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Browsing by Author "Zunt, Susan L., 1951-"
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Item Antibacterial efficacy of 0.12-percent and 2.0-percent chlorhexidine gluconate at 37˚C and 46˚C against enterococcus faecalis(2010) Thiessen, Craig B.D., 1978-; Vail, Mychel Macapagal, 1969-; Spolnik, Kenneth Jacob, 1950-; Zunt, Susan L., 1951-; Gregory, Richard L.; Legan, Joseph J.The purpose of this study was to investigate the antibacterial efficacy of 0.12-percent and 2.0-percent chlorhexidine gluconate (CHX) on eliminating Enterococcus faecalis from dentinal tubules, and whether this antibacterial effect was enhanced by heat. To date there have been no published articles that describe the heating of 2.0-percent CHX and its antimicrobial efficacy and clinical relevance towards E. faecalis within dentinal tubules in root canal systems. Ninety-five human extracted, single rooted, maxillary, anterior teeth were used to prepare dentin disk specimens. After proper sterilization, a 2.5-mm ISO-sized diameter lumen was prepared, and then the canals were filled with brain-heart infusion (BHI) broth infected with E. faecalis. The BHI was removed and the specimens in equally divided groups were rinsed with sterile saline and filled with saline, or 0.12 percent CHX or 2.0 percent CHX at ambient temperature (24°C) or experimental temperature (46°C) and incubated at oral temperature (37°C) or the experimental temperature (46°C), respectively. The specimens were frozen to -70˚C and pulverized in liquid nitrogen. Serial dilutions were prepared of 1:100 and 1:1000 and spiral plated on BHI agar plates in duplicate. They were incubated, and the number of bacterial colonies was recorded 24 hours later for data analysis. A two-way analysis of variance (ANOVA), with factors for solution, solution temperature, and the solution-by-temperature interaction was used to determine antibacterial efficacy. Pair-wise comparisons between groups were examined for significance using the Fisher’s Protected Least Significant Differences Method. The E. faecalis CFU were log-transformed to satisfy the assumptions required for the ANOVA. The results of this investigation demonstrated no statistically significant difference with the addition of heat to either test irrigation solution regarding the elimination of E. faecalis from dentinal tubules within the root canal system. There was a statistically significant difference in the antibacterial efficacy of CHX against E. faecalis in comparison with the concentration tested. A higher concentration of 2.0-percent CHX demonstrated a significantly higher antibacterial efficacy against E. faecalis compared with 0.12-percent CHX, and likewise with the saline control. It can be concluded that the use of a higher concentration of 2.0-percent CHX is advantageous as a final irrigation solution after copious amounts of NaOCl and EDTA have been utilized for effective antimicrobial efficacy and substantivity.Item Biodegradability of resilon, a resin based root canal obturating material, by typical endodontic pathogens(2012) Rexford, Ashleigh M.; Spolnik, Kenneth Jacob, 1950-; Vail, Mychel Macapagal, 1969-; Hara, Anderson T.; Ehrlich, Ygal; Zunt, Susan L., 1951-; Gregory, Richard L.; Legan, Joseph J.Root canal therapy is a recommended treatment for apical periodontitis. Root canal failure can occur as a result of microbial leakage. Resilon, a resin based root canal obturating cone material introduced in 2004 attempts to minimize leakage by a unique bonding method of the resin sealer to both the core material and to the dentin of the canal walls. Resilon has no bactericidal or antimicrobial effect15. Furthermore, it has been shown that Resilon is susceptible to alkaline and enzymatic hydrolysis as well as bacterial degradation.73, 184-186 It has been suggested that Resilon may be susceptible to degradation by microorganisms found in the infected root canal space. This work focuses on the susceptibility of root canal obturating materials to be degraded by endodontic pathogens seen in root canal treated teeth with apical periodontitis. The aim of this study was to determine if Resilon could be degraded by