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Item Curcumin—A Natural Medicament for Root Canal Disinfection: Effects of Irrigation, Drug Release, and Photoactivation(Elsevier, 2019-11) Sotomil, Julian M.; Münchow, Eliseu A.; Pankajakshan, Divya; Spolnik, Kenneth J.; Ferreira, Jessica A.; Gregory, Richard L.; Bottino, Marco C.; Prosthodontics, School of DentistryIntroduction Curcumin incorporation into polymeric fibers was tested for its antimicrobial properties and potential use in root canal disinfection. Methods Curcumin-modified fibers were processed via electrospinning and tested against a 7-day old established Actinomyces naeslundii (An) biofilm. The medicaments tested were as follows: curcumin-modified fibers at 2.5 and 5.0 mg/mL, curcumin-based irrigant at 2.5 and 5.0 mg/mL, saline solution (negative control), and the following positive controls: 2% chlorhexidine, 1% NaOCl, and triple antibiotic paste (TAP, 1 mg/mL). All medicaments, except for the positive controls, were allocated according to the light exposure protocol: photoactivation with an LED every 30 s for 4 min or without photoactivation. After treatment, the medicaments were removed and 1 mL of saline solution was added; the biofilm was scraped from the well and used to prepare a 1:2000 dilution. Spiral plating was done using anaerobic blood agar plates. After 24 h, colony-forming units (CFU/mL, n=11/group) were counted to determine the antimicrobial effects. Results Data exhibited significant antimicrobial effects on positive control groups, followed by the curcumin irrigants, and lastly, the photoactivated curcumin-modified fibers. There was a significant reduction of viable bacteria in curcumin-based irrigants, which was greater than the TAP-treated group. Curcumin-free fibers, saline, and the non-photoactivated curcumin-modified fibers did not display antimicrobial activity. Conclusions Curcumin seems to be a potential alternative to TAP when controlling infection, but it requires a minimal concentration (2.5 mg/mL) to be effective. Photoactivation of curcumin-based medicaments seems to be essential to obtain greater antibiofilm activity.Item Effectiveness of ozonated water irrigation against an established Enterococcus faecalis biofilm in root canal treated teeth in vitro(2020) Broady, Adam B.; Spolnik, Kenneth J.; Duarte, Simone; Gossweiler, Ana; Bringas, Josef S.; Ehrlich, YgalIntroduction: One of the main objectives of endodontic therapy is to reduce microbes and remove inflamed pulpal tissue within the root canal system (RCS). This is accomplished through chemomechanical debridement of the RCS using hand and rotary instrumentation along with an antimicrobial irrigant. Today, the most commonly used irrigant is sodium hypochlorite (NaOCl), often at concentrations toxic to human cells. The use of ozone as an endodontic irrigant is a novel technique that has been proven to be antimicrobial against several microorganisms. However, independent research is lacking on ozone’s efficacy against an established endodontic biofilm. If ozone’s efficacy against biofilms is confirmed, the use of toxic and potentially dangerous sodium hypochlorite could be replaced in some clinical situations (i.e., regeneration, immature teeth, resorption) with a safer and effective alternative. Objective: The aim of the current study was to evaluate the anti-biofilm activity of different concentrations of ozonated water compared to various concentrations of NaOCl against an established endodontic biofilm of Enterococcus faecalis in root canal treated teeth in vitro. Materials and Methods: The crowns of similarly sized, maxillary anterior teeth were removed, and the roots cut to a standard length (12 mm). All root canals were instrumented to a standard size. Specimens were sterilized and then inoculated with E. faecalis, which were allowed to grow for two weeks to form an established biofilm. There were six treatment groups: 1) 6% NaOCl; 2) 1.5% NaOCl; 3) 16µg/mL ozonated water; 4) 25µg/mL ozonated water; 5) 50µg/mL ozonated water, and 6) saline. Following treatment, samples were collected, plated, and incubated for two days. The number of CFU/mL were determined, and samples visualized using confocal imaging. The effect of treatment group on bacterial counts was made using one-way ANOVA followed by pair-wise comparisons. Null Hypothesis: Endodontically treated teeth irrigated with ozonated water will not demonstrate a statistically significant decrease in the E. faecalis biofilm compared to those treated with sodium hypochlorite Results: CFUs were converted