimicrobial protein which plays an important role in regulating bacteria that are associated with aggressive periodontitis. Lactoferrin kills directly (via its strongly cationic N-terminal region) and indirectly, through sequestering the iron that bacteria require for growth. As aggressive periodontitis has a strong heritable component, we hypothesized that genetic variation within the lactoferrin gene may play a role in susceptibility to this condition. We have identified and examined a novel, functional, single-point A/G nucleotide mutation causing a threonine/alanine substitution at position 11 (T11A) of the secreted lactoferrin protein. In a pilot case-controlled study of aggressive periodontitis, analysis of 46 African-American patients and 78 controls showed that patients were twice as likely to express the G nucleotide (alanine) allele over controls (60.3 vs 30.4%; P=0.0007, odds ratio=2.564, 95% CI=1.475-4.459). A Caucasian population of 77 patients and 131 controls showed no such association (P=0.5201, odds ratio=0.862, 95% CI=0.548-1.356). The data presented provide a new insight into the genetic susceptibility to aggressive periodontitis.

Topics: Alanine; Amino Acid Substitution; Black or African American; Case-Control Studies; Conserved Sequence; Female; Humans; Lactoferrin; Male; Periodontitis; Point Mutation; Polymorphism, Single Nucleotide; Threonine; White People

The aim of the study was to verify (i) if crevicular fluid defence variables reflect the changes after surgical periodontal treatment and (ii) if they are in correspondence with changes of these variables in the unstimulated and stimulated whole saliva.. For 12 male and 13 female volunteers with chronic periodontitis lactoferrin concentration as well as the lysozyme and peroxidase activities were determined in crevicular fluid as well as in unstimulated and stimulated saliva before and 14 days after surgical periodontal treatment by a minimal invasive flap technique.. The lactoferrin concentrations decreased significantly in the crevicular fluid eluting solution from 1.63 to 1.23 mg/l reflecting a decrease in the total amount collected, in unstimulated saliva from 10.54 to 8.96 mg/l, and in stimulated saliva from 9.00 to 7.11 mg/l after treatment. No significant change could be found for lysozyme. Peroxidase activity was significantly reduced from 269.06 to 186.15 U/l only in the crevicular fluid.. The results of this study suggest that (i) the defence factor lactoferrin is suitable for monitoring of periodontal treatment results and (ii) changes of the lactoferrin concentration in crevicular fluid are related with significant changes in unstimulated and stimulated saliva.

Topics: Biomarkers; Chronic Disease; Female; Gingival Crevicular Fluid; Humans; Immunoenzyme Techniques; Lactoferrin; Male; Middle Aged; Muramidase; Periodontitis; Peroxidase; Saliva; Statistics, Nonparametric

Human periodontal diseases are inflammatory disorders that are the result of complex interactions between periodontopathogens and the host's immune response. Two important and interrelated factors are involved in the pathophysiological progression of periodontal diseases, i.e. the activation of immune system and the production of oxygen radicals and their related metabolites. Increased production of oxygen radicals may contribute to oxidative stress, which is reported to be involved in many diseases, including periodontal diseases.. The objective of this study was to investigate glutathione peroxidase, lactoferrin and myeloperoxidase, which play an essential role in free radical production and defenses, and the proinflammatory cytokine interleukin-1beta (IL-1beta), which is important in the regulation of immunological and inflammatory reactions in human periodontal diseases.. Gingival crevicular fluid (GCF) samples were collected from 27 subjects, 19 periodontitis patients and eight healthy controls, ranging in ages from 24 to 62 years. Clinical parameters were recorded. GCF glutathione peroxidase, lactoferrin, myeloperoxidase and IL-1beta were analyzed by enzyme-linked immunosorbent assays (ELISA).. The periodontitis sites exhibited significantly greater total amount of glutathione peroxidase, lactoferrin, myeloperoxidase and IL-1beta than healthy sites. Total amount of glutathione peroxidase, lactoferrin, myeloperoxidase and IL-1beta was positively correlated with plaque index, gingival index, probing depth and probing attachment level (p < 0.05).. The imbalance between the levels of myeloperoxidase/IL-1beta and glutathione peroxidase/lactoferrin could result in tissue damage of reactive oxygen species (ROS) in periodontitis which is initiated and perpetuated by the chronic insults of periodontopathogens.

Topics: Adult; Analysis of Variance; Case-Control Studies; Female; Gingival Crevicular Fluid; Gingivitis; Glutathione Peroxidase; Humans; Interleukin-1; Lactoferrin; Male; Middle Aged; Oxidative Stress; Periodontal Index; Periodontitis; Peroxidase; Statistics, Nonparametric

The aim of the present study was to characterise microbiota and inflammatory host response around implants and teeth in patients with peri-implantitis. We included 17 partly edentulous patients with a total of 98 implants, of which 45 showed marginal bone loss of more than three fixture threads after the first year of loading. Nineteen subjects with stable marginal tissue conditions served as controls. Oral hygiene, gingival inflammation, and probing pocket depth were evaluated clinically at teeth and implants. Microbiological and crevicular fluid samples were collected from five categories of sites: 1) implants with peri-implantitis (PI), 2) stable implants (SI) in patients with both stable and peri-implantitis implants, 3) control implants (CI) in patients with stable implants alone, 4) teeth in patients (TP) and 5) controls (TC). Crevicular fluid from teeth and implants was analysed for elastase activity, lactoferrin and IL-1 beta concentrations. Elastase activity was higher at PI than at CI in controls. Lactoferrin concentration was higher at PI than at SI in patients with peri-implantitis. Higher levels of both lactoferrin and elastase activity were found at PI than at teeth in patients. The concentrations of IL-1 beta were about the same in the various sites. Microbiological DNA-probe analysis revealed a putative periodontal microflora at teeth and implants in patients and controls. Patients with peri-implantitis harboured high levels of periodontal pathogens, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus and Treponema denticola. These findings indicate a site-specific inflammation rather than a patient-associated specific host response.

