Gingival tissue, an extrasynovial source of malondialdehyde-acetaldehyde adducts, citrullinated and carbamylated proteins.

BACKGROUND AND OBJECTIVE: Postranslational modification of proteins can lead to the production of autoantibodies and loss of immune tolerance. This process has been hypothesised to be a critical factor in the pathogenesis of rheumatoid arthritis. The objective of this study was to demonstrate that inflamed human gingival tissue provides an extrasynovial source of malondialdehyde-acetaldehyde adducts, citrullinated and carbamylated proteins all of which are considered to be linked to the development of rheumatoid arthritis. Identification of such modified proteins in inflamed gingiva may explain, in part, how inflammation of the periodontal tissues may influence the development of rheumatoid arthritis. MATERIAL AND METHODS: Gingival biopsies of healthy, mild and moderate periodontitis were triple stained with antibodies against malondialdehyde-acetaldehyde adducts, citrullinated and carbamylated proteins. RESULTS: Assessment of healthy gingival tissue revealed negligible staining for carbamylated, malondialdehyde-acetaldehyde (MAA), or citrullinated proteins. Mild periodontitis was positive for all three modifications. Furthermore, there was an increase in staining intensity for carbamylated, citrullinated and MAA-modified proteins in moderate periodontitis. Negative staining results were observed for the isotype controls. CONCLUSION: This study provides evidence for the presence of citrullinated, carbamylated and MAA adduct modified proteins in inflamed periodontal tissues. The potential for these proteins to play a role in autoimmunity in a multi-system inflammatory syndromic disease model now needs to be determined.

Filth Fly Activity Associated With Composted and Noncomposted Beef Cadavers and Laboratory Studies on Volatile Organic Compounds.

Commercial livestock facilities are faced with the challenge of managing large amounts of waste including manure and animal mortalities. One method of disposing of dead animals is composting. The cadavers are enveloped in carbon material that creates a barrier between the dead tissue and the surrounding environment. Dead tissue can release materials that not only contaminate the soil but also the groundwater and nearby surface water. Animal cadaver composting is designed to facilitate decomposition without the aid of carrion-feeding insects and reduce the presence of common pathogens associated with animal waste and dead tissue. The aim of this study was to evaluate insect activity associated with composted and exposed beef cadavers, specifically filth flies that can serve as mechanical vectors of important human pathogens such as E. coli 0157:H7. Greater numbers of all types of arthropods were trapped overall at the exposed animal site than the composted animal site. Most importantly, the number of filth flies was significantly lower at the composted site (P = 0.0009). Laboratory analysis of volatile organic compounds from composted and noncomposted rats indicated that known fly attractants such as dimethyl disulfide may be inhibited by the composting process. Implementing composting programs at livestock facilities could reduce the risk of flies spreading harmful pathogens to surrounding areas, including farms that grow fresh produce.

Salivary and serum procalcitonin and C-reactive protein as biomarkers of periodontitis in United States veterans with osteoarthritis or rheumatoid arthritis.

Serum procalcitonin (ProCT) is elevated in response to bacterial infections, whereas high sensitivity C-reactive protein (hsCRP) is a nonspecific inflammatory marker that is increased by excess adipose tissue. We examined the efficacy of ProCT and hsCRP as biomarkers of periodontitis in the saliva and serum of patients with arthritis, which is characterized by variable levels of systemic inflammation that potentially can confound the interpretation of inflammatory biomarkers. Blood and unstimulated whole saliva were collected from 33 patients with rheumatoid arthritis (RA) and 50 with osteoarthritis (OA). Periodontal status was assessed by full mouth examination and patients were categorized as having no/mild, moderate or severe periodontitis by standard parameters. Salivary and serum ProCT and hsCRP concentrations were compared. BMI, diabetes, anti-inflammatory medications and smoking status were ascertained from the patient records. Differences between OA and RA in proportionate numbers of patients were compared for race, gender, diabetes, adiposity and smoking status. Serum ProCT was significantly higher in arthritis patients with moderate to severe and severe periodontitis compared with no/mild periodontitis patients. There were no significant differences in salivary ProCT or salivary or serum hsCRP in RA patients related to periodontitis category. Most of the OA and RA patients were middle aged or older, 28.9% were diabetic, 78.3% were overweight or obese, and slightly more than half were either current or past smokers. The OA and RA groups differed by race, but not gender; blacks and males were predominant in both groups. The OA and RA groups did not differ in terms of controlled or uncontrolled diabetes, smoking status or BMI. The RA patients had been prescribed more anti-inflammatory medication than the OA patients. Our results demonstrate that circulating ProCT is a more discriminative biomarker for periodontitis than serum hsCRP in patients with underlying arthritis. Any elevation in salivary and serum hsCRP due to periodontitis apparently was overshadowed by differences among these patients in factors that influence CRP, such as the extent of inflammation between RA and OA, the extent of adipose tissue, the use of anti- inflammatory medications and smoking status. Although our study showed no differences in salivary ProCT related to severity of periodontitis, this biomarker also may be useful with further refinement.

