Browsing by Author "Spolnik, Kenneth"

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    ACCURACY OF GUIDED ENDODONTICS IN ENDODONTIC MICROSURGERY
    (2024-06) Grayson, Michelle Sarah; Spolnik, Kenneth; Ehrlich, Ygal; Dutra, Vinicius; Hine, Charlie; Warner, Ned
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    Accuracy of Guided Endodontics in Simulated Perforated Teeth with Calcified Canals
    (2023) Eidelstein, Dyana M.; Spolnik, Kenneth; Ehrlich, Ygal; Dutra, Vinicius; Hine, Charles; Warner, Ned
    Introduction: Endodontic treatment in teeth following iatrogenic accidents can be challenging and difficult to correct. Guided Endodontic Access (GEA) has been used as an adjunct in endodontic treatment to in in treating teeth with complex anatomy and may assist in treating teeth with procedural accidents. The accuracy of GEA stents in teeth with a simulated procedural accident will be assessed. Objective: This in vitro study will utilize a 3D model of a tooth with a simulated procedural accident: deviation and perforation during root canal treatment(RCT). GEA stents will be fabricated designed to overcome and compare the previous ledge formation and perforation. The influence on the degree of deviation of the designed access path from the prepared path will be determined by assessing the degree of angle of deviation and amount of deviation in millimeters. Materials and Methods: A 3-D printed maxillary model of an anonymous patient will be used. The deviated path and perforation will be simulated in a 3D printed tooth #4 using the meshmixer software tooth at two levels: apical and mid-root. The stent extends from tooth #3 to tooth #14 and was designed using coDiagnostiX software over tooth #4 with a simulated perforation. 15 GEA stents will have guides for the mid root perforation, 15 GEA stents will have guides for the canals not in contact with the perforation, and 15 GEA stents will have guides for canals apical to the perforation. All cases will be accessed with a 1.0 mm drill that is planned to fit the access sleeve. Results: The distal( base), the distal(tip) and the vestibular (tip) for the guided access in which the canal was in contact with the deviated path had no significant difference in deviation. The angle was significantly deviated in all models, but the model in which the canal was in contact with the deviated canal had a significantly lower angle, distal base, and vestibular base that both models in which the canal was not in contact with the deviated path and at different heights. The degree of deviation for all samples ranged from 1.40° to 10.60°. The largest degree of deviation corresponds with the increased depth of the original canal system. Conclusion: In conclusion, our study revealed that the utilization of GEA in calcified teeth with PCO and a deviated path demonstrated greater effectiveness in canals located in closer proximity compared to those located farther away from the deviated path. Therefore, we can infer that the use of a GEA stent in calcified teeth with PCO and a deviated path is favorable only under certain conditions.
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    Buffered vs. Unbuffered Local Anesthesia in Mandibular Molars Diagnosed with Symptomatic Irreversible Pulpitis: A Controlled, Randomized, Double-Blind Study
    (2022) Alena, Peter; Spolnik, Kenneth; Ehrlich, Ygal; Warner, Ned
    Introduction: Profound pulpal anesthesia is not always adequate in mandibular teeth after the administration of local anesthesia, especially in the presence of irreversible pulpitis. Failure to achieve anesthesia has been seen in 30–80% of patients in teeth with a diagnosis of irreversible pulpitis. Onpharma™ developed an FDA-approved device that uses sodium bicarbonate to buffer a standard local anesthetic (LA) solution so that its pH may become closer to its pKa. Claims have been made that buffering a local anesthetic increases the anesthetic’s effect. Previous studies on the anesthetic efficacy of Onpharma’s Onset buffering system were inconclusive and may be dependent on the techniques used. Objectives: The aim of this study is to determine whether a buffered local anesthetic can lead to more profound and faster pulpal anesthesia in mandibular molars diagnosed with symptomatic irreversible pulpitis as compared to a standard, unbuffered local anesthetic. Materials and Methods: 40 total subjects completed the study. Screened and eligible subjects with a mandibular molar diagnosed with symptomatic irreversible pulpitis were randomly allocated into 2 groups so 1 group received a total of 3 cartridges of a standard, unbuffered 2% lidocaine with 1:100,000 epinephrine via inferior alveolar nerve block (IANB) followed by supplemental buccal and lingual infiltrations, while the other received the equivalent yet buffered formulation. An electronic pulp tester (EPT) was used to objectively determine baseline pulpal status of the affected tooth, followed by 2-minute interval testing following the administration of all local anesthesia. The onset of pulpal anesthesia was defined by the first of 2 