Topics: Aged; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Bacteroides; Dental Implants; Dental Plaque Index; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Interleukin-1; Jaw, Edentulous, Partially; Lactoferrin; Male; Middle Aged; Pancreatic Elastase; Periodontal Index; Periodontal Pocket; Periodontitis; Porphyromonas gingivalis; Prevotella intermedia; Statistics, Nonparametric; Treponema

Porphyromonas gingivalis possesses a hemoglobin receptor (HbR) protein on the cell surface as one of the major components of the hemoglobin utilization system in this periodontopathogenic bacterium. HbR is intragenically encoded by the genes of an arginine-specific cysteine proteinase (rgpA), lysine-specific cysteine proteinase (kgp), and a hemagglutinin (hagA). Here, we have demonstrated that human lactoferrin as well as hemoglobin have the abilities to bind purified HbR and the cell surface of P. gingivalis through HbR. The interaction of lactoferrin with HbR led to the release of HbR from the cell surface of P. gingivalis. This lactoferrin-mediated HbR release was inhibited by the cysteine proteinase inhibitors effective to the cysteine proteinases of P. gingivalis. P. gingivalis could not utilize lactoferrin for its growth as an iron source and, in contrast, lactoferrin inhibited the growth of the bacterium in a rich medium containing hemoglobin as the sole iron source. Lactoferricin B, a 25-amino acid-long peptide located at the N-lobe of bovine lactoferrin, caused the same effects on P. gingivalis cells as human lactoferrin, indicating that the effects of lactoferrin might be attributable to the lactoferricin region. These results suggest that lactoferrin has a bacteriostatic action on P. gingivalis by binding HbR, removing it from the cell surface, and consequently disrupting the iron uptake system from hemoglobin.

Topics: Anti-Bacterial Agents; Bacterial Proteins; Culture Media; Hemoglobins; Humans; Lactoferrin; Models, Biological; Mutation; Peptides; Periodontitis; Porphyromonas gingivalis; Protein Binding; Receptors, Cell Surface

Our aim was to compare the levels of laminin and interleukin-8 (IL-8) in GCF from inflamed shallow (GP) and deep pockets (PP) in patients with periodontitis to these levels in GCF from inflamed shallow pockets (GG) in subjects with gingivitis alone.. Lactoferrin was used as a marker for the number of neutrophils in the sites sampled. The periodontitis group consisted of 13 subjects, having at least 6 sites with a pocket depth > or = 5 mm. The healthy control group consisted of 12 subjects with no clinical signs of periodontal destruction. GCF was collected with paper strips and the volume was measured immediately after sampling. Laminin, lactoferrin and IL-8 were measured using specific antibodies with an ELISA.. The total amount of laminin was significantly higher in PP than in GG, but no significant difference was seen between GP and PP. The concentration and the total amounts of lactoferrin were similar in the three groups. The ratio between laminin and lactoferrin was higher in the samples from the patient, suggesting that neutrophils in patients are more activated and degrades more of the membrane per cell. The total amounts of IL-8 were very similar in the 3 groups, while the concentration also tended to be higher in the GP.. Our study showed higher amounts of laminin in GCF from patients with periodontitis suggesting the presence of hyperactive neutrophils during the transmigration process through the endothelium/epithelium.

Topics: Adult; Basement Membrane; Chemotaxis, Leukocyte; Female; Gingival Crevicular Fluid; Humans; Interleukin-8; Lactoferrin; Laminin; Male; Neutrophil Activation; Periodontitis; Statistics, Nonparametric

This investigation was designed to determine and compare the distribution pattern of anti-neutrophil cytoplasmic antibodies (ANCA) in patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) in the presence or absence of periodontal disease.. Sera of 30 patients with SLE and 30 with RA were tested for ANCA utilizing an indirect enzyme immunosorbent assay (ELISA) directed to a neutrophil granular extract and 6 neutrophil granule proteins. A control group of 20 healthy individuals showing neither evidence of periodontal disease nor systemic compromise was also included in this study.. For RA, the number of ANCA-positive sera was very low but was evenly distributed among patients with and without periodontitis. Conversely, a high number of ANCA-positive sera in SLE was found mostly in individuals presenting periodontal compromise. A statistically significant association between ANCA and periodontitis in SLE patients was found (P <0.005, chi square test).. A marked difference in the number and distribution of ANCA with respect to periodontitis between RA and SLE was found. Hyperresponsiveness of B cells and polyclonal B activation to periodontopathic bacteria in SLE might be accountable for the high numbers of ANCA and the close association observed between those autoantibodies and periodontitis in SLE.

