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Item Bioactive nanofibrous scaffolds for regenerative endodontics(SAGE, 2013-11) Bottino, M.C.; Kamocki, K.; Yassen, G.H.; Platt, J.A.; Vail, M.M.; Ehrlich, Y.; Spolnik, K.J.; Gregory, R.L.; Endodontics, School of DentistryHere we report the synthesis, materials characterization, antimicrobial capacity, and cytocompatibility of novel antibiotic-containing scaffolds. Metronidazole (MET) or Ciprofloxacin/(CIP) was mixed with a polydioxanone (PDS)polymer solution at 5 and 25 wt% and processed into fibers. PDS fibers served as a control. Scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), tensile testing, and high-performance liquid chromatography (HPLC) were used to assess fiber morphology, chemical structure, mechanical properties, and drug release, respectively. Antimicrobial properties were evaluated against those of Porphyromonas gingivalis/Pg and Enterococcus faecalis/Ef. Cytotoxicity was assessed in human dental pulp stem cells (hDPSCs). Statistics were performed, and significance was set at the 5% level. SEM imaging revealed a submicron fiber diameter. FTIR confirmed antibiotic incorporation. The tensile values of hydrated 25 wt% CIP scaffold were significantly lower than those of all other groups. Analysis of HPLC data confirmed gradual, sustained drug release from the scaffolds over 48 hrs. CIP-containing scaffolds significantly (p < .00001) inhibited biofilm growth of both bacteria. Conversely, MET-containing scaffolds inhibited only Pg growth. Agar diffusion confirmed the antimicrobial properties against specific bacteria for the antibiotic-containing scaffolds. Only the 25 wt% CIP-containing scaffolds were cytotoxic. Collectively, this study suggests that polymer-based antibiotic-containing electrospun scaffolds could function as a biologically safe antimicrobial drug delivery system for regenerative endodontics.Item Attachment and proliferation of dental pulp stem cells on dentine treated with different regenerative endodontic protocols(Wiley, 2017) Alghilan, M. A.; Windsor, L. Jack; Palasuk, Jadesada; Yassen, Ghaeth H.; Department of Endodontics, School of DentistryAim To investigate the attachment and proliferation of dental pulp stem cells (DPSC) on dentine treated with various endodontic regeneration protocols. Methodology Standardized dentine samples were irrigated with sodium hypochlorite (1.5% NaOCl) and ethylenediaminetetraacetic acid (17% EDTA) and randomized into four treatment groups and two control groups. The treatment groups were treated with a clinically used concentration of triple antibiotic paste (TAP), double antibiotic paste (DAP), calcium hydroxide (Ca(OH)2) or diluted TAP in a methylcellulose system (DTAP) for 1 week. Each sample in the treatment groups was then irrigated with EDTA. The two control groups were treated with EDTA or received no treatment. Dental pulp stem cells were seeded on each dentine specimen (10 000 cells). Lactate dehydrogenase activity assays were then performed to evaluate the attached DPSC after 1 day of incubation. Water-soluble tetrazolium assays were used to determine DPSC proliferation after three additional days of incubation. Friedman's test followed by least significant difference were used for statistical analyses (α = 0.05). Results Triple antibiotic paste and DTAP regeneration protocols, as well as EDTA-treated dentine, caused significant increases in DPSC attachment to dentine. Triple antibiotic paste, DAP and Ca(OH)2 regeneration protocols caused significant reductions in DPSC proliferation on dentine. However, the DTAP regeneration protocol did not have any significant negative effects on DPSC proliferation. Conclusions The clinically used endodontic regeneration protocols that include the use of TAP, DAP or Ca(OH)2 medicament negatively affected DPSC proliferation on dentine. However, the use of DTAP medicament during regenerative endodontic treatment may not adversely affect the proliferation of DPSC.Item Effect of angiotensin receptor blockade on insulin sensitivity and endothelial function in abdominally obese hypertensive patients with impaired fasting glucose(Portland Press, 2012-02) Perlstein, Todd S.; Henry, Robert R.; Mather, Kieren J.; Rickels, Michael R.; Abate, Nicola I.; Grundy, Scott M.; Mai, Yabing; Albu, Jeanine B.; Marks, Jennifer B.; Pool, James L.; Creager, Mark A.; Department of Endodontics, IU School of DentistryAngII (angiotensin II) may contribute to cardiovascular risk in obesity via adverse effects on insulin sensitivity and endothelial function. In the present study, we examined the effects of ARB (angiotensin receptor blocker) therapy (losartan, 100 mg/day) on insulin sensitivity and endothelial function in 53 subjects