consecutive EPT=80 readings, and the endodontic treatment could begin. Profound pulpal anesthesia was ultimately determined if the patient reported a comfortable pulpotomy as reflected on the Wong-Baker FACES Visual Analog Scale. Null Hypothesis 1: Subjects possessing mandibular molars diagnosed with symptomatic irreversible pulpitis will not achieve pulpal anesthesia more profoundly using buffered 2% lidocaine w/ 1:100,000 epinephrine in comparison to the standard, unbuffered anesthetic formulation. Null hypothesis 2: Subjects possessing mandibular molars diagnosed with symptomatic irreversible pulpitis will not achieve pulpal anesthesia faster using buffered 2% lidocaine w/ 1:100,000 epinephrine in comparison to the standard, unbuffered anesthetic formulation. Results: We observed a local anesthetic success rate of 45% in the buffered group, 70% in the unbuffered group, and ultimately 57.5% between both groups. The findings further indicate that the VAS scores after pulpotomy is significantly different between the 2 groups (p=0.019), with the unbuffered group having a more profound mean VAS score of 1.2 (as opposed to a buffered mean of 3.1). Regarding the time of onset for pulpal anesthesia, there was no statistically significant difference noted between the buffered and unbuffered groups. Conclusion: Based on the findings of this study, the null hypothesis 1 cannot be rejected since unbuffered 2% lidocaine with 1:100,000 epinephrine had a statistically significant increase in profound pulpal anesthesia compared to the buffered equivalent. The null hypothesis 2 cannot be rejected since there was no evidence of a significant difference in the time to pulpal anesthesia between the buffered and unbuffered groups.
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    Comparing Brief Relaxation Period to Virtual Reality Period in Reducing Dental Anxiety Prior to Root Canal Treatment: A Randomized Control Trial
    (2023-06) Mintz, Caley Faith; Spolnik, Kenneth; Ehrlich, Ygal; Schrader, Stuart; Flowers, Leslie; Warner, Ned
    Introduction: Anxiety is a debilitating and difficult sensation many people face on a daily basis. Up to 20% of American adults experience dental anxiety2. Dental anxiety can present both psychologically and physiologically as a barrier to starting, completing and/or finishing dental treatment.2,4 Catastrophizing the procedure, which is common practice in anxious patients, will alter and increase the perception of pain66. Approaching and understanding dental anxiety has shown to produce more positive treatment outcomes and overall increased patient satisfaction2, 4, 6. Non-pharmacological approaches to reducing dental anxiety can be a quick, non-invasive, method to put patients at ease and can save chair time for both practitioner and patient, as well as a more pleasant experience for the patient. Objectives: The goal of this study is to investigate non-pharmacologic approaches to reducing dental anxiety prior to non-surgical root canal treatment. This will be done by comparing an Auditory Alone Brief Relaxation period (ABR) to a Relaxation Virtual Reality period (RVR). Materials and Methods: 60 subjects who need non-surgical root canal treatment was randomly allocated into 2 groups. One group received earphones to listen to a guided brief relaxation recording, focusing on breathing and a body scan. The other group received virtual reality goggles and choose a scene of their liking to experience. State Trait Anxiety Indicator (STAI), Visual Analog Scale (VAS), and vitals were measured to objectively determine baseline anxiety score followed by the same metrics after brief relaxation or virtual reality experience. A student will perform root canal treatment, and VAS and vitals were again be recorded at the end of the appointment. Results: Both RVR and ABR showed a statistically significant decrease in anxiety in STAI-S (p value <.001 for both), STAI-T (p value 0.025 ABR; <.001 RVR). VAS scores also significantly reduced from T1 to T2 to T3 time frame. Discussion: Our study investigated and compared two different interventions in reducing anxiety prior to endodontic therapy. The results showed that both ABR and RVR reduced anxiety prior to endodontic therapy and had an effect even after the endodontic therapy was completed. Additionally, when comparing the time points T1, T2, and T3, heart rate decreased throughout the procedure and more importantly, after the intervention (ABR or RVR) was delivered. Both anxiety scales, STAI and VAS showed a statistical significant decrease in anxiety throughout the appointment. Conclusion: Non-pharmacological techniques like guided meditation and virtual reality are a valid and unique approach to reducing anxiety prior to endodontic therapy. Both ABR and RVR interventions reduced the feelings of anxiety throughout the entire endodontic appointment. This study displays the ease of incorporating both interventions to reduce anxiety in an economical and noninvasive fashion.
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    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.