Topics: Adult; Antibodies, Antineutrophil Cytoplasmic; Arthritis, Rheumatoid; Autoantigens; B-Lymphocytes; Bacteria; Cathepsin G; Cathepsins; Chi-Square Distribution; Enzyme-Linked Immunosorbent Assay; Female; Humans; Lactoferrin; Leukocyte Elastase; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Middle Aged; Muramidase; Myeloblastin; Neutrophils; Periodontitis; Peroxidase; Serine Endopeptidases

The main object of this study was to determine if there was a difference between patients with adult periodontitis and healthy controls in the release of elastase. We also wanted to test the release of alpha-1-antitrypsin and lactoferrin from in vitro-activated peripheral neutrophils. A leukocyte-rich preparation from venous blood was made by lysing the red blood cells. The leukocytes were stimulated for 1 h at 37 degrees C with opsonized Staphylococcus aureus and the released elastase was measured with a chromogenic substrate. The release of elastase after stimulation with bacteria was significantly higher in patients than in controls. The amounts of elastase from unstimulated cells, i.e., both released extracellularly and extracted from the pellet, were similar in the 2 groups. However, after stimulation, the amount of elastase in the patient group, but not in the control group, was significantly increased. Similar releases of alpha-1-antitrypsin (AIAT) and lactoferrin were found in both groups of subjects. In conclusion, this study shows that peripheral neutrophils from patients with adult periodontitis release more active elastase after in vitro activation compared to healthy controls. The release of A1AT and lactoferrin showed no differences, indicating that the increased elastase activity was not due to a impaired inhibition by A1AT and that the differences in degranulation were limited to the primary granula.

Topics: Adult; Aged; alpha 1-Antitrypsin; Cell Degranulation; Cells, Cultured; Chromogenic Compounds; Female; Flow Cytometry; Humans; Lactoferrin; Leukocyte Elastase; Leukocytes; Male; Middle Aged; Neutrophil Activation; Neutrophils; Oligopeptides; Periodontitis; Pyrrolidonecarboxylic Acid; Staphylococcus aureus

Concentrations and output of lactoferrin and of low-Mr mucin MG2 were determined in saliva of subjects suffering from Actinobacillus actinomycetemcomitans-associated periodontal disease and healthy subjects. Periodontal patients were clinically examined and a microbiological sample was taken from the deepest bleeding pockets in each quadrant. The number of viable A. actinomycetemcomitans was determined in the sampled sites of each patient. The MG2 output in the diseased subjects (13.6 microg protein/min) was decreased at least by a factor three compared to periodontal healthy subjects (44.3 microg protein/min). On the other hand, output of lactoferrin was not significantly different in healthy (9.5 microg/min) and diseased subjects (7.6 microg/min). Western analyses demonstrated a higher iron-saturation of lactoferrin in diseased subjects in comparison with the healthy subjects. Lactoferrin degrading enzymes, probably derived from microbial sources, could be detected in saliva of the periodontally diseased subjects, but not in saliva of healthy subjects. The combination of iron-saturation and degradation of lactoferrin suggests that anti-microbial properties of lactoferrin are diminished in periodontitis patients. Moreover, the low concentration of mucin MG2 suggests a decline in mucin defence and consequently a higher susceptibility for oral infection. A negative correlation (r= -0.4, p < 0.05) between the number of subgingival A. actinomycetemcomitans and lactoferrin in saliva suggested that low concentrations of lactoferrin favour the growth of the bacterium. These data indicate that a decline in the salivary defence system might increase the risk for oral infection by A. actinomycetemcomitans.

Topics: Actinobacillus Infections; Adolescent; Adult; Aggregatibacter actinomycetemcomitans; Albumins; Blotting, Western; Colony Count, Microbial; Cystatins; Cysteine Proteinase Inhibitors; Electrophoresis, Polyacrylamide Gel; Female; Humans; Immunoblotting; Iron; Lactoferrin; Male; Middle Aged; Mucins; Periodontal Pocket; Periodontitis; Risk Factors; Saliva; Salivary Proteins and Peptides

The present study was designed to determine in a cross-sectional study whether there was any relationship between the levels of lactoferrin in gingival crevicular fluid and clinical periodontal parameters. Crevicular fluid was collected from individual sites using standardized filter paper strips (clinically healthy sites, N = 23; periodontitis sites, n = 66) and evaluated for lactoferrin by enzyme-linked immunosorbent assay. The data showed that: (1) the total amounts of lactoferrin were 0.003-0.021 ng (30 second sample) (average 0.009 +/- 0.005 ng) in a clinically healthy periodontium group and 0.016-3.847 ng (30 second sample) (average 0.575 +/- 0.069 ng) in adult periodontitis patients (statistically significantly higher in adult periodontitis patients); and (2) the total amounts of lactoferrin were significantly correlated with clinical parameters, especially a strong positive correlation with gingival crevicular fluid volume (r = 0.85, p < 0.01) and with probing depth (r = 0.71, p < 0.01). These results indicated that quantification of lactoferrin in gingival crevicular fluid may be a more sensitive indicator of periodontal pathology than traditional clinical indices.