TLR4 Asp299Gly polymorphism may be protective against chronic periodontitis.

BACKGROUND AND OBJECTIVE: Periodontitis results from interplay between genetic and environmental factors. Single nucleotide polymorphisms (SNPs) in the coding region of the toll-like receptor 4 gene (TLR4) may be associated with periodontitis, although previous studies have been inconclusive. Moreover, the interaction between environmental factors, such as cigarette smoking (a major risk factor for periodontitis), and Porphyromonas gingivalis (a major periodontal pathogen) with the TLR4 coding region Asp299Gly SNP (rs4986790; a SNP associated with lipopolysaccharide-mediated inflammatory responses in periodontitis), have been largely ignored in previous reports. Therefore, the objective of this study was to examine the association between TLR4 Asp299Gly (rs4986790) with alveolar bone height loss (ABHL) and periodontitis, accounting for interactions between this SNP with smoking and P. gingivalis prevalence. The CD14/-260 SNP (rs2569190) served as a control, as a recent meta-analysis suggested no relationship between this SNP and periodontitis. MATERIAL AND METHODS: This multicenter study included 617 participants who had rheumatoid arthritis or osteoarthritis. This report presents a secondary outcome from the primary case-control study examining the relationship of periodontitis with established rheumatoid arthritis. The Centers for Disease Control/American Academy of Periodontology case definitions of periodontitis were used for this analysis. Participants received a full-mouth clinical periodontal examination and panoramic radiograph. Percentage ABHL was measured on posterior teeth. The TLR4 Asp299Gly and CD14/-260 SNPs were selected a priori and genotypes were determined using the ImmunoChip array (Illumina(®) ). Minor allele frequencies and associations with periodontitis and ABHL did not differ according to rheumatoid arthritis vs. osteoarthritis status; therefore, data from these two groups were pooled. The presence of P. gingivalis was detected in subgingival plaque by PCR. Multivariate ordinal logistic regression examined associations between the SNPs and periodontitis or ABHL. SNP interactions with smoking and P. gingivalis were analyzed. RESULTS: A significant, negative interaction was observed between the TLR4 SNP and the presence of P. gingivalis (p = 0.045) with respect to periodontitis. The TLR4 minor variant was also associated with less ABHL: 16.8% of individuals with low ABHL, 9.0% with moderate ABHL and 11.2% with high ABHL had the minor allele [p = 0.029; odds ratio = 0.58 (95% confidence interval: 0.36-0.95)]. The interaction between the TLR4 SNP and smoking was not significant with respect to periodontitis or ABHL. The CD14 SNP was not associated with periodontitis or ABHL. CONCLUSION: The TLR4 Asp299Gly SNP significantly interacted with P. gingivalis in conferring a decreased risk of periodontitis and may be protective against ABHL, a feature of periodontitis. Agents blocking TLR4 signaling, a strategy currently under investigation for the treatment of other inflammatory conditions, may warrant investigation in the context of periodontitis related to the presence of P. gingivalis.

Interactions of microorganisms with polymer nanocomposite surfaces containing oxidized carbon nanotubes.