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    Differentiation and Activity of Murine Derived Stromal Osteoblasts After Electromagnetic Wave Stimulation
    (2022) Wu, Jennifer L.; Spolnik, Kenneth; Bruzzaniti, Angela; Ehrlich, Ygal; Warner, Ned
    Introduction: Elimination of bacteria and active infection within an infected root canal system is one of the primary objectives of nonsurgical root canal treatment. One of the measures of successful root canal treatment is subsequent bone healing of periapical lesions caused by previous infection. A previous study by Yumoto et al. showed that electromagnetic wave stimulation can increase proliferation of osteoblastic cells with no cytotoxicity, and it can also up-regulate growth factors such as vascular endothelial growth factor and platelet-derived growth factor.18 They also showed increased proliferation of an immortalized osteoblastic MC3T3-E1 cell line 3 days following electromagnetic stimulation (EMS).18 Previously, Pauly et al. found increased alkaline phosphatase (ALP) activity with 10 mA EMS application to primary murine calvaria-derived osteoblastic cells with 5 pulses at 1 second per pulse, but no significant differences were found for MTS proliferation nor mineral deposition compared to a negative control group.82 Optimization of the different variables including post-treatment incubation time, current delivery, and number of pulses per treatment may be necessary to improve osteogenic activity. The use of mesenchymal stem cells from murine bone marrow may also offer a physiologically relevant model for osteoblastic regeneration of periapical lesions. Objectives: The goal of this study was to investigate and optimize the effects of electromagnetic wave stimulation (EMS) on murine bone marrow mesenchymal stem cells (MSCs) by evaluating the proliferation and differentiation of the cells after exposure to different EMS treatment regimens. Materials and Methods: 5 x104 stromal osteoblasts (SOBs) were cultured in 24-well plates in α-MEM containing 10% fetal bovine serum. Cells were then subjected to pulsed EMS treatments of 1 mA, 10 mA, and 50 mA. EMS was generated using an electromagnetic apical treatment (EMAT) device created by J. Morita MFG Corp. Proliferation was assessed via MTS assay 1 days after treatment. For osteogenic differentiation, ascorbic acid and β-glycerol phosphate were added to the culture media, and SOBs were cultured for 14 days. Afterwards, alkaline phosphatase (ALP) activity and Alizarin-red S mineral deposition were quantified as measures of osteoblast activity. Cells grown in osteogenic media without EMS treatment served as the negative control. Results: Although MSC proliferation was unaffected by different EMS treatment regimens, 50 mA EMS resulted in a decrease in ALP activity and mineral deposition by osteoblasts. Conclusions: Our findings suggest bone healing by EMS may involve a different cellular mechanism, that is not reproduced in vitro in our studies. Utilizing different amperage and EMS regimens may improve osteogenic differentiation.
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    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.
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    The effects of electromagnetic wave stimulation (EMS) on osteoblast differentiation and activity
    (2020-06) Pauly, Katherine L.; Spolnik, Kenneth; Bruzzaniti, Angela; Ehrlich, Ygal; Bringas, Josef S.
    Introduction: The goal of nonsurgical root canal therapy is to reduce the bacterial load within an infected root canal system, and the subsequent objective is to prevent or treat apical periodontitis. Clinical studies have shown more expedient healing of apical periodontitis treated with electromagnetic wave stimulation (EMS) as compared to apical periodontitis not treated with EMS. Stimulation of osteoblasts and growth factors has been shown when EMS was applied to rat calvaria, resulting in increased bone healing. Objective: The purpose of this vitro study was to evaluate the effects of EMS on the proliferation and differentiation of osteoblasts. Using primary neonatal calvaria osteoblast-lineage cells, the effects of different EMS regimens on proliferation, alkaline phosphatase (ALP) activity, and mineral deposition were determined. Materials and Methods: EMS regimen included currents of 0mA, 0.1mA, 1mA, and 10mA delivered for five consecutive 1s pulses per day for one, two, and three days. Cell proliferation was assayed after 1 or 2 days using an MTS assay. Alkaline phosphatase activity and mineral deposition were assayed after culturing the cells in osteogenic media containing ascorbic acid and -glycerol phosphate for 7 days. Comparisons were performed using analysis of variance, with a 5% significance level. Results: There was no statistically significant differences noted in MTS proliferation and mineral deposition between the experiment EMS treatment groups of 0.1, 1.0, and 10.0 mA compared to the control group of 0 mA current on calvaria-derived osteoblast. While there were no statistically significant differences noted in ALP activity in the 0.1, and 1.0 mA EMS groups, compared to 0 mA control, alkaline phosphatase activity was significantly increased in the 10 mA EMS group. Conclusion: There was no significant differences in MTS proliferation and mineral deposition of the EMS group compared to the control group. However, 10 mA EMS favored increased ALP activity suggesting EMS can promote matrix maturation by osteoblasts. Additional in vitro experimental studies, including different stem cell populations, culture duration and EMS treatment regimens are needed to understand the mechanism of action of EMS for future applications in regenerative endodontics.
