Effect of the most common wound antiseptics on human skin fibroblasts.

BACKGROUND: Antiseptics are used for the cleansing of acute or chronic wounds to eliminate micro-organisms from the wound bed. However, they have effects on the skin cells. AIM: To determine the effects of hexetidine, povidone-iodine (PI), undecylenamidopropyl-betaine/polyhexanide (UBP), chlorhexidine, disodium eosin and hydrogen peroxide on human skin fibroblasts. METHODS: CCD-1064Sk cells were treated with hexetidine, PI, UBP, chlorhexidine, disodium eosin or hydrogen peroxide. Spectrophotometry was used to measure cell viability and flow cytometry was used to study apoptosis and necrosis after the treatment. In vitro wound scratch assays were performed to determine the gap closure. RESULTS: All antiseptics significantly reduced the viability of human skin fibroblasts compared with controls. The percentage wound closure was lower with hexetidine, PI and UBP. The scratch assay could not be measured after treatments with chlorhexidine, disodium eosin or hydrogen peroxide, owing to their cytotoxicity. The apoptosis/necrosis experiments evidenced a significant reduction in viable cells compared with controls. An increased percentage of apoptotic cells was observed after treatment with all antiseptics. Compared with controls, the percentage of necrotic cells was significantly increased with all antiseptics except for hexetidine. CONCLUSION: The proliferation, migration and viability of human skin fibroblasts are reduced by treatment with hexetidine, PI, UBP, chlorhexidine, disodium eosin and hydrogen peroxide.

Analysis of Antiviral Properties of Hexoral In Vitro against Some Viruses that Cause Acute Respiratory Infections and Herpes.

Antiviral properties of Hexoral (0.1% solution and 0.2% aerosol for local application) and its constituent hexetidine against viruses causing human respiratory tract infections and herpes virus were studied in vitro. It was found that non-cytotoxic concentrations of hexetidine (alone and as a component of Hexoral) attenuated infectious properties of highly virulent influenza virus A/H5N1, pandemic influenza virus A/H1N1pdm, respiratory syncytial virus, and herpes simplex virus type 1 after a short-term exposure (30 sec) by 100 or more times. It was found that hexidine mostly contributes to the virucidal effect of Hexoral.

Effect of oral antiseptic agents on phospholipase and proteinase enzymes of Candida albicans.

OBJECTIVE: Candida-associated denture stomatitis is the most prevalent form of oral candida infections among the denture wearers. Generally, antiseptic oral rinses used in the treatment of these infections are considered as an adjunct or alternative antifungal treatment. Studies have suggested that the intraoral concentrations of antiseptics decrease substantially to the sub-therapeutic levels on account of the dynamics of the oral cavity. This condition yields the question about the minimum antiseptic concentration that effect the character or pathogenesis of Candida during treatment. The extracellular phospholipase and proteinase enzymes of Candida albicans are regarded to have a crucial role in the pathogenesis of human fungal infections. Therefore, the aim of this study was to investigate the effect of different sub-therapeutic concentrations of chlorhexidine gluconate, hexetidine and triclosan on the production of these enzymes by C. albicans strains isolated from 20 patients with denture stomatitis. METHODS: Phospholipase test was done by using Sabouraud dextrose agar with egg yolk, proteinase test was done by using bovine serum albumin agar. METHODS: Phospholipase test was done by using Sabouraud dextrose agar with egg yolk, proteinase test was done by using bovine serum albumin agar. RESULTS: Exoenzyme production of 20 strains which were brief exposured to sub-therapeutic concentrations of three antiseptic agents decreased significantly compared with the strains that were not exposured with antiseptic values (p<0.05). There was significant difference between the sub-therapeutic concentrations of each of three antiseptics (p<0.05). When the same concentrations of each antiseptic was compared, there were no significant differences between enzymatic activities (p>0.05). CONCLUSIONS: The results of this study show that sub-therapeutic levels of each antiseptic may modulate candidal exoenzyme production, consequently suppressing pathogenicity of C. albicans.

An in vitro oral biofilm model for comparing the efficacy of antimicrobial mouthrinses.