In many environmental scenarios, the fate and impact of polymer nanocomposites (PNCs) that contain carbon nanotubes (CNT/PNCs) will be influenced by their interactions with microorganisms, with implications for antimicrobial properties and the long-term persistence of PNCs. Using oxidized single-wall (O-SWCNTs) and multi-wall CNTs (O-MWCNTs), we explored the influence that CNT loading (mass fraction≤0.1%-10%) and type have on the initial interactions of Pseudomonas aeruginosa with O-CNT/poly(vinyl alcohol) (PVOH) nanocomposites containing well-dispersed O-CNTs. LIVE/DEAD staining revealed that, despite oxidation, the inclusion of O-SWCNTs or O-MWCNTs caused PNC surfaces to exhibit antimicrobial properties. The fraction of living cells deposited on both O-SWCNT and O-MWCNT/PNC surfaces decreased exponentially with increasing CNT loading, with O-SWCNTs being approximately three times more cytotoxic on a % w/w basis. Although not every contact event between attached microorganisms and CNTs led to cell death, the cytotoxicity of the CNT/PNC surfaces scaled with the total contact area that existed between the microorganisms and CNTs. However, because the antimicrobial properties of CNT/PNC surfaces require direct CNT-microbe contact, dead cells were able to shield living cells from the cytotoxic effects of CNTs, allowing biofilm formation to occur on CNT/PNCs exposed to Pseudomonas aeruginosa for longer time periods.

Serum bone biomarkers and oral/systemic bone loss in humans.

UNLABELLED: We recently reported that subantimicrobial-dose doxycycline (SDD) significantly reduced serum bone-resorption biomarkers in subgroups of post-menopausal women. We hypothesize that changes in serum bone biomarkers are associated not only with systemic bone mineral density (BMD) changes, but also with alveolar bone changes over time. One hundred twenty-eight eligible post-menopausal women with periodontitis and systemic osteopenia were randomly assigned to receive SDD or placebo tablets twice daily for two years, adjunctive to periodontal maintenance. Sera were analyzed for bone biomarkers. As expected, two-year changes in a serum bone biomarker were significantly associated with systemic BMD loss at the lumbar spine (osteocalcin, bone-turnover biomarker, p = 0.0002) and femoral neck (osteocalcin p = 0.0025). Two-year changes in serum osteocalcin and serum pyridinoline-crosslink fragment of type I collagen (ICTP; bone-resorption biomarker) were also significantly associated with alveolar bone density loss (p < 0.0001) and alveolar bone height loss (p = 0.0008), respectively. Thus, we have shown that serum bone biomarkers are associated with not only systemic BMD loss, but with alveolar bone loss as well. CLINICAL TRIAL REGISTRATION INFORMATION: Protocol registered at ClinicalTrials.gov, NCT00066027.

Doxycycline effects on serum bone biomarkers in post-menopausal women.

We previously demonstrated that subantimicrobial-dose-doxycycline (SDD) treatment of post-menopausal osteopenic women significantly reduced periodontal disease progression, and biomarkers of collagen destruction and bone resorption locally in periodontal pockets, in a double-blind placebo-controlled clinical trial. We now hypothesize that SDD may also improve biomarkers of bone loss systemically in the same women, consistent with previous studies on tetracyclines (e.g., doxycycline) in organ culture and animal models of bone-deficiency disease. 128 post-menopausal osteopenic women with chronic periodontitis randomly received SDD or placebo tablets daily for 2 years adjunctive to periodontal maintenance therapy every 3-4 months. Blood was collected at baseline and at one- and two-year appointments, and sera were analyzed for bone resorption and bone formation/turnover biomarkers. In subsets of the study population, adjunctive SDD significantly reduced serum biomarkers of bone resorption (biomarkers of bone formation were unaffected), consistent with reduced risk of future systemic bone loss in these post-menopausal women not yet on anti-osteoporotic drugs.

Salmonella populations and prevalence in layer feces from commercial high-rise houses and characterization of the Salmonella isolates by serotyping, antibiotic resistance analysis, and pulsed field gel electrophoresis.

