Human saliva proteome and transcriptome.

This paper tests the hypothesis that salivary proteins and their counterpart mRNAs co-exist in human whole saliva. Global profiling of human saliva proteomes and transcriptomes by mass spectrometry (MS) and expression microarray technologies, respectively, revealed many similarities between saliva proteins and mRNAs. Of the function-known proteins identified in saliva, from 61 to 70% were also found present as mRNA transcripts. For genes not detected at both protein and mRNA levels, we made further efforts to determine if the counterpart is present. Of 19 selected genes detected only at the protein level, the mRNAs of 13 (68%) genes were found in saliva by RT-PCR. In contrast, of many mRNAs detected only by microarrays, their protein products were found in saliva, as reported previously by other investigators. The saliva transcriptome may provide preliminary insights into the boundary of the saliva proteome.

The Oral Fluid MEMS/NEMS Chip (OFMNC): diagnostic and translational applications.

The ability to monitor health status, disease onset and progression, and treatment outcome through non-invasive means is a most desirable goal in health-care promotion and delivery. There are three prerequisites for this goal to be realized: specific biomarkers associated with a health or disease state, a non-invasive approach to detect and monitor the biomarkers, and the technologies to discriminate between and among the biomarkers. We present a roadmap to achieve these goals using oral fluids as the diagnostic medium to scrutinize the health and/or disease status of individuals. This is an ideal opportunity to bridge state-of-the-art micro-/nano-electromechanical system (MEMS/NEMS) sensors to oral fluid for diagnostic applications. As the "mirror of body", oral fluid is a perfect medium to be explored for health and disease surveillance. The translational applications and opportunities are enormous.

The significance of prior mechanical therapy for changes of periodontal status achieved by local delivery of a doxycyclinehyclate containing gel.

AIM: The purpose of this retrospective analysis was to examine what effect, if any, the time elapsed between an individual's last episode of scaling and root planing and subsequent treatment with a sustained-release doxycycline hyclate gel (DH) alone or scaling and root planing alone (SRP) would have upon adult periodontitis. METHODS: A total of 207 subjects were included in the DH group and 210 patients in the SRP group. Periodontitis was defined as those sites which presented with pocket depths > or =5 mm and exhibited bleeding upon probing. In both DH and SRP groups, 3 sub-groups of subjects were identified according to their last episode of scaling and root planing prior to the study baseline: within 2 to 6 months, >6 but < or =12 months, and one or more times in their life but not within the last 12 months. Each study site was treated twice over a 9-month study period, once at baseline and again at 4 months. Data from the study sites at 4, 6, and 9 months were then evaluated for changes in probing depth, clinical attachment level, and bleeding upon probing. RESULTS: At the 9-month evaluation, all sub-groups in the DH and SRP treatment arms presented with improvement in the measured clinical parameters, as compared to baseline. No significant differences were observed in the measured periodontal indices among the study sites between the three sub-groups for either treatment. CONCLUSIONS: It is concluded that the time interval since the last episode of scaling and root planing had no observable effect on the results achieved when treating periodontitis sites with locally delivered doxycycline hyclate alone or scaling and root planing alone. The treatment of periodontitis sites with locally delivered doxycycline hyclate resulted in clinical improvement comparable to scaling and root planing irrespective of the patient's prophylaxis frequency.

Controlling biofilm and microbial contamination in dental unit waterlines.

Despite the fact that the ADA had set the goal of less than 200 colony-forming units per milliliter of unfiltered output water from dental unit waterlines to be achieved voluntarily by the year 2000, there is much confusion and resistance within the profession with regard to waterlines. Many in the profession are still wondering what the most effective means are to predictably achieve the goal. It is a well-established fact that bacterial biofilm can readily form within dental unit waterlines and degrade the microbial quality of the water in dental units regardless of the water source. These biofilms are primarily formed by various microcolonies of bacteria that attach to surfaces over time within the waterlines. An increasing number of medically compromised and immunocompromised patients being treated in dental offices and increased public awareness have brought about renewed interest in this issue. There are generally four categories of products that are available to address this issue: independent water systems, sterile water delivery systems, filtration, and chemical treatment protocols. A recent study at the University of California at Los Angeles demonstrates that the Ultra chemical treatment protocol can be an effective means of controlling biofilm in dental unit waterlines.