Topics: Adult; Analysis of Variance; Biomarkers; Cross-Sectional Studies; Dental Plaque Index; Enzyme-Linked Immunosorbent Assay; Female; Gingival Crevicular Fluid; Humans; Lactoferrin; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Periodontium

The number of decayed, missed and filled permanent teeth (DMFT), the degree of periodontal inflammation (Periodontal Status Index, PSI), stimulated salivary flow rate and the concentrations of total protein, lactoferrin, lysozyme, myeloperoxidase, salivary peroxidase, calcium, potassium, sodium and thiocyanate in whole saliva of 26 adult asthma patients were compared with those of 33 non-asthmatic controls. The saliva was also analysed for mutans streptococci, lactobacilli, total anaerobic flora and Candida spp. The mean PSI (p < 0.05; 95% confidence interval for the difference between means (95% CI) 2.47-25.30) was higher and the mean stimulated salivary flow rate (p < or = 0.05; 95% CI 0.57-0.55) was lower in the asthmatic group than in the control group. No differences were found between the groups in non-immune defense factors, except for myeloperoxidase. The myeloperoxidase concentrations were higher in asthmatics than in non-asthmatics (p < 0.05; 95% CI 4.4-134.0 ng/ml). No differences in microbial counts were found. It was concluded that stimulated salivary flow rates decrease while myeloperoxidase concentrations increase in adult asthmatic patients compared with non-asthmatic adults. The higher concentrations of myeloperoxidase are explained by a higher PSI in asthmatics.

Topics: Adult; Asthma; Bacteria, Anaerobic; Calcium; Candida; Colony Count, Microbial; Confidence Intervals; DMF Index; Female; Humans; Lactobacillus; Lactoferrin; Male; Middle Aged; Muramidase; Periodontal Index; Periodontitis; Peroxidase; Peroxidases; Potassium; Saliva; Salivary Proteins and Peptides; Secretory Rate; Sodium; Streptococcus mutans; Thiocyanates

A 40-kDa lactoferrin-binding protein was identified in a strain of Prevotella nigrescens isolated from a patient with periodontitis. The protein was purified by affinity column chromatography using a Sepharose-lactoferrin column and detergent-solubilized membranes. The N-terminal sequence revealed no apparent similarities with any other sequenced bacterial protein. The native conformation of the 40-kDa protein was a condition to bind either iron-free or iron-saturated lactoferrin. A possible function of this Lf-binding protein could be related with an iron acquisition mechanism in P. nigrescens.

Topics: Bacterial Outer Membrane Proteins; Bacterial Proteins; Bacteroidaceae Infections; Carrier Proteins; Humans; Lactoferrin; Molecular Weight; Periodontitis; Prevotella; Protein Denaturation; Sequence Analysis

We have earlier reported hyperreactive peripheral neutrophils in adult periodontitis, measured as respiratory burst after Fc gamma receptor-mediated activation in vitro, but we have not been able to relate this increased activity to aberrations in the expression of relevant membrane molecules. Various types of inflammatory conditions involving the gingiva should affect membranes differently. We therefore collected crevicular neutrophils from three types of inflammatory sites: (i) with and (ii) without tissue destruction in the same periodontitis patients and (iii) inflamed sites in controls with gingivitis alone and compared the expression of membrane molecules by flow cytometry. The % of positively stained cells and their mean intensities of fluorescence (IFL) were similar in the three types of sites for CD15, CD11a, CD11b and CD16. Peripheral neutrophils studied with the same markers were not activated. This was verified by similar plasma concentrations of lactoferrin and L-selectins in the periodontal and control groups. Compared to peripheral cells, the crevicular neutrophils showed a significantly lower percentage of stained cells, while the stained cells increased their IFL. In conclusion, hyperreactive peripheral neutrophils in periodontitis show the same expression of membrane molecules after migration through different types of inflammatory lesions as do normal neutrophils in gingivitis.

Topics: Adult; Antigens, CD; CD11 Antigens; Cell Movement; Female; Flow Cytometry; Fluorescent Antibody Technique, Direct; Gene Expression Regulation; Gingival Crevicular Fluid; Gingivitis; Humans; L-Selectin; Lactoferrin; Lewis X Antigen; Male; Middle Aged; Neutrophil Activation; Neutrophils; Periodontitis; Receptors, IgG; Respiratory Burst

The objectives of this study is to determine if periodontitis-related ANCA hinder the accurate estimation of this kind of autoantibodies in systemic lupus erythematosus (SLE), due to the frequent coexistence of SLE and periodontitis, and the high incidence of antineutrophil cytoplasmic antibodies (ANCA) in this periodontal condition. Thirty SLE, thirty periodontitis lacking systemic involvement patients, and twenty healthy controls were utilized in this study. The periodontal condition and the presence of ANCA in sera of all individuals was carefully evaluated. For ANCA determination an EIA assay was utilized, directed to a neutrophil granular extract and six neutrophil granule proteins. Sixty percent of SLE patients had periodontitis, and sixty-five percent were ANCA positive. Eighty three percent of all ANCA cases were coexisting with periodontitis. A significant association (p > 0.005) between periodontitis and ANCA was found (Chi Square Test). Fifty percent of the patients with periodontitis lacking systemic involvement were ANCA positive. The results obtained in this study suggest that the figures of ANCA previously reported for SLE, might be overestimated due to the inadvertent presence of periodontitis.

