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Background: A common clinical observation following surgical periodontal therapy with an enamel matrix derivative (Emdogain) is the improved healing of the soft tissues and the limited inflammation of the operated areas. These clinical observations are empirical and difficult to explain. One of the factors influencing the early wound healing might be a potential antimicrobial effect of Emdogain., Aim: To investigate the effect of Emdogain on the vitality of ex vivo supragingival dental plaque and to compare this effect to that of a standard 0.2% chlorhexidine solution., Materials and Methods: 24 patients suffering from adult periodontitis were included in the study. At the beginning of the experiment, all participants were given a professional tooth cleaning. For the following 4 days, they had to refrain from any kind of oral hygiene measures. At day 5, from each of the volunteers, a voluminous plaque biofilm sample was taken with a sterile curette from the vestibular surfaces of the 1st lower molars and divided into 5 equal parts. Each part was mounted with 5 microl of the following solutions: (1) NaCl, (2) enamel matrix derivative dissolved in water (EMD), (3) enamel matrix derivative dissolved in the vehicle (Emdogain), (4) vehicle (propylene glycol alginate, PGA), (5) 0.2% chlorhexidine digluconate (CHX). After a reaction time of 2 min the test solutions were sucked off, and subsequently the biofilm was stained with a fluorescence dye. The vitality of the plaque flora after the treatments was evaluated under the fluorescence microscope (VF%)., Results: Plaque samples treated with NaCl showed a mean vitality of 76.8+/-8%. The EMD, Emdogain, PGA and CHX showed VF values of 54.4+/-9.2, 21.4+/-10.6%, 19.6+/-11.6% and 32.3+/-11.8%, respectively. Emdogain, PGA and CHX showed statistically highly significant reductions (p<0.0001) in terms of bacteria vitality when compared to water (negative control) and EMD. Both Emdogain and PGA were found to be statistically significantly different compared to CHX (p<0.001) (positive control)., Conclusion: The results of this study indicate that Emdogain might have an antibacterial effect on the vitality of the ex vivo supragingival dental plaque flora.

An experimental set-up allowing differentiation in vivo between antibacterial and antiadhesive properties of mouthrinses is described. The percentage of vital bacteria (= microbial vitality) and the bacterial counts were microscopically evaluated in saliva and in supragingival dental plaque both collected simultaneously at various times during de novo plaque formation. In a cross-over design, 12 healthy participants refrained from all oral hygiene for four separate periods of 2 x 4 h and 2 x 72 h after having rinsed with either an amine fluoride/stannous fluoride solution (Meridol) or 0.9% NaCl (placebo). Stimulated whole saliva was collected before and after the rinse. Together with whole-saliva samples, representative 4, 24 and 72-h-old plaque samples were separately taken from defined vestibular tooth surfaces that had been either exposed to the mouthrinse (unprotected sites) or temporarily covered with inert plastic films (protected sites) during rinsing. The pooled plaque and saliva were stained with fluorescent dyes to differentiate vital from dead micro-organisms which permitted the estimation of the percentages of vital bacteria. The total bacterial counts were quantified under the darkfield microscope. The Wilcoxon test was used for selected pairwise comparisons (alpha = 0.05). The percentage of vital bacteria in saliva fell significantly from 80-95% to about 50-60% as a result of the antibacterial activity of the test solution. These baseline values and those found in the presence of 4 and 24-h-old plaque were frequently lower than those recorded after the placebo rinse. In comparison to the placebo, microbial vitality was significantly reduced in early supragingival plaque formed on unprotected sites after applying the test solution. The similar total bacterial counts in 4-h-old plaque recorded after the use of the test solution on the unprotected and the protected areas did not point to an antiadhesive effect of the agent. It is concluded that this new experimental set-up allows decoding of the mode of action of a mouthrinse.

The object of this study was to monitor the proportion of vital bacteria (microbial vitality: VF in %) present in subgingival dental plaque following one single subgingival irrigation with saline (S), chlorhexidine (CHX) or povidone iodine (I2), but without any subgingival instrumentation. Its effect on the main composition of the microflora was also assessed. Seventeen patients with adult periodontitis took part in this investigation. In each patient four initially untreated pockets (pocket depth 5-11 mm) associated with bleeding were selected for the standardised pocket irrigation and plaque sampling at baseline (0 h) and after the following 1 h, 24 h, 7 days and 31 days. The subgingival irrigation was only performed once (0 h). One pocket per quadrant was irrigated using 0.9% prereduced S, 0.2% CHX or 0.05% I2 (Iso-Betadine Buccale). The remaining untreated pocket without any irrigation served as an additional control (C). Using an acrylic splint as a guide, paperpoints were inserted into the pocket precisely at the same site to collect subgingival plaque. The bleeding on sampling (BOS) was thereafter noted. The proportions of bacterial morphotypes were examined by darkfield microscopy. VF was evaluated using a vital fluorescence staining. The undisturbed subgingival dental plaque was composed of 86% (median value) vital bacteria. The sampling procedure alone and the saline irrigation led to a decrease in the number of spirochetes but had no influence on the vitality of the flora. Large variations in VF could be observed in the short-term (1 h, 24 h) irrigation effect of CHX and I2. The reduction of VF was still significant after 7 days (VF(CHX) 30-80%, VF(I2) 35-80%) but persisted up to 31 days only after I2 irrigation (VF(I2) 12-90%). The findings indicated that all single subgingival irrigations resulted in a temporary change of the subgingival microflora while povidone iodine produced the longest lasting antimicrobial effect. Any clinical advantage of this situation should be further investigated.

