Functionalized Conjugated Polymers for Signal Amplifying Biosensors and Sensor Arrays.

Conjugated polymers (CPs) are great alternatives to the conventional fluorescence dyes as signaling reporters in biosensor design due to the fluorescent signal amplification property of CPs. Two series of CPs, poly(p-phenyleneethynylene) (PPE) and poly(poxadiazole- co-phenylene) (POX) derivatives, have been systematically designed, developed, and studied in this thesis to devise highly sensitive and selective novel molecular biosensors and sensor arrays for the detection of clinically important biological molecules. The key concept developed in the thesis work was the molecular design principles to combine biological receptor molecules for specific detection of target oligonucleotides and CPs as the signal transduction and amplification unit. To achieve this goal, a series of completely water-soluble and highly emissive conjugated polyelectrolytes (CPEs) were first developed through systematic investigation on the xxii correlation between the polymer structure and its water-solubility. We also developed a method to bioconjugate CPEs to peptides and DNA by end-modification of the CPEs with a carboxylic acid group to develop hybrid bio/-synthetic sensory CPs and to achieve selective detection of target with amplified fluorescence signal in aqueous solution. DNA detection results using the CPE-DNA hybrid system confirmed large signal amplification by means of efficient Förster energy transfer from the energy harvesting CPEs to the fluorescent dye attached to the complementary analyte DNA. To apply the signal amplification scheme to practically more useful solid-state microarray novel conjugated polymers, POXs, having unique photochemical stabilities were developed. By applying on-chip DNA synthesis on the POXs and achieving efficient Förster energy transfer from POXs to the dye-labeled target DNA we successfully developed signal amplifying DNA microarrays. The signal amplifying scheme was combined with a self-signaling concept by means of introducing intercalating dyes and molecular beacon into the CPs for labelfree detection. As a result of sensitive and selective prostate specific antigen detection has been demonstrated. In addition to the biosensor development, the developed bioconjugation technique between biological molecules and CPEs was uniquely applied to the development of CPE-antibody for live cell imaging. Selective live cell imaging of human B-cell lymphoma and human T-cell leukemia having largely enhanced sensitivity and excellent selectivity was demonstrated by using the CPE-antibody.Ph.D.Materials Science and EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/60834/1/kangwonl_1.pd

Conjugated Polymers Combined with a Molecular Beacon for Label-Free and Self-Signal-Amplifying DNA Microarrays

A conjugated polymer (CP) and molecular-beacon-based solid-state DNA sensing system is developed to achieve sensitive, label-free detection. A novel conjugated poly(oxadiazole) derivative exhibiting amine and thiol functional groups ( POX-SH ) is developed for unique chemical and photochemical stability and convenient solid-state on-chip DNA synthesis. POX-SH is soluble in most nonpolar organic solvents and exhibits intense blue fluorescence. POX-SH is covalently immobilized onto a maleimido-functionalized glass slide by means of its thiol group. Molecular beacons having a fluorescent dye or quencher molecule as the fluorescence resonance energy transfer (FRET) acceptor are synthesized on the immobilized POX-SH layer through direct on-chip oligonucleotide synthesis using the amine side chain of POX-SH . Selective hybridization of the molecular beacon probes with the target DNA sequence opens up the molecular beacon probes and affects the FRET between POX-SH and the dye or quencher, producing a sensitive and label-free fluorescence sensory signal. Various molecular design parameters, such as the size of the stem and loop of the molecular beacon, the choice of dye, and the number of quencher molecules are systematically controlled, and their effects on the sensitivity and selectivity are investigated.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/64307/1/3317_ftp.pd

Suppression of magnetic ordering in XXZ-type antiferromagnetic monolayer NiPS3

How a certain ground state of complex physical systems emerges, especially in two-dimensional materials, is a fundamental question in condensed-matter physics. A particularly interesting case is systems belonging to the class of XY Hamiltonian where the magnetic order parameter of conventional nature is unstable in two-dimensional materials leading to a Berezinskii-Kosterlitz-Thouless transition. Here, we report how the XXZ-type antiferromagnetic order of a magnetic van der Waals material, NiPS3, behaves upon reducing the thickness and ultimately becomes unstable in the monolayer limit. Our experimental data are consistent with the findings based on renormalization group theory that at low temperatures a two-dimensional XXZ system behaves like a two-dimensional XY one, which cannot have a long-range order at finite temperatures. This work provides experimental examination of the XY magnetism in the atomically thin limit and opens new opportunities of exploiting these fundamental theorems of magnetism using magnetic van der Waals materials.Comment: 57 pages, 24 figures (including Supplementary Information

Bone regeneration via novel macroporous CPC scaffolds in critical-sized cranial defects in rats

