A critical analysis of research methods and experimental models to study tooth discolouration from endodontic materials.

As a range of materials used in endodontics may cause tooth discolouration, exploration of factors responsible for the darkening of the tooth crown or root is warranted. This narrative review paper discusses the range of technical factors that apply in laboratory studies that assess endodontic discolouration. As an example of how these factors operate, particular examples relating to discolouration caused by endodontic medicaments containing tetracycline antibiotics are used. Following the PRILE 2021 guideline approach, a summary of key variables to be addressed in the methodology for laboratory studies is presented, to inform future work.

Development and Reversal of Discolouration in Roots Filled with Ledermix or Doxymix Pastes Stored in the Dark for Three Months then Daylight for Twenty Four Months.

OBJECTIVE: To explore the long-term effects on discolouration by demeclocycline HCl (Ledermix, LED) or doxycycline hyclate (Doxymix, DOX) pastes placed in extracted human teeth over a 27-month period under different storage conditions. METHODS: The canals in 38 teeth were prepared carefully, to minimize exposure to contamination from irrigants, then either LED (Lederle Pharmaceuticals, Germany) or DOX (Ozdent, Australia) were placed. Samples were stored in the dark for 3 months followed by daylight for 24 months. The storage conditions varied as follows: Group 1: Open access, dry storage (OD); Group 2: Closed access, dry storage (CD); Group 3: Open access, wet storage (OW); Group 4: Closed access, wet storage (n=4 for each material). Additional teeth were used as controls: Polyethylene glycol only in a closed canal; and saline only irrigation with LED paste in a closed canal. Standardised digital photographs were taken over 27 months and evaluated for changes in luminosity. RESULTS: Darkening of tooth structure occurred in all LED groups and in the two DOX groups that were stored wet, during exposure to light, with a faster rate with LED. The most rapid staining occurred with LED in moist conditions with an open access cavity. The least staining occurred with DOX in samples stored dry. With prolonged exposure to light, a reversal in staining occurred with DOX at 3 months and LED at 9 months. CONCLUSION: Staining of tooth structure is influenced by the choice of medicament, and by exposure to moisture and air. Light has a bimodal effect, first driving staining, but later reversing it. This can be explained by different wavelengths of light causing photodegradation and photo-oxidation of tetracyclines and their complexes with tooth mineral.

Comparison of Commercial Calcium Hydroxide Pastes for Prolonged Antibacterial Effect using a Colourimetric Assessment.

The anti-microbial activity of calcium hydroxide pastes used in endodontics is dependent on establishing high levels of hydroxyl ions in dentine. This study investigated hydroxyl ion diffusion from different commercial calcium hydroxide pastes using a novel colourimetric method. In this method, human tooth roots were stained with anthocyanin dye, which changed their colour according to the local pH conditions. Prepared root canals were filled with pastes formulated with the vehicle of water (Pulpdent™, Calasept Plus™), polyethylene glycol (PEG) (Calmix™) or a mixture of water, PEG and ibuprofen (Odontocide™). The changes in dye colour at fixed distances from the canal wall were monitored using standardised digital photography over a period of 3 weeks. A repeated measures analysis tracked changes in each root from baseline. Release of hydroxyl ions varied between the different commercial compositions containing water or PEG as solvents. The colour changes in the dentine, due to released hydroxyl ions, were greatest and more prolonged for completely non-aqueous compositions, when using PEG 400 as the vehicle. When water was present in the product, the duration of the pH changes was shorter. This was attributed to the presence of hydroxyl ions in the water (the common-ion effect) and a more vigorous buffering of hydroxyl ions by dentine proteins.

Mechanisms of Staining with Demeclocycline and Doxycycline Root Canal Medicaments.

