Libyan Thymus capitatus essential oil: antioxidant, antimicrobial, cytotoxic and colon pathogen adhesion-inhibition properties.

AIMS: In the present work, the Libyan wild-growing Thymus capitatus essential oil (EO) was evaluated for its biological properties. METHODS AND RESULTS: Carvacrol (68.19%) and thymol (12.29%) were found to be the main compounds of the oil. Antioxidant properties, determined by 2,2-diphenylpicrylhydrazyl (DPPH) assay, revealed that IC50 values were 119, 403 and 105 µg ml(-1) for oil, thymol and carvacrol respectively. Microdilution method showed strong antibacterial and especially antifungal potential. Tetrazolium (MTT) colorimetric assay indicated moderate cytotoxicity towards human cell lines MRC-5, HCT 116 and HT-29 (IC50 = 30-150 µg ml(-1)). In adhesion-inhibition assay oil and main compounds reduced adhesion of Escherichia coli and Listeria monocytogenes on colon cells HT-29 (51 and 39% of inhibition against L. monocytogenes and E. coli respectively). CONCLUSIONS: Essential oil of Th. capitatus showed moderate cytotoxic activity, together with excellent antimicrobial effect, in particular against fungi, and significant potential to reduce pathogen colonization in colon. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report that EO of Th. capitatus could protect against colonization of pathogens to colon epithelium. Thymus capitatus from Libya should be recognized as possible new source of natural antioxidants, antimicrobials as well as possible source of new chemotherapeutics.

Micromorphology of trichomes of Thymus malyi (Lamiaceae).

Micromorphological, ultrastructural and morphometric investigations of the trichomes of Thymus malyi were carried out using a light microscope, a scanning electron microscope and a transmission electron microscope. Unbranched non-glandular trichomes, peltate and capitate glandular trichomes were described. The leaves of Thymus malyi bear non-glandular and glandular trichomes on both sides. Estimates of the volume density (i.e. their volume fraction per unit volume) of non-glandular trichomes were higher as compared to volume density of peltate and capitate glandular trichomes. Estimates of the number of these trichomes per area on sections showed that the capitate trichomes were the most abundant. Ultrastructural analyses of cell inner structure have shown numerous mitochondria, big nuclei and plastids with lipid globules and starch grains.

Antifungal activity of the essential oil of Thymus vulgaris L. and thymol on experimentally induced dermatomycoses.

The in vivo evaluation of the toxicological and antifungal activity of the essential oil of Thymus vulgaris L. and its main component thymol was made on 2-month-old male Wistar rats. We examined the therapeutic potency against experimentally induced dermatomycoses in rats, using the most frequent dermatomycetes, Trichophyton mentagrophytes, T. rubrum, and T. tonsurans. The therapeutic efficacy of a 1% solution of the essential oil of Thymus vulgaris and thymol as well as the commercial preparation bifonazole was evaluated. During the 37-day observation period the oil-treated animals were cured.

Evaluation of a combined thiourea and hydrogen peroxide regimen to bleach bloodstained teeth.

BACKGROUND: Current techniques for intra-coronal bleaching of stained root-filled teeth employ oxidative bleaching with hydrogen peroxide. However, concern over the potential for invasive cervical resorption following the use of hydrogen peroxide has been expressed by many researchers, and recommendations have been made to limit the use of this agent. A reductive-oxidative bleaching process using a thiourea and hydrogen peroxide regimen is proposed as an effective and safer bleaching combination. The efficacy of this novel bleaching regimen is evaluated in this study. METHODS: The study involved a quantitative and qualitative spectrophotometric assessment of the ability of two amine (bleaching) agents, aqueous thiourea and acidified thiourea, to alter the absorption spectra of haemoglobin and methaemoglobin compared to hydrogen peroxide. In addition, extracted premolar teeth discoloured by blood were subjected to different bleaching regimens using amine reducing agents and hydrogen peroxide. The change in the colour of the bloodstained dentine samples was measured at each stage of the bleaching process with a Photometer and Reflectance Densitometer. Comparisons of different treatments were made using a method of least significant difference and/or analysis of variance. RESULTS: Spectrophotometric studies showed that acidified thiourea solution greatly reduced the colour of the haemoglobin and methaemoglobin in the visible range (330-760 nm). Aqueous thiourea had no effect on the presence of haemoglobin and methaemoglobin. Reflection Densitometer and Photometer scores indicate that the greatest bleaching effect was achieved by the combined acidified thiourea and hydrogen peroxide regimen. CONCLUSION: The recognition that bleaching discoloured teeth is a chemical process, which can be achieved by both reducing and oxidizing agents, offers the possibility of developing new and safer clinical bleaching protocols. It is concluded that the bleaching regimen which employs the sequential use of 0.1 M acidified thiourea and 30% w/v hydrogen peroxide is as effective at bleaching bloodstained dentine as 30% w/v hydrogen peroxide alone. However, the addition of thiourea to the bleaching regimen has the potential benefit of reducing the level of damaging hydroxyl radicals and achieving a safer bleaching process.