The ability of commercial mouthrinses to reduce total viable counts of mixed microbial populations was examined using a previously developed in vitro model of supragingival plaque. Exploratory experiments aimed at fine-tuning the model indicated that optimal correspondence between in vitro and clinical results for chlorhexidine-containing formulations were obtained at a saliva:medium ratio of 70:30 (v/v); moreover, expanding the microbial population from 5 bacterial species to 5 bacterial species + Candida albicans had no noticeable impact on overall results. The efficacies of 12 different mouthrinse proprietary products containing chlorhexidine, hexetidine, octenidine, Triclosan, plant extracts, or aminefluoride/stannous fluoride vis-à-vis biofilm clearance were compared. All mouthrinses promoted a statistically significant reduction in microbial load compared to distilled water. The herbal- and phenolic-based products were substantially less effective than most chlorhexidine-containing mouthrinses, or mouthrinses containing hexetidine or octenidine. No significant difference between the plaque-clearing plaque-clearing abilities of Listerine and Meridol was observed. This polyspecies biofilm model can be a valuable tool for preclinical testing of antiplaque formulations, particularly during the product development stage.

[Hexetidine--an oral antiseptic]. / Heksetidin--oralni antiseptik.

Hexetidine is very safe oral antiseptic with broad antibacterial and antifungal activity in vivo and in vitro. It has local-anesthetics, astringent and deodorant activity. Also, it has very strong antiplac effects. Resistention of microorganisms on hexetidine is short and transient. These characteristics give important therapeutic role in treatment of oral infections.

In vitro activities of antimicrobial agents against Candida species.

OBJECTIVE: Antimicrobial mouthrinses may represent a valid alternative to topical antifungal agents. However, the action of antimicrobials could be affected by the different ingredients incorporated into mouthrinse products. The purpose of the present study was to investigate the in vitro antifungal and fungicidal activities of antimicrobials alone. STUDY DESIGN: A broth macrodilution method was used to determine the minimum inhibitory concentration of 4 antimicrobial agents against Candida species. Minimum fungicidal concentration was also determined. RESULTS: All antimicrobials showed antifungal activity against all tested organisms, but cetylpyridinium chloride received significantly lower minimum inhibitory concentrations (P < .005). Cetylpyridinium chloride also showed a greater fungicidal activity than chlorhexidine digluconate and hexetidine (P< .005), whereas sanguinarine chloride appeared to be less fungicidal against most of the isolates tested. CONCLUSIONS: These findings suggest that cetylpyridinium chloride may be used as a topical antifungal agent. Clinical trials are now required to assess its value in the management of oral candidosis.

[Antiseptic efficacy and acceptance of Octenisept computed with common antiseptic mouthwashes]. / Antiseptische Wirksamkeit und Akzeptanz von Octenisept im Vergleich zu ausgewählten herkömmlichen Mundhöhlenantiseptika.

The efficacy of common antiseptic mouth rinses was evaluated in a study with healthy volunteers. Octenisept and Cetylpyridiniumchlorid had a significant stronger impact on the microbial burden of the oral cavity than Chlorhexidin (Corsodyl) immediately and 10 min after the application. Dobendan (contains Cetylpyridiniumchlorid) showed a better antimicrobial effect than Corsodyl immediately after application, after 10 min an increasing loss of efficacy was noted. The antiseptic efficacy of hexetidin-based Gurfix was very similar to the efficacy observed with Corsodyl from the start of the antiseptic treatment until 1 h later. Acriflavine-solution (0.2% m/v) was significantly less efficient after 10, 30 and 60 min compared to Corsodyl. The antimicrobial impact of Fluomint-Lysoform was very similar to the effect of rinsing with dest. sterile water. A subjective assessment of taste and smell of the mouth rinse solutions concludes the evaluation. Further methodological aspects for a common test guideline for antiseptic mouth rinse solutions are discussed.

In vitro antifungal properties of mouthrinses containing antimicrobial agents.

The purpose of this study was to investigate the in vitro antifungal properties of seven commercial mouthrinses containing antimicrobial agents. These included cetylpyridinium chloride (CPC), chlorhexidine digluconate (CHX), hexetidine (HEX), sanguinarine (SNG), and triclosan (TRN). The minimum fungicidal concentration (MFC) against six species of yeasts was determined by a broth macrodilution method. The kill-time of mouthrinses at half the concentration of the commercial formulations was also determined. MFCs were achieved with each mouthrinse, except the SNG-containing mouthrinse, against all the organisms being tested. However, the CPC-containing mouthrinse appeared more active than the other products (P < 0.001). There were no significant differences in MFC values among CHX mouthrinse products, once adjusted for initial concentration differences (P = 0.1). Kill-times of mouthrinses containing either CHX or CPC were less than or equal to 180 seconds with all the species of yeasts, and no significant differences were found among these products (P = 0.18). On the other hand, mouthrinses containing either TRN or HEX did not show a lethal effect on Candida albicans, Candida parapsilosis, or Candida guilliermondii. No kill-times were achieved with the SNG-containing mouthrinse. These results suggest that mouthrinses containing antimicrobial agents might represent an appropriate alternative to conventional antifungal drugs in the management of oral candidiasis. However, the effectiveness of antimicrobial mouthrinses as antifungal agents needs to be evaluated in further clinical trials.

