IU Indianapolis ScholarWorks :: Browsing by Author "Bringas, Josef"

Browsing by Author "Bringas, Josef"

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The ability of new intracanal medicaments to prevent the formation of multi-species biofilm on radicular dentin
(2017) Jacobs, Jordon C.; Spolnik, Kenneth J.; Ehrlich, Ygal; Gregory, Richard L.; Yassen, Ghaeth; Bringas, Josef
The residual antibacterial effects of antimicrobials used in endodontic regeneration against biofilm bacteria obtained from immature and mature teeth Jordon C. Jacobs DDS, Richard L Gregory PhD, Ygal Ehrlich DMD, Kenneth Spolnik DDS, MS, Josef S. Bringas DMD, DDS, MS, and Ghaeth Yassen BDS, MSD, PhD We explored the residual antibacterial properties of dentin pretreated with low concentrations of double antibiotic paste (DAP) against biofilm bacteria obtained from different clinical sources. Dentin blocks were sterilized and randomized into 4 treatment groups and 2 control groups (n=20). Blocks from treatment groups were pretreated with DAP (1 or 5 mg/ml) loaded into methylcellulose, calcium hydroxide (Ca(OH)2), or methylcellulose paste. After one week, the treatment pastes were removed and all blocks were immersed in PBS. The dentin blocks from treatment groups and one of the control groups were then inoculated with bacterial isolates obtained from immature or mature teeth with pulpal necrosis(n=10). The remaining control group received no bacteria and was used as a sterile control. Blocks were then incubated anaerobically for 3 weeks. Biofilm disruption assays were conducted for all samples. Two-way ANOVA and pair-wise comparisons were used for statistical analyses. The residual antibacterial effect of dentin pretreated with 5 mg/ml of DAP was significantly higher than all other groups regardless of the source of biofilm. Dentin pretreated with 1 mg/ml of DAP demonstrated significantly higher residual antibacterial effects in comparison to dentin pretreated with placebo paste and Ca(OH)2 only in bacterial isolates obtained from mature teeth with pulpal necrosis. Dentin pretreated with Ca(OH)2 did not demonstrate any residual antibacterial effects. Dentin pretreated with 1 or 5 mg/ml of DAP demonstrated significantly better residual antibacterial effects against biofilm bacteria obtained from mature teeth with pulpal necrosis in comparison to bacterial isolates obtained from immature teeth with pulpal necrosis.
The antibacterial effect of a radiopaque double antibiotic paste against both an established multispecies and a single enterococcus faecalis biofilm
(2019) Haslam, Bryce S.; Spolnik, Kenneth J.; Ehrlich, Ygal; Gregory, Richard L.; Bringas, Josef; Warner, Ned A.
For regenerative endodontic procedures (REPs) to be successful an elimination of bacteria from the root canal system must be accomplished. Many different medicaments with antibacterial properties have been used to obtain complete disinfection. Double antibiotic paste (DAP) containing a mixture of ciprofloxacin and metronidazole has been shown to be a promising intracanal medicament. The addition of a radiopaque filler such as zirconium oxide to DAP may affect the antibacterial properties of DAP as well as allow precise placement and radiographic visualization of its position in the canal system. The aim of the proposed study was to evaluate the direct antibacterial properties of zirconium oxide radiopacifier combined with DAP (RoDAP) against a multispecies biofilm from a bacterial isolate from an infected immature tooth with a necrotic pulp and a known single species biofilm. 4x4 mm radicular dentin specimens (n = 112) obtained from human extracted teeth were used prepared and sterilized prior to use. A multispecies clinical bacterial isolate from an immature tooth with a necrotic pulp and a single species Enterococcus faecalis isolate were obtained. These bacterial isolates were used to inoculate dentin slabs and grown for 3 weeks. The dentin slabs were treated for 1 week with 1.0-mg/mL and 10- mg/mL RoDAP, 1.0-mg/mL DAP, and two placebo pastes consisting of methyl cellulose (MC) and methyl cellulose combined with zirconium oxide (RoMC), respectively, as well as two no-treatment controls. Following treatment, the grown biofilm was detached and spiral plated. The plated biofilm cells were cultured for 24 hours and each group examined using a colony counter to determine bacterial numbers (CFUs/mL). Data analysis, using a 5.0-percent significance level was conducted using one-way ANOVA followed by pair-wise group comparisons. Both 1.0-mg/mL and 10 mg/mL RoDAP demonstrated significant antibacterial effects against bacterial isolates from an immature tooth with a necrotic pulp as well as an E. faecalis isolate. The precise application of RoDAP confirmed radiographically with its direct antibacterial properties may be beneficial for intracanal disinfection during REPs.