The aim of this double-blind study was to enumerate the total number of living and dead bacteria on defined tooth areas during the application of antibacterial mouthrinses. After prophylaxis, 40 students refrained from all oral hygiene measures for 3 d, during which they rinsed with a phenolic compound (Listerine), an amine fluoride/stannous fluoride solution (Meridol), 0.2% chlorhexidine (CHX) or a control solution (0.02% quinine-hydrochloride). The plaque index (P1I) was recorded at the start and the end of the investigation. Total bacterial counts (BC) and colony-forming units (CFU) of 1d-, 2d- and 3d-old dentogingival plaque were determined. The plating efficiency (PE) was calculated as a percentage of CFU/BC and the portion of vital microflora estimated by a vital fluorescence technique (VF). All groups started with a P1I approximating 0.1. On day 3, the P1I values were 1.21 in the control group and 0.51, 0.37 and 0.14 after Listerine, Meridol and CHX use, respectively. A tremendous variation existed between the numbers of viable bacteria found per mm2 on the enamel surface and day 3 (CHX: 0.2; Meridol: 300; Listerine; 6x10(4); control: 2x10(6)), while higher total numbers of bacteria were concomitantly present (CHX and Meridol: 1-2x10(4); Listerine: 2x10(5); control: 2x10(6)). Both vitality parameters PE and VF reached 92% in the control group at day 3, but only 7% after CHX use. With Meridol and Listerine, the corresponding PE values were 3% and 43%, respectively, while the VF values reached 48% and 54%. The PII, BC, CFU and PE values of the CHX and the Meridol groups differed significantly from those of the control group. In contrast, Listerine showed no difference as compared to the control rinse. Due to the strong antibacterial action of CHX and Meridol during their use, almost only dead or non-proliferating bacteria were found on the tooth surfaces. Thus, only a thin plaque could develop. As a clinical consequence, both substances showed retardation of plaque development as reflected by significantly reduced plaque indices.

The objective of this investigation was to determine the microbial generation time during early human supragingival plaque formation. On 2 occasions, 13 participants refrained from all oral hygiene measures for periods of 1, 2, 4, 8, 24 and 72 h. Plaque was sampled from defined areas of the teeth 13, 14, 15, 23, 24 and 25. Two independent methods were used to estimate the total number of viable cells: colony-forming units BCVF, total bacterial counts (BC) as enumerated by microscopic examination, and the percentage of vital bacteria using a vital fluorescence technique (VF) were recorded. The total number of vital microorganisms was calculated by BCVF (= BC x VF). The generation time based on either colony-forming units or BCVF increased significantly with plaque age. During the first interval (1-4 h), the microbial multiplication took slightly less than 1 h. After the establishment of a more mature plaque (24-72 h), the replicating time was between 12.5 h (BCVF) and 14.8 h (colony-forming units). It was concluded that the microbial generation time recorded at the initial stages of plaque formation is considerably lower than that found in older plaque.

The effect of chlorhexidine (CHX) on the viability of Streptococcus sanguinis was investigated in a preclinical biofilm model separately on cells in the planktonic or attached life form. Saliva-coated human enamel and glass slides were exposed to the streptococci suspended in sterile saliva for 30 min and 60 min in the flow chamber system. The CHX exposition was performed in two parts: pretreatment of the planktonic bacteria before their attachment to enamel or glass, and treatment of bacteria already attached to enamel. The susceptibility measured by vitality percentages was determined by fluorescence microscopy using vital/dead cells. After CHX pretreatment of planktonic cells, the mean values of the vitality percentages after adhesion were 14–18% (enamel) and 24–25% (glass). In contrast, the mean vitality percentages of untreated attached streptococci reached 70–75% (enamel) and 68% (glass). The vitality percentages of CHX-exposed bacteria dropped markedly to 2–5%, whereas those of untreated attached cells remained at 65–66%. The exposure of initially attached streptococci to CHX resulted in greater reduction of bacterial viability than with the planktonic counterparts. This preclinical biofilm model allows the investigation of various bacterial life forms and can furthermore be used to select efficient antiplaque therapeutics which might be beneficial for clinical plaque control. [ABSTRACT FROM AUTHOR]

The experimental gingivitis model was used to compare the antigingivitis, antiplaque, and antimicrobial efficacies of two commercially available (a phenolic and a plant alkaloid) compounds used as mouthrinses with those of a mouthrinse containing chlorhexidine digluconate. Thirty-one male and female volunteers with healthy gingivae and clean teeth ceased all oral hygiene procedures for 21 days during which they rinsed twice daily with: a) a phenolic compound (Listerine®), b) sanguinarine (Viadent®), c) 0.12% chlorhexidine digluconate, or d) a placebo mouthrinse. After 21 days of rinsing, virtually no signs of clinical gingivitis were observed in the chlorhexidine group. Subjects rinsing with Listerine, Viadent, or placebo developed clinical signs of gingivitis as measured by gingivitis occurrence, severity, and the proportion of gingival bleeding sites. At the same time, plaque accumulation was only slightly higher than at baseline in the chlorhexidine group, while the Listerine, Viadent, and placebo groups had significantly greater plaque accumulations, particularly during the initial period of treatment between day 0 and day 7. Microbiological enumeration of supragingival plaque collected at the end of the treatment period showed that chlorhexidine digluconate reduced plaque bacteria by 62-99% compared to the placebo group. No significant reductions in plaque bacteria were found among subjects using Listerine or Viadent. This study demonstrated that 0.12% chlorhexidine digluconate was superior to Listerine and Viadent in its ability to maintain optimal gingival health during the entire three weeks of mouthrinse use. It was of particular interest to note these effects during the final 14 to 21-day period of extreme challenge in this experimental model when gingivitis severity and bleeding site occurrence are most pronounced in the placebo group. [ABSTRACT FROM AUTHOR]

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