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Objectives: Although the enhancement of plasma protein adsorption to titanium ( Ti ) following wetting has been recognized, the relationship between wettability and electrostatic forces has remained unclear. Thus, we have carried out a series of studies to determine the role of wettability and electrostatic forces on protein adsorption., Methods: Titanium disks with different surfaces were wetted with a range of solutions, two of which contained divalent positive ions ( Ca and Mg ). Unwetted disks served as a control. Subsequently, the wetted disks were subjected to three treatment regimes: (1) incubation in human serum albumin (HSA) or human serum fibronectin (HSF); (2) drying the wetted disks, followed by incubation in HSA or HSF; and (3) following protein adsorption, the Ca originating in the wetting solutions was removed by divalent positive ions chelator treatment (EGTA), and the remaining quantities were assessed. The quantity of the adsorbed proteins was determined by ELISA., Results: It was found that in the case of HSA, adsorption was enhanced by the wettability, the presence of Ca and Mg in the wetting solution, and the existence of rough surfaces. For HSF, the wettability and rough surfaces enhanced adsorption., Conclusion: The results demonstrate that in addition to wettability, the composition of the wetting solution affects the protein adsorption. While wetting reduces the time for the HSA and HSF adsorption to reach saturation, the electrostatic forces enhance the amount of HSA adsorption. Thus, the protein adsorption capacity of titanium rough surfaces can be selectively manipulated by changing of the wetting solution., (© 2012 John Wiley & Sons A/S.)

Objectives: Modifications of titanium (Ti) implant surfaces have a significant effect on early biofilm formation and the outcome of implant procedures. The aim of this study was to examine the role of plasma proteins and electrostatic forces in the adhesion mechanism of oral bacteria to modified Ti surfaces., Materials and Methods: Ti discs with three different types of surface modifications, machined, acid-etched, and acid-etched and blasted, were examined for adhesion of oral bacteria: Streptococcus mutans, Porphyromonas gingivalis, and Fusobacterium nucleatum. Following pretreatment of the Ti with ion rich solutions or coating by human serum albumin or fibronectin, bacterial adhesion was examined by scanning electron microscopy and assessed quantitatively by DNA analysis. Ti coating by proteins as well as bacterial adhesion and their interrelationships were further investigated through confocal scanning laser microscopy., Results: Acid-etched and blasted Ti surfaces exhibited significantly higher amounts of bacteria adhesion than the other two surfaces. Calcium was found to serve as a bridging agent in the adhesion process of S. mutans and F. nucleatum to Ti surfaces. Although albumin coating of the Ti reduced the adhesion of S. mutans to all surfaces, it had no influence on the adhesion of P. gingivalis or F. nucleatum. Coating the Ti with fibronectin enhanced P. gingivalis and F. nucleatum adhesion., Conclusions: Bacterial adhesion to Ti surfaces is roughness-dependent, and the adhesion mechanism is influenced by ions and proteins of the initial coating derived from the blood., (© 2011 John Wiley & Sons A/S.)

Background: Localized aggressive periodontitis (LAgP) is an inflammatory disease associated with specific bacteria, particularly Aggregatibacter actinomycetemcomitans, which can result in early tooth loss. The bacteria grow as a biofilm known as subgingival plaque. Treatment includes mechanical debridement of the biofilm, often associated with empirical antibiotic treatment., Objective: The aims of this study were to test in vitro the sensitivity of A. actinomycetemcomitans JP2 during planktonic and biofilm growth to doxycycline and to the combination of metronidazole and amoxicillin, which are two antibiotic protocols commonly used in clinical practice., Design: Two in vitro biofilm models were used to test the effects of the antibiotics: a static 96-well plate assay was used to investigate the effect of these antibiotics on biofilm formation whilst a flow chamber model was used to examine the effect on established biofilms., Results: Of the antibiotics tested in this model system, doxycycline was most efficacious with a minimal inhibitory concentration (MIC) against planktonic cells of 0.21 mg/L and minimal biofilm inhibitory concentration (MBIC) of 2.10 mg/L. The most commonly prescribed antibiotic regimen, amoxicillin + metronidazole, was much less effective against both planktonic and biofilm cells with an MIC and MBIC of 12.0 mg/L and 20.2 mg/L, respectively. A single treatment of the clinically achievable concentration of 10 mg/L doxycycline to sparse A. actinomycetemcomitans biofilms in the flow chamber model resulted in significant decreases in biofilm thickness, biovolume, and cell viability. Dense A. actinomycetemcomitans biofilms were significantly more resistant to doxycycline treatment. Low concentrations of antibiotics enhanced biofilm formation., Conclusion: A. actinomycetemcomitans JP2 homotypic biofilms were more susceptible in vitro to doxycycline than amoxicillin + metronidazole.

A novel method of collecting in vivo plasma proteins of humans from osteotomies prepared during insertion of an oral implant is described. A rod containing a collecting portion with a predetermined surface is introduced into the osteomy, removed, and transferred for enzyme-linked immunosorbent assay analysis. Two experiments were used to examine the feasibility of the method. In the first, titanium (Ti) rods with different roughness were exposed for 10 min to the blood. Blasted and acid-etched surfaces adsorbed four times more and acid-etched surfaces adosorbed two times more plasma proteins as compared to machined surfaces. In the second experiment, blasted and acid-etched rods were wetted for 10 s prior to the insertion. The adsorption for fibronectin, albumin, fibrinogen, and IgG was enhanced significantly compared with nonwetted rods. These results are discussed in the light of previous methods used in studies on adsorption. Thus, use of the collecting instrument enables aspects of human plasma-implant interface to be studied in a more realistic manner.

Host defense peptides are innate immune effectors that possess both bactericidal activities and immunomodulatory functions. Deficiency in the human host defense peptide LL-37 has previously been correlated with severe periodontal disease. Treponema denticola is an oral anaerobic spirochete closely associated with the pathogenesis of periodontal disease. The T. denticola major surface protein (MSP), involved in adhesion and cytotoxicity, and the dentilisin serine protease are key virulence factors of this organism. In this study, we examined the interactions between LL-37 and T. denticola. The three T. denticola strains tested were susceptible to LL-37. Dentilisin was found to inactivate LL-37 by cleaving it at the Lys, Phe, Gln, and Val residues. However, dentilisin deletion did not increase the susceptibility of T. denticola to LL-37. Furthermore, dentilisin activity was found to be inhibited by human saliva. In contrast, a deficiency of the T. denticola MSP increased resistance to LL-37. The MSP-deficient mutant bound less fluorescently labeled LL-37 than the wild-type strain. MSP demonstrated specific, dose-dependent LL-37 binding. In conclusion, though capable of LL-37 inactivation, dentilisin does not protect T. denticola from LL-37. Rather, the rapid, MSP-mediated binding of LL-37 to the treponemal outer sheath precedes cleavage by dentilisin. Moreover, in vivo, saliva inhibits dentilisin, thus preventing LL-37 restriction and ensuring its bactericidal and immunoregulatory activities.