The antibacterial effect of new intracanal medicaments against established mutlispecies biofilm
(2017) Troxel, Alex; Spolnik, Kenneth J.; Gregory, Richard; Ehrlich, Ygal; Bringas, Josef; Zunt, Susan L.; Yassen, Ghaeth
We investigated the antibacterial effect of low concentrations of double antibiotic paste (DAP) loaded into a methylcellulose system against bacterial biofilms obtained from mature and immature teeth with necrotic pulps. Standardized radicular dentin specimens were randomly divided into six experimental groups (n = 20). Group 1: 5mg/mL DAP treatment. Group 2: 1mg/mL DAP treatment. Group 3: Calcium hydroxide (Ca(OH)2) treatment. Group 4: Methylcellulose. Group 5: No treatment. Group 6: No bacteria or treatment. Clinical bacterial isolates were obtained from mature and immature teeth with necrotic pulps indicated for endodontic regeneration or routine endodontic treatment, respectively. Specimens in each group were inoculated with either bacterial isolates (n = 10) and incubated anaerobically for 3 weeks. Specimens were then treated for one week with the assigned group treatment. Treatments were rinsed with sterile saline and biofilms were detached and spiral plated using biofilm disruption assays. Wilcoxon Rank Sum tests followed by pair-wise comparisons were used for statistical analyses. Treatment of infected dentin with 1 mg/ml of DAP, 5 mg/mL of DAP, and Ca(OH)2 demonstrated significant and substantial antibiofilm effects in comparison to untreated control groups or groups treated with placebo paste. Furthermore, 1 mg/mL of DAP caused complete eradication of biofilm obtained from mature tooth with necrotic pulp. However, the same concentration was not able to completely eradicate biofilm obtained from the immature tooth with necrotic pulp. Low concentrations of DAP (1-5 mg/mL) loaded into a biocompatible methylcellulose system demonstrated significant antibacterial effects against biofilm obtained from both mature and immature teeth with necrotic pulps.
The antibacterial effects of radiopaque double antibiotic pastes against clinical bacterial isolates from mature and immature teeth with necrotic pulps
(2018) Ibrahim, Carolin Francis; Spolnik, Kenneth J.; Ehrlich, Ygal; Gregory, Richard L.; Zunt, Susan; Bringas, Josef; Yassen, Ghaeth
Low concentrations (1-10mg/mL) of double antibiotic paste (DAP) have demonstrated antibacterial properties in regenerative endodontics. The aim of this study was to evaluate if DAP made radiopaque (RoDAP) with barium sulfate has antibacterial effects against bacterial isolates from a mature and immature tooth with necrotic pulp. Clinical bacterial isolates were obtained from the canals of mature and immature teeth with necrotic pulps during root canal therapy or a regenerative procedure, respectively. Bacterial isolates were grown anaerobically for three weeks on 4x4mm dentin specimens prepared from extracted human teeth (n=48 per biofilm type). The dentin specimens were allocated into six groups and treated as follows: 1mg/mL RoDAP, 10mg/mL RoDAP, calcium hydroxide (UltraCal), placebo (barium sulfate in methylcellulose), no treatment, and no bacteria or treatment (sterile control). After one week of treatment the biofilm was detached and biofilm disruption assays were conducted to determine the bacterial numbers (CFUs/mL). The data was analyzed using Wilcoxon Rank Sum tests followed by pairwise comparisons. 1 and 10 mg/mL RoDAP as well as calcium hydroxide demonstrated significant antibacterial effects against the tested bacterial isolates. The placebo paste did not demonstrate any significant antibacterial effects. No significant difference in antibacterial effects was found against isolates from both mature and immature teeth regardless of the type of treatment. Both 1 and 10 mg/mL RoDAP demonstrated significant antibacterial effects against bacterial isolates from mature and immature teeth with necrotic pulps. RoDAP can be beneficial clinically since its adequate placement within the canal system can be confirmed radiographically.