Salmonella species are recognized as a major cause of foodborne illnesses that are closely associated with the consumption of contaminated poultry and egg products. The objectives of this study were to evaluate the Salmonella populations and prevalence in layer feces during the laying cycle and molting of the hen and to characterize the layer fecal Salmonella isolates by serotyping, antibiotic resistance analysis, and pulsed field gel electrophoresis. Fecal samples were collected from a commercial layer complex consisting of 12 houses. Composite fecal samples across each row were collected as a function of bird age [18 wk (at placement), 25 to 28 wk (first peak of production cycle), 66 to 74 wk (molting), and 75 to 78 wk (second peak of production cycle)]. Bird ages and molting practice did not significantly affect (P > 0.05) Salmonella populations with an average of 1.25, 1.27, 1.20, and 1.14 log most probable number/g for the 18-, 25- to 28-, 66- to 74-, and 75- to 7-wk birds, respectively. However, the 18-wk birds had the highest prevalence of Salmonella (55.6%), followed by the 25- to 28-wk birds (41.7%), 75- to 78-wk birds (16.7%), and 66- to 74-wk birds (5.5%). Of the 45 Salmonella isolates characterized, the most predominant serovar was Salmonella Kentucky (62%). Thirty-five percent of the Salmonella isolates were resistant to at least 1 antibiotic. As expected, considerable genetic diversity was observed within and across the different serovars.

Modeling the growth and death kinetics of Salmonella in poultry litter as a function of pH and water activity.

Contaminated poultry litter, serving as a reservoir for Salmonella, can be linked to both food safety concerns when contaminated birds enter processing plants and environmental concerns when used as a fertilizer. Predictive modeling allows for the estimation of microbial growth or inactivation as a function of controlling environmental growth factors. A study was conducted to observe the combined effects of pH and water activity (A(w)) at a constant temperature on Salmonella populations in used turkey litter to predict microbial response over time. Litter, first pH-adjusted and then inoculated with a 3-strain Salmonella serovar cocktail to an initial concentration of approximately 10(7) cfu/g, was placed into individual sealed plastic containers with saturated salt solutions for controlling A(w). A balanced design including 3 A(w) values (0.84, 0.91, 0.96), 3 pH values (4, 7, 9), and a constant temperature of 30 degrees C was used, with litter samples periodically removed and analyzed for Salmonella populations, pH, and A(w). At each combination of environmental factors, the Churchill or exponential inactivation mathematical models were used to describe the growth and death of Salmonella over time. Salmonella populations exhibited growth (approximately 2 log) with little decline up to 42 d in litter environments of pH 7 and 9 and a A(w) of 0.96. As litter A(w) and pH levels were reduced, populations declined, with the most drastic reductions (approximately 5 log in 9 h) occurring in low-pH (4) and low-A(w) (0.84) environments. Generalized models for bacterial growth and death under grouped pH environments were successfully developed to predict Salmonella behavior in litter over time. These findings suggest that the best management practices and litter treatments that lower litter A(w) to < or =0.84 and pH to < or =4 are effective in reducing Salmonella populations. The use of a single equation to predict the growth and decline of Salmonella populations as a function of pH and A(w) has potential application for use in the development of effective pathogen control strategies at the farm level.

The association of cigarette smoking with alveolar bone loss in postmenopausal females.

BACKGROUND, AIMS: The purpose of this 2-year longitudinal clinical study was to determine the impact of smoking on alveolar bone height and density changes in postmenopausal females. METHODS: 59 postmenopausal women completed this study, including 38 non-smokers and 21 smokers. All subjects had a history of periodontitis, participated in 3- to 4-month periodontal maintenance programs and were within 5 years of menopause at the study outset. 4 vertical bite-wing radiographs of posterior sextants were taken at baseline and 2-year visits. Radiographs were evaluated using computer-assisted densitometric image analysis (CADIA); changes in interproximal alveolar bone density and changes in alveolar bone height were determined. Relative clinical attachment levels (RCAL) and presence/absence of plaque and bleeding on probing were recorded. RESULTS: Smokers exhibited a higher frequency of alveolar bone height loss (p<0.05) and crestal (p<0.03) and subcrestal (p<0.02) density loss relative to non-smokers. Smokers exhibited a trend (p<0.08) toward a higher frequency of > or =2.0 mm RCAL loss over the 2-year period. Plaque and bleeding on probing did not differ between smokers and non-smokers. A significant interaction, determined by repeated measures ANOVA, was noted between systemic bone mineral density (BMD) at the lumbar spine and smoking on alveolar bone density change (p<0.05). Only non-smoking patients with normal BMD realized a mean net gain in alveolar bone density; osteoporotic/osteopenic subjects (n=25) and smokers lost alveolar bone density. CONCLUSION: Postmenopausal female smokers were more likely to lose alveolar bone height and density than non-smokers with a similar periodontitis, plaque and gingival bleeding experience. In addition, both smoking and osteoporosis/osteopenia provided a negative influence on alveolar bone.