selected pathogenic bacteria found in the infected root canal system, and if this degradation is more severe than with gutta-percha, a conventional obturating material. P. intermedia, E. faecalis and P. aeruginosa, known endodontic pathogens were inoculated on discs of obturating material (Resilon or gutta-percha) mounted on a platform and placed in wells containing TSB incubated at 37°C under aerobic conditions. The discs were polished, examined by SEM, profilometry, and elemental analysis prior to inoculation to establish a baseline, and were then re-examined by these methods one month after inoculation. The overall results were inconclusive; and using these methods it cannot be determined that the selected bacteria can degrade Resilon. An ideal future study would utilize SEM with gold coated samples as well as atomic force microscopy to evaluate for changes in topographical features of these obturating materials. A notable finding was that Resilon turns black when exposed to bacteria, and the significance of this finding should be addressed in future studies.Item Comparison of electrosurgical and formocresol pulpotomy procedures(1997) Fulkerson, Bradley Todd; Dean, Jeffrey A.; Sanders, Brian J.; Zunt, Susan L., 1951-; Legan, Joseph E.; Avery, David R.Formocresol is the most commonly used pharmacologic pulpotomy agent. Concerns over its safety have led investigators to search for new pulpotomy medicaments. This study compared the electrosurgical pulpotomy with the formocresol pulpotomy in teeth requiring pulp therapy after carious involvement. There were 25 pulpotomies performed in each group. The teeth were evaluated for clinical and radiographic success after at least six months. In the electrosurgical group, the clinical and radiographic success rates were 96 percent and 84 percent, respectively. The age range at the time of treatment was 26 to 97 months, with a mean treatment age of 63.6 months. The postoperative observation time range was six to 31 months, with the mean being 10.9 months. In the formocresol group, the clinical and radiographic success rates were 100 percent and 92 percent, respectively. The age range at the time of treatment was 32 to 126 months, with a mean treatment age of 68.2 months. The postoperative observation time ranged from five to 25 months, with the mean being 11.5 months. The electrosurgical and forrnocresol groups were compared for differences in the percentage of successes by using a Fisher's Exact test. There were no statistical differences between the two groups at the p < 0.05 level. Therefore, this study failed to demonstrate a statistically significant difference in the success rate between the electrosurgical and formocresol pulpotomy techniques and supports the use of the electrosurgical pulpotomy as a viable and safe alternative to formocresol.Item Effect of Antibiotic Pastes on Chemical Structure and Microhardness of Radicular Dentin(2014) Prather, Blake; Spolnik, Kenneth Jacob, 1950-; Legan, Joseph J.; Zunt, Susan L., 1951-; Ehrlich, Ygal; Platt, Jefrey A., 1958-Introduction: Regenerative endodontic therapy in immature teeth with necrotic pulps triggers continued root development, thereby improving the prognosis of these teeth. Disinfection of the canal is accomplished with an intracanal medicament, such as triple antibiotic paste (TAP) composed of metronidazole, ciprofloxacin, and minocycline. A modified triple antibiotic paste (MTAP) that replaces minocycline with clindamycin has recently been suggested to avoid the tooth discoloration and potential demineralization from minocycline. The effect these pastes have on radicular dentin is unknown. Objectives: The aim of this study was to investigate the effects of two intracanal medicaments used during endodontic regeneration, TAP and MTAP, at concentrations of 1 g/mL and 1 mg/mL, on the microhardness and chemical structure of radicular dentin. Materials and Methods: Roots from extracted, unrestored, non-carious human premolar teeth were sectioned. An antibiotic paste (MTAP or TAP) or sterile water (control) was applied to treatment groups and stored for four weeks in 80-percent humidity at 37 °C. The effect of each paste on the microhardness of radicular dentin