The antibacterial stability of a new radiopaque double antibiotic paste
(2018-06) Epkey, Kathryn Eileen; Spolnik, Kenneth J.; Ehrlich, Ygal; Gregory, Richard L.; Zunt, Susan; Bringas, Josef; Yassen, Ghaeth
We evaluated the antibacterial stability (shelf life) of a new radiopaque double antibiotic paste (RoDAP) loaded in a methylcellulose system with 30% w/v barium sulfate against biofilm collected from an immature tooth with necrotic pulp. Uniform radicular dentin specimens were infected with bacterial biofilm obtained from an immature tooth with a necrotic pulp and incubated anaerobically for three weeks. These samples were randomly divided into 6 experimental groups (n=7) and treated for 1 week at three time points of aged radiopaque DAP: 0 months, 3 months, and 6 months. Group 1: 1mg/mL RoDAP treatment. Group 2: 10 mg/mL RoDAP treatment. Group 3: Calcium hydroxide (Ca(OH)2) treatment. Group 4: Methylcellulose with barium sulfate. Group 5: No treatment. Group 6: No bacteria or treatment. The samples were rinsed with sterile saline to detach biofilms and then spiral plated using a biofilm disruption assay. Statistical analyses were performed using Wilcoxon rank-sum tests and Wilcoxon signed rank tests with fixed effects for treatment, time, and the treatment-by-time interaction. Treatment of infected dentin with 1 mg/mL RoDAP, 10 mg/mL RoDAP, and Ca(OH)2 demonstrated significant and substantial antibiofilm effects in comparison to untreated control groups or groups treated with placebo paste after 0, 3, and 6 months of aging. Calcium hydroxide, however, showed slightly less antibiofilm activity after 6 months of aging when compared to 0 months and 3 months of aging. This difference was statistically significant (p > 0.05). In conclusion, both concentrations of RoDAP maintained full antibacterial efficacy after 6 months of aging, while calcium hydroxide lost some antibacterial activity after a shelf life of 6 months.
Caries Presentation in Illict Drug Users and Excessive Soft Drink Consumers.
(04/13/15) Parker, Eric; Ehrlich, Ygal; Pfotenhauer, David; Bringas, Josef; Warner, Ned; Parker, Eric; Ehrlich, Ygal; Endodontics
Advanced cervical caries in anterior teeth is common to two different patient behaviors. Rampant cervical caries (“meth mouth”) is a common presentation in drug users. Extensive consumption of soft drinks also presents a similar pattern of cervical caries. Two cases are presented and dental treatment considerations are discussed. Case One: excessive soft drink consumption. A young male presented to IUSD for evaluation and treatment of a painful #9. Clinical exam: #9 had extensive facial and cervical caries with a pulp exposure. Cervical caries on teeth #6, 7,8,10,11,12,13. Patient’s history revealed frequent daily consumption of multiple sweet soft drinks. #9 had become increasingly painful and that caused him to seek treatment. Pulpal and Percussion/Palpation testing and radiographic exam of #9 were conducted. Diagnosis: #9 Pulpal Necrosis with Symptomatic Apical Periodontitis. Treatment: Oral hygiene instruction to reduce the frequency of soft drink consumption. #9 was endodontically treated and restored. Caries in #6, 7,8,10,11,12,13 were treated and teeth restored. Case Two: cocaine abuse. A young female presented to IUSD for evaluation and treatment of a painful #9. Clinical exam: #9 had extensive cervical caries. Cervical caries on #7, 8, 9 and dental attrition. Patient reports having used cocaine for many years. Pulpal and Percussion/Palpation testing and radiographic exam of #9 were conducted. Diagnosis: #9 Pulpal Necrosis with Symptomatic Apical Periodontitis. Treatment considerations: Local anesthesia was achieved without the use of epinephrine due to possible occasional unreported use of cocaine. #9 was endodontically treated and restored. Patient did not return for continued treatment at IUSD. Conclusion: Cervical caries in anterior teeth associated with soft drinks demonstrate similar characteristics to those observed in patients with reported cocaine abuse. This should alert dental providers dentists to be aware of both illicit drug use and soft drink intake as part of the patient’s medical, dental, and social history information.