Soft tissue root coverage as treatment for cervical abrasion and caries.

Root exposure caused by gingival recession may result in cervical abrasion, root caries, root sensitivity, and compromised esthetics. Although cervical root lesions can be treated with Class V restorations, there may be advantages in treating them with soft tissue grafts for root coverage since they restore the dento-gingival unit to its prerecession condition. The rationale for root coverage with soft tissue grafts is reviewed and two cases are presented of successful root coverage procedures in which gingival recession associated with cervical abrasion and caries was present.

The effect of locally delivered controlled-release doxycycline or scaling and root planing on periodontal maintenance patients over 9 months.

BACKGROUND: This research report evaluates clinical changes resulting from local delivery of doxycycline hyclate (DH) or traditional scaling and root planing (SRP) in a group of patients undergoing supportive periodontal therapy (SPT). METHODS: In all, 141 patients received either DH (67) or SRP (74) treatment in sites > or =5 mm on one-half of their dentition at baseline and month 4. RESULTS: Clinical results were determined at month 9. Baseline mean probing depth recordings were similar between the two groups (DH = 5.9 mm; SRP = 5.9 mm). Mean month 9 results showed similar clinical results for attachment level gain (DH 0.7 mm; SRP 0.8 mm) and probing depth reduction (DH 1.3 mm; SRP 1.1 mm). Percentage of sites showing > or =2 mm attachment level gain at month 9 was 24.7% in the DH group and 21.2% in the SRP group. Thirty-nine percent (39%) of DH sites and 38% of SRP sites showed > or =2 mm probing depth reduction. When treated sides of the dentition were compared to untreated sides, DH showed a difference in disease activity (> or =2 mm attachment loss) from 19.3% (untreated) to 7.2% (treated); and SRP from 14.3% (untreated) to 8.1% (treated). CONCLUSIONS: Results show that both DH without concomitant mechanical instrumentation and SRP were equally effective as SPT in this patient group over the 9-month study period.

A controlled re-entry study on the effectiveness of bovine porous bone mineral used in combination with a collagen membrane of porcine origin in the treatment of intrabony defects in humans.

AIM: The purpose of this study was to evaluate the clinical effectiveness of a bovine porous bone mineral used in combination with a porcine derived collagen membrane as a barrier in promoting periodontal regeneration in intrabony defects in humans. MATERIAL AND METHODS: The study employed a split-mouth design. 22 paired intrabony defects were treated and surgically re-entered 6 months after treatment. Experimental sites were grafted with bovine porous bone mineral and received a collagen membrane for guided tissue regeneration. Control sites were treated with an open flap debridement. RESULTS: Preoperative pocket depths, attachment levels and trans-operative bone measurements were similar for control and experimental sites. Post surgical measurements revealed a significantly greater reduction in pocket depth (differences of 1.89 +/- 0.31 mm on buccal 0.88 +/- 0.27 mm on lingual measurements) and more gain in clinical attachment (differences of 1.51 +/- 0.33 mm on buccal and 1.50 +/- 0.35 mm on lingual measurements) in experimental sites. Surgical reentry of the treated defects revealed a significantly greater amount of defect fill in favor of experimental sites (differences of 2.67 +/- 0.91 mm on buccal and 2.54 +/- 0.87 mm on lingual measurements). CONCLUSIONS: The results of this study indicate that clinical resolution of intrabony defects can be achieved using a combination of bovine porous bone mineral and an absorbable, porcine derived collagen membrane when employing a technique based on the principles of guided tissue regeneration. The nature of the attachment between the newly regenerated tissue and the root surfaces needs to be evaluated histologically to confirm the presence of new attachment.