The use of distraction osteogenesis to induce new suprabony periodontal attachment in the beagle dog.

Using a technique called distraction osteogenesis (DO), orthopedic surgeons are capable of reconstructing 4 to 5 cm of bone and soft tissue without bone grafts or pedicle flap procedures. DO has been used recently to generate maxillofacial bone as an alternative to maxillary and mandibular osteotomies. Using DO methodology, this study attempted to regenerate supraalveolar buccal periodontium on the mandibular second, third, and fourth premolars of beagle dogs. A small but statistically significantly greater amount of new cementum was generated in experimental teeth, with no difference found between experimental and control groups with respect to regenerating alveolar bone. Further studies using modifications of this technique are warranted to explore the potential of DO as an alternative to conventional regenerative approaches.

Longitudinal alveolar bone loss in postmenopausal osteoporotic/osteopenic women.

The purpose of this 2-year longitudinal clinical study was to investigate alveolar (oral) bone height and density changes in osteoporotic/osteopenic women compared with women with normal lumbar spine bone mineral density (BMD). Thirty-eight postmenopausal women completed this study; 21 women had normal BMD of the lumbar spine, while 17 women had osteoporosis or osteopenia of the lumbar spine at baseline. All subjects had a history of periodontitis and participated in 3- to 4-month periodontal maintenance programs. No subjects were current smokers. All patients were within 5 years of menopause at the start of the study. Four vertical bitewing radiographs of posterior sextants were taken at baseline and 2-year visits. Radiographs were examined using computer-assisted densitometric image analysis (CADIA) for changes in bone density at the crestal and subcrestal regions of interproximal bone. Changes in alveolar bone height were also measured. Radiographic data were analyzed by the t-test for two independent samples. Osteoporotic/osteopenic women exhibited a higher frequency of alveolar bone height loss (p<0.05) and crestal (p<0.025) and subcrestal (p<0.03) density loss relative to women with normal BMD. Estrogen deficiency was associated with increased frequency of alveolar bone crestal density loss in the osteoporotic/osteopenic women and in the overall study population (p<0.05). These data suggest that osteoporosis/osteopenia and estrogen deficiency are risk factors for alveolar bone density loss in postmenopausal women with a history of periodontitis.

Influence of estrogen and osteopenia/osteoporosis on clinical periodontitis in postmenopausal women.

BACKGROUND: In Western societies, more than one-third of the female population above age 65 suffers from signs and symptoms of osteoporosis, a disorder characterized by low bone mass. Estrogen deficiency is the dominant pathogenic factor for osteoporosis in women. The impact of estrogen deficiency and osteopenia/osteoporosis on periodontitis is unclear, partially due to the lack of longitudinal studies evaluating clinical signs of gingival inflammation and periodontitis progression. The purpose of this investigation was to analyze prospectively the influence of serum estradiol levels and osteopenia/osteoporosis on common clinical measurements of periodontal disease over a 2-year period. METHODS: Fifty-nine moderate/advanced adult periodontitis patients and 16 non-periodontitis subjects, all within 5 years after menopause at baseline, completed the study. Serum estradiol levels (E2) were measured yearly by 125I radioimmunoassay, and osteopenia/osteoporosis was determined by dual energy x-ray absorptiometry of the lumbar spine. Posterior interproximal clinical measurements were obtained every 6 months for the periodontitis patients, including explorer-detectable supragingival plaque, bleeding on probing (BOP) and relative clinical attachment level (RCAL). Baseline probing depths, smoking history, and demographic data also were collected. RESULTS: Data indicated that baseline demographic measurements and bone mineral density (BMD) of the lumbar spine were not different between E2-deficient and E2-sufficient subjects. Smoking activity (packs smoked/day, years smoked) was higher in periodontitis patients (P=0.0001). E2-sufficient periodontitis subjects had a higher frequency of supragingival plaque without increasing gingival inflammation. E2 status did not influence the percentage of sites losing RCAL for either periodontitis or non-periodontitis groups, but when non-smoking osteopenic/osteoporotic periodontitis patients were evaluated, E2-deficient subjects had more BOP (43.8% versus 24.4%, P<0.04) and a trend toward a higher frequency of > or =2.0 mm RCAL loss (3.8% versus 1.2%, P<0.1) than E2-sufficient subjects. CONCLUSIONS: These data suggest that E2 supplementation (serum E2>40 pg/ml) is associated with reduced gingival inflammation and a reduced frequency of clinical attachment loss in osteopenic/osteoporotic women in early menopause.