Previous studies have shown that whole cells of several periodontal pathogenic bacteria including Porphyromonas gingivalis may degrade the clinically used regeneration membranes Biomend Extend and Bio-Gide. Fractionation of P. gingivalis cells revealed that cell membrane-associated proteases are responsible for the in vitro degradation of the collagen membranes. In the present study, the specific role of extracellular vesicles and the purified Arg-gingipain enzyme of P. gingivalis in the degradation of three differently cross-linked collagen membranes (Ossix; Bio-Gide and Biomend Extend) was examined. In addition, the inhibitory effect of antibacterial agents and antibiotics used in local periodontal therapy on the enzymatic degradation was evaluated. The data presented show that while all tested collagen membranes, are prone to lysis by oral bacterial proteases, cross-linked membranes are more resistant to proteolysis. Furthermore, therapeutical concentrations of the antibacterial and antibiotic agents chlorhexidine, cetylpyridiniumchloride, minocycline and doxycycline were found to partially inhibit the enzymatic breakdown of the membranes, while metronidazole had no such effect. These results suggest that the presence of P. gingivalis cells, extracellular vesicles and enzymes in the vicinity of regeneration membranes in the periodontium, may change their physical structure and therefore alter their biological properties. Furthermore, the use of cross-linked collagen membranes and antibacterial agents may significantly inhibit this proteolytic process.

Coagreggation of Treponema denticola with either Porphyromonas gingivalis or Fusobacterium nucleatum was characterized and the role of the major outer sheath protein (MSP) in the coaggregation process of these bacteria was evaluated. The MSP of T. denticola was found to be able to bind to P. gingivalis and F. nucleatum cells and this binding could be inhibited by MSP in a concentration-dependent manner. While sodium dodecyl sulfate polyacrylamide gel electrophoresis and Periodic acid-Schiff (PAS) staining of MSP revealed that it is a glycoprotein, monosaccharide analysis showed that MSP contains: Glc (44.4), Gal (20.4%) GlcN (1.3%), GalN (31.6%) and Fuc (9.2%). Peptide N-glycosidase F deglycosylation of MSP was found to inhibit its binding to F. nucleatum but not to P. gingivalis cells. Sugar-binding studies showed that the requirements for the binding of both T. denticola and MSP to F. nucleatum cells are similar to those of the F. nucleatum galactose-binding lectin. These data suggest that MSP acts as an adhesin during the coaggregation process of T. denticola with P. gingivalis and F. nucleatum through its protein and carbohydrate moieties, respectively.

Objectives: The aim of this study was to examine the effect of modified titanium (Ti) surfaces on the initial events of plasma proteins adsorption., Materials and Methods: 'Ti disks' with three types of surface modifications were compared: machined, acid-etched and acid-etched and blasted. Physical and chemical characterizations of the surfaces were performed via scanning electron microscopy (SEM), atomic force microscopy (AFM) used for analysis of surface topography, characterization of the titanium oxide (TiO2) layer was carried out by X-ray photoelectron spectroscopy (XPS) and characterization of surface energy by the determination of contact angles. Evaluation of plasma proteins' adsorption to the treated Ti surfaces was performed by mass spectrometry, confocal laser scanning microscopy and XPS. Quantitative proteins' assessment was carried out by enzyme-linked immunosorbent assay., Results: SEM images revealed major differences in the topography of the examined surfaces. Acid-etched and blasted Ti surfaces were found to have higher roughness values and a thicker TiO2 layer as compared with acid-etched and machined surfaces. Moreover, acid-etched and blasted surfaces showed high surface area differentiation, pointing to a high increase in the three-dimensional (3D) surface area over the 2D surface area compared with the other surfaces. Adsorption of plasma proteins to the acid-etched and blasted Ti surfaces was both qualitatively and quantitatively more intense compared with the machined and acid-etched surfaces. This was shown for each examined protein, total proteins and by the removal degree of the protein coat., Conclusions: The preferential adsorption of plasma proteins to the acid-etched and blasted Ti surfaces may be explained by its topographical characteristics and by the increase of the 3D surface area of this modified surface.

Previous reports have shown that coaggregation between Porphyromonas gingivalis and Fusobacterium nucleatum, two important periodontopathogens, is mediated by a galactoside on the surface of P. gingivalis and a lectin on F. nucleatum. In the present study, purified capsular polysaccharide (CPS) and lipopolysaccharide (LPS) of P. gingivalis PK 1924 (serotype K5) were found to be able to bind to F. nucleatum cells and to inhibit binding of F. nucleatum to P. gingivalis serotype K5. Sugar binding studies showed that the requirements for binding of P. gingivalis serotype K5 CPS and LPS to the F. nucleatum lectin are: the presence of a metal divalent ion, an axial free hydroxyl group at position 4 and free equatorial hydroxyl groups at position 3 and 6 of d-galactose. These data suggest that P. gingivalis serotype K5- CPS and LPS act as receptors mediating coaggregation between P. gingivalis and fusobacteria.

A 65 kDa protease was partially purified from extracellular vesicles of Fusobacterium nucleatum cultures by preparative SDS-PAGE followed by electroelution. The pH optimum of the protease is 7.5-8.0 and its activity could be inhibited by serine protease inhibitors. The protease was found to degrade the extracellular matrix proteins fibrinogen and fibronectin as well as collagen I and collagen IV which were degraded at 37 degrees C but not at 28 degrees C, indicating the presence of a gelatinase activity in these bacteria. The 65 kDa protease was also able to digest the alpha-chains of immunoglobulin A but not immunoglobulin G. The 65 kDa F. nucleatum protease, capable of degrading native proteins, may play an important role in both the nutrition and pathogenicity of these periodontal microorganisms. The degradation of extracellular matrix proteins by bacterial enzymes may contribute to the damage of periodontal tissues, and degradation of IgA may help the evasion of the immune system of the host by the bacteria., (Copyright Blackwell Munksgaard, 2004.)

