Browsing by Author "Spolnik, Kenneth Jacob, 1950-"
Now showing 1 - 10 of 20
Results Per Page
Sort Options
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 Antimicrobial properties of drug-containing electrospun scaffolds(2012) Jeppson, John; Spolnik, Kenneth Jacob, 1950-; Vail, Mychel Macapagal, 1969-; Erhlich, Ygal; Bottino, Marco C.; Gregory, Richard L.; Legan, Joseph J.; Zunt, Susan L., 1950-Endodontic treatment of the infected immature tooth has undergone a dramatic change. Conventional endodontic treatment can control infection, but root development usually remains impaired. A novel regenerative endodontic procedure, the revascularization method, can now control the infection and enable such teeth to continue root development. This is done by creating a fibrin-matrix scaffold in the antibiotic treated root canal space (RCS). Dental stem cells and growth factors have been able to continue root development in such an environment. The fibrin-matrix scaffold is dependent on the induction of a blood clot into the RCS, and this cannot always be predictably induced. PDS is a biocompatible material that can be electrospun to provide a matrix for cells and growth factors and perhaps improve on the blood clot induced fibrin scaffold by incorporating metronidazole as an adjuvant antimicrobial. A metronidazole containing electrospun PDS scaffold was examined in vitro using a turbidimetric test, the modified direct contact test. This scaffold significantly inhibited growth of an anaerobic primary endodontic pathogen Porphyromonas gingivalis. This scaffold may improve the treatment of the infected immature tooth by providing a designed matrix for root regeneration while serving simultaneously as an antibiotic drug delivery device to disinfect the RCS. The aim of this study is to evaluate in vitro the property of a synthetic scaffold to function as an antibacterial drug delivery device. PDS*II (polydioxanone) suture was obtained from Ethicon, INC. (Somerville, NJ) and was dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol, HFIP (Sigma Aldrich). Three different scaffolds were electrospun onto an aluminum foil background; (1) control scaffold with no antibiotic incorporated, (2) scaffold with 5.0-wt % metronidazole incorporated, and (3) 25-wt % metronidazole incorporated. All scaffolds were cut using a 4-mm diameter biopsy punch under aseptic conditions and removed from foil, control scaffold (n = 64), scaffold containing 5.0-wt % metronidazole (n = 32), and scaffold containing 25-wt % metronidazole (n=32). Experimental scaffolds were placed in a 96- well sterile flat bottom microtiter plate. Porphyromonas gingivalis a known primary endodontic pathogen was grown in 5 ml of BHI + YE with 0.25 μl of vitamin K with incubation at 37°C under anaerobic conditions for 48 hours. Microplates were sterilized before inoculation with Pg with 400 μl of 70-percent EtOH for a minimum of 30 minutes then pipetted out. After sterilization the microwells were washed with 400 μl of sterile water and pipetted out. Group 1 (negative control) microwells (n = 8) contained control scaffold and 190 μl of broth only. Group 2 (positive control) microwells (n = 8) contained 190 μl of broth and Pg only. Group 3 microwells (n = 8) contained control scaffold, 190 μl of broth, and 10 μl of Pg inoculum. Group 4 microwells (n = 8) contained scaffold with 5 wt % metronidazole, 190 μl of broth, and 10 μl of Pg inoculum. Group 5 microwells (n = 8) contained scaffold with 25 wt % metronidazole, 190 μl of broth, and 10 μl of Pg inoculum. Group 6 contained 190 μl of uninoculated broth for spectrophotometer calibration. Sterile microplate lids were used to isolate microwells from the surrounding environment. Microplates were incubated at 37°C under anaerobic conditions for 48 hours. After 48 hours the microplates were read by using an endpoint reading in the spectrophotometer. This was repeated four times. Comparisons among the groups for differences in optical density as a measure of bacterial growth were made using mixed-model ANOVA, with a fixed effect for group and a random effect for experimental run. Pair-wise group comparisons were performed using Tukey's multiple comparisons procedure to control the overall significance level at 5 percent. The analyses were performed using the ranks of the data. Broth had significantly lower OD than all other groups (p < 0.0001). Broth+Pg and Broth+Pg+Scaffold had significantly higher OD than 5-wt % Metro (p < 0.0001) and 25-wt % Metro (p < 0.0001), but Broth+Pg and Broth+Pg+Scaffold were not significantly different from each other (p = 0.97). 