to log10 and compared using Fisher’s Exact tests or one-way ANOVA followed by pair-wise tests. In all observations utilizing NaOCl irrigation, no colonies formed following treatment. The two NaOCl groups, with 0 CFU/mL, were significantly different than the other four groups (p=0.009). Saline showed a trend towards higher CFU/mL than 50 µg/ml O3 (p=0.068). None of the other comparisons approached statistical significance (p=0.453 25 µg/ml O3, p=0.606 16 µg/ml O3, p=0.999 25 µg/ml O3 vs 50 µg/ml O3, p=0.990 16 µg/ml O3 vs 50 µg/ml O3, p=1.000 16 µg/ml O3 vs 25 µg/ml O3). Confocal imaging helped illustrate effects of irrigation and confirm CFU findings. Conclusion: The results of this study failed to reject the null hypothesis. There was a statistically significant difference in the E. faecalis biofilm remaining in the groups treated with ozonated water compared to those treated with NaOCl. However, there was a trend towards higher CFU/mL in the saline group compared to the 50µg/mL ozonated water group. According to this finding, future studies should evaluate the effects of higher concentrations of ozonated water against an established E. faecalis biofilm. In addition, other follow-up studies might include ozonated water’s effect on human cells, such as the stem cells of the apical papilla that are so critical to the success of regenerative endodontic procedures. Due to university and laboratory closures caused by the COVID-19 pandemic, this project was stopped short and an insufficient sample size did not allow for proper statistical power. Additional occasions should be run upon the university’s re-opening to allow for proper statistical power.Item An in-vitro comparison of working length determination between a digital system and conventional film when source-film/sensor distance and exposure time are modified(2009) Ley, Paul J. (Joseph), 1980-; Vail, Mychel Macapagal, 1969-; Parks, Edwin T. (Edwin Thomas), 1955-; Spolnik, Kenneth Jacob, 1950-; Legan, Joseph J.; Zunt, Susan L., 1951-Accurate determination of working length during endodontic therapy is a crucial step in achieving a predictable outcome. This is determined by the use of electronic apex locators, tactile perception, and knowledge of average tooth lengths and/or dental radiography whether digital or conventional is utilized. It is the aim of this study to determine if there is a difference between Schick digital radiography and Kodak Insight conventional film in accurately determining working lengths when modifying exposure time and source-film/sensor distance. Twelve teeth with size 15 K-flex files at varying known lengths from the anatomical apex were mounted in a resin-plaster mix to simulate bone density. Each tooth was radiographed while varying the source-film/sensor distance and exposure 122 time. Four dental professionals examined the images and films independently. Ten images and 10 films were selected at random and re-examined to determine each examiner?s repeatability. The error in working length was calculated as the observed value minus the known working length for each tooth type. A mixed-effects, full-factorial analysis of variance (ANOVA) model was used to model the error in working length. Included in the ANOVA model were fixed effects for type of image, distance, exposure time, and all two-way and three-way interactions. The repeatability of each examiner for each film type was assessed by estimating the intra-class correlation coefficient (ICC). The repeatability of each examiner on digital film was good with ICCs ranging from 0.67 to 1.0. Repeatability on the conventional film was poor with ICCs varying from -0.29 to 0.55.We found there was an overall difference between the conventional and digital films (p < 0.001). After adjusting for the effects of distance and exposure time, the error in the working length from the digital image was 0.1 mm shorter (95% CI: 0.06, 0.14) than the error in the working length from the film image. There was no difference among distances (p = 0.999) nor exposure time (p = 0.158) for film or images. Based on the results of our study we conclude that although there is a statistically significant difference, there is no clinically significant difference between digital radiography and conventional film when exposure time and source-film/sensor distance are adjusted.Item An in-vitro SEM study comparing the debridement efficacy of the EndoVac® system versus the Canal CleanMax® following hand-rotary istrumentation(2010) Ricketts, Benjamin P. (Benjamin Paul), 1980-; Vail, Mychel; Legan, Joseph