Comparison of guided endodontic access with and without pin fixation in 3D printed teeth with simulated pulp canal obliteration
(2021-06) Long, Jacob Daniel; Spolnik, Kenneth; Ehrlich, Ygal; Dutra, Vinicius; Bringas, Josef; Warner, Ned
Introduction: In order to successfully treat an infected root canal system (RCS), it is required to locate all root canals and have an access path to the apex of each canal. This can be challenging in teeth with pulp canal obliteration (PCO), often leading to increased chair time and increased risk of iatrogenic errors. Guided endodontic access (GEA) combines information from a cone-beam computed tomography (CBCT) scan with an intra-oral scan to create a stent. GEA stents with or without fixation pins have been shown to be successful in accurately negotiating a RCS with PCO. Objective: Compare the degree of deviation and difference in 3D offset at the base to apical tip of the drill from the designed access path when a GEA stent with and without pin fixation is used to access tooth #8 with PCO. Materials and Methods: A 3-D printed maxillary model of an anonymous patient had a GEA stent designed using coDiagnonstiX software. The stent extended from tooth #3 to tooth #14 with the guide sleeve over tooth #8. Tooth #8 with no calcification, calcification to the cervical third, and calcification to the middle third of the RCS were designed in the coDiagnostiX software. Tooth #8 will be accessed using a 1.3 mm drill that fits a 1.3 mm sleeve used for both access and pin fixation. 15 of the 30 GEA samples will utilized pin fixation, while the other 15 samples did not utilize pin fixation. Following GEA in all 30 samples a CBCT was taken of each sample. Each post-operative CBCT was aligned with the pre-operative CBCT in the coDiagnostiX software. The coDiagnostiX software was able to calculate the degree of deviation and difference in 3D offset between the base and apical tip of the drill during GEA. Paired t-tests were used to test each group for significant differences in 3D offset between base and tip. Two-way ANOVA was used to evaluate the effects of pin fixation and calcification on the degree of deviation and the deviation of 3D offset of the entry point and tip. Results: There was a significant interaction between use of pin fixation and calcification level on the degree of deviation of GEA. GEA with pin fixation had a significantly larger degree of deviation than GEA without pin fixation with calcification extending to the middle third of the RCS. GEA with and without pin fixation did not have a significant difference when calcification extended to the cervical third of the RCS or no calcification was present. There was a significant interaction between use of pin fixation and calcification level on 3D offset difference. GEA with pin fixation had a significantly larger 3D offset difference than GEA with no pin fixation for calcification in the middle third of the RCS. For GEA with and without pin fixation there was no significant difference when calcification extended to the cervical third of the RCS or no calcification was present. Conclusion: The use of pin fixation did not result in a decrease of degree of deviation or difference in 3D offset during GEA access. It can be concluded that the use of pin fixation is not necessary for GEA of teeth with PCO when a full dentition is present to provide stability and retention of the stent.