A comparative pilot study of the effects of a dentifrice containing green tea bioflavonoids, sanguinarine or triclosan on oral bacterial biofilm formation.

The aim of the present study was to evaluate the effect of a green tea bioflavonoid/zinc ascorbate-containing dentifrice (Natural Antioxidant Toothpaste) on bacterial plaque biofilm accumulation. Plaque biofilms were formed and observed on saliva-conditioned synthetic hydroxyapatite disks (HA) over 4 days. Two test treatments were performed daily over a 4-day experimental period for a total of 8 treatments. Four test dentifrices were evaluated: a 0.1% bioflavonoid/0.2% zinc ascorbate; a non-active control test dentifrice; a 0.075% sanguinarine/2.0% zinc chloride dentifrice; and a 0.3% triclosan dentifrice. The repeated treatment of the HA with the green tea bioflavonoid/zinc ascorbate, the sanguinarine/zinc chloride, or the triclosan dentifrice resulted in significant reductions in total viable plaque biomass when compared to a non-active placebo control dentifrice.

Effects of smoking on local delivery of controlled-release doxycycline as compared to scaling and root planing.

This paper examines the effects of smoking on the treatment outcomes of two nonsurgical therapies: (1) scaling and root planing alone (SRP) or (2) controlled-release of subgingivally delivered doxycycline hyclate in a polylactic acid based polymer gel. Subjects from 2 9-month multicenter studies were classified as nonsmokers (never smoked: 100 subjects), former smokers (137 subjects), and current smokers (> or = 10 cigarettes/day: 121 subjects). Clinical parameters were analyzed for treated sites with baseline probing depths > or = 5 mm and for a subset of treated sites with baseline probing depths of > or = 7 mm. Clinical parameters (plaque levels, clinical attachment levels, pocket depths, and bleeding on probing) were analyzed at baseline, 4, 6, and 9 months. In the doxycycline treated group in general, there were neither marked significant differences in clinical attachment gain nor differences in probing depth reduction among the 3 smoking groups. On the other hand, in the scaling and root planing treated group in general, there were significant differences in clinical attachment gain and pocket depth reduction, with non-smokers responding better than former smokers and current smokers at 6 and 9 months. These differences in clinical response between scaling and root planing alone versus controlled-release of locally-delivered doxycycline hyclate among these 3 smoking groups are discussed in relation to treatment implications for smokers.

Two multi-center studies evaluating locally delivered doxycycline hyclate, placebo control, oral hygiene, and scaling and root planing in the treatment of periodontitis.

BACKGROUND: The clinical efficacy and safety of doxycycline hyclate (8.5% w/w) delivered subgingivally in a biodegradable polymer (DH) was compared to placebo control (VC), oral hygiene (OH), and scaling and root planing (SRP) in 2 multi-center studies. METHODS: Each study entered 411 patients who demonstrated moderate to severe periodontitis. Patients had 2 or more quadrants each with a minimum of 4 qualifying pockets > or =5 mm that bled on probing. At least 2 of the pockets were > or =7 mm. Treatment with DH, VC, OH, or SRP was provided at baseline and again at month 4. Clinical parameters were recorded monthly. RESULTS: DH and SRP resulted in nearly identical clinical changes over time in both studies. Mean 9 month clinical attachment level gain (ALG) was 0.8 mm for the DH group and 0.7 mm for the SRP group in Study 1, and 0.8 mm (DH) and 0.9 mm (SRP) in Study 2. Mean probing depth (PD) reduction was 1.1 mm for the DH group and 0.9 mm for the SRP group in Study 1 and 1.3 mm for both groups in Study 2. Frequency distributions showed an ALG > or =2 mm in 29% of DH sites versus 27% of SRP sites in Study 1 and 31% of DH sites versus 34% of SRP sites in Study 2. PD reductions > or =2 mm were seen in 32% of DH sites versus 31% of SRP sites in Study 1 and 41% of DH sites versus 43% of SRP sites in Study 2. Comparisons between DH, VC, and OH treatment groups showed DH treatment to be statistically superior to VC and OH. Safety data demonstrated a benign safety profile with use of the DH product. CONCLUSIONS: Results of this trial demonstrate that treatment of periodontitis with subgingivally delivered doxycycline in a biodegradable polymer is equally effective as scaling and root planing and superior in effect to placebo control and oral hygiene in reducing the clinical signs of adult periodontitis over a 9-month period. This represents positive changes resulting from the use of subgingivally applied doxycycline as scaling and root planing was not limited regarding time of the procedure or use of local anesthesia.