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    The effects of radicular dentine treated with double antibiotic paste and ethylenediaminetetraacetic acid on the attachment and proliferation of dental pulp stem cells
    (Wiley, 2015-10) Kim, Ki Wan; Yassen, Ghaeth H.; Ehrlich, Ygal; Spolnik, Kenneth; Platt, Jeffrey A.; Windsor, L. Jack; Department of Biomedical and Applied Sciences, IU School of Dentistry
    Aim This study explored the effects of dentine treated with two concentrations of double antibiotic paste (DAP) and ethylenediaminetetraacetic acid (EDTA) on the attachment and proliferation of dental pulp stem cells (DPSCs). Materials and Methods Radicular dentine samples were prepared with identical dimensions and randomized into six groups (n = 4). Four groups were treated with double antibiotic paste (DAP) at concentrations of 500 mg ml−1 or 1 mg ml−1 with or without EDTA. The other two groups were treated with EDTA only or received no treatment. DPSCs were seeded on each dentine sample (10 000 cells per sample). Lactate dehydrogenase activity assays were used to calculate the attached DPSCs after 1 day of incubation. Water soluble tetrazolium assays were performed to investigate DPSCs proliferation on the treated dentine samples after three additional days of incubation. Two-way anova followed by Tukey–Kramer tests was used for statistical analyses (α = 0.05). Results Dentine treated with 1 or 500 mg ml−1 of DAP followed by EDTA caused significant increases in DPSCs attachment compared to the dentine treated with the DAP alone. The 500 mg ml−1 of DAP with or without EDTA caused significant reductions in DPSCs proliferation. However, the treatment of dentine with 1 mg ml−1 of DAP did not have significant negative effects on DPSCs proliferation regardless of the use of EDTA. Conclusion The use of 1 mg ml−1 of DAP followed by 10 min of irrigation with EDTA in endodontic regeneration procedure may have no negative effects on the attachment and proliferation of DPSCs.
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    The impact of hydroxyapatite on alkaline phosphatase activity and mineral deposition of dental pulp stem cells using a double antibiotic paste loaded methylcellulose carrier
    (2020) Fischer, Benjamin I.; Bruzzaniti, Angela; Spolnik, Kenneth; Ehrlich, Ygal; Bringas, Josef; Gregory, Richard
    Introduction: Regenerative endodontic procedures (REPs) are a type of endodontic treatment aimed at replacing damaged tooth structures, including dentin and root structures, as well as cells of the pulp-dentin complex. Double antibiotic paste (DAP) has been shown to be efficacious in achieving disinfection of the root canal system while minimizing cytotoxicity to dental pulp stem cells (DPSCs). Hydroxyapatite (HA) is an extracellular, mineralized component of bone that has shown much promise as a scaffold in the field of regenerative medicine. Objective: The objective of this study was to evaluate the effects of HA in a DAP loaded methylcellulose (MC) carrier on the differentiation and mineral deposition of DPSC over time. Materials and Methods: DPSCs were plated in 24-well plates with culture media. The following day, semi-permeable 0.1 m chambers were inserted into the wells to separate the reservoirs and permit delivery of medicaments. 100 L treatment paste composed of MC with 1% DAP and either 0.5% or 1.0% nano-HA was added, followed by additional culture media. After 3 days of treatment, medicaments were removed and DPSCs were cultured for an additional 9 days with replacement of media every 3-4 days. At Day 12, DPSCs were evaluated for alkaline phosphatase (ALP) activity using a biochemical assay and mineral deposition using an Alizarin Red S Ca2+ staining assay (4 wells/group). Comparisons between groups were performed using one-way analysis of variance (ANOVA) with a 5% significance level used for all tests. Results: A trend towards increased ALP and mineral deposition activity was noted among the groups with HA added to DAP with MC. Although these trends were not statistically significant, a trend towards increased ALP and mineral deposition was observed after 3-day medicament exposure. The results were similar to previous findings using 7-day medicament treatments. Conclusion: The addition of HA showed a trend towards improved differentiation and mineral deposition of DPSCs compared to DAP with MC. Although additional studies are required, these results showed suggest that even with a shortened treatment time, increased differentiation and mineral deposition of DPSCs may be possible. This study provides additional support that low concentration DAP in a MC carrier has potential application in regenerative endodontic procedures. The novel addition of HA may provide additional osteogenic potential.