was measured using a Vickers Microhardness Tester (n = 17) to take three pretreatment and post-treatment measurements at both 500 µm and 1000 µm from the pulp-dentin interface. The chemical structure was assessed from dentin specimens treated with the same medicaments or sterile water for four weeks. After treatment, three measurements were taken on each specimen using Attenuated Total Reflection Fourier Transform Infrared Spectroscopy to measure the phosphate/amide I ratios of dentin (n = 7). Results: The 1 g/mL of TAP or MTAP and the 1 mg/mL methylcellulose-based TAP caused significant reduction in microhardness of roots compared with untreated control roots at 500 µm and 1000 µm from the pulp-dentin interface. Furthermore, the methylcellulose-based 1 mg/mL TAP and MTAP caused significantly less reduction in microhardness compared with 1 g/mL TAP and MTAP. The 1 g/mL of TAP and DAP caused significantly lower phosphate/amide I ratios compared with other groups. Conclusion: The use of methylcellulose based 1 mg/mL of TAP and MTAP may minimize the reduction in microhardness of roots compared with the currently used 1 g/mL concentration of these antibiotics.Item Effect of HA-coating and HF etching on experemental zirconia implant evaluation using in vivo rabbit model(2010) Huang, Sung-En; Chu, Tien-Min Gabriel; John, Vanchit (Vanchit Kurien), 1965-; Kowolik, Michael J.; Zunt, Susan L., 1951-; Blanchard, Steven B.The objective of this study was to evaluate the in vivo performance of the hydroxyapatite (HA) coating and hydrofluoric acid (HF) etching zirconia (ZrO) implants and to compare the result with titanium (Ti) implants treated in a similar manner. A total of four different implant types were tested in this study. Threaded zirconia implants with HA coating (Test 1) and zirconia implants with HF-treated surfaces (Test 2) were used to compare to the same size of titanium implants treated in identical fashion (control 1 and control 2). All implants measured about 3.5 mm at the thread diameter and 7.0 mm in total length. Each rabbit received two zirconia and two titanium implants treated in the same manner (either HA-coated or HF-etched). The samples were implanted into the rabbit tibias and retrieved at 6 weeks. Upon retrieval, 24 specimens (6 samples for each group) were fixed and dehydrated. The samples were then embedded undecalcified in PMMA for histomorphometry to quantify the bone-to-implant contact (BIC). Another 24 samples were kept in 0.9% saline and were evaluated using removal torque (RT) analysis to assess the strength of the implant-to-bone interface. The histomorphometric examination demonstrated direct bone-to-implant contact for all four groups. HA particle separation from the implants surface was seen in a majority of the HA-coated samples. No signs of inflammation or foreign body reaction were found during examination. Due to the HA particle smear contamination in the ZrO-HA group, no data was collected in this group. The mean BIC at the first three threads of the Ti-HA, Ti-HF and ZrO-HF were 57.78±18.22%, 46.41±14.55% and 47.41±14.05%, respectively. No statistically significant difference was found pair-wise among these three groups. When comparing the BIC data with the machined-surface implants, a statistically significant difference was found between the Ti-HA versus Ti implant group and the Ti-HF versus Ti implant group. The mean bone area (BA) at the first three threads for Ti-HA, Ti-HF and ZrO-HF showed statistically significant difference (p<0.05) between the ZrO-HF and Ti-HA groups, favoring the ZrO-HF group. The value of the peak removal force could only be collected from the Ti-HA group during the removal torque test. The mean RT value for the Ti-HA group was 24.39±2.58 Ncm. When comparing the RT result with our pilot study using machined-surface implants, the Ti-HA group showed statistically significant (p<0.05) higher values than the machined-surface Ti implants. The result of this study proves the in vivo biocompatibility of all four implant types tested. In the three measurable implant groups, the histomorphologic analysis showed comparable osseointegration properties in this animal model.Item The effect of triple antibiotic