Gingival fluid IL-1beta in postmenopausal females on supportive periodontal therapy. A longitudinal 2-year study.

Posterior interproximal alveolar bone in 59 women, within 5 years after menopause, was assessed at baseline and after 2 years of supportive periodontal therapy (history of moderate/advanced periodontitis) using digitized image analysis. Baseline lumbar spine bone mineral density, smoking status, and yearly serum estradiol (E2) levels also were obtained to group subjects. An additional 16 non-periodontitis postmenopausal women were followed 2 years for clinical and estrogen status. 2-min GCF IL-1beta levels averaged from 2 baseline periodontal pockets (in periodontitis subjects) and 2 non-periodontitis sites (in non-periodontitis and periodontitis subjects) were determined with an enzyme immunoassay. A progressive and stable site were also monitored every 6 months for GCF IL-1beta in 15 patients. Results after 2 years indicated that 17 subjects had no posterior interproximal sites losing > or =0.4 mm of alveolar crest bone height, while 13 subjects had > or =3 such sites. Using analysis of variance, none of the above clinical groupings resulted in a significant difference in mean baseline or longitudinal GCF IL-1beta levels. However, when subjects who lost alveolar crest bone height were considered, E2-sufficient subjects had significantly depressed baseline GCF IL-1beta (in past-periodontitis sites) compared to E2-deficient patients (9.1+/-2.1 versus 31.7+/-10.2 pg/2-min sample, p<0.05), suggesting E2 influences gingival IL-1beta production in progressive periodontitis patients.

Histological alterations following short-term smokeless tobacco exposure in humans.

The purpose of this study was to investigate the histological changes following short-term smokeless tobacco application in humans. Sixteen smokeless tobacco-using subjects participated in this trial. Each subject had used at least 3 cans of snuff per week for the previous 2 yr and had an existing lesion at the site of habitual snuff placement. The experimental design included subject placement of moist snuff (University of Kentucky reference tobacco brand 1S3) at a new site in the mandibular arch. At either 2 or 7 d, biopsies were taken from the new lesions and from a non-placement site in the opposing arch. The volume density of inflammatory cells was determined by point counting. Keratin and epithelial thickness were evaluated by digitizing morphometry. Data were analyzed by repeated measures analysis of variance. In 7-d lesions, increased keratin thickness was observed at the new sites compared to the non-placement sites (p = 0.05). Increased volume density of fibroblasts (p = 0.027) and decreased volume densities of macrophages (p = 0.0083) and mast cells (p = 0.05) were observed at 2 d in new versus non-placement sites. Clinically, the new sites showed erythema, erythema plus ulceration, or white striations. This study demonstrated histological and clinical changes at new snuff placement sites in as few as 2-7 d, underscoring the rapidity of tissue alterations following snuff use.

Development of smokeless tobacco-induced oral mucosal lesions.

