Gloria J. Kim

Nanotechnology in cancer therapeutics: bioconjugated nanoparticles for drug delivery

Nanotechnology refers to the interactions of cellular and molecular components and engineered materials—typically, clusters of atoms, molecules, and molecular fragments into incredibly small particles—between 1 and 100 nm. Nanometer-sized particles have novel optical, electronic, and structural properties that are not available either in individual molecules or bulk solids. The concept of nanoscale devices has led to the development of biodegradable self-assembled nanoparticles, which are being engineered for the targeted delivery of anticancer drugs and imaging contrast agents. Nanoconstructs such as these should serve as customizable, targeted drug delivery vehicles capable of ferrying large doses of chemotherapeutic agents or therapeutic genes into malignant cells while sparing healthy cells. Such “smart” multifunctional nanodevices hold out the possibility of radically changing the practice of oncology, allowing easy detection and then followed by effective targeted therapeutics at the earliest stages of the disease. In this article, we briefly discuss the use of bioconjugated nanoparticles for the delivery and targeting of anticancer drugs. [Mol Cancer Ther 2006;5(8):1909–17]

HFT-T, a Targeting Nanoparticle, Enhances Specific Delivery of Paclitaxel to Folate Receptor-Positive Tumors

Nonspecific distribution of chemotherapeutic drugs (such as paclitaxel) is a major factor contributing to side effects and poor clinical outcomes in the treatment of human head and neck cancer. To develop novel drug delivery systems with enhanced efficacy and minimized adverse effects, we synthesized a ternary conjugate heparin-folic acid-paclitaxel (HFT), loaded with additional paclitaxel (T). The resulting nanoparticle, HFT-T, is expected to retain the antitumor activity of paclitaxel and specifically target folate receptor (FR)-expressing tumors, thereby increasing the bioavailability and efficacy of paclitaxel. In vitro experiments found that HFT-T selectively recognizes FR-positive human head and neck cancer cell line KB-3-1, displaying higher cytotoxicity compared to the free form of paclitaxel. In a subcutaneous KB-3-1 xenograft model, HFT-T administration enhanced the specific delivery of paclitaxel into tumor tissues and remarkably improved antitumor efficacy of paclitaxel. The average tumor volume in the HFT-T treatment group was 92.9 +/- 78.2 mm(3) vs 1670.3 +/- 286.1 mm(3) in the mice treated with free paclitaxel. Furthermore, paclitaxel tumors showed a resurgence of growth after several weeks of treatment, but this was not observed with HFT-T. This indicates that HFT-T could be more effective in preventing tumors from developing drug resistance. No significant acute in vivo toxicity was observed. These results indicate that specific delivery of paclitaxel with a ternary structured nanoparticle (HFT-T) targeting FR-positive tumor is a promising strategy to enhance chemotherapy efficacy and minimize adverse effects.

Multiresponse strategies to modulate burst degradation and release from nanoparticles.

Logic gate nanoparticles, where two chemical transformations take place one after the other, were successfully formulated from a newly synthesized random co-polymer. This polymer, poly([2,2′-(propane-2,2-diylbis(oxy))bis(ethane-2,1-diyl) diacrylate ]-co-[hexane-1,6-diyl diacrylate]-4,4′ trimethylene dipiperidine), (poly-β-aminoester ketal-2) contains two pH responsive moieties within its backbone. As nanoparticles they function akin to an AND logic gate. The β-aminoester backbone moiety provides a pH triggered solubility switch, only when this switch is “ON” does the ketal moiety also turn “ON” to undergo rapid acid catalyzed hydrolysis. These AND logic gate polymeric nanoparticles were prepared using an oil in water emulsion method. Their degradation in the pH range of 7.4−5 was monitored by dynamic light scattering and showed excellent stability at pH 7.4 and rapid degradation at pH 5. Our results indicate that the prepared logic gate nanoparticles may prove valuable in delivering therapeutics and diagnostics to cells and diseased tissue.

Inflammation responsive logic gate nanoparticles for the delivery of proteins

Oxidative stress and reduced pH are important stimuli targets for intracellular delivery and for delivery to diseased tissue. However, there is a dearth of materials able to deliver bioactive agents selectively under these conditions. We employed our recently developed dual response strategy to build a polymeric nanoparticle that degrades upon exposure to two stimuli in tandem. Our polythioether ketal based nanoparticles undergo two chemical transformations; the first is the oxidation of the thioether groups along the polymer backbone of the nanoparticles upon exposure to reactive oxygen species (ROS). This transformation switches the polymeric backbone from hydrophobic to hydrophilic and thus allows, in mildly acidic environments, the rapid acid-catalyzed degradation of the ketal groups also along the polymer backbone. Dynamic light scattering and payload release studies showed full particle degradation only in conditions that combined both oxidative stress and acidity, and these conditions led to higher release of encapsulated protein within 24 h. Nanoparticles in neutral pH and under oxidative conditions showed small molecule release and swelling of otherwise intact nanparticles. Notably, cellular studies show absence of toxicity and efficient uptake of nanoparticles by macrophages followed by cytoplasmic release of ovalbumin. Future work will apply this system to inflammatory diseases.