Purified Actinobacillus actinomycetemcomitans serotype b lipopolysaccharide (LPS) was found to be able to bind Fusobacterium nucleatum cells and to inhibit binding of F. nucleatum to A. actinomycetemcomitans serotype b. Sugar binding studies showed that the requirements for binding of A. actinomycetemcomitans serotype b LPS to the F. nucleatum lectin are the presence of a metal divalent ion, an axial free hydroxyl group at position 4, and free equatorial hydroxyl groups at positions 3 and 6 of D-galactose, indicating that the beta-N-acetyl-D-galactosamine in the serotype b LPS trisaccharide repeating unit is the monosaccharide residue recognized by the F. nucleatum lectin. These data strongly suggest that A. actinomycetemcomitans serotype b LPS is one of the receptors responsible for the lactose-inhibitable coaggregation of A. actinomycetemcomitans to fusobacteria.

Bacterial infection in the vicinity of guided tissue regeneration barrier membranes was shown to have a negative effect on the clinical outcomes of this increasingly used technique. Several oral and specifically periodontal bacteria were shown to adhere to such membranes in vivo and in vitro with a higher affinity to membranes constructed from collagen. The present study examined the role of periodontal bacteria and their enzymes in the degradation of commercially used collagen membranes. Degradation of two collagen membranes [Biomend (Calcitek, Colla-Tec Inc., Plainsboro, NJ) and Bio-Gide (Geistlich Biomaterials, Wolhousen, Switzerland)] labeled by fluorescein isothiocyanate was examined by measuring soluble fluorescence. Porphyromonas gingivalis, Treponema denticola and Actinobacillus actinomycetemcomitans and their enzymes were evaluated. Collagenase from Clostridium hystolyticum was used as a positive control. While whole cells of P. gingivalis were able to degrade both types of membranes, T. denticola could degrade Bio-Gide membranes only and A. actinomycetemcomitans whole cells could degrade none of the membranes. Fractionation of P. gingivalis cells revealed that cell membrane associated proteases were responsible for the degradation of the two collagen membranes. In T. denticola, the purified major phenylalanine protease was found to be responsible for the degradation of Bio-Gide membranes. These results suggest that proteolytic bacterial enzymes may take part in the degradation of collagen barrier membranes used for guided tissue regeneration.

The present study compares periodontal parameters, caries, and levels of colony forming units (CFU) of bacteria from subgingival plaque of permanent teeth, to those of primary teeth examined 4 years previously. Six children who had periodontitis and 5 who had no periodontitis in primary teeth (groups A and B respectively) were examined. The microbial examination included the number of CFU of the total anaerobic count, Actinobacillus actinomycetemcomitans, Streptococcus mutans and Porphyromonas gingivalis. The differences in CFU values for the permanent teeth between groups A and B were not significant. Group A had significantly higher gingival inflammation values in the permanent teeth than group B. Permanent teeth had significantly higher CFU values of P. gingivalis than the primary teeth. Based on the present limited sample, the number of CFU from bacteria of the subgingival plaque of primary teeth are not an adequate predictor of periodontal disease or caries in the permanent teeth.

A Porphyromonas endodontalis ATCC 35406 protease was purified from Triton X-114 cell extracts by preparative SDS-PAGE followed by electroelution. The purified enzyme exhibits a molecular size of 88 kDa and was dissociated into two polypeptides of 43 and 41 kDa upon heating in the presence of sodium dodecyl sulfate with or without a reducing agent. The protease (pH optimum 7.5-8.0) degraded the extracellular matrix proteins fibrinogen and fibronectin. Collagen IV was also degraded at 37 degrees C but not at 28 degrees C. The protease also cleaved the bioactive peptide angiotensin at amino acid residue phenylalanine-8 and tyrosine-4 but failed to hydrolyze bradykinin, vasopressin and synthetic chromogenic substrates with phenylalanine or tyrosine at the P1 position. In addition, two peptidases were detected in P. endodontalis cells: a proline aminopeptidase that remained associated with the cell pellet after detergent extraction and peptidase/s that partitioned into the Triton X-114 phase after phase separation and degraded the bioactive peptides bradykinin and vasopressin. These P. endodontalis peptidases and proteases may play an important role in both the nutrition and pathogenicity of these assacharolytic microorganisms. The inactivation of bioactive peptides and degradation of extracellular matrix proteins by bacterial enzymes may contribute to the damage of host tissues accompanied with endodontic infections.

Among periodontal anaerobic pathogens, the oral spirochetes, and especially Treponema denticola, have been associated with periodontal diseases such as early-onset periodontitis, necrotizing ulcerative gingivitis, and acute pericoronitis. Basic research as well as clinical evidence suggest that the prevalence of T denticola, together with other proteolytic gram-negative bacteria in high numbers in periodontal pockets, may play an important role in the progression of periodontal disease. The accumulation of these bacteria and their products in the pocket may render the surface lining periodontal cells highly susceptible to lysis and damage. T. denticola has been shown to adhere to fibroblasts and epithelial cells, as well as to extracellular matrix components present in periodontal tissues, and to produce several deleterious factors that may contribute to the virulence of the bacteria. These bacterial components include outer-sheath-associated peptidases, chymotrypsin-like and trypsin-like proteinases, hemolytic and hemagglutinating activities, adhesins that bind to matrix proteins and cells, and an outer-sheath protein with pore-forming properties. The effects of T. denticola whole cells and their products on a variety of host mucosal and immunological cells has been studied extensively (Fig. 1). The clinical data regarding the presence of T. denticola in periodontal health and disease, together with the basic research results involving the role of T. denticola factors and products in relation to periodontal diseases, are reviewed and discussed in this article.

The purpose of the present study was to develop a DNA probe for Porphyromonas endodontalis. Pure cultures of P. endodontalis were grown in TYP medium, in an anaerobic chamber. DNA was extracted from the P. endodontalis and labeled using the Genius System by Boehringer Mannheim. The labeled P. endodontalis DNA was used in dot-blot hybridization reactions with homologous (P. endodontalis) and unrelated bacterial samples. To determine specificity, strains of 40 other oral bacterial species (e.g. Porphyromonas gingivalis, Porphyromonas asaccharolytica, and Prevotella intermedia) were spotted and reacted with the P. endodontalis DNA probe. None of the panel of 40 oral bacteria hybridized with the P. endodontalis probe, whereas the blot of the homologous organism showed a strong positive reaction. To determine the sensitivity of the probe, dilutions of a P. endodontalis suspension of known concentration were blotted onto a nylon membrane and reacted with the probe. The results of our investigation indicate that the DNA probe that we have prepared specifically detects only P. endodontalis and can detect at least 3 x 10(4) cells.