The effects of hexetidine (Oraldene) on the adherence of Candida albicans to human buccal epithelial cells in vitro and ex vivo and on in vitro morphogenesis.

PURPOSE: This study reports the effects of hexetidine (Oraldene) on two virulence attributes of Candida albicans, namely, in vitro and ex vivo adherence of yeast cells to buccal epithelial cells (BEC) and in vitro morphogenesis. METHODS: The effects of hexetidine treatment of either yeast cells (stationary and exponential phases) or BEC on Candidal adherence, in terms of viable and non-viable adherent yeast cells, were evaluated using an acridine orange stain in conjunction with fluorescence microscopy. Ex vivo anti-adherence effects were determined by rinsing BEC in vivo with hexetidine (0.1%), removal of BEC after defined periods and inclusion in the adherence assay. The effects of hexetidine on morphogenesis were evaluated using light microscopy. Yeast cell viability following exposure to a range of concentration of hexetidine (0.005-0.1% v/v) for defined periods was determined following serial dilution and enumeration on solid media. RESULTS: Treatment of stationary and exponential phase yeast cells or BEC with hexetidine (0.1%) for a range of times (10-300 s) or, alternatively, with a range of concentrations of hexetidine (0.005-0.1%) for a fixed time (30s) significantly decreased the resultant Candidal/ epithelial adhesion. No correlations were observed between reduced adherence and either time of treatment or hexetidine concentration. In vivo treatment of BEC with hexetidine (0.1%) for 30s resulted in prolonged and significant reductions in the ex vivo adherence of both viable and non-viable yeast cells for periods of up to (and including) four hours post-rinsing. Treatment of C. albicans blastospores with hexetidine (0.05, 0.1% v/v) for 10s and 30s totally inhibited Candida morphogenesis, whereas treatment with lower antiseptic concentrations significantly reduced the extent of Candida morphogenesis and the rate of hyphal development. The effects of hexetidine on yeast cell viability were both concentration and time-dependent. CONCLUSIONS: The reduced adherence of C. albicans to BEC and the modification or inhibition of morphogenesis following exposure to hexetidine suggests a clinical role for hexetidine in the prophylaxis of both superficial candidosis and the systemic complications resulting from invasion of sub-epithelial tissue.

An approach to efficacy screening of mouthrinses: studies on a group of French products (I). Staining and antimicrobial properties in vitro.

There is a large and increasing number of oral hygiene products available to the general public. As such, it is difficult to demonstrate efficacy for all in long-term home-use studies. The aim of this project was to determine whether screening studies could position the activity and efficacy of a number of antiseptic mouthrinses, available in France, by comparison with an established product. This study reports the experiments in vitro. The products under test were, 4 containing chlorhexidine (Eludril, Hibident, Parodex and Prexidine) with Hibident considered the positive control, one containing cetylpyridinium chloride (Alodont) and one containing hexetidine (Hextril). The 1st study determined the antibacterial profile of the chlorhexidine products against a panel of oral bacteria using an agar dilution method. The 2nd study recorded, by optical density, the propensity of all products to induce tea staining on clear acrylic specimens. The maximum inhibitory dilution (MID) of the chlorhexidine products against the test organisms, once adjusted for initial concentration differences, were essentially similar. One exception was Eludril which had an increased MID against Capnocytophaga sputigena, an organism normally less sensitive to chlorhexidine. Hibident and Prexidine produced expected levels of tea staining; that produced by Parodex was lower than the expected. Staining by Eludril was little different than water controls suggesting markedly reduced availability of chlorhexidine in this product.(ABSTRACT TRUNCATED AT 250 WORDS)

Dietary staining in vitro by mouthrinses as a comparative measure of antiseptic activity and predictor of staining in vivo.

