Indiana University-Purdue University Indianapolis (IUPUI) Endodontic Regeneration Graduate Endodontic Department / 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.

Indiana University School of Dentistry Department of Endodontics / Introduction: Triple antibiotic paste (TAP, 1000 mg/ml) is composed of equal portions of ciprofloxacin, metronidazole and minocycline and is used as an intracanal dressing to disinfect the infected immature root canal during endodontic regeneration procedures. Lower concentrations of TAP have been recommended to minimize detrimental effects on pulp stem cells. TAP can be retained within the dentin matrix and its continual release confers an antibacterial effect to the dentin. Objective: The aim of this in vitro study was to investigate the residual antibacterial effect of dentin treated with various concentrations of TAP loaded into a gel system. Materials and Methods: Radicular dentin slabs were prepared from human teeth after obtaining IRB approval. The slabs were sterilized and treated with methylcellulose-based TAP of 100 mg/mL, 10 mg/mL, 1 mg/mL, 1.5% NaOCl, placebo paste with no TAP, or a positive control group with pure 1000 mg/mL TAP. Samples in each group were treated with the assigned TAP concentration for three weeks or immersed in 1.5% NaOCl for five minutes (n =18 per group). All samples were then irrigated with sterile water followed by 17% EDTA and incubated in phosphate buffered saline for either 2 or 4 weeks. Samples were then inoculated with Enterococcus faecalis and incubated for an additional 3 weeks. Biofilm formed on each sample was then dislodged and spiral plated to evaluate the bacterial colony-forming units. Data were analyzed using Fisher’s Exact tests and Wilcoxon rank sum tests (α = 0.05). Results: Dentin treated with 10, 100, or 1000 mg/mL of TAP demonstrated significant residual antibacterial effects up to four weeks. However, only 100 mg/mL TAP was able to completely prevent bacterial colonization after four weeks. No considerable residual antibacterial effect was observed in dentin treated with placebo gel, 1 mg/ml TAP or 1.5% NaOCl. Conclusion: At least 10 mg/mL of TAP loaded into a methylcellulose system is required to achieve a substantial residual antibacterial effect for four weeks.

Indiana University-Purdue University Indianapolis (IUPUI) Indiana University School of Dentistry Indiana University Endodontic Department / 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.

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.

Indiana University-Purdue University Indianapolis (IUPUI); Graduate Endodontics, Masters degree thesis / 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.

Indiana University-Purdue University Indianapolis (IUPUI) / Introduction: Regenerative endodontic procedures require adequate disinfection of immature teeth with necrotic pulps. Regeneration endodontic procedures have been shown to increase width and length of dentin after disinfection of the root canal system with various antibiotic pastes such as double antibiotic paste (DAP). DAP is composed of an equal ratio of metronidazole and ciprofloxacin. After the root canal system is disinfected with DAP, it is later flushed out with 17-percent EDTA. There is a need for a radiopaque component in the DAP to facilitate proper placement. Following disinfection with DAP, a residual antibacterial effect is formed in dentin. It is not known if this effect is modified by making DAP radiopaque. Objectives: The residual antibacterial effects of a radiopaque double antibiotic paste against a bacterial isolate obtained from an immature tooth with necrotic pulp on radicular dentin will be explored utilizing various strengths of DAP (1 mg/mL, 10 mg/mL, and 25 mg/mL) after 1 week and 4 weeks. Materials and Methods: Dentin samples of 4x4-mm will be prepared from previously extracted human posterior teeth. The samples will be assigned to seven treatment groups in a random fashion (G1 will be treated with 25-, G2-10, and G3-1 mg/mL of radiopaque DAP. G4 will be treated with the radiopaque filler without DAP. G5 will be treated with Ca(OH)2 and G6 sterile water with bacterial biofilm, and G7 will be sterile water). The groups will be treated for both 1 week 4 weeks. These samples will then be stored in a phosphate buffered saline (PBS) solution for 1 week and then inoculated with the cultured bacterial isolate from an immature tooth with necrotic pulp. The specimens will be incubated for three weeks to permit adequate formation of a biofilm. The biofilm will be detached, diluted and spirally plated onto blood agar plates and incubated for a total of 48 hours in 5-percent CO2 at 37°C. The number of CFUs/mL will be counted using an automated colony counter. A two way ANOVA and Fisher’s Protected Least Significant Differences test using 5-percent significance level will be used to evaluate the resulting data. Expected outcome: A residual antibacterial effect will be observed on radicular dentin samples when treated with radiopaque DAP. Conclusion: Using a radiopaque DAP that provides residual antibacterial effects, further awareness into regenerative endodontic procedures will be gained leading to alterations in treating such cases. / 2020-06-30