J.; Moore, B. Keith; Spolnik, Kenneth Jacob, 1950-; Zunt, Susan L., 1951-This in-vitro, prospective, randomized study microscopically compared the debridement efficacy of negative pressure irrigation with the EndoVac (Discus Dental, Culver City, CA) versus the Canal CleanMax (Maximum Dental, Inc., Secaucus, NJ). Sixty extracted human canines were instrumented using a combination of hand-instrumentation with Lexicon K-type files and rotary instrumentation with ProTaper files. All canals were irrigated with 6.0-percent sodium hypochlorite and 17- percent ethylenediaminetetraacetic acid (EDTA). However, the irrigation/aspiration techniques differed among three groups of 20 randomly selected teeth. Group one (control) was irrigated with only a 12-ml Monoject syringe via 30-gauge side-vented, closed-end needle. Group two was irrigated with the EndoVac system. Group three was irrigated similar to group one, but with the adjunct of the Canal CleanMax system. All teeth were sectioned longitudinally, and the more intact sections were divided into coronal, middle, and apical thirds. Each portion of the canal was photographed with a scanning electron microscope (SEM). The photographs were scored by two independent examiners according to relative amount of debris and/or smear layer present, as well as relative number of patent dentinal tubules. These scores were statistically analyzed using a Krustal-Wallis test and Wilcoxon Rank Sum tests to determine differences between groups. The coronal aspect of root canal walls irrigated with the EndoVac system exhibited significantly less debris and/or smear layer present when compared to the coronal aspect of root canals irrigated with only a standard 12-ml Monoject syringe equipped with 30-gauge ProRinse side-vented, closed-end needle (control). There were no other significant differences in scores between any groups at any location. For all locations combined, the EndoVac system produced significantly cleaner root canal walls as compared to the control. No significant differences were seen between the Canal CleanMax and Control or Canal CleanMax and EndoVac. This study suggested negative pressure irrigation delivery with the EndoVac system during and after hand-rotary instrumentation is more effective in removal of debris and smear layer from the coronal third and combined thirds of root canal walls compared to irrigation with a standard 12-ml Monoject syringe equipped with 30-gauge ProRinse side-vented, closed-end needle.Item A novel patient-specific three-dimensional drug delivery construct for regenerative endodontics(Wiley, 2018-10-03) Bottino, Marco C.; Albuquerque, Maria T. P.; Azabi, Asma; Münchow, Eliseu A.; Spolnik, Kenneth J.; Nör, Jacques E.; Edwards, Paul C.; Oral Pathology, Medicine and Radiology, School of DentistryEvoked bleeding (EB) clinical procedure, comprising a disinfection step followed by periapical tissue laceration to induce the ingrowth of undifferentiated stem cells from the periodontal ligament and alveolar bone, is currently the only regenerative-based therapeutic approach to treating pulp tissue necrosis in undeveloped (immature) permanent teeth approved in the United States. Yet, the disinfection step using antibiotic-based pastes leads to cytotoxic, warranting a biocompatible strategy to promote root canal disinfection with no or minimal side-effects to maximize the regenerative outcomes. The purpose of this investigation was to develop a tubular three-dimensional (3D) triple antibiotic-eluting construct for intracanal drug delivery. Morphological (scanning electron microscopy), chemical (Fourier transform infrared spectroscopy), and mechanical (tensile testing) characteristics of the polydioxanone-based triple antibiotic-eluting fibers were assessed. The antimicrobial properties of the tubular 3D constructs were determined in vitro and in vivo using an infected (Actinomyces naeslundii) dentin tooth slice model and a canine method of periapical disease, respectively. The in vitro data indicated significant antimicrobial activity and the ability to eliminate bacterial biofilm inside dentinal tubules. In vivo histological findings demonstrated that, using the EB procedure, the tubular 3D triple antibiotic-eluting construct allowed the formation of an appropriate environment that led to apex closure and the ingrowth of a thin layer of osteodentin-like tissue into the root canal. Taken together, these findings indicate that our novel drug delivery construct is a promising biocompatible disinfection strategy for immature permanent teeth with necrotic pulps.