Topics: Adolescent; Adult; Antibodies, Antineutrophil Cytoplasmic; Autoantigens; Chi-Square Distribution; Female; Humans; Immunoenzyme Techniques; Lactoferrin; Lupus Erythematosus, Systemic; Male; Middle Aged; Myeloblastin; Periodontitis; Pilot Projects; Serine Endopeptidases

The 19 patients included in this study had all been successfully treated for total or partial edentulism with single crystal sapphire implants as retention for overdentures or fixed bridges. As there is a need for more reliable non-invasive parameters for detecting changes surrounding endosseous implants, the aim of this study was to assess the content and the activity of neutrophils in crevicular fluid samples from 3 categories of sites: (1) crevices around implants from edentulous patients (2) crevices around implants in partially edentulous patients, and (3) crevices surrounding teeth. Fluid samples were taken with paper strips from 9 partially edentulous and 10 edentulous patients and the volume measured with a Periotron 6000. Elastase activity was measured as a marker of neutrophil activity and lactoferrin concentration as a marker of the number of neutrophils. Elastase activity per microliter and lactoferrin concentration was, despite similar clinical and radiographic signs, significantly higher in samples from crevices surrounding teeth and implants in the partially edentulous patients compared to samples from crevices around implants in the totally edentulous patients. There were no differences between teeth and implants in partially edentulous patients. The increased elastase activity and lactoferrin concentration indicates a higher neutrophil activity in patients with remaining teeth.

Topics: Adult; Aged; Aged, 80 and over; Aluminum Oxide; Biomarkers; Dental Implantation, Endosseous; Dental Implants; Enzyme-Linked Immunosorbent Assay; Female; Gingival Crevicular Fluid; Humans; Jaw, Edentulous, Partially; Lactoferrin; Leukocyte Count; Leukocyte Elastase; Male; Middle Aged; Mouth, Edentulous; Neutrophil Activation; Neutrophils; Periodontitis; Proteins; Statistics, Nonparametric

The release of free oxygen radicals and degranulation was studied in neutrophils from 14 patients with adult periodontitis and 14 age- and sex-matched healthy controls. The neutrophils were activated by Fc gamma-receptor stimulation, using Staphylococcus aureus opsonized with gamma globulin. Release of oxygen radicals was measured as luminol-enhanced chemiluminescence. Degranulation was assessed as release of elastase, measured with a specific substrate and as release of lactoferrin measured with ELISA. The neutrophils from the patients showed a significantly higher chemiluminescence and a slightly higher release of elastase, whereas the release of lactoferrin was the same in both groups. In contrast, the ratio between the 2 degranulation products, elastase and lactoferrin, was significantly higher in the group with periodontitis. A flow cytometric analysis of the membrane expression of the adhesion molecules CD 11a, CD 11b, CD 15, CD 16, CD 35 and Mel 14 showed no differences in the median immunofluorescence between the 2 groups. This study showed a more than 2-fold higher release of free oxygen radicals from Fc-gamma-receptor stimulated neutrophils compared with healthy controls, which indicates a specific neutrophil-associated host response in adult periodontitis.

Topics: Adult; Case-Control Studies; Cell Degranulation; Female; Flow Cytometry; Fluorescent Antibody Technique, Direct; Free Radicals; gamma-Globulins; Humans; Lactoferrin; Leukocyte Elastase; Lewis X Antigen; Luminescent Measurements; Luminol; Lymphocyte Function-Associated Antigen-1; Macrophage-1 Antigen; Male; Middle Aged; Neutrophils; Opsonin Proteins; Pancreatic Elastase; Periodontitis; Reactive Oxygen Species; Receptors, Complement 3b; Receptors, IgG; Staphylococcus aureus

The aim of the present study was to investigate whether incipient periodontal disease breakdown could be associated with changes in gingival crevicular fluid (GCF) acute-phase protein levels. In addition, the potential of clinical indices to act as predictors of significant attachment level (AL) change was investigated. AL measurements were taken at baseline and 3 months using the Florida Probe stent handpiece from a total of 384 sites in 38 patients. The average standard deviation of duplicate AL measurements was 0.423. When the tolerance method was used to detect significant AL change, 3.9% of the sites lost attachment. When a less stringent criterion of AL change of > or = 1 mm was used 9.9% of the sites lost attachment during the 3-month period. With the exception of probing depth, baseline clinical parameters failed to predict AL change. Fourteen active periodontitis sites that demonstrated significant attachment loss were paired to stable periodontitis sites within the same patient. The levels of four acute-phase proteins, namely alpha 2-macroglobulin (alpha 2-M), alpha 1-antitrypsin (alpha 1-AT), transferrin (TF) and lactoferrin (LF), and also albumin (Alb) were assessed in the same gingival crevicular fluid sample using sandwich ELISAs. Results were expressed either as ng/30 s and ng/microgram Alb. Acute-phase protein levels in GCF failed to differentiate between active and stable periodontitis sites at baseline. In conclusion, the degree of gingival inflammation of the tissues adjacent to the crevice/pocket seems to influence the levels of protease inhibitors and iron-binding proteins in GCF to a greater extent than probing attachment loss.

