Master of Science, University of Toledo, 2019, Bioengineering

Musculoskeletal tissue injuries affect around 1 in 3 Americans and 1.7 billion people worldwide. This is a huge economic burden, costing an estimated $120 billion in the US alone. With limited success from surgery or subcutaneous injections of medicine, where only temporary relief or complications can occur, alternative measures should be explored. Injectable biologically-loaded hydrogels are one avenue and act as drug delivery systems. They provide a minimally invasive approach to release biologics in a sustained and controlled manner to provide long-lasting relief without toxic effects and with less risk of surgical complications. In this study, the immunological application of a previously-developed nanofibrous PCL-interspersed collagen hydrogel, (PNCOL) was explored by loading PNCOL with the cytokine IL-4 and identifying its effect upon macrophages. Furthermore, the effect of a simulated digestive inflammatory environment (DIE) had upon protein release kinetics as well as scaffold integrity were characterized. Genipin crosslinking was then explored to improve scaffold resistance to degradation, and an optimal genipin concentration was identified to impart sufficient scaffold crosslinking, increased mechanical strength, and a prolonged release profile, with minimal cytotoxic effects. Lastly, the immunomodulatory effect of IL-4 released from crosslinked and uncrosslinked scaffolds were investigated through identifying the impact of IL-4 on macrophage differentiation. The IL-4 released from PNCOL polarized macrophages toward an anti-inflammatory, pro-healing state, while genipin crosslinking with and without IL-4's presence appeared to lower macrophage activity.

Committee: Eda Yildirim-Ayan PhD (Advisor); Eda Yildirim-Ayan PhD (Committee Chair); Halim Ayan PhD (Committee Member); Arun Nadarajah PhD (Committee Member) Subjects: Biology; Biomedical Engineering; Biomedical Research; Cellular Biology; Materials Science

Doctor of Philosophy, The Ohio State University, 2022, Pharmaceutical Sciences

In recent decades, lipid-based nanovesicles (LNPs) have been developed for the therapeutic delivery of chemotherapeutics nucleic acid therapeutics respectively. Liposomes can be used to modulate the pharmacokinetics of drug molecules and change their tissue biodistribution. This can lead to increased therapeutic efficacy and reduced systemic toxicity, which are key to the clinical application of anticancer drugs. Bortezomib (BTZ) has been used for the treatment of multiple myeloma for the last 20 years. BTZ has a narrow therapeutic window, which limits its clinical application. To improve the pharmacokinetics of BTZ, a novel liposomal formulation based on remote loading driven by a Tiron gradient was developed. The AUC of liposomal BTZ was shown to be much higher than that of free BTZ. The anti-tumor effect of liposomal BTZ was shown in murine xenograft tumor models (Chapter 2). Nucleic acids are more challenging to deliver into cells due to their high molecular weight and high electrical charge compared to small molecular components. However, conventional liposomes have limited capacity in the delivery of nucleic acids, with low encapsulation efficiency, poor cellular uptake, and lack of endosomal escape. LNPs containing cationic and/or ionizable lipids have been developed for RNA delivery. Lipid nanoparticles consisting of ionizable lipids can entrap nucleic acids by electrostatic self-assembly. Typically, LNPs are composed of a phospholipid, a cationic or an ionizable lipid, cholesterol and PEG-lipid. LNP encapsulation can prevent RNA degradation by RNase in plasma and improve intracellular delivery of RNA through endosomal escape. Ionizable lipid-based LNPs have been found to be more effective in RNA delivery in vivo, exemplified by the FDA approval of drugs such as Patisiran and the two COVID-19 vaccines. For further improvement of delivery efficiency and reduction of the liver distribution of nucleic acids after IM injections, a new formulation of LNP compo (open full item for complete abstract)

Committee: Robert Lee (Advisor); Emanuele Cocucci (Committee Member); Dan Shu (Committee Member); Yizhou Dong (Committee Member) Subjects: Pharmaceuticals

Master of Science in Pharmaceutical Science (MSP), University of Toledo, 2021, Pharmaceutical Sciences (Industrial Pharmacy)