with stage I hypertension, abdominal obesity and impaired fasting glucose. The study design was a randomized double-blinded parallel design placebo-controlled multi-centre trial of 8 weeks duration. We used the hyperinsulinaemic-euglycaemic clamp technique to measure insulin sensitivity (expressed as the 'M/I' value) and RH-PAT (reactive hyperaemia-peripheral arterial tonometry) to measure endothelial function. Additional measures included HOMA (homoeostasis model assessment)-B, an index of pancreatic β-cell function, and markers of inflammation [e.g. CRP (C-reactive protein)] and oxidative stress (e.g. F2-isoprostanes). ARB therapy did not alter insulin sensitivity [5.2 (2.7) pre-treatment and 4.6 (1.6) post-treatment] compared with placebo therapy [6.1 (2.9) pre-treatment and 5.3 (2.7) post-treatment; P value not significant], but did improve the HOMA-B compared with placebo therapy (P=0.05). ARB therapy also did not change endothelial function [RH-PAT, 2.15 (0.7) pre-treatment and 2.11 (0.7) post-treatment] compared with placebo therapy [RH-PAT, 1.81 (0.5) pre-treatment and 1.76 (0.7) post-treatment; P value not significant]. Markers of inflammation and oxidative stress were not significantly changed by ARB therapy. In conclusion, ARB therapy did not alter peripheral insulin sensitivity or endothelial function in this cohort of patients with essential hypertension, abdominal obesity and impaired fasting glucose, but did improve pancreatic β-cell function.Item Endodontic management of an Infected Immature Tooth with Spontaneous Root Closure and Type II Dens Invaginatus: A Case Report(2015) Baker, Ryan W.; Spolnik, Kenneth J.; Ghoneima, Ahmed; Ehrlich, YgalApical periodontitis in an immature maxillary lateral incisor (#10) with arrested root development and a natural closure of the open apex in 22-year-old male was endodontically treated in a manner that conserved the existing hard tissue barrier (HTB). A dens invaginatus Oehlers II was also present. The patient reported no symptoms, but did recall an incident of dental trauma as a child. A 3D image showed the nature of the HTB closing the open foramen as well as the anatomy of the immature root with the dens invaginatus. Porosities were seen in the HTB and that is consistent with the histological “Swiss Cheese” appearance known to occur in apexification. Endodontic treatment was performed in a single-visit. Access to the root canal system (RCS) included penetration through the dens invaginatus. The canal was not mechanically cleaned, but only irrigated with sodium hypochlorite, ethylenediaminetetra-acetic acid (EDTA), chlorhexidine using the EndoVac® system. The barrier was maintained and covered with a 5mm layer of mineral trioxide aggregate (MTA). A sterile damp sponge was placed on the MTA. On the following visit the MTA had set, and an EndoSequence fiber post was placed in the canal and EndoSequence dual-cure core build-up material was used to close the access and restore the tooth. A clinical and radiographic follow-up, 30 months after the initial treatment revealed resolution of the radiolucency and apical trabecular bone deposition and the patient was asymptomatic.Item Susceptibility of methacrylate-based root canal filling to degradation by bacteria found in endodontic infections(Quintessence International, 2014-07-10) Whatley, Jenny D.; Spolnik, Kenneth J.; Vail, Mychel M.; Adams, Benjamin H.; Huang, Ruijie; Gregory, Richard L.; Ehrlich, YgalObjectives: To present a case of endodontic failure obturated with a methacrylate-based root filling material, Resilon/ RealSeal (RS). To determine if RS is susceptible to biodegradation by endodontically relevant microbes by a method known to show RS degradation. Method and Materials: Emulsions of RS were dispersed in agar with minimal bacterial nutrients in culture plates. Lipase PS served as a positive control. Pseudomonas aeruginosa, Fusobacterium nucleatum, Prevotella intermedia, Porphyromonas gingivalis, Porphyromonas asaccharolytica, Enterococcus faecalis, Streptococcus sanguis, Streptococcus mutans, Staphylococcus aureus, and Staphylococcus epidermidis were tested for their ability to biodegrade RS. The bacteria were inoculated in the plates and examined daily for RS degradation for 7 days. Results: Degradation of the emulsified RS manifested in the formation of clear zones around P aeruginosa, P intermedia, P asaccharolytica, S aureus, and S epidermidis. No degradation was seen with the other tested bacteria or in plates that did not contain RS emulsion. Conclusion: Endodontic pathogenic bacteria can degrade RS. These findings complement other work and suggest that the seal and integrity of root canal fillings obturated with RS may be impaired by a microbial insult.