Topics: Acute-Phase Proteins; Adult; Aged; Albumins; alpha 1-Antitrypsin; alpha-Macroglobulins; Carrier Proteins; Enzyme-Linked Immunosorbent Assay; Female; Forecasting; Gingival Crevicular Fluid; Gingivitis; Humans; Iron-Binding Proteins; Lactoferrin; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Protease Inhibitors; Receptors, Transferrin; Transferrin; Transferrin-Binding Proteins

Topics: Adult; Antibodies, Antineutrophil Cytoplasmic; Antibody Specificity; Antibody-Dependent Cell Cytotoxicity; Cathepsin G; Cathepsins; Cytoplasmic Granules; Enzyme-Linked Immunosorbent Assay; Female; Fluorescent Antibody Technique, Indirect; Humans; Lactoferrin; Leukocyte Elastase; Male; Middle Aged; Neutrophil Activation; Neutrophils; Periodontitis; Peroxidase; Serine Endopeptidases

The degradation of human lactoferrin by putative periodontopathogenic bacteria was examined. Fragments of lactoferrin were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and measured by densitometry. The degradation of lactoferrin was more extensive by Porphyromonas gingivalis and Capnocytophaga sputigena, slow by Capnocytophaga ochracea, Actinobacillus actinomycetemcomitans and Prevotella intermedia, and very slow or absent by Prevotella nigrescens, Campylobacter rectus, Campylobacter sputorum, Fusobacterium nucleatum ssp. nucleatum, Capnocytophaga gingivalis, Bacteroides forsythus and Peptostreptococcus micros. All strains of P. gingivalis tested degraded lactoferrin. The degradation was sensitive to protease inhibitors, cystatin C and albumin. The degradation by C. sputigena was not affected by the protease inhibitors and the detected lactoferrin fragments exhibited electrophoretic mobilities similar to those ascribed to deglycosylated forms of lactoferrin. Furthermore a weak or absent reactivity of these fragments with sialic acid-specific lectin suggested that they are desialylated. The present data indicate that certain bacteria colonizing the periodontal pocket can degrade lactoferrin. The presence of other human proteins as specific inhibitors and/or as substrate competitors may counteract this degradation process.

Topics: Aggregatibacter actinomycetemcomitans; Bacteria; Bacteroides; Campylobacter; Capnocytophaga; Enzyme Inhibitors; Fusobacterium nucleatum; Humans; Iodoacetamide; Lactoferrin; Leupeptins; Peptostreptococcus; Periodontitis; Phenylmethylsulfonyl Fluoride; Porphyromonas gingivalis; Prevotella intermedia; Serine Proteinase Inhibitors; Tosyllysine Chloromethyl Ketone; Tosylphenylalanyl Chloromethyl Ketone

The aim of the present study was to evaluate the salivary secretion rate and composition in a group of 16 children and young adults (6-27 years) with Papillon-Lefèvre Syndrome (PLS), and to compare the findings with a group (n = 16) of healthy controls. Unstimulated and stimulated whole saliva was collected at least 2 h after meals and the secretion rate determined. The stimulated saliva was assessed for buffer capacity, total protein, peroxidase and hexosamine, while the unstimulated samples were evaluated for total protein, lysozyme, thiocyanate, lactoferrin and salivary IgA. Both the unstimulated (p < 0.01) and stimulated (p < 0.05) saliva secretion rates were significantly lower among the PLS patients compared with the controls. Furthermore salivary buffer capacity was significantly (p < 0.01) lower in the PLS patients. The total protein content in saliva was comparatively high in the study group, while the concentrations of immunoglobulins and non-immunoglobulins were within normal ranges. When calculating the output of the assessed antimicrobial factors, the mean peroxidase level in stimulated whole saliva was found to be significantly (p < 0.01) lower in the PLS patients than in the healthy controls. In conclusion, the present study indicates an impaired water secretion and a somewhat altered saliva gland function in children and young adults with PLS.

Topics: Adolescent; Adult; Case-Control Studies; Child; Child, Preschool; Female; Hexosamines; Humans; Immunoglobulin A, Secretory; Lactoferrin; Male; Muramidase; Papillon-Lefevre Disease; Periodontitis; Peroxidase; Saliva; Salivary Glands; Salivary Proteins and Peptides; Secretory Rate; Statistics, Nonparametric; Thiocyanates; Water-Electrolyte Balance

To compare the relative amounts of elastase (primary polymorphonuclear leucocyte granule constituent) and lactoferrin (secondary PMN granule constituent) in the gingival crevicular fluid (GCF) of healthy, gingivitis and periodontitis sites.. This cross-sectional study looked at the two GCF constituents in three categories of disease status within the same subject.. Patients with chronic adult periodontitis were screened and those exhibiting all three types of sites ie periodontally healthy, gingivitis and periodontitis sites were recruited (n=10) and had GCF collected from the three sites. Lactoferrin and elastase were measured in eluates of GCF by enzyme-linked immunosorbent assay.. The absolute amount of lactoferrin measured in ng per 30 s samples was significantly lower in healthy and gingivitis sites as compared to periodontitis sites; however this difference failed to reach significance when the concentration of lactoferrin in GCF was used as the analytical unit. No significant differences were found for elastase levels at any sites when expressed as either absolute amounts or concentrations. Secondary granule release, as evidenced by lactoferrin levels, occurs during cell migration and the process is independent of primary granule release, which is thought to correlate with PMN activation. The relationship between granule constituents in the samples showed significant differences, the highest lactoferrin/elastase ratio being at periodontitis sites (P<0.001).. These findings imply a change in the relative amounts of elastase and lactoferrin released at different disease level sites, wth an almost 10-fold increase in the proportion of lactoferrin to elastase in periodontitis sites over healthy and gingivitis sites. This variation in the release by PMNs of primary and secondary granule constituents may indicate alterations in PMN function in different disease environments.