paste and EDTA on the surface loss and surface roughness of radicular dentin(2014) Nerness, Andrew; Spolnik, Kenneth Jacob, 1950-; Zunt, Susan L., 1951-; Platt, Jeffrey A., 1958-; Ehrlich, YgalIntroduction: Regenerative endodontic therapy in immature teeth with necrotic pulps triggers continued root development thereby improving the prognosis of these teeth. Several agents are under consideration for the disinfection and conditioning phases of this therapy. Triple antibiotic paste (TAP, i.e. equal parts of ciprofloxacin, metronidazole, minocycline) is used for canal disinfection and 17% EDTA solution is used for dentin conditioning. However, TAP and EDTA cause demineralization and their effect on surface loss and surface roughness of radicular dentin during regenerative procedures has not been quantified. Surface loss may be correlated with reduced tooth strength and surface roughness may be correlated with stem cell attachment. Objectives: The aim of this in vitro study was to quantitatively investigate the surface loss and surface roughness on human radicular dentin after treatment with two concentrations of TAP followed by EDTA. Materials and Methods: Human radicular dentin specimens were prepared from extracted human anterior teeth and randomized into six experimental groups. Group 1: saline control; Group 2: 17% EDTA; Group 3: TAP 1 mg/mL; Group 4: TAP 1 mg/mL and 17% EDTA; Group 5: TAP 1,000 mg/mL; Group 6: TAP 1,000 mg/mL and 17% EDTA for 5 minutes. After TAP is applied to Groups 3-6, all groups were incubated for 4 weeks. Then, groups 2, 4, and 6 were treated with EDTA for 5 minutes. Dentin surface loss (μm) and surface roughness (Ra, μm) were quantified after various treatments using non-contact and contact profilometry, respectively. Data were analyzed by one-way analysis of variance (α = 0.05) Hypothesis: It was hypothesized that there would be a significant difference in surface loss or surface roughness between at least two treatment groups. Results: All treatment groups showed significantly higher surface loss compared to untreated control. Dentin treated with 1g/mL TAP caused significant increase in surface loss and surface roughness compared to dentin treated with 1 mg/mL TAP. However, only 1g/mL TAP treated dentin showed significantly higher surface roughness compared to untreated control. The use of EDTA after both concentrations of TAP did not have significant additive effect on surface loss and surface roughness of dentin. Conclusion: The use of 1 mg/mL TAP can minimize surface loss and surface roughness of radicular dentin compared to higher concentrations. The use of EDTA after TAP may not cause additional surface loss and surface roughness of dentin.Item Effects of DynaMatrix® Membrane on Angiogenic Cytokine Expression From Human Dental Pulp Stem Cells(2013) Baker, Ryan William; Spolnik, Kenneth Jacob, 1950-; Ehrlich, Ygal; Vail, Mychel Macapagal, 1969-; Song, Fengyu; Legan, Joseph J.; Zunt, Susan L., 1951-; Windsor, L. JackThe aim of this current study was to determine if the exposure of human dental pulp stem cells (HDPSC) to the DynaMatrix membrane will result in an increased production of angiogenic cytokines that are critical for pulp/root regeneration. Angiogenesis cytokine arrays have been established as a viable method for assessing expression of cytokines.20 HDPSC were chosen as they are expected to be found in the apical papilla and the infected immature root canal system of teeth that current regenerative endodontic techniques are designed to treat.Item Effects of DynaMatrix® on angiogenic cytokine expression from human dental pulp fibroblasts : an in vitro study(2015) Adams, Joseph Benjamin; Spolnik, Kenneth Jacob, 1950-; Erhlich, Ygal; Bringas, Josef; Warner, Ned A. (Ned Alan); Zunt, Susan L., 1951-; Windsor, L. JackEFFECTS OF DYNAMATRIX® ON ANGIOGENIC CYTOKINE EXPRESSION FROM HUMAN DENTAL PULP FIBROBLASTS: AN IN VITRO STUDY by Joseph Benjamin Adams Indiana University School of Dentistry Indianapolis, IN Introduction: An exogenous scaffold may lead to more predictable pulp tissue regeneration and continued root formation in a regenerative endodontic procedure. DynaMatrix® is a natural membrane scaffold made of porcine small