Guided tissue regeneration (GTR) techniques are increasingly used for the treatment of periodontal defects, or in conjunction with dental implant procedures. As adhesion of bacteria to barrier membranes used in these techniques may lead to failure, a prerequisite for treatment success is an infection-free healing process. The present study examined the adhesion of 3 periodontal pathogenic bacteria: Actinobacillus actinomycetemcomitans, Treponema denticola and Porphyromonas gingivalis, to 3 barrier membranes: Collagen, (Biomend) PTFE, (TefGen-FD) and e-PTFE, (Gore-Tex). The membranes were incubated with 3[H]-thymidine labeled bacteria, and the number of adherent bacteria was calculated using a scintillation counter. The effect of albumin coating on bacterial adherence to the membranes was also studied. Bacterial adherence to the membranes was further examined by scanning electron microscopy (SEM). The results show that the adherence of all bacterial strains to collagen membranes was significantly higher than to the other membranes tested. Precoating of the membranes with albumin did not change the bacterial adherence significantly. These findings are of importance in evaluating the ability of periodontal bacteria to colonize and infect different types of barrier membranes.

The 160, 190 and 270 kDa outer sheath proteases of Treponema denticola ATCC 35404 were found to be multiple forms of the major 91 kDa phenylalanine protease (PAP) by immunoblotting using anti-91 kDa specific antibodies. Multiple forms of the phenylalanine protease were also found in 2 other T. denticola strains studied, ATCC 33520 and the clinical isolate GM-1. Protein, proteolytic and Western blot analyses using antibodies against the PAP and the major outer sheath protein (MSP) indicated that the 190 and 270 kDa proteases were protein complexes formed by the MSP and the PAP. These complexes dissociated by storage in 0.3% or higher SDS concentrations. The purified PAP was found to completely degrade keratin, but was unable to degrade native actin either in its monomeric or polymerized form. The association of the MSP adhesin with a protease capable of degrading host native proteins may benefit the obtention of protein-based nutrients necessary to support the growth of these treponemes. These complexes may also play a role in the structural organization of T. denticola outer sheath.

We have recently demonstrated that the periodontopathogenic oral spirochete Treponema denticola possesses membrane-associated lipoproteins in addition to lipooligosaccharide (LOS). The aim of the present study was to test the potential of these oral spirochetal components to induce the production of inflammatory mediators by human macrophages, which in turn may stimulate tissue breakdown as observed in periodontal diseases. An enriched lipoprotein fraction (dLPP) from T. denticola ATCC 35404 obtained upon extraction of the treponemes with Triton X-114 was found to stimulate the production of nitric oxide (NO), tumor necrosis factor alpha (TNF-alpha), and interleukin-1 (IL-1) by mouse macrophages in a dose-dependent manner. Induction of NO by dLPP was at 25% of the levels obtained by Salmonella typhosa lipopolysaccharide (LPS) at similar concentrations, while IL-1 was produced at similar levels by both inducers. dLPP-mediated macrophage activation was unaffected by amounts of polymyxin B that neutralized the induction produced by S. typhosa LPS. dLPP also induced NO and TNF-alpha secretion from macrophages isolated from endotoxin-unresponsive C3H/HeJ mice to an extent similar to the stimulation produced in endotoxin-responsive mice. Purified T. denticola LOS also produced a concentration-dependent activation of NO and TNF-alpha in LPS-responsive and -nonresponsive mouse macrophages. However, macrophage activation by LOS was inhibited by polymyxin B. These results suggest that T. denticola lipoproteins and LOS may play a role in the inflammatory processes that characterize periodontal diseases.

Several implant materials used in dental and orthopedic surgery were placed in rat tibial bones to study their effects on mineralization. The implants consisted of bone bonding and non-bonding materials. Changes in mineralization were defined by morphometric analysis of matrix vesicle distribution at the implant interface and in normal bone healing following marrow injury. Bone-bonding materials induced an increase in matrix vesicle activity. This finding was supported by study of the biochemical changes in the same model that manifested high correlations to the morphometrical observations with regard to enhancement or delay of primary mineralization. In addition, the study of healing using nuclear methods indicated that implants alter bone healing as shown by the different uptakes of 99mTc and 32P in the different bone compartments. Decreased 32P uptake by the organic phase in the presence of bone-bonding implants suggested that cleavage of 99mTc-MD32P into its technetium and methylene diphosphonate moieties was inhibited by administration of implants. Further studies on the effect of bacterial infection on the peri-implant tissues revealed a decrease in woven bone formation due to infection.

In this study, the adhesion of radioactively labeled Actinomyces viscosus (A. viscosus), Actinobacillus actinomicetemcomitans (Aa) and Porphyromonas gingivalis (P. gingivalis) to titanium (Ti) and Ti-6-Al-4V alloy (Ti-alloy) coated with albumin or human saliva was investigated. All the tested bacteria displayed greater attachment to Ti-alloy than to Ti. P. gingivalis exhibited less adhesion to Ti and Ti-alloy than did the other bacterial strains. Adhesion of A. viscosus and Aa was greatly reduced when Ti or Ti-alloy were coated with albumin or saliva. P. gingivalis demonstrated a lesser reduction in adhesion to albumin or saliva-coated surfaces. The results show that oral bacteria have different adhesion affinities for Ti and Ti-alloy and that both albumin and human saliva reduce bacterial adhesion.

The aim of the in vitro study was to examine the accumulation of Streptococcus mutans on light-cured composite materials and amalgam. Bacteria cultures were grown in a brain heart infusion medium, and their growth rate was determined through turbidity measurements. The data, so obtained, were evaluated statistically by analysis of variance (ANOVA) followed by Scheffe test. Experiments on amalgam showed better results compared to those on composite materials. There were no statistically significant differences in plaque accumulation on different composite materials after finishing and polishing procedures, compared to plaque accumulation on composite materials against a Mylar strip.

Our previous studies have shown that human albumin is one of the main salivary proteins that adsorb to titanium (Ti). The goal of the present study was to investigate the role of electrostatic interactions in the adsorption of human albumin to Ti-oxide (TiO2) in vitro. The binding profile of human albumin to Ti was analyzed according to an adsorption isotherm. Purified human serum albumin (HSA) was suspended with native, calcium-, magnesium-, or potassium-treated commercially pure Ti powders, at pH 3.0 and 7.0. The amount of unadsorbed protein in the supernatant fluid was measured. The maximum amount of adsorbed albumin was 0.13 mg/1.0 g Ti. The albumin-Ti association constant was 2.77 mL/mg. Pretreatment of Ti with calcium, or magnesium alone, or combined with increasing pH values (3.0-7.0) resulted in augmented adsorption of HSA to Ti. No increase in adsorption was observed following pretreatment of Ti with potassium. These results point to the involvement of electrostatic interactions in the adsorption of HSA to TiO2.