Topics: Adult; Analysis of Variance; Biomarkers; Chronic Disease; Cross-Sectional Studies; Cytoplasmic Granules; Dental Plaque Index; Disease Progression; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Lactoferrin; Leukocyte Elastase; Male; Middle Aged; Neutrophils; Pancreatic Elastase; Periodontal Index; Periodontitis; Statistics, Nonparametric

Periodontitis affects a limited number of susceptible humans. The aim of this study was to determine whether there is a difference in the inflammatory reaction between patients with gingivitis and those with periodontitis. For this purpose the levels of elastase and lactoferrin were measured in gingival crevicular fluid (GCF) from three types of sites: i) inflamed sites in patients with gingivitis alone, inflamed sites both ii) with and iii) without tissue destruction in patients with periodontitis. Elastase activity, measured with a chromogenic substrate was significantly higher in the two types of sites in periodontitis patients. Lactoferrin levels, measured with ELISA were the same in the three types of sites. In vitro activation of granulocytes from healthy volunteers with Fc-receptor stimulation showed that the entire release of lactoferrin occurred immediately. In contrast, elastase release was time-dependent and continued throughout the experiment. Thus, the degranulation of the specific (lactoferrin) and azurophil granule (elastase) are under separate control and the two parameters can be combined in a ratio in order to characterize the granulocytes of a given patient. Assuming an immediate release of lactoferrin from the activated granulocytes in vivo, similar amounts of lactoferrin in the three types of sites can be regarded as reflecting similar numbers of granulocytes in the three types of sites. Consequently, a higher elastase activity in GCF from patients with periodontitis indicates a higher rate of release from the cells per se and a granulocyte-associated specific host response.

Topics: Adult; Cell Degranulation; Clinical Enzyme Tests; Diagnosis, Differential; Disease Susceptibility; Gingival Crevicular Fluid; Gingivitis; Granulocytes; Humans; Lactoferrin; Middle Aged; Pancreatic Elastase; Periodontal Attachment Loss; Periodontal Index; Periodontitis; Receptors, Fc; Statistics, Nonparametric

The heritability of saliva protein concentrations was investigated in stored samples of clarified stimulated whole saliva from adult twins participating in a study of periodontal disease genetics. Saliva was obtained from 29 monozygous and 20 dizygous twin pairs. Visits were scheduled so that both twins in a pair donated saliva at the same time of day. Flow rate was determined, and frozen samples later assayed for lactoferrin, lysozyme, secretory IgA, total peroxidase, myeloperoxidase and total protein. Pairs were always assayed together. Within- and between-pair variances were used to estimate twin intraclass correlations. Pearson correlations were used to estimate associations between saliva variables and clinical indices of gingivitis, dental plaque, periodontal attachment loss, and probing depth. Significant genetic contributions to variance were seen for total protein, lactoferrin, and total peroxidase. Total protein showed a significant positive correlation with gingivitis. There were no other correlations with clinical indices, and intraclass correlations for saliva variables did not change after adjustment for gingivitis. Dizygous twin correlations were higher than monozygous twin correlations for flow rate, lysozyme, and secretory IgA. That may be an artefact due to small numbers of pairs. It seems unlikely that a common environmental factor would strongly affect saliva in twins living apart as adults. Present findings, taken as sib correlations, support a genetic contribution to saliva protein concentrations. Problems with the twin model in saliva might be resolved by longitudinal studies of large numbers of twins.

Topics: Adult; Aged; Diseases in Twins; Female; Gingivitis; Humans; Immunoglobulin A, Secretory; Lactoferrin; Male; Middle Aged; Muramidase; Periodontitis; Peroxidase; Peroxidases; Saliva; Salivary Proteins and Peptides; Secretory Rate; Twins; Twins, Dizygotic; Twins, Monozygotic

This study examined lactoferrin (LF) levels in gingival crevicular fluid (GCF) and set out to test the hypothesis that LF could act as a marker of crevicular polymorphonuclear leucocytes (PMN). Therefore, 2 experiments were conducted: (a) to quantify total LF (ng/30 s sample) in GCF; (b) to correlate LF levels (ng/microliters) and PMN numbers (PMNs/microliters) in gingival crevicular washings (GCW). GCF was collected from 71 sites in a total of 22 patients. These sites were classified on the basis of clinical indices of gingivitis (GI) and pocket depth (PD) into three clinical groups: 'healthy', 'gingivitis' and 'periodontitis'. GCWs were obtained from an additional 63 sites in 21 patients. LF in GCF and GCWs was assayed by a sandwich ELISA. Total leucocyte and differential counts were performed on the GCWs. GCF LF (ng/30 s) correlated positively with GI (r = 0.418, p < 0.001), PD (r = 0.415, p < 0.001) and GCF volume (r = 0.624, p < 0.001). Gingivitis (n = 21) and periodontitis sites (n = 24) demonstrated significantly higher (p < 0.05) total GCF LF than healthy (n = 26) sites. In GCWs LF (ng/microliters) showed stronger correlations with clinical indices (GI: r = 0.452, PD: r = 0.513, p < 0.001) than did PMN numbers (PMNs/microliters) (GI: r = 0.279, PD: r = 0.388, p < 0.05). LF correlated strongly with PMNs in GCWs (r = 0.531, p < 0.001) and provides a simple and effective marker of crevicular PMN numbers.