intestine, currently used in periodontal regenerative surgeries. Objective: The purpose of this study was to investigate if human dental pulp fibroblasts (HDPFs) seeded on DynaMatrix® membrane would result in an increase in the expression of angiogenic cytokines. Materials and Methods: HDPFs (75,000 per well) were seeded in 6-well plates. Three groups were tested: Group 1 (C): HDPFs in 70 media only; Group 2 (M): DynaMatrix® (Cook Biotech, Indianapolis, IN) alone in media; and Group 3 (C+M): HDPFs seeded on DynaMatrix® membranes. After 72 hours of incubation in serum positive, the conditioned media were collected and analyzed for the expression of 20 angiogenic cytokines utilizing RayBiotech Inc., arrays per the manufacturer’s instruction. The data were analyzed by ANOVA. Results: Group M was significantly higher than C for bFGF (p = 0.0023). C+M was significantly higher than M for ANG (p = 0.0104); GRO (p = 0.0003); IFN-γ (p = 0.0023); IL-6 (p = 0.0003); IL-8 (p = 0.0003); Leptin (p = 0.0003); MCP-1 (p = 0.0104); TIMP-1 (p = 0.0190); TIMP-2 (0.0123). C was significantly higher than C+M for ANG (p = 0.0104); MCP-1 (p = 0.0104); and THPO (p = 0.0308). Cytokines such as b-FGF, ANG, and leptin promote angiogenesis, and stimulate migration and proliferation of cells. Conclusion: The cytokine expression profile from the cells seeded on DynaMatrix® suggests that it might be a suitable scaffold for regenerative endodontic procedures. It could improve vascularization by increasing angiogenic cytokines in the microenvironment of the treated root canal and supporting tissue regeneration.Item Effects of kinetic cavity preparation vs. conventional handpiece preparation on the human dental pulp(1998) Collins, Julie M.; Sanders, Brian J.; Dean, Jeffrey A.; Zunt, Susan L., 1951-; El-Kafrawy, Abdel Hady, 1935-The purpose of this investigation was to compare the histopathologic effects of kinetic cavity preparation to the histopathologic effects of conventional high-speed handpiece preparation on the human dental pulp. The objective was to test the following hypothesis: kinetic cavity preparation results in significantly fewer pulpal effects than does conventional preparation using the high-speed handpiece. Class V cavity preparations were made in 26 teeth of seven patients who required extraction of these teeth for orthodontic purposes. Thirteen teeth were prepared using kinetic cavity preparation, using 27-um aluminum oxide particles at 160 pounds per square inch pressure. Thirteen were prepared using the high-speed handpiece and 330bur. Glass ionomer restorations were placed in all teeth. Extractions were done 10 to 15days after preparation. On teeth with closed apices, the apical one-third of the root was removed. All teeth were placed in 10 percent formalin solution. Teeth were sectioned and selected slides stained with hematoxylin and eosin for histologic evaluation. Microscopic findings indicated that the amount of remaining dentin was of significant thickness to be protective to the pulp. Pulpal responses ranged from no response in 22 specimens to a mild response in 4 specimens. Based on the results of this study, it was concluded that shallow preparation into the dentin does not cause pulpal damage at 10 to 15 days post-preparation, when using either kinetic cavity preparation or high-speed handpiece preparation. The hypothesis that kinetic cavity preparation causes significantly fewer pulpal effects than does conventional preparation with the high-speed handpiece was rejected.Item The Effects of Nicotine on the Proteolytic Activity of Periodontal Pathogens(2011) Kaeley, Janice,1976-; Gregory, Richard L.; Blanchard, Steven B.; Kowolik, Michael J.; Windsor, L. Jack; Zunt, Susan L., 1951-Periodontal disease is the leading cause of tooth loss in adults. Bacterial biofilm on tooth surfaces is the primary initiator of periodontal disease. Various factors contribute to the severity of periodontal disease including the different virulence factors of the bacteria within the biofilm. In the progression of periodontal disease, the microflora evolves from a predominantly Gram positive microbial population to a mainly Gram negative population. Specific gram negative bacteria