The presence of lipoproteins and lipooligosaccharides in Treponema denticola, an oral spirochaete associated with periodontal diseases, was investigated. T. denticola ATCC 35404 and the clinical isolate GM-1 were metabolically labeled with [3H]-cis-9-octadecenoic acid and extracted with the non-ionic detergent Triton X-114. The extract was phase separated, precipitated with acetone and delipidated to remove non-covalently bound lipid (dLPP). In T. denticola ATCC 35404, sodium dodecyl sulfate polyacrylamide electrophoretic separation followed by autoradiography showed [3H]-cis-9-octadecenoic acid incorporation in bands with apparent molecular masses of 14, 20, 26, 31, 38, 72 and 85 kDa and a broad band running from 113 kDa to the top of the gel. This last band resolved into a 53 kDa [3H]-cis-9-octadecenoic acid band upon heating for 10 min, at 100 degrees C. The structural relationship of the outer sheath major oligomeric polypeptide of strain ATCC 35404 and the 53 kDa protein was demonstrated immunologically. Antibodies against the 113 kDa component of the oligomer cross-reacted with the 53 kDa protein. Proteinase K degraded the [3H]-cis-9-octadecenoic acid bands with the exception of the 14 kDa. The 14 kDa was also the major [3H]-fatty acid labeled compound found in the water phase following phenol-water extraction of whole T. denticola ATCC 35404 cells. This compound was purified from the water phase by gel filtration followed by hydrophobic chromatography. Chemical analysis showed that hexadecanoic acid was the predominant fatty acid bound to T. denticola lipoproteins. In the GM-1 strain [3H]-cis-9-octadecenoic acid incorporation was observed in the 116 kDa and 14 kDa bands. dLPP from strain ATCC 35404 caused an enhanced (0.8-8 micrograms/ml) luminol dependent chemiluminiscence (LDCL) effect in human polymorphonuclear neutrophils (PMN) which could be related to protein concentration. The addition of dLPP to PMN together with FMLP at submaximal concentration (1 microM) resulted in a synergistic activation of LDCL. At 21 micrograms/ml, dLPP also induced lysozyme release by the PMN at approximately 30% of the release induced by the chemotactic peptide at 1 microM. In addition, dLPP (21 micrograms/ml) increased additively the release of lysozyme caused by 1 microM FMLP. The release of beta-glucuronidase was not affected. The modulation of neutrophil activity was abolished by preincubation of dLPP with proteinase K. The purified 14 kDa had no effect on either LDCL or exocytosis of lysosomal enzymes of PMN. These data strongly suggest that T. denticola possesses several lipoproteins including outer sheath major oligomeric polypeptides (113-234 kDa) and a lipooligosaccharide of molecular mass of 14 kDa. In addition, an enriched lipoprotein fraction from this oral spirochaete modulates oxygen dependent and independent mechanisms for controlling microorganisms by human PMN.

Amine fluoride (AmF)- and stannous fluoride (SnF2)-containing products were found to have a therapeutic effect on gingivitis and periodontitis. This effect was suggested to correlate with the antibacterial activity of the fluoride compounds. However, their effect on inflammatory cell function can also play a role in the therapeutic effect on gingival inflammation. The present study was designed to test the effects of AmF, SnF2, and an AmF/SnF2 combination on the function of human peripheral blood neutrophils, as compared with effects of chlorhexidine and salicylic acid. Neutrophils were isolated from human blood by ficoll centrifugation followed by dextran sedimentation. The neutrophils were pre-incubated with AmF, SnF2, or AmF/SnF2, followed by stimulation with fMLP. Cell vitality was verified by trypan-blue exclusion (> 95% vitality at all tested concentrations). Superoxide production was measured by cytochrome C reduction and the enzymatic activity of lysozyme and beta-glucoronidase by optical density measurement of substrate conversion. The results showed that AmF, SnF2, or AmF/SnF2 enhanced by two- to three-fold the superoxide release from fMLP-stimulated human neutrophils. Furthermore, the effective concentration of the AmF/SnF2 combination was several-fold lower than that of AmF or SnF2 alone (10 nM for AmF, 0.5 microM for SnF2, and 3 pM for SnF2/AmF). On the other hand, chlorhexidine and salicylic acid were found to reduce superoxide production by the cells. All the tested compounds had no effect on granular enzyme release by the stimulated neutrophils. The results suggest that AmF and SnF2 enhance the oxygen-dependent antibacterial activity of neutrophils. This effect may contribute to a more efficient elimination of bacteria from the periodontal environment, resulting in improvement in gingival health.

The is study examined the relationships between the microbial composition of the subgingival plaque, contact loss caused by caries and alveolar bone loss (ABL) in primary molars. The study included 10 children with contact loss in at least two sites, one with ABL and one without ABL, and 10 children without ABL with sites with or without contact loss. The microbial composition of subgingival plaque was examined by dark-field microscopy and by cultures of total anaerobic bacteria, Actinobacillus actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg). Dark-field microscopy confirmed that spirochetes and motile rods may be part of the indigenous flora of the oral cavity. More spirochetes and motile rods were observed in sites with ABL than in control sites in the same subject and control subjects without ABL. Lower numbers of cocci were seen in sites with ABL than in sites in children without ABL, but a significant difference was not observed between sites with ABL and healthy sites within the same subjects. No significant differences in the dark-field values were evident in sites without ABL, with or without contact loss. Aa and Pg were found in children and sites with or without ABL. In sites with Aa, larger proportions of spirochetes, lower values of cocci, and more colonies of Pg were evident. No significant differences in anaerobic bacteria were evident between sites with or without contact loss or with or without ABL. ABL in the primary dentition was found to be related to the microbial composition of the subgingival plaque, but not related to contact loss per se.

In vivo adsorption of salivary proteins onto prosthetic titanium components was analyzed after exposure of titanium abutments to the oral environment for a period of 2 to 6 weeks. Gel electrophoresis and Western immunoblotting were used to separate and identify the proteins, which were mainly alpha-amylase and serum albumin. Selective adsorption of proteins enables attachment of specific oral bacteria and thus may alter the composition of the dental plaque formed on titanium surfaces.