Poly(d,llactide–co-ethyl ethylene phosphate)s as new drug carriers

Many biodegradable polymers have been developed for controlled drug delivery. The plethora of drug therapies and types of drugs demand different formulations, fabrications conditions and release kinetics. No one single polymer can satisfy all the requirements. To extend the properties of poly(D,L-lactide) (PLA), we synthesized copolymers of PLA and poly(ethylethylene phosphate) (PEEP) by ring-opening polymerization using Al(Oipr)3 as the initiator. The copolymers were structurally characterized by IR and 1H NMR. DSC data confirmed the formation of random microphase structure in all the copolymers, and showed a decrease of Tg from 43.2 to -22.6 degrees C when the molar content of ethylethylene phosphate (EEP) increased from 5 to 40%. The hydrophilicity of the copolymers increased with EEP content. In contrast to the degradation behavior of PLA, disc samples made of PLAEEP90 showed a linear weight loss profile in PBS (pH 7.4) at 37 degrees C. BSA microspheres using PLAEEP90 were prepared by double-emulsion method, yielding a loading level of 4.3% and a loading efficiency of 75%. The BSA release profile consisted of an initial burst (9%) on the first day, followed by a daily 4% release for the following 40 days, resulting in 91% of the BSA release in a near linear manner. The released BSA remained intact according to SDS-PAGE data. Cytotoxicity and histopathology studies showed low toxicity in HeLa cells and good tissue biocompatibility in mouse brain, respectively. PLAEEP is a promising biodegradable polymer for controlled drug delivery.

Targeted cancer nanotherapy

Significant progress has been made in the development of new agents against cancer and new delivery technologies. Proteomics and genomics continue to uncover molecular signatures that are unique to cancer. Yet, the major challenge remains in targeting and selectively killing cancer cells while affecting as few healthy cells as possible. Nanometer-sized particles have novel optical, electronic, and structural properties that are not available from either individual molecules or bulk solids. When linked with tumor-targeting moieties, such as tumor-specific ligands or monoclonal antibodies, these nanoparticles can be used to target cancer-specific receptors, tumor antigens (biomarkers), and tumor vasculatures with high affinity and precision.

Perspective on Flipping Circuits I

A flipped-classroom approach was implemented in a Circuits I class for electrical and computer engineering majors to lower its high attrition and failure rate. Students were asked to watch online lectures and then come to class prepared to work problems in small groups of four. The attitude, retention, and performance of students in the flipped group in Spring 2013 were compared to those for the traditionally taught group in Fall 2012. The Fall 2012 lectures were recorded, so that each group saw the same lectures. Student retention and test performance was significantly higher in the flipped course. In Fall 2012, 56% of the initially enrolled students received a C or better. In Spring 2013, this improved to 83%. The first exam scores were significantly better in Spring 2013, and this helped with student success. The authors believe that it was the alignment of online lectures, face-to-face student/teacher and peer/peer interactions, combined with the active learning component of the flipped classroom that led to these improvements .

Fabrication of an all SU-8 electrospun nanofiber based supercapacitor

Supercapacitors (SCs) as energy storage devices are advantageous in their rapid charge/discharge capabilities and their immense charge storage capacity. Two important components of a SC are the electrically conductive electrodes (anode and cathode) and an electrically non-conductive separator between the two electrodes. This paper details a fabrication process for nanofibrous carbon electrodes and a nanoporous polymer separator using all SU-8 based electrospinning and post electrospinning processes, such as lithographical patterning, conversion of the nanofibrous polymer to carbon structures using heat treatment (carbonization) and their assembly to complete a SC. The process produces immensely porous electrodes with good conductivity; it is scalable and economical compared with the carbon nanotube electrode approach. High throughput tube nozzle electrospinning for nanofiber (NF) production and its photolithographical patterning have been employed to facilitate manufacturability. The dependence of the NF morphology on the carbonization temperatures is studied. Also, SC testing and characterization are discussed.

Fabrication of nanoporous membrane and its nonlithographic patterning using Electrospinning and Stamp-thru-mold (ESTM)

The Electrospinning and Stamp-thru-mold (ESTM) technique, an integrated fabrication process which incorporates the versatility of the electrospinning process for nanofiber fabrication with the non-lithographic patterning ability of the stamp-thru-mold process is introduced. In-situ multilayer stacking of orthogonally aligned nanofibers, ultimately resulting in a nanoporous membrane, has been demonstrated using orthogonally placed collector electrode pairs and an alternating bias scheme. The pore size of the nanoporous membrane can be controlled by the number of layers and the deposition time of each layer. Nonlithographic patterning of the fabricated nanoporous membrane is then performed by mechanical shearing using a pair of pre-fabricated micromolds. This patterning process is contamination free compared to other photo lithographical patterning approaches. The ability to pattern on different substrates has been tested with and without oxygen plasma surface treatment. In vitro tests of ESTM poly-lactic-co-glycolic acid (PLGA) nanofibers verify the biocompatibility of this process. Simulation by the COMSOL Multiphysics tool has been conducted for the analysis of electrospun nanofiber alignment.

Fabrication of carbon nanofibrous microelectrode array (CNF-MEA) using nanofiber immersion photolithography

Microelectrode arrays (MEAs) are widely used for stimulating and receiving electrical signals between human and machines and for in vitro neural study. This work demonstrates the fabrication process of nanofibrous 3D microelectrodes using immersion lithography. Oil immersion negates the diffraction effects intrinsic in the photopatterning of electrospun nanofibers to give increased aspect ratio microarchitectures. Nanofiber electrode resistivity is characterized and its performance compared to that of carbon thin film. In vitro testing of electrodes are performed using E18 cortical neurons and analyzed for cell density and cell viability.