The Effects of a Pyk2 Kinase Inhibitor on the Proliferation and Differentiation of Human Dental Pulp Stem Cells
(2021) McIntyre, Patrick; Bruzzaniti, Angela; Ehrlich, Ygal; Bringas, Josef; Spolnik, Kenneth
Introduction: Regenerative endodontic procedures are an effective treatment option for immature teeth with infected necrotic pulps to allow for healing and potential continued root development, yet challenges to ideal treatment outcomes remain. Consistent development of root length and width of dentin remains a challenge, as does development of the pulp-dentin complex. Previous in vitro studies have assessed the role of different growth factors and bioactive molecules in combination with scaffolds to potentially facilitate continued development of the pulp-dentin complex using dental pulp stem cells (DPSCs). The proline-rich tyrosine kinase 2 (Pyk2) is linked with osteoblast activity and the regulation of bone mass. Further, the Pyk2 inhibitor PF-4618433 (PF-46) has been shown in previous studies to enhance osteoblast activity and mineral deposition in vitro. However, whether Pyk2 targeting promotes the osteogenic differentiation of DPSCs remains unknown. Objective: The purpose of this study was to investigate the effect of a Pyk2 inhibitor, PF-46, on the proliferation, differentiation, and mineralization of human DPSCs. Materials and Methods: Human DPSCs were cultured in 24-well plates with α-MEM with 10% FBS, and containing 0 μM (vehicle control) or 0.1 μM, 0.3 μM, or 0.6 μM PF-46. Fresh media and treatments were replaced every 2-3 days. After 1 day incubation, cytotoxic effects were evaluated by using an MTS proliferation assay. After 4 days of treatment, direct cell counting was performed. To induce osteogenic differentiation, ascorbic acid and β-glycerol phosphate were added to the culture media and the DPSCs were cultured with PF-46 for 14 days. Then, an alkaline phosphatase (ALP) assay and mineral deposition assay were performed. Differences between treatment groups were analyzed by a one-way ANOVA followed by pair-wise tests conducted using Tukey’s multiple comparisons procedure with a 5% significance level. Results: The 0.6 μM PF-46 group had a significantly higher cell count, ALP activity and mineral deposition when compared to 0 μM PF-46. The 0.1 and 0.3 μM PF-46 groups also had significantly higher ALP activity compared to the 0 μM PF-46 group after 14 days of incubation. There was a general trend of increased differentiation and mineral deposition as the concentration of PF-46 increased from 0.1 μM to 0.6 μM. Conclusion: There was a general concentration-dependent increase in cell count, differentiation, and mineral deposition by human DPSCs as the concentration of PF-46 increased from 0 μM up to 0.6 μM, with the highest activity observed with 0.6 μM PF-46. Although further research is needed, these results suggest that strategies that target Pyk2 may potentially be used to improve the osteogenic differentiation of DPSCs to aid endodontic regeneration.
The effects of concentration and treatment time on the residual antibacterial properties of DAP
(2016) Jenks, Daniel Brent; Spolnik, Kenneth J.; Bringas, Josef; Gregory, Richard L.; Yassen, Ghaeth H.; Ehrlich, Ygal; Warner, Ned A.