Self-micro-emulsifying drug delivery systems (SMEDDS) have been proven to have improved drug stability, lower toxicity, and increase bioavailability of insoluble drugs. It is a drug delivery design that can prevent physical and chemical drug degradation. The goal of this study was to develop a solid formulation incorporating a self-micro-emulsifying drug delivery system (SMEDDS) for the oral delivery of Monodora myristica essential oil (MMEO). MMEO was extracted from the blended seeds of Monodora myristica using the hydro-distillation method. MMEO was characterized by evaluating the physicochemical properties to ascertain the quality and purity of the essential oil by comparing with MMEO data in the literature. The design of the experiment was done by using Fusion Pro by S-Matrix (Fusion Pro Software Version 9.9.0 Build690, S-Matrix Corporation (www.smatrix.com)) to compare a combination of MMEO/Tween 80/Transcutol HP and MMEO/ Kolliphor/ Labrasol 12 formulations. MMEO (10.92%) / Tween 80 (48%) /Transcutol HP (41.8%) was predicted to be the best formulation with desirable characteristics such as a mean particle size of 112.7 nm, the zeta potential of +5.10 mv, and a transparent emulsion. The emulsion formed was stable over 90 days without any form of emulsion instability or oil precipitation. The liquid-SMEDDS was adsorbed unto Neusilin US2 to form solid-SMEDDS. The solid-SMEDDS was added to cellulose, lactose, starch, talc, magnesium stearate to directly compress type 1 and type 2 tablets while the solid-SMEDDS was directly compressed to formulate type 3 tablets. Type 3 tablets had the highest drug loading capacity unlike type 1 and type 2 tablets. Also, type 3 had the highest breaking force and longest disintegration time. Using one-way ANOVA, the P-value obtained was below 0.05 for tablet thickness, tablet breaking force, and disintegration tests. Therefore, there was a statistically significant difference between type 1, type 2, and type 3 tablets properties su (open full item for complete abstract)

Committee: Jerry Nesamony Dr (Committee Chair); Gabriella Baki Dr (Committee Member); Liyanaaratchige Tillekeratne Dr (Committee Member) Subjects: Health Sciences; Intellectual Property

Doctor of Philosophy, University of Akron, 2019, Chemical Engineering

In the study of this Ph.D. dissertation, two research topics related to zwitterionic materials have been investigated. Even though the applications have different objectives, the unique properties that existed in zwitterionic materials have, including charge property and antifouling property, have been used in both of research. The first topic is the development of zwitterionic-peptides gene delivery system. The gene delivery system, with high efficacy, low toxicity, long blood circulation time and targeting the specific cancer cell, is investigated. The second topic is the functionalization of protein therapeutics with zwitterionic polymers. The protein therapeutics with better solubility, stability, and activity is developed. The non-viral gene delivery system is under research due to their low toxicity, low immunogenic and large DNA loading size in gene therapy. Peptides gene delivery system is reported with the cationic charge and buffering effect which overcomes the barrier and delivery DNA into the nucleus. In our group, the economic dextran-peptide hybrid gene delivery system was developed with high transfection efficiency and low toxicity. The first topic of my research was expanded as a continuous work under the same research interest. The effect of the design of peptides length, zwitterionic group and targeting group was studied for the optimization objects on achieving low toxicity, transfection efficiency and blood circulation time, which was summarized into three research projects under this topic. The system was adjusted by the peptides length for toxicity and economic purpose. The system was functionalized with the zwitterionic group for improved stability, enhanced endosomal escape and longer blood circulation time. The system was also conjugating with targeting ligand for targeting gene delivery. It was found that the shorter length of peptides will not provide enough charge to form stable micelle with report DNA. The zwitterionic functionalized (open full item for complete abstract)

Committee: Gang Cheng (Advisor); Jie Zheng (Advisor); George Chase (Committee Member); Lingyun Liu (Committee Member); Ge Zhang (Committee Member); Coleen Pugh (Committee Member) Subjects: Biomedical Engineering; Chemical Engineering; Polymers