The mining industry is a major economic driver and job creator for many countries. However, mining is associated with geo-hazards and environmental issues, such as the disposal of large volumes of waste, acid mine drainage, and ground subsidence. As a result, efficient mining waste management is crucial for sustainable development. The geotechnical, economic, and environmental benefits of cemented paste backfill (CPB) have piqued the interest of researchers and academicians worldwide, making it an essential aspect of underground mining management. CPB is a thickened cementitious combination of dewatered tailings (70 - 85 wt.%), binders (usually 3 to 8% wt.%), and water used to backfill mine waste into underground mining stopes. Despite being used in small amounts, the cost of cement makes up to 80% of the cost of backfilling operations. In addition, clinker production accounts for 5-8% of global human created carbon dioxide (CO₂) emissions. This predicament necessitates the development of a viable alternative to cement. Partially substituting cement with supplementary cementitious materials like fly ash, blast furnace slag, natural pozzolans, and other materials has been increasingly prevalent in CPB. It is evident that the addition of slag to cement can increase the mechanical strength of CPB at the advanced ages but decreases the strength and suction development due to the slow reaction kinetics in the CPB at the early ages, which may negatively affect the mechanical stability of the CPB, mining cycle, and safety of mineworkers. Moreover, the supply of these materials is limited and may not be enough for the future needs of the industry. Furthermore, there has been a surge in interest in using limestone powder (LS) owing to its abundance, low cost, and lack of environmental costs which are associated with Portland cement - Type 1 (PCI). The addition of LS accelerates hydration at the early ages, thus resulting in high early strength, but the dilution effect can reduce the late strength. The combination of LS and slag in a ternary blended cement can be potentially used as a binder for CPB with acceptable strength development at the early and advanced ages while lowering the cost of the CPB and the carbon footprint of the mining industry. Nevertheless, the rheology, mechanical strength, and stability are important key performance quality criteria for CPB; however, the effect of ternary cement blends on these parameters is not well known. In this research program, the impact of the binary and ternary cement blends on (i) the fresh properties of CPB, such as the rheological properties (yield stress, viscosity) and setting time, and (ii) the strength and suction development of CPB are investigated. To understand the effect of substituting slag with LS in the binary binder in the first phase of the study, binary binders with two differ-ent PCI: Slag proportions of 50/50 and 80/20 are examined with no limestone, followed by replacing slag with an increasing amount of LS from 0 to 20 wt. % of the total binder, with a constant cement content, over a period of 4 hrs (0, 0.25, 1, 2, and 4 hrs) of curing at room temperature. In the second phase, the effect of a ternary binder (PCI-Slag- LS) with varying proportions on the suction development and the mechanical behavior of hardened CPB is investigated over a curing period of up to 90 days. The changes in strength of these binary and ternary binders on the CPB sample are tested for 1, 3, 7, 28, 60, and 90 days. An unconfined compression test (UCS) is conducted to evaluate the strength development. The microstructure of the mixes is examined through mercury intrusion porosimetry (MIP) for changes are validated through monitoring for the development of hydration and suction, electrical conductivity (EC), and temperature, which is carried out for up to 30 days. This is followed by a microstructure analysis with a thermogravimetric/differential thermogravimetric test on fresh and hardened samples. The results of the first phase show that an increase in the percentage of substituted cement in the binary binder (from PCI/Slag 80/20 to 50/50) increases the yield stress of the CPB but decreases the viscosity of the mix. However, the addition of LS as a substitution for slag shows a considerable decrease in the yield stress of the control mix with an increase in viscosity with increasing dosages of LS, thus indicating an improvement in the flowability of CPB. The second phase results indicate that the slow hydration kinetics of slag influences early age suction and strength changes in the binary sample with a high slag content (50/50); however, its latent hydraulic and pozzolanic properties enhance strength gain after 28 days. The addition of 5% limestone to the ternary blend increases the strength gain by up to 7 days compared to the binary control samples. Indeed, the presence of LS influences the rate of hydration of cement and slag through both physical (filler, nucleation, dilution) and chemical (hydrate) effects. However, substituting more than 10% LS for slag affects the mechanical performance at all ages. Overall, up to 50 wt.% slag and 10 wt.% limestone with cement as a ternary binder can be used without significant compressive strength loss. This study demonstrates that the partial substitution of ordinary Portland cement (OPC) with varying percentages of slag and LS is complementary, and overall, the interaction of slag and LS is observed. The optimal use of LS and slag in a ternary system may serve as a sustainable alternative to the commonly used OPC and PCI/Slag binders, thereby reducing the energy consumption and carbon footprint associated with cement. The findings of this study will ultimately help to develop a better understanding of the impact of ternary blends with increasing percentages of LS on the rheology and setting time of CPB mixes and mechanical strength changes in designing an efficient mixing plant, particularly its transport system.