5-wt % Metro and 25-wt % Metro were not significantly different from each other (p = 0.24). From the results of our study, we concluded that the 5.0-wt % and 25-wt % metronidazole containing scaffolds significantly inhibited bacterial growth and could be effectively utilized for the endodontic regeneration procedure.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 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 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 The effects of radicular dentin treated with double antibiotic paste and EDTA on dental pulp stem cell proliferation : an in-vitro study(2014) Kim, Ki Wan; Spolnik, Kenneth Jacob, 1950-; Ehrlich, Ygal; Platt, Jeffrey A., 1958-; Zunt, Susan L., 1951-; Windsor, L. JackIntroduction: Regenerative endodontic therapy in immature teeth promotes continuation of root development and likely increases the prognosis of these teeth. The use of double antibiotic paste (DAP), equal parts of ciprofloxacin and metronidazole, followed by the dentin conditioner, ethylenediaminetetraacetic acid (EDTA), has been suggested for canal disinfection and facilitation of stem cell attachment/proliferation, respectively. However, the effect is unknown when all these agents are used on on radicular dentin surfaces to facilitate the level of stem cell proliferation. Objectives: The aim of this in-vitro study is to compare the proliferation of human dental pulp stem cells (hDPSCs) on human radicular dentin treated with two different concentrations of DAP followed by EDTA. Materials and Methods: Human premolars and incisors were prepared into standardized polished 4 mm x4 mm radicular dentin specimens. Groups of specimens were treated with DAP 500 mg/mL, DAP 1 mg/mL, DAP 500 mg/mL followed by 17-percent EDTA, DAP 1 mg/mL followed by 17-percent EDTA; 17% EDTA, or no treatment. All groups treated with antibiotics were incubated with DAP at 37°C for one week. All specimens were washed with distilled water. The hDPSCs were seeded across all specimens and unattached cells were collected after 24 hours. LDH assay was completed on unattached cells for quantification. Three days after attachment, WST viability and LDH cytotoxicity assays were performed. Hypothesis: There is no significant difference in hDPSC viability, unattachment, and cytotoxicity on dentin specimens treated with DAP and 17-percent EDTA. Clinical Significance: These results can be used to help identify the best treatment concentrations when using DAP and/or EDTA to promote endodontic regeneration. Results: The results demonstrated significantly less viability of hDPSCs on specimens treated with 500 mg/mL DAP with and without 17-percent EDTA. Groups treated with 1 mg/mL DAP, 1 mg/mL DAP and 17-percent EDTA, and 17-percent EDTA alone had no statistically significant difference in viability compared with control untreated dentin. The results of the unattached cells from the LDH demonstrated that cells from the specimens treated with solely 500 mg/mL and 1 mg/mL DAP had significantly higher levels of unattached cells when compared with all other groups. The LDH assays in summation with the WST assays showed a trend of a lack of proliferation on groups treated with 500 mg/mL DAP with and without 17-percent EDTA. Conclusions: Paste-like concentrations (500 mg/mL) of DAP are detrimental to hDPSC viability, whereas the present study supports the use of low-concentration antibiotics consistent with current recommendations for intracanal medicaments used during endodontic regenerative procedures.Item Efficacy of propolis against fusobacterium nucleatum biofilm(2013) Griglione, Anthony Leonard; Spolnik, Kenneth Jacob, 1950-; Gregory, Richard L.; Vail, Mychel Macapagal, 1969-; Legan, Joseph J.; Zunt, Susan L., 1951-; Eckert, George J.; Ehlich, YgarThe primary goal of root canal treatment is to eliminate microbes from the root canal system, which is the cause of pulpal and periapical infections. Research shows that after a single visit of chemomechanical debridement microbes continue to remain within the canal system. An interappointment medication step has been advocated to maximize potential elimination of microbes within the root canal system. Previous studies have shown propolis to be antibacterial against common endodontic microbes. Studies have shown trends in different microbes being present in primary verus secondary