with pronounced virulence factors have been implicated in the etiology and pathogenesis of periodontal disease, namely Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola which form the red complex of bacteria. The orange complex bacteria become more dominant in the maturation process of dental plaque and act to bridge the early colonizers of plaque with the later more dominant red complex bacterial and consists of such bacteria as Campylobacter showae, Campylobacter rectus, Fusobacterium nucleatum and Prevotella intermedia. Perhaps the most investigated contributing factor is the relationship between smoking and periodontal disease. When examining the association between cigarette smoking and interproximal bone loss, greater bone loss is associated with higher cigarette consumption, longer duration (i.e., pack year history) and higher lifetime exposure. The presence of various virulence factors such as the production of a capsular material, as well as the proteolytic activity of the various periopathodontic bacteria has been associated with the pathogenesis of periodontitis. Even though many different enzymes are produced in large quantities by these periodontal bacteria, trypsin-like enzymes, chymotrypsin-like enzymes and elastase-like enzymes, as well as dipeptidyl peptidase-like enzymes, have been thought to increase the destructive potential of the bacterium and mediate destruction of the periodontal apparatus. More specifically, it is hypothesized that the proteolytic activity of other clinically important periodontal pathogens, such as Fusobacterium nucleatum, Prevotella intermedia and Porphyromonas assacharolyticus, is increased in the presence of nicotine. The purpose of this study was to determine the effects of nicotine on F. nucleatum, P. intermedia and P. assacharolyticus proteolytic activity. Cultures were maintained on anaerobic blood agar plates containing 3% sheep blood. Bacterial cells were harvested from the plates and washed. Washed F. nucleatum, P. intermedia and P. assacharolyticus cells were incubated with 1 mg/ml of nicotine. Bacterial cells not incubated with nicotine were used as positive controls. Secreted enzymatic activity was measured using the synthetic chromogenic substrates glycyl-L-proline-p-nitroanilide (GPPNA), N-succinyl-L-alanyl-L-alanyl-L-alanyl-p-nitroanilide (SAAAPNA), N-succinyl-alanine-alanine-proline-phenylalanine-p-nitroanilide (SAAPPPNA) and N-α-benzoyl-L-arginine-p-nitroanilide (L-BAPNA) (Sigma-Aldrich Products, St. Louis, MO, USA). Appropriate means and standard deviations were determined for each of the enzymatic activities measured and analysis of variance (ANOVA) was used to compare the groups utilizing a 5% significance level for all comparisons. Results demonstrated that after 60 minutes of incubation of F. nucleatum, P. intermedia and P. assacharolyticus cells with 1 mg/ml of nicotine and the various synthetic substrates, had the following proteolytic activity for GPPNA: 0.83 ± 0.14, 0.72 ± 0.03 and 0.67 ± 0.10, respectively; SAAAPNA: 0.82 ± 0.06, 0.76 ± 0.05 and 0.68 ± 0.08, respectively; SAAPPPNA: 0.90 ± 0.13, 0.85 ± 0.17 and 0.72 ± 0.03, respectively; and BAPNA: 0.81 ± 0.15, 0.74 ± 0.13 and 0.74 ± 0.16, respectively. In conclusion, the results indicate that in the presence of 1 mg/ml of nicotine, the proteolytic activity of F. nucleatum and P. assacharolyticus was increased with all of the synthetic substrates (with statistical significance seen only in the increases with F. nucleatum and GPPNA, SAAAPNA and BAPNA). The proteolytic activity exhibited an increasing trend in activity for P. intermedia with SAAPPPNA and BAPNA but a decreasing trend in activity with GPPNA and SAAAPNA when incubated with 1 mg/ml of nicotine, once again demonstrating no statistical significance for any of the substrates. Therefore, it could be concluded that based on these results nicotine at a concentration of 1 mg/ml may increase the proteolytic activity of periodontal pathogens and thus may increase periodontal disease activity and subsequent periodontal breakdown. Further studies are needed to validate these results utilizing different concentrations of nicotine.
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