Bacteriostatic effects of hyaluronic acid.

BACKGROUND: This investigation is one of a series of projects seeking to ascertain whether hyaluronic acid (HA) is therapeutically effective in tissue regeneration procedures. The rationale for these investigations is to test the hypothesis that HA can serve as a bioabsorbable carrier for other substrates as well as itself actively promote the regeneration of tissue. METHODS: In this paper, we report on the bacteriostatic and bactericidal properties of 3 molecular weight formulations of recombinant HA (low, 141 kD; medium, 757 kD; and high, 1,300 kD) on selected oral and non-oral microorganisms in the planktonic phase. Three concentrations of each HA formulation were screened, 0.5, 1.0, and 2.0 mg/ml, using a standard broth culture assay. RESULTS: Recombinant HA exerted varied bacteriostatic effects on all the bacterial strains tested depending on its molecular weight (MW) and concentration. The high concentrations of the medium MW HA had the greatest bacteriostatic effect, particularly on the Actinobacillus actinomycetemcomitans, Prevotella oris, Staphylococcus aureus, and Propionibacterium acnes strains. The 1.0 mg/ml concentration of high MW HA had the greatest overall bacteriostatic effect, inhibiting the growth of all 6 bacterial strains tested. Among the bacterial strains studied, HA was found to have no bactericidal effects, regardless of concentration or molecular weight. CONCLUSIONS: The results of this study suggest that HA in the MW range of 1,300 kD may prove beneficial in minimizing bacterial contamination of surgical wounds when used in guided tissue regeneration surgery.

An in vitro assessment and a pilot clinical study of electrical resistance of demineralized enamel.

Electrical resistance measurement was evaluated in vitro and in vivo as a method for comparing the remineralizing performance of toothpastes. In the in vitro study, areas of sound enamel on 12 unrestored, mature, extracted human molars and bicuspids, with electrical resistance readings greater than 99.99 M omega, were demineralized to an electrical resistance of 1 to 4 M omega. The teeth were divided into three groups. The change in electrical resistance was measured following a 15 cycle regimen of treatment, demineralization and salivary soaking. Treatments were five-minute exposures to either a 1:2 slurry in saliva of Enamelon (E), a remineralizing fluoride-toothpaste also containing soluble calcium and phosphate ions, a 1:2 slurry in saliva of Crest (C), a conventional fluoride toothpaste (P), or to saliva alone. Demineralization was performed with a 30-minute exposure to 0.1 M lactic acid 50% saturated with calcium hydroxyapatite. The salivary soaking was 1 hour in duration. The mean electrical resistance of the E, C and the saliva treated sites was 63.9 +/- 4.3, 37.6 +/- 9.5 and 2.1 +/- 0.7 M omega, respectively. The final resistance was statistically different for each group (p < 0.05). A pilot clinical study was then conducted to assess the electrical resistance technology in vivo. Eighteen adult subjects with at least one site of early enamel caries with an electrical resistance between 1.0 M omega and 20.00 M omega were randomly assigned to either Enamelon, Crest, or a non-fluoride placebo toothpaste and asked to brush twice daily. After three months, the mean resistance of the test sites was 23.57 M omega for E, 9.61 M omega for C and 13.24 M omega for P. However, the mean resistance changes did not proceed consistently over time. At the end of the study, the electrical resistance measurements on four sites out of twelve in Group E were suggestive of remineralization, whereas measurements on one site out of ten were suggestive of remineralization in Group C and two or three sites out of twelve were suggestive of remineralization in Group P. Progression of demineralization was possibly indicated in only one site, which was in Group C. There were insufficient subjects for statistical significance in the pilot clinical study. There were apparent problems with the variability of some measurements between visits in the in vivo study. Overall, however, the results of both studies indicate that with modifications to the equipment, electrical resistance measurements may be a means of comparing the remineralization performance of toothpastes.