Topics: Adult; Biomarkers; Cell Movement; Female; Gingiva; Gingival Crevicular Fluid; Gingivitis; Humans; Lactoferrin; Leukocyte Count; Leukocytes, Mononuclear; Male; Middle Aged; Neutrophils; Periodontal Diseases; Periodontal Pocket; Periodontitis

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

Topics: Adult; Aged; Analysis of Variance; Arylsulfatases; Biomarkers; Clinical Enzyme Tests; Enzyme-Linked Immunosorbent Assay; Gingival Crevicular Fluid; Humans; L-Lactate Dehydrogenase; Lactoferrin; Middle Aged; Periodontal Diseases; Periodontitis; Peroxidase

Human isolates of Porphyromonas gingivalis (n = 16), Prevotella intermedia n = 82) and Prevotella melaninogenica (n = 18) from diseased periodontal pockets were examined for interaction with human lactoferrin (HLf) in a standardized 125I-labeled protein binding assay. The highest HLf binding was found in P. intermedia strains, followed by P. gingivalis and P. melaninogenica. Further characterization of the interaction was performed with 1 representative strain from each species. HLf binding to P. gingivalis reached a saturation instantly and was optimal at pH 5.0-6.5. The corresponding values for P. melaninogenica were 90 min and pH 3.0-5.5. The HLf binding to the 2 strains seem to be nonspecific. In contrast, P. intermedia demonstrated specific binding, and a time-saturability within 60 min with an optimal uptake at pH 6.0-7.5. Scatchard analysis implied 45,000 receptors per cell with an affinity constant of 5.5 x 10(-7) M on P. intermedia strain 4H. The binding capacity in all 3 strains was affected by the culture medium. HLf binding components in these strains were susceptible to heat or proteases. Binding was eliminated in P. gingivalis and was enhanced in P. intermedia and P. melaninogenica by periodate treatment. Unlabeled HLf or bovine lactoferrin effectively displaced labeled HLf binding. Various proteins and carbohydrates did not inhibit HLf binding. Our data suggest that HLf binds to these periodontitis-associated species and that this mechanism is distinct from the previously known ligand interactions in oral bacteria.

Topics: Acute-Phase Proteins; Bacteroides; Binding, Competitive; Humans; Lactoferrin; Periodontitis; Porphyromonas gingivalis; Prevotella melaninogenica; Protein Binding; Salivary Proteins and Peptides

Fourteen subjects were examined for lactoferrin content in PMNs of venous blood. Eight of the subjects were diagnosed localized juvenile periodontitis (LJP) and four adult periodontitis (AP), all having subgingival occurrence of Actinobacillus actinomycetemcomitans (A.a.). Two subjects had healthy gingival conditions and no detectable A.a. Deficiency or low PMN lactoferrin amounts were found in six of the eight subjects with LJP and in two of the subjects with AP. The reduced lactoferrin content in the PMNs was suggested to be depending on a cytotoxic factor produced by A.a. adding to an intrinsic PMN defect.

Topics: Actinobacillus; Adolescent; Adult; Aggressive Periodontitis; Chemotactic Factors; Humans; Lactoferrin; Middle Aged; Neutrophils; Periodontitis

Lysosomal enzyme release from viable human neutrophils occurs during phagocytic activity in vivo. Phagocytosis of a strain of T. denticola, an oral spirochete, was indicated by the finding of lactoferrin in the extracellular medium of neutrophils challenged with this organism. The extracellular release of lactoferrin was dependent on duration of bacterial challenge, but peak concentration appeared at a later stage than seen in phagocytosis experiments with Escherichia coli, which served as a control. Neutrophil phagocytosis of T. denticola may be of importance as a defence factor in periodontal disease.

Topics: Escherichia coli; Humans; In Vitro Techniques; Lactoferrin; Lactoglobulins; Neutrophils; Periodontitis; Phagocytosis; Treponema

This study was designed to determine if quantitation of lysosomal products in crevicular fluid may be useful as a diagnostic test to evaluate clinical status in periodontal disease. Levels of lysozyme and lactoferrin were quantitated in crevicular fluid from patients with gingivitis, generalized adult periodontitis, localized juvenile periodontitis and normals. Crevicular fluid (CF) was collected from each patient by standardized filter paper strips and evaluated for lysozyme and lactoferrin by rocket immunoelectrophoresis. Levels of lysozyme (micrograms of protein per microliter of CF) were significantly higher in localized juvenile periodontitis patients as compared to gingivitis and adult periodontitis. On the other hand, levels of lactoferrin (micrograms of protein per microliter of CF) did not show significant differences between gingivitis, adult periodontitis and localized juvenile periodontitis. These results indicate that a lysozyme to lactoferrin ratio could be of value as a diagnostic test for localized juvenile periodontitis patients.

Topics: Adolescent; Adult; Child; Gingival Crevicular Fluid; Gingivitis; Humans; Immunoelectrophoresis; Lactoferrin; Lactoglobulins; Middle Aged; Muramidase; Periodontal Diseases; Periodontitis