Master of Science in Chemical Engineering, Cleveland State University, 2015, Washkewicz College of Engineering

Elastin-like polypeptides (ELPs) are a class of biopolymers with the potential to function as a novel drug delivery platform. These protein based polymers are composed of the repeating pentapeptide sequence (GαGβP)n where n is the number of pentapeptide repeats while α and β are guest amino acid residues. ELP constructs have been designed to respond to various external stimuli including temperature, pH, and ionic strength where their response to these stimuli results in the separation of the ELP from solution. This phase separation results in a two-phase system consisting of a protein poor supernatant phase and a protein rich coacervate phase. Under certain conditions select ELP constructs are able to self assemble into micellar structures of nanometer scale when raised above their transition temperature. The micellar architecture consists of an inner hydrophobic core, with a composition like that of the protein rich coacervate phase, surrounded by hydrophilic head groups. For the use of ELP micelles as a drug delivery platform these particles should possess the ability to encapsulate solute molecules. In this study, solute solubility within the micelle core was investigated by measuring the partition coefficients of several solutes in five different ELP two-phase systems, then all data was fit to a linear free energy relationship (LFER) model to provide insight into the dominate interactions governing solute partitioning in ELP systems. From the LFER it is shown that the cavity formation energy, solute size, and the solvents hydrogen bond acidity are important parameters governing solute partitioning in the ELP solvents investigated. Additionally, the partition coefficients provide a measurement of ELP phase hydrophobicity from which the development of future ELP constructs is possible.

Committee: Nolan B Holland PhD (Advisor); Rolf Lustig PhD (Committee Member); David Anderson PhD (Committee Member) Subjects: Analytical Chemistry; Chemical Engineering; Chemistry; Engineering; Pharmaceuticals; Polymer Chemistry; Polymers

MS, University of Cincinnati, 2015, Pharmacy: Pharmaceutical Sciences

Stimulus-induced drug delivery systems and nanotechnology allow selective targeting of disease-affected tissues in order to improve pharmacokinetic properties of therapeutics while minimizing adverse events associated with systemic exposure. The objective of this research was to explore Eudragit L-100, a pH sensitive co-polymer comprised of methacrylic acid and methyl methacrylate, in the fabrication of nanoparticles (NPs) that efficiently accumulate in alveolar macrophages and preferentially release the payload in Mycobacteria tuberculosis (TB)-infected lysosomes. The lipophilic fluorescent dye Rhodamine123 (Rh123) was selected as a surrogate for the anti-TB agent rifampicin. Eudragit L-100 NPs were prepared using the nanoprecipitation technique, which afforded colloids with a hydrodynamic diameter between 238-270 nm and a zeta potential ranging from –21.8 to –33.1 mV. Subsequent in vitro release experiments performed in phosphate-buffered saline at pH 5.5, 6.2 and 7.4 demonstrated pH-dependent release kinetics of Rh123 from NPs. In comparison to pH 5.5, the cumulative amount of Rh123 released was approximately 3-fold greater at pH 6.2 and 6-fold increased at pH 7.4. Regression analyses using various mathematical models suggested predominant contribution of passive diffusion in Rh123 release at pH 5.5, with increasing assistance of carrier erosion at pH 6.2 and 7.4. Incubation of the mouse alveolar macrophage (MH-S) cell line with Rh123-containing NPs revealed a 50% decrease in cellular viability at NP concentrations of 2.8 mg/mL. Consequently, cellular uptake experiments were performed using a NP concentration of 1.62 mg/mL where viability > 99.9% was guaranteed. To simulate less acidic lysosomal pH conditions of TB-infected macrophages, MH-S cells were exposed to NH4Cl concentrations up to 100 mM. Only the highest NH4Cl concentration increased the lysosomal pH of MH-S cells from resting pH 5.5 to the desired pH 6.2. Cellular uptake of Rh123-containing Eudragit L- (open full item for complete abstract)

Committee: Giovanni Pauletti Ph.D. (Committee Chair); Gary Thompson Ph.D. (Committee Member); Kevin Li Ph.D. (Committee Member) Subjects: Pharmaceuticals