Disinfection and monitoring of dental unit waterlines.

Recent increased awareness of dental unit waterline contamination has prompted investigations into potential protocols for improvement in the quality of water delivered to patients. When the protocol was followed (n = 12), contamination levels were reduced to below measurable colony forming units (CFU) defined as < 1 CFU/mL within three weeks and maintained thereafter. When adherence to the protocol was not strictly followed after initial disinfection, contamination quickly returned and remained until additional disinfection was performed.

Chemotaxis in Borrelia burgdorferi.

Borrelia burgdorferi is a motile spirochete which has been identified as the causative microorganism in Lyme disease. The physiological functions which govern the motility of this organism have not been elucidated. In this study, we found that motility of B. burgdorferi required an environment similar to interstitial fluid (e.g., pH 7.6 and 0.15 M NaCl). Several methods were used to detect and measure chemotaxis of B. burgdorferi. A number of chemical compounds and mixtures were surveyed for the ability to induce positive and negative chemotaxis of B. burgdorferi. Rabbit serum was found to be an attractant for B. burgdorferi, while ethanol and butanol were found to be repellents. Unlike some free-living spirochetes (e.g., Spirochaeta aurantia), B. burgdorferi did not exhibit any observable chemotaxis to common sugars or amino acids. A method was developed to produce spirochete cells with a self-entangled end. These cells enabled us to study the rotation of a single flagellar bundle in response to chemoattractants or repellents. The study shows that the frequency and duration for pausing of flagella are important for chemotaxis of B. burgdorferi.

The inhibiting effect of aqueous Azadirachta indica (Neem) extract upon bacterial properties influencing in vitro plaque formation.

The purpose of this investigation was to examine the inhibitory effects of aqueous extracts derived from the bark-containing sticks (Neem stick) of Azadirachta indica upon bacterial aggregation, growth, adhesion to hydroxyapatite, and production of insoluble glucan, which may affect in vitro plaque formation. Neem stick extracts were screened for minimal bacterial growth inhibition (MIC) against a panel of streptococci by means of a broth dilution assay. Initial bacterial attachment was quantified by the measurement of the adhesion of 3H-labeled Streptococcus sanguis to saliva-conditioned synthetic hydroxyapatite. The effect of the Neem stick extract upon insoluble glucan synthesis was measured by the uptake of radiolabeled glucose from 14C-sucrose. Aggregating activity of the Neem stick extracts upon a panel of streptococci was also examined. No inhibition of bacterial growth was observed among the streptococcal strains tested in the presence of < or = 320 micrograms/mL of the Neem stick extract. The pre-treatment of S. sanguis with the Neem stick extract or the gallotannin-enriched extract from Melaphis chinensis at 250 micrograms/mL resulted in a significant inhibition of the bacterial adhesion to saliva-conditioned hydroxyapatite. Pre-treatment of saliva-conditioned hydroxyapatite with the Neem stick or gallotannin-rich extract prior to exposure to bacteria yielded significant reductions in bacterial adhesion. The Neem stick extract and the gallotannin-enriched extract from Melaphis chinensis inhibited insoluble glucan synthesis. Incubation of oral streptococci with the Neem stick extract resulted in a microscopically observable bacteria aggregation. These data suggest that Neem stick extract can reduce the ability of some streptococci to colonize tooth surfaces.