Objectives. Calcium phosphate cement (CPC) is promising for dental and craniofacial applications due to its ability to be injected or filled into complex-shaped bone defects and molded for esthetics, and its resorbability and replacement by new bone. The objective of this study was to investigate bone regeneration via novel macroporous CPC containing absorbable fibers, hydrogel microbeads and growth factors in critical-sized cranial defects in rats. Methods. Mannitol porogen and alginate hydrogel microbeads were incorporated into CPC. Absorbable fibers were used to provide mechanical reinforcement to CPC scaffolds. Six CPC groups were tested in rats: (1) control CPC without macropores and microbeads; (2) macroporous CPC + large fiber; (3) macroporous CPC + large fiber + nanofiber; (4) same as (3), but with rhBMP2 in CPC matrix; (5) same as (3), but with rhBMP2 in CPC matrix + rhTGF-beta 1 in microbeads; (6) same as (3), but with rhBMP2 in CPC matrix + VEGF in microbeads. Rats were sacrificed at 4 and 24 weeks for histological and micro-CT analyses. Results. The macroporous CPC scaffolds containing porogen, absorbable fibers and hydrogel microbeads had mechanical properties similar to cancellous bone. At 4 weeks, the new bone area fraction (mean +/- sd; n = 5) in CPC control group was the lowest at (14.8 +/- 3.3)%, and that of group 6 (rhBMP2 + VEGF) was (31.0 +/- 13.8)% (p < 0.05). At 24 weeks, group 4 (rhBMP2) had the most new bone of (38.8 +/- 15.6)%, higher than (12.7 +/- 5.3)% of CPC control (p < 0.05). Micro-CT revealed nearly complete bridging of the critical-sized defects with new bone for several macroporous CPC groups, compared to much less new bone formation for CPC control. Significance. Macroporous CPC scaffolds containing porogen, fibers and microbeads with growth factors were investigated in rat cranial defects for the first time. Macroporous CPCs had new bone up to 2-fold that of traditional CPC control at 4 weeks, and 3-fold that of traditional CPC at 24 weeks, and hence may be useful for dental, craniofacial and orthopedic applications. (C) 2014 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved

Acute Hemorrhagic Conjunctivitis caused by Coxsackievirus A24 Variant, South Korea, 2002

In summer 2002, a nationwide outbreak of acute hemorrhagic conjunctivitis occurred in South Korea. The etiologic agent was confirmed as coxsackievirus A24 variant (CA24v) by virus isolation and sequencing of a part of the VP1 gene. Phylogentic analysis, based on the protease 3C sequences, showed that the Korean isolates were clustered into a lineage distinct from the CA24v isolates reported in previous outbreaks in Asia

Matrix elasticity of void-forming hydrogels controls transplanted-stem-cell-mediated bone formation

The effectiveness of stem cell therapies has been hampered by cell death and limited control over fate. These problems can be partially circumvented by using macroporous biomaterials that improve the survival of transplanted stem cells and provide molecular cues to direct cell phenotype. Stem cell behaviour can also be controlled in vitro by manipulating the elasticity of both porous and non-porous materials, yet translation to therapeutic processes in vivo remains elusive. Here, by developing injectable, void-forming hydrogels that decouple pore formation from elasticity, we show that mesenchymal stem cell (MSC) osteogenesis in vitro, and cell deployment in vitro and in vivo, can be controlled by modifying, respectively, the hydrogel’s elastic modulus or its chemistry. When the hydrogels were used to transplant MSCs, the hydrogel’s elasticity regulated bone regeneration, with optimal bone formation at 60 kPa. Our findings show that biophysical cues can be harnessed to direct therapeutic stem cell behaviours in situ

Matrix elasticity of void-forming hydrogels controls transplanted-stem-cell-mediated bone formation

The effectiveness of stem cell therapies has been hampered by cell death and limited control over fate. These problems can be partially circumvented by using macroporous biomaterials that improve the survival of transplanted stem cells and provide molecular cues to direct cell phenotype. Stem cell behaviour can also be controlled in vitro by manipulating the elasticity of both porous and non-porous materials, yet translation to therapeutic processes in vivo remains elusive. Here, by developing injectable, void-forming hydrogels that decouple pore formation from elasticity, we show that mesenchymal stem cell (MSC) osteogenesis in vitro, and cell deployment in vitro and in vivo, can be controlled by modifying, respectively, the hydrogel's elastic modulus or its chemistry. When the hydrogels were used to transplant MSCs, the hydrogel's elasticity regulated bone regeneration, with optimal bone formation at 60 kPa. Our findings show that biophysical cues can be harnessed to direct therapeutic stem cell behaviours in situ