The ability of thiourea to scavenge hydrogen peroxide and hydroxyl radicals during the intra-coronal bleaching of bloodstained root-filled teeth.

BACKGROUND: Hydrogen peroxide, an agent used in the intra-coronal bleaching of root-filled teeth for over a century, has been shown to diffuse from the pulp chamber to the outer root surface. Furthermore, it has been demonstrated that destructive hydroxyl radicals, the by-products of the bleaching process, have been detected on the external root surface. The control of such diffusion may be of importance in minimizing the risk of invasive cervical resorption (ICR) which has been linked to intra-coronal bleaching of discoloured root-filled teeth using hydrogen peroxide. The aims of the present in vitro study are to quantify the diffusion of hydrogen peroxide and hydroxyl radicals to the outer root surface following intra-coronal bleaching, and to evaluate the ability of thiourea incorporated into the bleaching protocol to scavenge residual hydrogen peroxide and hydroxyl radicals. METHODS: Thirty-five single rooted premolar teeth with intact cementum at the cemento-enamel junction were used in this project. Thirty teeth were stained with red blood cells and root-filled with gutta-percha and AH26. The five unstained teeth were root-filled and constituted a negative control (Group 1). The stained teeth were divided equally into the following experimental groups and subjected to various intra-coronal bleaching regimes: Group 2--'walking bleach' with 20 microl 30 per cent w/w hydrogen peroxide; Group 3--20 microl 30 per cent w/w hydrogen peroxide and thermocatalytically activated; Group 4--20 microl acidified thiourea; Group 5--20 microl acidified thiourea and 20 microl 30 per cent w/w hydrogen peroxide; Group 6--20 microl acidified thiourea and 20 microl one per cent sodium hypochlorite; Group 7--20 microl acidified thiourea, 20 microl one per cent sodium hypochlorite and 20 microl 30 per cent w/w hydrogen peroxide. The reaction products of the bleaching process were quantified at the outer root surface using high performance liquid chromatography and electrochemical detection (HPLC-ECD). RESULTS: Results showed that hydrogen peroxide used alone in Groups 2 and 3 was able to be detected at the outer root surface in 100 per cent of the samples, and that the presence of the hydroxyl radical generated in both groups was detected in equal amounts (P < 0.05). When thiourea was incorporated into the bleaching protocols in Groups 5-7, it was shown to scavenge both hydrogen peroxide and hydroxyl radicals to a significant degree (P < 0.05). CONCLUSIONS: Acidulated thiourea is an effective scavenger of residual hydrogen peroxide and hydroxyl radicals generated during the intra-coronal bleaching of bloodstained root-filled teeth.

Microbiological evaluation of endodontic files after cleaning and steam sterilization procedures.

BACKGROUND: Infection control procedures are essential for modern dental practice and they are continually evolving to meet the dental profession's high standards. The present study evaluated the efficacy of two cleaning procedures to reduce bacterial numbers on endodontic files, and evaluated the effect of biological debris on the subsequent sterilization of files. METHODS: Stainless steel and nickel-titanium (NiTi) files were examined upon removal from the manufacturer's packaging, after instrumentation in root canals of human teeth inoculated with a broth containing two anaerobic species and one facultative anaerobic species of bacteria, and after instrumentation and cleaning with either an ultrasonic bath or a thermal disinfector. For each file, the bacterial numbers were quantified using routine microbiological techniques in an anaerobic chamber. RESULTS: No bacteria were detected from files direct from their packets. The size, taper and type of file did not affect the ability of either of the cleaning procedures to reduce bacterial numbers. However, an absence of bacteria was more likely when files were cleaned in the thermal disinfector. No bacteria were detected from files that were-subjected to steam sterilization irrespective of the type of prior cleaning procedure. CONCLUSIONS: Steam sterilization eliminated all bacteria from the endodontic files irrespective of the presence of biological debris. The majority of bacteria were eliminated from endodontic files after either ultrasonic cleaning or using a thermal disinfector.