The determination of ionic bonding interactions of N-phenyl glycine and N-(2-hydroxy-3-methacryloxypropyl)-N-phenyl glycine as measured by carbon-13 NMR analysis.

The purpose of the present investigation was to determine whether high-resolution carbon-13 nuclear magnetic resonance could be utilized for detection of ionic bonding interactions of NPG and NPG-GMA with selected inorganic cations. The C1-carbonyl carbon of NPG and NPG-GMA were labeled with 13C to help magnify the spectral response signal. The labeled monomers were allowed to react with Ca(OH)2 and LiOH. The C-13 carbonyl resonance was observed by solution- and solid-state NMR. The interaction of the cations in solution and on a powered glass support resulted in a downfield chemical shift of about 2-5 ppm, indicating the presence of a chemical interaction between the monomer and the cations. The reaction of the monomers with the solid substrates of hydroxyapatite, and bovine dentin did not result in a significant downfield shift in the carbonyl resonance. The results provide evidence that high-resolution nuclear magnetic resonance can be used for evaluation of chemical bonding interactions of dental adhesives with inorganic ions.

Detachment of oral bacteria from saliva-coated hydroxyapatite by human polymorphonuclear leukocytes.

Human polymorphonuclear leukocytes (PMN) demonstrated the ability to detach Actinomyces viscosus, A. naeslundii and Streptococcus sanguis from saliva-coated hydroxyapatite beads (SHA). Between 60 to 80% of bacteria were detached within 1 hour at PMN-to-bacteria ratios between 1:10 to 1:22. Detachment was enhanced by treating bacteria with fresh but not heat-inactivated normal human serum. Detachment of serum-treated A. viscosus was inhibited by cytochalasin B, L-1-tosylamide-2-phenylethylchloromethyl ketone (TPCK), and deoxyglucose but not colchicine, phenylmethylsulfonyl fluoride (PMSF), N-carbobenzoxy-L-phenylalanine chloromethyl ketone (ZPCK), and sodium azide. In the absence of serum treatment, the detachment of A. viscosus was insensitive to lactose, galactose, and mannose. We conclude that PMN can efficiently detach bacteria from SHA, this detachment is enhanced by serum, and this enhancement is probably dependent upon complement. Additionally, detachment of A. viscosus bound to SHA by PMN (1) does not appear to involve bacterial lectin activity, (2) seems to be dependent upon glycolytic metabolism, microfilament formation, and the activity of a TPCK-sensitive serine protease, and (3) is not sensitive to inhibitors of tubulin polymerization or heme-protein activity.

A study of in vitro attachment of Streptococcus sanguis and Actinomyces viscosus to saliva-treated titanium.

This study examined the initial attachment of Streptococcus sanguis G9-B and Actinomyces viscosus T14V to saliva-treated powdered enamel and titanium surfaces. Using an in vitro adherence model, significantly lower numbers of Actinomyces viscosus T14V bound to the saliva-treated titanium surface when compared to that of the similarly treated enamel. The binding of Streptococcus sanguis G9-B to titanium or enamel did not vary significantly. A comparison of the percentage of cells bound to the titanium surface revealed that S sanguis cells attached in significantly higher numbers when compared to the A viscosus cells.

Dental professional's role in monitoring and maintenance of tissue-integrated prostheses.

The long-term success of an implant-supported prosthesis is dependent not only on osseointegration of the implant to the surrounding osseous structures, but also the integrity and health of the surrounding peri-implant tissues. This paper defines and discusses the dental professional's role in (1) examining and assessing the peri-implant tissues (2) providing clinical maintenance protocols, and (3) recommending effective home-care instructions for a patient with implant-supported prostheses.