Extrinsic staining of teeth is a side-effect of some antiseptic mouthrinses. However, few of the many rinse products available to the general public have been investigated for their propensity to cause staining. Dietary factors play an aetiological role in staining and have been used in vitro to study and compare the activity of rinses. The aim of this study was to assess rinse products for staining in vitro and, through the staining reaction, to compare the activity of products containing the same ingredients. Perspex blocks, with or without saliva pretreatment, were soaked in rinses for 2 min, washed and placed in a standard tea solution for 60 min and then the optical density (OD) read on a spectrophotometer. The cycle was repeated 10 times for saliva and 17 times for no saliva specimens or until the maximum OD was exceeded. A series of three separate experiments was performed by this method. The maximum OD was not exceeded by any product before seven passages and therefore data were compared at six passages. For most products OD increased with saliva pretreatment. Some cetylpyridinium chloride (CPC) rinses stained comparably to a chlorhexidine rinse. CPC rinses, most of which contained the same concentration of the antiseptic, varied considerably in their propensity to induce staining and one was little different to water controls. A 0.1% chlorhexidine rinse stained slightly more than a 0.2%. A phenolic/essential oil product produced some staining but zinc, triclosan and other essential oil rinses did not stain.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of subinhibitory concentrations of chemical agents on hydrophobicity and in vitro adherence of Streptococcus mutans and Streptococcus sanguis.

The aim of this investigation was to determine whether sublethal concentrations of chlorhexidine (Cx), hexetidine (Hx), cetylpyridinium chloride (Cc), sodium dodecyl sulfate (SDS), sanguinarine (Sg), sodium fluoride (NaF), and ammonium fluoride (NH4F) could affect hydrophobicity and adhesion of Streptococcus sanguis and Streptococcus mutans to saliva-coated hydroxyapatite (S-HA). Determination of the minimum inhibitory concentrations (MICs) showed that both species were susceptible to all agents tested. Growth in the presence of sub-MIC concentrations of Cx, SDS, Cc, NaF, or NH4F did not change significantly the hydrophobicity of S. sanguis cells when compared to the control which lacked any agent. However, growth in the presence of Hx or Sg resulted in a significant reduction in their hydrophobicity. Sub-MIC levels of SDS or Sg in the growth medium resulted in S. mutans cells with increased affinity for hexadecane compared with the control. The adherence of S. sanguis was changed significantly only by Hx or Sg, resulting in less cells adhering to S-HA. However, S. mutans cells previously incubated with NaF, NH4F, or Sg showed a higher adherence to S-HA than the control. The mechanisms of interference with adherence are at present not completely understood. Thus, antimicrobial agents at sub-MIC concentrations can interfere selectively with hydrophobicity and/or adhesion of oral streptococci.

[Stability and new formulations of hexetidine. 2. Formulations and in vitro bioavailability]. / Stabilität und neue Formulierungen von Hexetidin. 2. Mitteilung: Formulierungen und in vitro-Verfügbarkeit.

The solubilization of the water insoluble antimicrobial compound hexetidine is achieved by tensides and cyclodextrins. Using tensides the active ingredient is associated with the lipophilic part of the emulsifier. Tensides with a hydrophilic element in the lipophilic part of the molecule show better solubilizing properties. In contrast a higher stability is achieved with emulsifiers without hydrophilic elements in the lipophilic part of the molecule. The antimicrobial activity, determined by an in vitro test, is not influenced by the type of emulsifier. Cyclodextrins form inclusion complexes with hexetidine. Hydroxy propyl-beta-cyclodextrin is preferred due to its high water solubility. This inclusion complex shows compared with tenside solubilization a better taste and stability. The in vitro antimicrobial activity was comparable to tenside solubilized products.

Antiseptic activity of some antidental plaque chemicals on Streptococcus mutans biofilms.

S. mutans is one of the bacterial species involved in the initiation of cariogenic dental plaque. In situ, antiseptic activity evaluation of dental plaque chemicals is fairly difficult. A dynamic biofilm model of S. mutans on Tygon support is suitable for the quantification of antiseptic activity and the comparison of commercial chemical formulations, submitted or not to the french standards and Pharmacopea. Some of these chemicals, especially quaternary ammonia solutions, do not reduce living bacteria counts by 10E5 as AFNOR specifies. In the case of dental plaque, antiseptic activity is probably not the only relevant parameter: biofilm dissociation is rather more relevant because the dead or living bacteria adhere to the teeth, favouring the establishment of a new active biofilm. Two proceedings can be followed: support surface can be modified, or bacteria exopolymers synthesis can be inhibited to prevent cell adherence; the disruption of the biofilm glycocalyx can be achieved with enzymes or surfactants.

Correlation between the in vivo and in vitro antimicrobial properties of commercially available mouthwash preparations.