A SEM evaluation of debris removal from endodontic files after cleaning and steam sterilization procedures.

BACKGROUND: In recent times, it has been proposed to classify endodontic files as single-use items due to a perceived inability to adequately clean the instruments. The purpose of the present study was to quantify the surface debris on files removed from the manufacturer's packaging, and after cleaning using an ultrasonic bath or a thermal disinfector. METHODS: Stainless steel and rotary nickel-titanium files were examined after removal from the manufacturer's packaging, after instrumentation in broth-contaminated human teeth, and after various cleaning procedures. The cleaning procedures consisted of either a thermal disinfector cycle, ultrasonication with the files placed in a perforated container or ultrasonication with the files loosely placed in a beaker. The presence of manufacturing debris and biological debris was evaluated using scanning electron microscopy and quantified using image analysis software. RESULTS: The effectiveness of cleaning was not affected by variation in the size or taper of the files when an effective cleaning procedure was used. Cleaning the files in a thermal disinfector or by ultrasonication within a container did not consistently achieve complete removal of biological debris. Placing the files loosely in the ultrasonic bath achieved the most effective cleaning, an average of 98.33 per cent of the file surface area was freed of any biological debris. CONCLUSIONS: A conventional cleaning method is capable of effectively removing biological debris from endodontic files. The efficacy of ultrasonic cleaning was impaired when the files were placed within a perforated container.

The chemical composition of some Lauraceae essential oils and their antifungal activities.

The antifungal activity of Aniba rosaeodora, Laurus nobilis, Sassafras albidum and Cinnamomum zeylanicum essential oils were investigated against 17 micromycetes. Among the tested fungal species were food poisoning, spoilage fungi, plant and animal pathogens. In order to determine fungistatic and fungicidal concentrations (MIC and MFC) macrodilution and microdilution tests were used. Linalool was the main component in the essential oil of A. rosaeodora, while 1.8-cineole was dominant in L. nobilis. In sassafras essential oil safrole was the major component and in the oil of C. zeylanicum the main component was trans-cinnamaldehyde. The essential oil of cinnamon showed the strongest antifungal activity.

Activity of essential oils of three Micromeria species (Lamiaceae) against micromycetes and bacteria.

The chemical composition of essential oils from three Micromeria species: M. dalmatica Benth., M. albanica (Griceb. ex K. Mal) Silic and M. thymifolia (Scop.) Fritsch were investigated by GC and GC-MS and their antibacterial and antifungal activities against seven fungal and six bacterial species were evaluated. Biological assays showed strong fungitoxicity of oils from all three Micromeria spp., particularly M. albanica, against all fungi tested. Essential oils of these species also exerted antibacterial effect against Streptococcus aureus, Escherichia coli, Bacillus subtilis and Micrococcus luteus at low concentrations. Higher concentrations of essential oil of M. albanica and M. dalmatica were active against Gram-negative Pseudomonas aeruginosa, which could be due to the high content of piperitenone oxide.

Acacetin glycosides as taxonomic markers in Calamintha and Micromeria.

A new acetylated flavone glycoside, acacetin 7-O-[6""-O-acetylglucosyl(1""-->2")]rhamnosyl(1"'-->6")glucoside, has been isolated from the leaves of Calamintha glandulosa together with the known compound acacetin 7-O-rhamnosyl(1"'-->6")glucoside. The occurrence of these flavonoids in the closely related genera Satureja, Micromeria, Acinos and Clinopodium indicates that their distribution may be of taxonomic significance.

Antimicrobial activity of essential oils and ethanol extract of Phlomis fruticosa L. (Lamiaceae).