Titanium (Ti) is among the most widely used implant materials in dentistry today. The success of Ti implants is associated with their interactions with the surrounding tissues and biological fluids. In the present study, the adsorption of salivary proteins to Ti and the effect of calcium (Ca) on this process were investigated. Untreated and Ca-treated Ti powders were suspended in human clarified whole saliva. After incubation, the supernatant fluid was collected for protein analysis. The powders were then washed and resuspended in EDTA to desorb proteins from Ti surfaces. Sodium dodecylsulphate polyacrylamide gel electrophoresis and Bradford protein assay were conducted to determine the concentration and type of proteins that adsorbed onto Ti surfaces. The presence of Ca ions enhanced the adsorption of salivary proteins to Ti. A 66 kDa protein, identified by immunoblotting as albumin, was found as the main adsorbed salivary protein. Adsorption of albumin to Ti pretreated with Ca was significantly greater than to native Ti. The Ca-dependent adsorption process was reversed by EDTA. The data suggest that salivary albumin is one of the main constituents of a salivary biofilm formed on Ti dental implants and its adsorption to Ti surfaces is Ca-dependent. The presence of albumin on Ti dental implants may affect plaque accumulation on the implants and the biocompatibility of Ti implants.

Electron microscopical observations of the oral periodontopathogen Treponema denticola show the presence of extracellular vesicles bound to the bacterial surface or free in the surrounding medium. Extracellular vesicles from T. denticola ATCC 35404, 50 to 100 nm in diameter, were isolated and further characterized. Protein and proteolytic patterns of the vesicles were found to be very similar to those of isolated T. denticola outer sheaths. They were enriched with the major outer sheath polypeptides (molecular sizes, 113 to 234 kDa) and with outer sheath proteases of 91, 153, 173, and 228 kDa. These findings indicate that treponemal outer sheath vesicles contain the necessary adhesins and proteolytic arsenal for adherence to and damage of eucaryotic cells and mammalian matrix proteins. The major outer sheath- and vesicle-associated protease of T. denticola ATCC 35404 was purified and characterized. The purified enzyme had a molecular size of 91 kDa, and it dissociated into three polypeptides of 72, 38, and 35 kDa upon heating in the presence of sodium dodecyl sulfate with or without a reducing agent. The activity of the enzyme could be inhibited by diisopropylfluorophosphate, phenylmethylsulfonyl fluoride, and phenylboronic acid. The value of the second-order rate constant of the protease inactivation by phenylmethylsulfonyl fluoride was 0.48 x 10(4) M(-1) min-1. Inhibition of the enzyme by phenylboronic acid was rapid (< 1 min) and pH dependent. These data strongly suggest that this major surface proteolytic activity belongs to a family of serine proteases.

Bone marrow-transplanted patients can suffer from severe life-threatening infections. Oral bacterial cultures were collected from a group of 40 patients prior to and following bone marrow transplantation every 3 days, following initial preparation and eradication of oral infections. The samples were grown on the Titertek-Enterobac kit specific for Enterobacteriaceae. In 426 oral cultures 30.5% grew gram-negative bacteria, 76.6% of them were Enterobacteriaceae. young male patients had 8.3% positive cultures at the study start, a percentage which constantly increased during later periods. Older patients did not follow the same pattern. Also, the allogeneic transplantation group had a higher percentage of Enterobacteriaceae than the autologous group (49.0 versus 19.5%). In blood cultures 18 out of the 94 positive ones showed the presence of Enterobacteriaceae. The most commonly found microorganisms in oral cultures were Klebsiella oxytoca (23%), Enterobacter cloacae (18%) and Klebsiella pneumoniae (15%). The decrease in the positive oral cultures from 35.0% during the pretransplantation period to 5.4% close to the transplantation, demonstrates that the preparatory protocol used for the prevention of oral infections was highly effective.

Purpose: To evaluate the long-term effect of subgingival irrigations with amine fluoride mixed with stannous fluoride on the repopulation of bacteria in deep periodontal pockets (> 5 mm) following the active phase of non-surgical mechanical treatment., Materials and Methods: A split-mouth design was utilized in eight periodontal patients and the effect of the treatment was evaluated by microbiological culturing and clinical parameters., Results: Slight reduction in total anaerobe counts were detected in both treated and control sides, up to the first 5 weeks, followed by an increase in the counts reaching a plateau after 30 weeks. In contrast, black-pigmented bacteroides species (BPB) were significantly suppressed in the treated sides as compared to the control sides over the whole study period. The difference between the clinical parameters and between the tested and control sides, although in some points were found to be statistically significant, seem to have no clinical significance. The use of amine fluoride-stannous fluoride gel for irrigations of deep periodontal pockets as an adjunctive to conventional treatment, can prevent the repopulation of the pockets by BPB, which are considered to be a major group of periodontal pathogens.

Local applications of sustained-released varnishes of chlorhexidine and arginine were used in a controlled pilot study of 34 mentally retarded patients, ages 18-45, assigned to one of these groups: chlorhexidine (C), arginine (A), or placebo (P). A professional scaling followed by four weeks of professional brushing to reach a Plaque Index (PII) and Gingival Index (GI) of 1.0 at baseline preceded eight weeks of daily varnish application to the buccal and labial surfaces of all teeth. Clinical parameters (PII and GI) and bacterial samples from selected teeth were collected at predetermined intervals. Four and eight weeks following the baseline, the PII was significantly different among the groups, with the lowest score in the chlorhexidine group. No significant differences among the three groups were noted for the GI. The chlorhexidine and arginine groups showed significant reductions (p < 0.05 and p < 0.01, respectively) in the number of S. mutans. The arginine group showed a nonsignificant increase in the number of S. sanguis. These results suggest that the topical antimicrobial agents may have some relevance to plaque control among patients with mental retardation.

Pathogenic bacteria constitute the primary extrinsic agent in the etiology of early onset periodontitis. However, the risk of developing periodontal disease is not equal for all individuals, suggesting host factors are involved in determining an individual's disease susceptibility. In this report, a case of an otherwise healthy female, who exhibited prepubertal periodontitis (PPP) at age 10, juvenile periodontitis (JP) at age 13, and rapidly progressive periodontitis (RPP) at age 29 years, is presented. Microbial, immunological, and genetic features of the case are presented. PPP, JP, and RPP are considered distinct disease entities, albeit with similar pathology and pathogenesis, yet all were manifest sequentially in the same individual. This report presents the idea that certain individuals are predisposed to early-onset periodontal diseases and the early identification of risk factors is important in the management of these individuals.