Tetracyclines are a unique class of antibiotics which also have additional effects including anti-inflammatory, anti-resorptive and substantive within the root canal. There has been a long-held view that tetracycline medicaments discolour teeth and should be avoided. The evidence base around this topic was explored, including a review of the methodology used in laboratory studies. A search of PubMed, Medline and Scopus databases was conducted to identify studies of demeclocycline and doxycycline medicaments used in root canal therapy. An analysis of the methodology used in these studies was performed to determine if these replicate current clinical practice. The related literature on mechanisms of tetracycline stability and the effects of light, oxidation, moisture and chemical interactions was examined. Studies investigating the effects of Ledermix paste on segments of bovine dentine and avulsed or reimplanted teeth as well as combinations with other antibiotics were excluded from this review. Even though demeclocycline medicament pastes were introduced in 1962, the first laboratory studies of discolouration were not done until 2000. All later studies followed a similar approach, which included exposure to sodium hypochlorite for up to 30 minutes and storage in moist conditions with 100% humidity. Staining during dark storage and enhanced staining on exposure to light were reported, indicating multiple pathways of degradation of demeclocycline and its reaction products. Light, moisture and oxidation are the key factors which drive discolouration from demeclocycline. Clinical issues from tooth staining can be prevented by removal of medicament pastes from the access cavity, and placement of a sound interim restoration. Use of a doxycycline paste obviates concerns of staining. Laboratory assessments of the potential for staining should replicate in vivo conditions.

Aspects of Solvent Chemistry for Calcium Hydroxide Medicaments.

Calcium hydroxide pastes have been used in endodontics since 1947. Most current calcium hydroxide endodontic pastes use water as the vehicle, which limits the dissolution of calcium hydroxide that can be achieved and, thereby, the maximum pH that can be achieved within the root canal system. Using polyethylene glycol as a solvent, rather than water, can achieve an increase in hydroxyl ions release compared to water or saline. By adopting non-aqueous solvents such as the polyethylene glycols (PEG), greater dissolution and faster hydroxyl ion release can be achieved, leading to enhanced antimicrobial actions, and other improvements in performance and biocompatibility.

Sealing Ability of Alkaline Endodontic Cements versus Resin Cements.

Penetration of oral bacteria through root fillings leads to their long term failure. Dimensionally stable alkaline cements have been developed. A saliva challenge model was used to compare resistance to bacterial penetration of these alkaline cements to conventional root fillings that combine gutta percha (GP) with epoxy resin sealers. A sample of 140 human roots with single straight canals prepared to standard length and canal size were obturated with mineral trioxide aggregate (MTA) (Nex MTA or MTAmix), with an alkaline calcium hydroxide hard setting cement (Supercal), or with GP and a resin cement (either AH-Plus or Zirmix). Negative control roots were sealed with wax, while positive controls were left open. The test assemblies were gamma sterilised, then the coronal root face was exposed daily to fresh stimulated human saliva diluted in broth. Bacterial penetration was determined by assessing growth in sterile brain-heart infusion (BHI) medium in contact with the root apex. Using Kaplan-Meier survival analysis, in order of performance from highest to lowest: Negative control, Supercal, Nex MTA, Zirmix, MTAmix, GP + AH-Plus, and the positive control. In addition, statistically significant differences were noted between Supercal and AH-Plus, and between the two MTA cements. It can be concluded that alkaline cements, particularly Supercal, can show considerable resistance to bacterial penetration from constant saliva challenge, and provide superior sealing ability in comparison to resin cements. While this property is due mostly to dimensional stability, the release of hydroxide ions could be a contributing factor to impaired bacterial survival, and this aspect should be explored further.

Tooth discolouration: staining effects of various sealers and medicaments.

The aim of this investigation was to quantify the discolouration of tooth roots caused by various medicaments and sealers. The roots of extracted teeth were chemo-mechanically prepared and the smear layer removed. The roots were filled with either Multi-Cal (Pulpdent Corporation, Watertown, MA, USA), Ledermix (Lederle Laboratories, Wolfratshausen, Germany), 2% chlorhexadine acetate in polyethylene glycol (PEG), 5% clindamycin in PEG, PEG alone (as a vehicle control), sealers AH26 (De Trey Dentsply, Zurich, Switzerland), AH Plus (De Trey Dentsply), an experimental epoxy resin with zirconium dioxide (Millenium Chemicals, Rockingham, Western Australia) or Araldite (Selleys Pty Ltd, Sydney, Australia). The roots were maintained at 37°C in dark humid conditions and standardised images of the coronal surfaces recorded over 12 months. Ledermix caused the most darkening of the introduced medicaments, while calcium hydroxide and the materials in PEG did not cause darkening. AH26 caused more darkening than did the other sealers. It was concluded that the use of alternatives to Ledermix and AH26 should be considered when aesthetics are important.