The effectiveness of six commercially available mouthwashes against common buccal organisms was studied. The minimum inhibitory concentrations (MIC) for two of the studied mouthwashes (Corsodyl and Oraldene) against buccal organisms were determined in Todd Hewitt medium with or without 5% serum. The concentration of the active substance in these two mouthwashes was in excess of the corresponding MIC. When the medium was supplemented with serum, lower MIC values were observed. Kill-time determinations, used at half the concentration of the normal preparation, revealed a rapid lethal effect for all tested mouthwashes. The slowest lethal effect was observed with Fluocaril mouthwash. When mouthwashes were tested in volunteers, an immediate significant fall in salivary bacterial counts was produced by all except Fluocaril. With the latter mouthwash the decrease was significant 2-30 minutes after rinsing. The bacterial levels returned to pre-rinse levels after 30 minutes for Listerine, after 90 minutes for both Oraldene and Mint and after 180 minutes for Corsodyl, Fluocaril and Sansilla mouthwashes. The results indicate that there is a good correlation between in vivo efficacy and in vitro determination of all mouthwash preparations.

[In vitro bacteriocidal activity of 5 oral antiseptics against the principal microorganisms implicated in oral disease]. / Activité bactéricide in vitro de 5 antiseptiques buccaux vis-à-vis des principaux germes impliqués dans les affections bucco-dentaires.

Bactericidal activities of five mouthrinses containing cetylpyridinium chloride, hexetidine or chlorhexidine have been tested in vitro, against the main microorganisms involved in buccal affections. Mouthrinses containing hexetidine or chlorhexidine are effective, but chlorhexidine activity appears more extensive and homogeneous.

Dentine hypersensitivity--effects of some proprietary mouthwashes on the dentine smear layer: a SEM study.

Exposed dentine may be sensitive to stimuli depending on the patency of the dentinal tubules. Most abrasive elements tend to produce a smear layer which obturates the dentinal tubules. This layer is easily removed by a number of erosive agents, particularly dietary acids. Mouthwashes are increasingly used and largely investigated for possible benefits to dental health. Rarely do studies assess potential detrimental activity. The aim of the present study was to examine effects of mouthwashes on dentine. Smear layers artificially produced on dentine sections were exposed, for time periods ranging from 3 to 300 min, to mouthrinses alone, or with subsequent brushing for 2 min using water or a fluoride toothpaste. Assessments were made by scanning electron microscopy. Of the nine rinses tested, six produced no consistently observable changes compared to water treated controls at any time period with or without brushing. A hexetidine rinse and a fluoride/antiseptic rinse both exposed tubules after exposure times of 2 h and longer, an effect enhanced by post-treatment brushing. A phenolic antiseptic rinse consistently removed the smear layer from specimens, an effect enhanced by brushing such that after 10 min exposure and 2 min brushing many tubules were open at the surface. The results indicate that the use of some mouthrinses could predispose to excessive tooth substance loss and dentine hypersensitivity, particularly if used prior to toothbrushing. There is a need to determine whether the intermittent use of some mouthrinses produces cumulative effects on dentine.

Synergistic antibacterial effects of copper and hexetidine against Streptococcus sobrinus and Streptococcus sanguis.

The aim of this study was to determine whether a combination of copper and hexetidine had a synergistic antibacterial effect against Streptococcus sobrinus OMZ 176 and S. sanguis 10556. Concentration ranges of the test agents alone and in combination were prepared by serial dilutions in microtiter trays with brain-heart infusion (BHI) broth as the bacterial growth medium. After incubation at 37 degrees C for 24 h, the minimum inhibitory concentration (MIC), corresponding to the lowest concentration showing no visible growth, was determined. Evaluated by the fractional inhibitory concentration index, a strong synergistic effect ranging from 0.39 to 0.40 was observed. A similar effect was also demonstrated by growth curves, which were constructed on the basis of growth in BHI broth with addition of MIC/4 of each agent alone or MIC/8 of each agent in combination. A probable explanation for these findings is that the surface-active hexetidine molecule alters the bacterial cell surfaces and thereby enables an increased amount of copper to be transported into the cell.

Decontamination of the oral cavity. Effect of six local anti-microbial preparations in comparison to water and parafilm as controls.

Six preparations (four liquid and two solid) were tested in a double-blind crossover design for their anti-bacterial effect on aerobic and anaerobic bacteria in saliva of 12 volunteers. The four liquid preparations contained either tyrothricin, hexetidine, hydrogen peroxide or ethanol and were tested against a rinse with water as control. The two solid preparations, in the form of lozenges, contained tyrothricin in doses of 4 or 10 mg and were tested against parafilm. A single rinse with either tyrothricin or hexetidine resulted in a significant reduction of aerobic and anaerobic bacteria in saliva which lasted for the whole 1 h observation period and was considerably more effective than hydrogen peroxide or ethanol. Similar results were seen with the solid preparations. The lozenge containing 10 mg tyrothricin was the most effective and was the only formulation capable of reducing oral aerobic bacterial counts by a factor of approximately 100.