The Moroccan Jewish community living in Israel shows a relatively large genetic distance from other North African Jewish communities. In this work the polymorphism of HLA class I and class II determinants, as defined by serology and oligotyping, is analyzed in 113 healthy unrelated Jews of Moroccan stock. The class I antigens HLA-A1, -B44, and -Cw7 showed the highest frequency, while the most prevalent class II variants were DRB1*0701 and *1104, DQA1*0501, and DQB1*0201 and *0301. HLA A1-B13-DR7, A2-B51-DR10, and A1-B44-DR13 were the most typical three-locus haplotypes. Although the antigen frequency distribution of the Moroccan Jews falls within the Caucasian diversity range, this community has a unique pattern in terms of antigen, gene, and haplotype frequencies. Thus, in the Moroccan Jews DRB1*1305, an allele believed to be the result of a recombination event between DRB1*1301-1302 and DRB1*1101, is represented to a much larger extent than in all the other population groups studied at the 11th IHWS. This allele may therefore be a typical Jewish variant. A particular finding was the high frequencies of HLA-B13, B52, and DR10, alleles common among some Oriental populations. The answer to this enigmatic phenomenon probably must be sought in the tortuous history of this community.

Several fibrinolytic activities of Treponema denticola, an oral spirochete associated with gingivitis and periodontal disease, were identified and characterized following phase partitioning with the nonionic detergent Triton X-114. The apparent molecular masses of the proteases ranged from 91 to 228 kDa when analyzed in sodium dodecyl sulfate-polyacrylamide gels containing fibrinogen as the protease substrate. A qualitative analysis of zymograms showed that the proteases were highly enriched in the detergent phase, although the 91-, 173-, and 228-kDa proteases were also found in the aqueous phase. Zymograms of crude outer sheaths prepared by repeated freezing-thawing revealed that the proteases may be associated with this subcellular compartment. The proteases displayed substrate specificity towards fibrinogen, were susceptible to sulfhydryl group reagents, and had a pH optimum between 7 and 8. The similarities in their sensitivity to inhibitors, temperature stability, pH optimum, and laddered protein profiles suggest that these hydrolytic enzymes may be part of a family of oligomeric proteases that may play an important role in the invasiveness of and tissue damage caused by the spirochete.

HLA proteins are genetically determined, and account in part for individual immune response. Several studies have been performed seeking an association between HLA antigens and various forms of periodontitis with no conclusive results. The aim of the present study was to determine the frequency of HLA antigens of patients suffering from the localized (LJP) and the generalized (SGP) forms of early-onset periodontitis (EOP). Twenty-six EOP patients from the same ethnic group were studied in comparison to 113 race-matched controls. The EOP group included 11 LJP and 15 SGP patients. HLA-A9 and B15 antigens were found to be significantly elevated in the patient group. These differences were found to be due to the high frequency of A9 and B15 antigens in the SGP patients, with the LJP patient group showing no significant difference from the control group. The results are in agreement with previous studies in which A9 and B15 were found to be associated with EOP. However, previous studies did not differentiate between the localized and the generalized form of EOP. These results support the hypothesis that the generalized and the localized forms of EOP are under different genetic control.

The secretion of prostaglandin E2 (PGE2), tumor necrosis factor alpha (TNF alpha), interleukin 1 beta (IL-1 beta), and interleukin 6 (IL-6) by adherent mononuclear cells (AMNC) from 28 patients with early-onset periodontitis was studied. The early onset-periodontitis patients consisted of 12 patients with localized juvenile periodontitis (LJP) and 16 patients with severe generalized periodontitis (SGP). The AMNC responses to different concentrations of lipopolysaccharide (LPS) (E. coli) were determined in these 28 patients and compared to 14 healthy controls. Mediator levels in the supernatant were measured using radioimmunoassays for PGE2, IL-1 beta, and IL-6 determination and an enzyme linked immunosorbent assay for TNF alpha levels. The mean age of the patients was 19.9 years for the LJP group, 30.4 years for SGP, and 28.0 years for the controls. The mean number of teeth per patient with attachment loss of > 6 mm was 4.75 in the LJP patients and 17.3 in the SGP group. In the absence of LPS, LJP AMNC secreted significantly more PGE2 than unstimulated control or SGP AMNC, while similar baseline amounts of IL-1 beta, IL-6, and TNF alpha were secreted by AMNC from the 3 patient groups. LPS stimulation resulted in the dose-dependent secretion of significantly higher levels of PGE2 by LJP AMNC compared to SGP AMNC which in turn secreted significantly more than controls. TNF alpha secretion by LJP monocytes was significantly greater than the SGP and the control groups while IL-1 beta secretion by the SGP AMNC was depressed compared to the other two patient groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Dental research has emphasized the importance of understanding the epidemiology and etiology of periodontal disease, the second most prevalent dental pathology. The World Health Organization includes periodontal health prevalence, according to the Community Periodontal Index of Treatment Needs (CPITN), in its Global Oral Data Bank. Recent immigrants to Israel from Ethiopia offer a unique and important opportunity for investigation. Seventy adults aged 35-45 years were examined. Dividing the dentition into six sextants (according to CPITN), on average, two sextants were found to be healthy and only 1.18 sextants revealed shallow or deep pockets. These results demonstrate a relatively healthy periodontal status, as compared both to Israeli adults and data from the WHO Data Bank. Periodontal health has traditionally been related to regular dental visits (professional teeth cleaning) and home oral hygiene maintenance (toothbrushing and flossing). This population of immigrants had never visited dentists or hygienists nor used toothbrushes or dental floss. The periodontal health of this population is therefore of particular interest.

The present study describes the prevalence of alveolar bone loss (ABL) in children in relation to caries, contact and space loss. In addition, the microbial composition of the subgingival plaque of 20 sites, from 5 children, is presented. Bite-wing radiographs from 500 children were examined. ABL was evident in: 99 sites from 60 children; > 1 site in 27 children; the maxilla only in 34 children; the mandible only in 17 children; both arches in 9 children; 37 sites with no caries; 4.9 percent of all sites with proximal caries; 15.8 percent of all sites with contact loss; and 20.5 percent of all sites with mesial drift. Anaerobic bacteria were cultured from all 20 sites. No significant differences in the percentages of colony forming units of Actinobacillus actinomycetemcomitans and black pigmented Bacteroides were found among sites with/without bone loss, with/without caries or probing depths smaller/equal or larger than 2.5 mm.

Adults (35-45 yr old) and children (5 and 12 yr old) recently arrived in Israel from rural areas in Ethiopia were examined. Caries levels were low: e.g. DMFT = 0.31 among 12-yr-olds, and 1.27 amongst adults. The 5-yr-olds were 86.8% caries-free, while 12-yr-olds were 81.8% and adults 54% caries-free respectively. According to interview data, the diet in Ethiopia had been based on local agricultural products and was almost sugar-free. The mean total count of salivary bacteria, as determined on blood agar, was 3.4 x 10(8); mean count of Streptococcus viridans, on mitis salivarius, was 6.7 x 10(7); and mean count of S. mutans, as determined on mitis salivarius with bacitracin, was 1.7 x 10(7). These levels were all high and were not significantly different from a control group of 20 Israelis. The mean number of lactobacilli, on Rogosa agar, was 2.75 x 10(4), which was significantly higher than among the controls (3.6 x 10(3). Salivary pH levels were generally similar between the Ethiopian group and the controls. Salivary flow was significantly higher for the Ethiopians (1.93 ml/min) than for controls (1.16 ml/min). Low levels of caries in this population can be attributed to an almost sugar-free diet and high salivary flow, but not to the composition of oral microflora.