Cemented paste backfill (CPB) is a highly advantageous method of backfill that has been increasing in use in recent decades as it provides many environmental, economic, and practical benefits. When combined with cement and water, it recycles a portion of the dewatered tailings produced from mines as backfill for underground stopes. CPB is transported from the plant on the surface through pipes to the stopes, sometimes over several kilometers, and then placed in underground mining cavities (stopes) to support the ground or rock mass. Therefore, it must meet certain rheological, setting time, and strength gain performance requirements. Additionally, as many mines around the world are located in areas of freshwater scarcity, and societies are holding corporations to ever higher standards for humanitarian and environmental responsibility, many mines are seeking to utilize locally available, saline groundwater or seawater as mixing water in backfill. The impacts of this decision on the rheological, setting, and strength properties of CPB must be better understood to allow for the confident selection of this convenient solution, as the risks associated with improper design include huge costs due to pipeline clogging and death or injury due to backfill failure and ground subsidence. NaCl is a contributor to natural groundwater and seawater salinity and may be present in concentrations of up to 300 g/L. An additional cost-saving measure favoured by mines is to replace some of the costly Portland cement with much cheaper supplemental cementitious materials such as blast furnace slag. Therefore, this thesis examines the impacts of NaCl concentration and binder composition on the yield stress, viscosity, initial and final setting time, and strength development of CPB. A robust experimental program has been undertaken in which CPB was subjected to the above-mentioned tests in addition to pH and MIP testing, SEM, TG/DTG, XRD, and zeta potential analyses, and electrical conductivity, suction, and water content monitoring. CPB samples were made with synthetic silica tailings, Portland cement, and water with NaCl concentrations of 0 g/L, 10 g/L, 35 g/L, 100 g/L, and 300 g/L and CPB made with 35 g/L and slag replacement percentages of 0%, 25%, 50%, and 75%. Additional samples tested were made with natural gold tailings, Portland cement, and NaCl concentrations of 0 g/L and 35 g/L for verification. Rheological testing was conducted at 0 minutes, 15 minutes, 1 hour, and 2 hours after mixing, and UCS testing was conducted after 1 day, 3 days, 7 days, 28 days, and 60 days of curing. Additional tests or analyses were performed on selected mixes and curing times for optimum insight and monitoring was conducted from 0 to 28 days after curing. It was found that low concentrations of NaCl (10 g/L and 35 g/L) generally had favourable impacts on the UCS and setting times of CPB, while higher concentrations had negative impacts. The impacts of slag replacement on UCS development of saline CPB were also generally favourable. However, the impacts of slag replacement on initial setting time were generally negative, and favourable at higher replacements (50% or more) for final setting time. Low NaCl concentration led to slightly negative impacts on yield stress, especially at longer curing times (1-2 hours), but high concentrations greatly reduced the yield stress. NaCl concentration had minor impacts to viscosity, with any concentration leading to a slightly higher initial viscosity but slightly lower viscosity at longer curing times. Slag replacement content had negligible effects on yield stress, but led to favourable decreases in viscosity over longer curing times. The combination of positive and negative impacts indicates that care must be taken to knowledgeably prioritize or balance critical properties in mix design, though there is indication of opportunities for overall improvement. Supplemental testing provided useful information to explain the mechanics behind the results which will allow designers to carefully select the required components for the desired properties.