endodontic infections. The majority of literature has focused on the efficacy of propolis against Enterococcus faecalis, a microbe commonly implicated in secondary endodontic 95 infections. The aim of this study was to demonstrate the efficacy of propolis against Fusobacterium nucleatum, a microbe commonly found in primary endodontic infections. This study aims to demonstrate the efficacy of propolis against a bacterium of primary endodontic infections (F. nucleatum) as well as against microbial biofilm to further support its potential use as a novel intracanal medicament. Dilutions of propolis were added to cultures of F. nucleatum in microtiter plates in a range from 390 μg/ml to 50,000 μg/ml. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and the minimum biofilm inhibitory concentration (MBIC) were determined. The MIC was determined of the total solution (biofilm+planktonic), planktonic, and biofilm (MBIC) after a 48-hour incubation period. The MBIC was determined by fixing biofilm to the wells and using crystal violet staining with spectrophotometry. The MBC was examined by plating solution from each concentration test well and reading the plates after 48 hours of incubation. The results show that the MIC of the total (biofilm+planktonic) appears to occur at a concentration of 6250 μg/ml. The MBIC appears to occur at the concentration of 1562.5 μg/ml. The planktonic results exhibit no significant difference in test and control wells. There was no MBC at any of the test concentrations. The propolis appears to inhibit bacterial growth and biofilm formation but does not appear to be bactericidal at any of the tested concentrations. The results of this study indicate that propolis has an MIC and MBIC when tested in vitro against F. nucleatum, although it does not show an MBC. There appears to be potentially significant interaction of propolis with biofilm as displayed by the lower concentration needed to exibit inhibitory effects on biofilm formation. This information 96 may contribute to the ability to develop a proper concentration of propolis to use in vivo when treating endodontic infections.Item Evaluating the use of 3D imaging in creating a canal-directed endodontic access(2015-06-09) Maru, Avni Mahendra; Spolnik, Kenneth Jacob, 1950-; Ghoneima, Ahmed; Bringas, Josef; Warner, Ned A. (Ned Alan); Zunt, Susan L., 1951-; Ehrlich, YgalIntroduction: During root canal treatment (RCT), an opening is made through the crown of the tooth to access and to disinfect the root canal system (RCS). Traditional endodontic access (TEA) may sacrifice tooth structure and weaken the tooth. Cone beam computed tomography (CBCT) provides information about the exact location of the root canals. This information can be used for the design of a canal-directed endodontic access (CDEA). It may also be used for the 3D printing of an acrylic endodontic stent that could help to create a conservative CDEA. Objective: 1) Evaluate the ability of the Dolphin 3D imaging software to assist in creating a CDEA; 2) Compare tooth structure loss in a CDEA to that in a TEA by measuring the volume of remaining tooth structure, surface area of the access opening at the occlusal, and remaining dentin thickness at the CEJ. Materials and Methods: Thirty extracted human mandibular premolars were used. Teeth with large, wide canals were excluded. CBCT images will be taken for all teeth using Kodak 9000. Fifteen teeth were randomly assigned to the TEA group and 15 teeth were assigned to the CDEA group. The CDEA path was mapped using Dolphin 3D imaging software. Acrylic access stents were designed using Rhino 3D software and printed using a 3D printer. The teeth were accessed through the corresponding stents. The 15 teeth that are part of the traditional access group were accessed without a stent. A CBCT scan was taken post-access for all 30 teeth. Wilcoxon Rank Sum Tests were performed to compare the following outcomes for the two groups: the volume of remaining tooth structure, the surface area of the access opening at the occlusal, and remaining dentin thickness at the CEJ. Results: The remaining dentin thickness (percent loss) was not significantly larger for TEA than for CDEA. The surface area (post-treatment) was significantly larger for TEA than for CDEA, and volume (percent loss) was significantly larger for TEA than for CDEA. Conclusion: The use of the CBCT and Dolphin 3D imaging provided an accurate and more conservative CDEA with the guide of an acrylic stent.