We examined changes in the bacterial flora in hospitalized patients with leukemia. This study placed special emphasis on enteric microorganisms and their relation to the general status of the patient. One hundred thirty bacterial cultures from 16 leukemia patients and 16 control subjects, were obtained. The organisms were isolated on MacConkey agar and identified by the API-20E system. Enteric microorganisms were isolated from 62.2% of the leukemia patients as compared with 28% from the control group (p < 0.001). The enteric positive cultures were identified as Klebsiella (42.7%), Enterobacter (18.8%), and Pseudomonas (15.6%). In contrast to the negative cultures (1342), enteric microorganisms were cultured from 2948 specimens (p < 0.005).

The aim of this in-vitro study was to evaluate the effect of chlorhexidine in solution and in a sustained-release device as an intracanal medication. Hollow cylindrical dentine specimens prepared from bovine incisors were incubated with Streptococcus faecalis for a period of 3 weeks. The intracanal medicaments tested were 0.2% chlorhexidine gluconate solution (CH), chlorhexidine in a sustained-release device (1.2 mg) (SBD), camphorated paramonochlorophenol (CMCP), and a control (no medication). Each medicament was introduced into the lumen of one dentine specimen and incubated for 5 min, 24 h, 48 h or 7 days at 37 degrees C. The bacteriological samples were taken by shaving the dentine inside the lumen with dental burs ranging in size from ISO 023-033. The dentine powder was collected in test-tubes containing growth medium and incubated for 24 h. The optical density of the medium was recorded by means of a spectrophotometer at a wavelength of 540 nm. There was a statistically significant difference between the control group and all the medicaments tested.

This study was conducted to evaluate the efficacy of the antibacterial activity of Ca(OH)2 and a sustained-release device containing chlorhexidine (SRD) in both sterilization and prevention of secondary infection of the root canal system. Bovine root dentine specimens previously incubated with Streptococcus faecalis were used in this experiment. Two different formulations of the SRD (differing in their cross-linkage), Ca(OH)2 and normal saline (control) were evaluated. The degree of bacterial infection of the root canal was tested after incubation periods of 24 h, 72 h and 7 days with these medicaments. Their efficacy in preventing secondary infection after recontamination was tested after 72 h and 7 days. The results demonstrated that both formulations of the SRD significantly reduced the bacterial population in the primary infected groups, as well as preventing secondary infection of the dentinal tubules in the recontaminated group. By contrast, Ca(OH)2 did not show any antibacterial activity, and failed to sterilize the dentinal tubules or prevent secondary infection after recontamination at the time periods examined.

The prevalence of dental caries and periodontal treatment needs in an institutionalized Down syndrome (DS) population was examined as well as the relationship between caries prevalence, salivary pH and salivary levels of Streptococcus mutans. Twelve patients with DS, aged 20 to 48, were compared with two similar age and gender control groups: healthy, and non-Down institutionalized mentally retarded patients. The pH levels did not differ significantly among the three groups. As to periodontal needs expressed by the CPITN, the institutionalized groups had the highest needs compared with the healthy group. Down adults, who were cariesfree, had significantly lower S. mutans counts compared with the patients with caries. In addition, the cariesfree patients with DS had much lower CPITN scores compared with the patients with caries.

The effect of local applications of sustained-release delivery polymer containing chlorhexidine as an adjunct to mechanical plaque removal was studied in institutionalized children with Down's syndrome. Thirty children, ages 8-13, participated in the study. Clinical parameters (P1I, GI, papillary bleeding) were recorded and bacterial samples from selected permanent teeth were collected and processed before and following treatment. Following the registration of clinical findings and collection of plaque, all children had their teeth scaled and polished and were randomized into three treatment groups of ten subjects each: Subjects in group I had their teeth coated with ethyl cellulose containing chlorhexidine; those in group II had their teeth coated with placebo polymer and those in group III received no further treatment. Individual oral hygiene habits were not interfered with. The application of the solutions to the respective groups was done every 3 days for 21 days. Chlorhexidine treatment significantly reduced the PLI, GI total aerobic counts and S. viridans counts compared to no treatment. The placebo group showed similar effects to that of the chlorhexidine-treated group except for the total aerobic counts which were not affected. The results suggest that the use of chlorhexidine in a sustained-release dosage form applied to the tooth surfaces may prove useful in the control of plaque and its sequela in children with Down's syndrome.

The release of chlorhexidine from an ethyl cellulose-based dosage form (SRD) has been shown to be effective in the reduction of the flora associated with periodontal pockets as well as in reducing probing depths. In this study, treatment with this dosage form was compared to routine maintenance therapy (RMT) in a 2-year, split mouth clinical trial. Ten patients with 84 pockets greater than or equal to 5 mm who had not received any periodontal therapy or systemic antibiotics over the last 6 months were included. The patients all received a full mouth scaling and root planing together with through oral hygiene instruction. Two months later (baseline) plaque index (P1I), bleeding on probing (BOP), pocket probing depth (PD), and attachment levels (AL) were assessed at all selected sites. Pockets on the control side then received RMT while the experimental pockets were treated with the SRD only. Treatment was repeated every 3 months for 2 years. The SRD treatment resulted in an improvement of greater than or equal to 3 mm in PD of at least 1 pocket in 8 of 10 patients, while RMT resulted in a similar improvement in only 1 of 10 patients (P = 0.012). Similarly a gain of attachment of greater than or equal to 3 mm was found in at least 1 SRD pocket in 8 of 10 patients. The RMT resulted in a similar improvement in only 2 patients (P = 0.012).(ABSTRACT TRUNCATED AT 250 WORDS)

The suppression of S. mutans is a predominate factor in preventing tooth decay. Sustained-release-delivery varnish of chlorhexidine was applied to dental plastic shells and administered to eight volunteers to be worn while sleeping. Salivary bacterial samples were taken on days 0, 7, 14, 21, 28, 35. A statistically significant reduction in S. mutans counts was recorded during the course of the study while a non-significant reduction in the total bacterial counts was found. Our results indicate that the application of chlorhexidine in the form of slow-release varnish in plastic shells is an effective intraoral drug delivery system resulting in reduction of S. mutans.