Introduction: Regenerative endodontic procedures are used to treat immature teeth with pulpal necrosis in order to control infection, enable continued root development and enhance formation of a pulp like tissue in the canal. Canal disinfection is an integral part the regenerative endodontic process. Double antibiotic paste (DAP; i.e., equal parts of ciprofloxacin and metronidazole) has been successfully used for canal disinfection in regenerative endodontics. A comparison of the residual antibacterial effect of dentin treated with various dilutions of DAP pastes on biofilm formation has not yet been investigated thoroughly. Objectives: The aims of this in-vitro study were to investigate how concentration and time of treatment affect the residual antibacterial properties of DAP in preventing E. faecalis biofilm formation on human dentin. Materials and Methods: Extracted human teeth were used to obtain 4x4mm radicular dentin specimens. Each specimen was pretreated for 1 or 4 weeks with the 77 clinically used concentration of DAP (500 mg/mL), low concentrations of DAP (1, 5 or 50 mg/mL) loaded into a methylcellulose system, calcium hydroxide (Ca(OH)2), or placebo paste. After treatment, samples were rinsed and placed in sterile phosphate buffered saline (PBS) for three weeks. Samples were then inoculated with cultured E. faecalis and incubated in anaerobic conditions for three weeks to allow mature biofilm formation. The dentin samples were rinsed and biofilms detached. The detached biofilm cells were then diluted and spirally plated for enumeration on blood agar plates. The plates were then incubated for 24 h and the number of CFUs/mL was determined using an automated colony counter. Data was analyzed using Fisher’s Exact and Wilcoxon rank sum tests were used for statistical comparisons (α=0.05). Results: Dentin pretreatment for 4 weeks with 5, 50 or 500 mg/mL of DAP demonstrated significantly higher residual antibacterial effects and complete eradication of E. faecalis biofilms in comparison to a 1 week pretreatment with similar concentrations. However, dentin pretreated with 1 mg/mL of DAP or Ca(OH)2 did not provide a substantial residual antibacterial effect regardless of the application time. Conclusion: Dentin treated with 500, 50, or 5 mg/mL of DAP for 4 weeks was able to completely prevent the colonization of bacterial biofilm. Four-week treatment of dentin with DAP offers superior residual antibacterial effect in comparison to a one-week treatment. Intracanal application of DAP for 4 weeks during endodontic regeneration may offer an extended residual antibacterial effect.
Effects of DynaMatrix on Angiogenic Cytokine and Matrix Metalloproteinase Expression from Human Endothelial Cells: An In-vitro Study
(2015) Hill, Scott Thomas; Spolnik, Kenneth J.; Warner, Ned; Zunt, Susan L.; Windor, L. Jack; Bringas, Josef; Ygal, Ehrlich
Introduction: Regenerative endodontics (RE) is a treatment alternative for the infected immature tooth to establish an environment in the canal that enables continued root development and the growth of pulp or pulp-like tissue within the canal. A scaffold created in the canal encourages the formation of vital tissue. The porcine sub-intestinal-submucosa (SIS) membrane, Dynamatrix®, has the potential to serve as an endodontic scaffold. Research at Indiana University School of Dentistry (IUSD) has shown that Dynamatrix® can support the growth of human dental pulp stem cells (HDPSC) and human pulp fibroblasts (HPF). Positive angiogenic cytokine profiles were seen after these cells were seeded on Dynamatrix®. Endothelial cells play an important role in the formation of blood vessels and are a source of angiogenic cytokines. Exposure of these cells to DynaMatrix® may result in a positive angiogenic profile for both cytokines and matrix metalloproteinases (MMPs). Objective: The aim of this in-vitro study was to investigate if the exposure of human endothelial cells to the DynaMatrix® membrane would result in differences in the expression of cytokines and MMPs that play roles in angiogenesis. Materials and Methods: Human endothelial cells (HUVECs) were obtained from American Type Culture Collection (ATTC, Manassas, VA) and used in this study. Groups were established as follows: (a) Group 1: HUVECs seeded in culture media only, (b) Group 2: DynaMatrix® membrane incubated alone in the serum-media without any cells, and (c) Group 3: HUVECs seeded on DynaMatrix® membranes. After 72 hours of incubation, the conditioned media were collected and analyzed for the expression of 20 angiogenic cytokines and MMPs utilizing cytokine and MMP protein arrays. The density of each cytokine and MMP expressed was measured, averaged, and statistically analyzed by ANOVA. Results: Exposure of human umbilical vein endothelial cells (HUVECs) to the DynaMatrix® membrane resulted in a positive angiogenic profile for both cytokines and MMPs. Conclusion: This work furthers the evidence for the potential of DynaMatrix® to serve as a more predictable scaffold in RE.

Browsing by Author "Bringas, Josef"

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