This study examined clinical and inflammatory mediator parameters during the development of snuff-induced mucosal lesions. Nineteen smokeless tobacco (ST) users placed moist snuff at designated new placement sites over either a 2- or 7-day period. By day 2, the predominant clinical alteration was an erythematous reaction, and one-third of the subjects demonstrated white striations in combination with erythema or ulceration. By 7 days, 56% of the subjects displayed white striated lesions. Tissue concentrations (pg/mg) of IL-1beta were 4.6+/-1.6 at new placement sites as compared with 0.7+/-0.4 at control sites (P<0.05). PGE2 and IL-1alpha concentrations also were significantly higher (P<0.05) at new placement sites as compared with control sites (9.4+/-2.2 vs 4.1+/-0.7 and 6.2+/-1.3 vs 3.2+/-0.7, respectively). In view of the recognized role of PGE2 and IL-1 in immune and inflammatory functions, these mediators may play a role in the pathogenesis of clinical alterations at ST placement sites.

Management of the gingival sulcus in fixed prosthodontics: a literature review and treatment protocol.

This review of the fixed prosthodontics and periodontology literature illustrates the influence of various factors on the health of the periodontium during and after fixed prosthodontic treatment. This article describes four categories of marginal finish lines for fixed prosthodontic restorations relative to the gingival margin and the epithelial attachment. The management of each category from both the periodontal and prosthodontic prespectives is also discussed.

Nicotine and smokeless tobacco effects on gingival and peripheral blood mononuclear cells.

The pathogenesis of tobacco-related periodontal diseases is not well understood. The purpose of this study was therefore to investigate smokeless tobacco extract (ST) and nicotine effects on prostaglandin E2 (PGE2) and interleukin-1beta (IL-1beta) secretion by peripheral blood mononuclear cells (PBMC, consisting of monocytes and lymphocytes) and gingival mononuclear cells (GMC). Both peripheral blood and gingival tissue adjacent to the alveolar crest were taken from non-smoking adult periodontitis patients. Gingival tissue was treated with collagenase and deoxyribonuclease and GMC and PBMC were isolated by Ficoll-Hypaque centrifugation. GMC and PBMC (100,000 cells/200 microl) were cultured for 24 hours in supplemented RPMI 1640 alone (control), or in supplemented RPMI 1640 containing 1% ST, 100 microg/ml nicotine, 1 microg/ml Porphyromonas gingivalis LPS, or 1 microg/ml P. gingivalis LPS and either 100 microg/ml nicotine or 1% ST. Enzyme immunoassays were used to quantify PGE2 and IL-1beta. Treatments were compared by repeated measures ANOVA. 100 microg/ml nicotine (7-fold, p<0.02) and 1% ST (3.5-fold, p<0.004) significantly increased secretion of PGE2 by PBMC relative to control cultures. 100 microg/ml nicotine and 1% ST, however, had no effect on IL-1beta secretion by PBMC. Enhanced PGE2 secretion also was seen when PBMC were treated with P. gingivalis LPS+ 100 microg/ml nicotine relative to P. gingivalis LPS alone (p<0.007). In contrast, 100 microg/ml nicotine significantly downregulated IL-1beta secretion by GMC relative to medium alone (p<0.008) and had no effect on PGE2 secretion by GMC. These data indicate that while nicotine and ST can stimulate PBMC to secrete PGE2, they cannot activate further mononuclear cells extracted from gingiva, possibly due to maximal previous stimulation in the periodontitis lesion.

Potential application of low-dose doxycycline to treat periodontitis in post-menopausal women.

The purpose of this paper is two-fold: (1) to review the evidence that osteoporosis and post-menopausal estrogen deficiency are associated with progressive alveolar bone loss and an elevated risk of tooth loss; and (2) to propose the use of tetracyclines, specifically low-dose doxycycline (LDD) (and, perhaps in the future, the chemically modified tetracyclines), to mitigate alveolar bone loss in post-menopausal osteoporotic/osteopenic women. Design concepts for a randomized clinical trial to study the effects of LDD on progressive alveolar bone loss in this patient population are reviewed. Since osteoporosis affects over 20 million people in the United States, progressive alveolar bone loss in this patient group represents a potentially significant public health problem unique from common adult periodontitis. Stopping progressive alveolar bone loss is essential to prevent both tooth loss and micro-architectural deterioration of alveolar bone.