Multiple view geometry is the foundation of an important class of computer vision techniques for simultaneous recovery of camera motion and scene structure from a set of images. There are numerous important applications in this area. Examples include video post-production, scene reconstruction, registration, surveillance, tracking, and segmentation. In video post-production, which is the topic being addressed in this dissertation, computer analysis of the motion of the camera can replace the currently used manual methods for correctly aligning an artificially inserted object in a scene. However, existing single view methods typically require multiple vanishing points, and therefore would fail when only one vanishing point is available. In addition, current multiple view techniques, making use of either epipolar geometry or trifocal tensor, do not exploit fully the properties of constant or known camera motion. Finally, there does not exist a general solution to the problem of synchronization of N video sequences of distinct general scenes captured by cameras undergoing similar ego-motions, which is the necessary step for video post-production among different input videos. This dissertation proposes several advancements that overcome these limitations. These advancements are used to develop an efficient framework for video analysis and post-production in multiple cameras. In the first part of the dissertation, the novel inter-image constraints are introduced that are particularly useful for scenes where minimal information is available. This result extends the current state-of-the-art in single view geometry techniques to situations where only one vanishing point is available. The property of constant or known camera motion is also described in this dissertation for applications such as calibration of a network of cameras in video surveillance systems, and Euclidean reconstruction from turn-table image sequences in the presence of zoom and focus. We then propose a new framework for the estimation and alignment of camera motions, including both simple (panning, tracking and zooming) and complex (e.g. hand-held) camera motions. Accuracy of these results is demonstrated by applying our approach to video post-production applications such as video cut-and-paste and shadow synthesis. As realistic image-based rendering problems, these applications require extreme accuracy in the estimation of camera geometry, the position and the orientation of the light source, and the photometric properties of the resulting cast shadows. In each case, the theoretical results are fully supported and illustrated by both numerical simulations and thorough experimentation on real data.

The low-temperature joining technique (LTJT) by silver sintering is being implemented by major manufacturers of power electronics devices and modules for bonding power semiconductor chips. A common die-attach material used with LTJT is a silver paste consisting of silver powder (micron- or nano-size particles) mixed in organic solvent and binder formulation. It is believed that the drying of the paste during the bonding process plays a critical role in determining the quality of the sintered bond-line. In this study, a model based on the diffusion of solvent molecules and viscous mechanics of the paste was introduced to determine the stress and strain states of the silver bond-line. A numerical simulation algorithm of the model was developed and coded in the C++ programming language. The numerical simulation allows determination of the time-dependent physical properties of the silver bond-line as the paste is being dried with a heating profile. The properties studied were solvent concentration, weight loss, shrinkage, stress, and strain. The stress is the cause of cracks in the bond-line and bond-line delamination. The simulated results were verified by complementary experiments in which the formation of cracks in bond-line and interface delamination was observed during the pressure-free drying of a die-attach nanosilver paste. Furthermore, the important drying parameters, such as drying pressure, low temperature drying time and temperature ramp rate of nanosilver LTJT process, are experimentally studied and analyzed with the numerical simulation. The simulated results were consistent with the experimental findings that the quality of sintered silver bond-line increases with increasing external drying pressure, with increasing low temperature drying time, and with decreasing temperature ramp rate. The insight offered by this modeling study can be used to optimize the process profile that enable pressure-free, low-temperature sintering of the die-attach material to significantly lower the cost of implementing the LTJT in manufacturing. / Ph. D.