The essential oils and an ethanol extract of Phlomis fruticosa L. were evaluated for antibacterial and antifungal activities. Seven bacterial and seven fungal species were used. Among them were human, animal and plant pathogens, food poisoning bacteria and fungi which are known as potential mycotoxin producers. The essential oils showed antibacterial activity against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Klebsiella pneumoniae and Micrococcus luteus. The essential oils extracted from the plants collected from two different localities showed similar antibacterial activities. The antifungal activity of the essential oils was positive against Aspergillus niger, A. ochraceus, Cladosporium cladosporioides, Fusarium tricinctum and Phomopsis helianthi. The ethanol extract showed antibacterial activity against Staphylococcus aureus and Bacillus subtilis and antifungal activity against Aspergillus niger, A. ochraceus, Cladosporium cladosporioides, Fusarium tricinctum and Phomopsis helianthi.

The avulsed tooth--the best implant.

Successful management of the avulsed tooth begins at the accident site. Correct first-aid care followed by proper management in the dental surgery will significantly improve the prognosis for the avulsed tooth. The major aim following avulsion is the prevention of further damage to the periodontal ligament and, if possible, the maintenance of pulp vascularity. Studies have indicated that variables such as the extra-oral period, the storage medium and the stage of root development of the avulsed tooth significantly influence the final outcome. Endodontic therapy and chemical conditioning of the root surface after an extended extra-oral period has the potential to delay the resorptive processes of a replanted tooth.

A quantitative comparison of traditional and non-peroxide bleaching agents.

Single-rooted premolar teeth, stained with blood utilizing the technique devised by Freccia & Peters (1981), were subjected to traditional and non-peroxide bleaching agents. Colour changes were recorded over a period of 7 days using a Speedmaster R75-CP Reflection Densitometer. The most efficient removal of staining occurred after the application of 30% hydrogen peroxide, with sodium perborate being 75% as effective. All bleaching agents realized their optimum efficacy within the first 3 days. A combination of three enzymes (amylase, lipase and trypsin) with disodium edetate was not as effective as the routine bleaching agents; however, the combination did have a modifying effect on the blood stains. It is suggested that other non-peroxide agents should be investigated to determine their efficacy in removing staining from experimentally induced blood-stained teeth.

Tooth discoloration by blood: an in vitro histochemical study.

An in vitro model, using a modification of a technique devised by Freccia & Peters, was developed to investigate tooth staining following pulpal haemorrhage. Samples of whole blood, erythrocytes, plasma and platelet concentrate and saline were individually placed in the pulp chambers of groups of five teeth and centrifuged twice daily for 25 min over a period of 3 consecutive days. This confirmed that the blood pigment responsible for the staining was found only in those samples containing erythrocytes. Teeth stained with packed red cells were then prepared for histological examination and subjected to four histochemical tests: (1) benzidine, (2) zinc leuco, (3) Perl's and (4) Turnbull Blue to analyse some of the biochemical changes following haemorrhage into the pulp chamber. These tests showed that, following haemolysis of erythrocytes within dentine, haemoglobin was found either intact or as one of the haematin molecules with no further breakdown of the haem structure and no evidence of any free ferric ions or haemosiderin.

Incidence of invasive cervical resorption in bleached root-filled teeth.

Invasive cervical resorption, a form of external root resorption, has been reported to be associated with intra-coronal bleaching of root-filled teeth and this has raised concerns about carrying out such bleaching procedures. The purpose of the present study was to examine the incidence of invasive cervical resorption in root-filled teeth which had been bleached using a standardized technique. Three observers examined records and radiographs from a total of 158 patients, whose bleaching treatment had been carried out in a specialist endodontic practice. The sample comprised 204 teeth with a review period of between 1 and 19 years. One-hundred-and-fifty-one teeth (77.94 per cent) had an associated history of traumatic injury. All teeth had been treated with a combination of thermocatalytic and 'walking bleach' procedures using 30 per cent hydrogen peroxide. In 54.41 per cent of teeth, gutta-percha and AH26 root-fillings were kept at the height of the cemento-enamel junction while 18.63 per cent were below and 26.96 per cent were above the CEJ. Sealing cement was not placed over the gutta-percha and AH26 root-fillings in any of the teeth in the study. It was found that a total of four teeth from the sample group (1.96 per cent) had developed invasive cervical resorption during the review period. All of these teeth had a history of traumatic injury and the level of gutta-percha was at the CEJ.(ABSTRACT TRUNCATED AT 250 WORDS)