Doctor of Philosophy, The Ohio State University, 2009, Pharmacy

Among various drug delivery systems, nanoparticles have shown some unique advantages. In this dissertation, a series of lipid and polymer-based nanoparticle systems were designed and prepared for the objective of improved drug delivery efficiencies and enhanced therapeutic efficacies. The development of nanoparticle formulation for nucleic acid drugs are described in Chapter 2, 3 and 4. In Chapter 2, a lipid-based, transferrin receptor (TfR)-targeted nanoparticle formulation containing protamine (Tf-LN) was developed to deliver antisense oligodeoxynucelotide G3139 against Bcl-2 to leukemia cells. Compared to free G3139 and non-targeted formulation (LN), Tf-LN showed increased cellular uptake and enhanced target gene downregulation. In Chapter 3, the effects of different components and composition on green fluorescence protein (GFP) gene delivery efficiency were investigated and provided useful information for further development of PEI and lipid-containing nanocrystal formulation of G3139 in Chapter 4. Chapter 4 described the development of a novel lipid-based nanoparticle formulation containing cationic lipid, PEI2000 and calcium, designated as nanocyrstal (NC), for delivery of antisense oligonucelotide G3139 to KB cells. ODN G3139 delivery by NC resulted in much higher cellular uptake and target gene downregulation in vitro. However, the downregulation was not observed in treated mice tumor, suggesting the other unknown factors in vivo may affect the antisense effect of G3139 nanocrystal. In this dissertation, we also developed nanoparticle delivery systems for chemotherapy drugs. In Chapter 5, a liposomal formulation of flavopiridol was developed to address the issues of solubility, high plasma protein-binding and side effects. Pharmacokinetic study in mice after i.v. bolus injection showed that the liposomal flavopiridol had an increased elimination phase half-life, decreased clearance and increased area under the plasma concentration-time curve compared to the (open full item for complete abstract)

Committee: Robert Lee (Advisor); Guido Marcucci (Committee Member); Kenneth Chan (Committee Member); L. James Lee (Committee Member) Subjects: Pharmaceuticals

Master of Science in Engineering, University of Akron, 2007, Biomedical Engineering

Dialysis access graft failure is a major complication in providing care to patients on hemodialysis therapy. The failure rates have been reported as high as 80% at one year for this procedure. The major cause of failure is intimal hyperplasia. Intimal hyperplasia is an exaggeration of the normal vascular wall healing response to injury resulting from the migration and proliferation of medial smooth muscle cells. To mitigate this disease condition, we developed a perivascular polymeric controlled drug delivery device which we hypothesized when applied locally, would be effective in preventing intimal hyperplasia by enabling a sustained release of the therapeutic agent to the anastamotic site. This novel polymeric device, named PolyRing, is a composite system consisting of poly (DL-lactide-co-glycolide) microspheres embedded in a poly (ethylene glycol) hydrogel. The drug for the treatment of intimal hyperplasia is encapsulated within the poly (DL-lactide-co-glycolide) microspheres. The device is snapped into place around the vessel to deliver the drug. This work focuses on evaluating the feasibility of releasing the drug Simvastatin from the device to provide localized, site specific, sustained drug delivery for the prevention of intimal hyperplasia in vascular tissue. Simvastatin, obtained in a pro-drug form, when hydrolyzed to its active form, Simvastatin Acid, acts as a potent competitive inhibitor of the 3-Hydroxy-3-methylglutaryl coenzyme A reductase. The inhibition of this enzyme suppresses the mevalonate pathway and thereby prevents the proliferation of smooth muscle cells. As the focus is on controlled localized delivery, we hypothesize the need of using the active form as against the lactone form. Therefore, we fabricated Simvastatin and Simvastatin Acid loaded poly (DL-lactide-co-glycolide) microspheres using oil-water and water-oil-water techniques respectively. The oil-water emulsion resulted in smooth surfaced microspheres (determined by Scanning Electron (open full item for complete abstract)

Committee: Stephanie Lopina (Advisor) Subjects: