Hongliang Cao

Single Cyclized Molecule Versus Single Branched Molecule: A Simple and Efficient 3D “Knot” Polymer Structure for Nonviral Gene Delivery

The large research effort focused on enhancing nonviral transfection vectors has clearly demonstrated that their macromolecular structure has a significant effect on their transfection efficacy. The 3D branched polymeric structures, such as dendrimers, have proved to be a very effective structure for polymeric transfection vectors; however, so far the dendritic polymers have not delivered on their promise. This is largely because a wide range of dendritic polymer materials with tailored multifunctional properties and biocompatibility required for such applications are not yet accessible by current routes. Herein, we report the design and synthesis of new 3D Single Cyclized polymeric gene vectors with well-defined compositions and functionalities via a one-step synthesis from readily available vinyl monomers. We observe that this polymer structure of a single chain linked to itself interacts differently with plasmid DNA compared to conventional vectors and when tested over a range of cell types, has a superior transfection profile in terms of both luciferase transfection capability and preservation of cell viability. This new knotted structure shows high potential for gene delivery applications through a combination of simplicity in synthesis, scalability, and high performance.

3D Single Cyclized Polymer Chain Structure from Controlled Polymerization of Multi-Vinyl Monomers: Beyond Flory–Stockmayer Theory

Controlled/living radical polymerization (CRP) is a widely used technique that allows the synthesis of defined polymer architectures through precise control of molecular weights and distributions. However, the architectures of polymers prepared by the CRP techniques are limited to linear, cross-linked, and branched/dendritic structures. Here, we report the preparation of a new 3D single cyclized polymer chain structure from an in situ deactivation enhanced atom transfer radical polymerization of multivinyl monomers (MVMs), which are conventionally used for the production of branched/cross-linked polymeric materials as defined by P. Flory and W. Stockmayer nearly 70 years ago. We provide new evidence to demonstrate that it is possible to kinetically control both the macromolecular architecture and the critical gelling point in the polymerization of MVMs, suggesting the classical Flory-Stockmayer mean field theory should be supplemented with a new kinetic theory based on the space and instantaneous growth boundary concept.

Thermoresponsive hyperbranched copolymer with multi acrylate functionality for in situ cross-linkable hyaluronic acid composite semi-IPN hydrogel

Thermoresponsive polymers have been widely used for in situ formed hydrogels in drug delivery and tissue engineering as they are easy to handle and their shape can easily conform to tissue defects. However, non-covalent bonding and mechanical weakness of these hydrogels limit their applications. In this study, a physically and chemically in situ cross-linkable hydrogel system was developed from a novel thermoresponsive hyperbranched PEG based copolymer with multi acrylate functionality, which was synthesized via an ‘one pot and one step’ in situ deactivation enhanced atom transfer radical co-polymerization of poly(ethylene glycol) diacrylate (PEGDA, Mnxa0=xa0258xa0gxa0mol−1), poly(ethylene glycol) methyl ether methacrylate (PEGMEMA, Mnxa0=xa0475xa0gxa0mol−1) and (2-methoxyethoxy) ethyl methacrylate (MEO2MA). This hyperbranched copolymer was tailored to have the lower critical solution temperature to form physical gelation around 37°C. Meanwhile, with high level of acrylate functionalities, a chemically cross-linked gel was formed from this copolymer using thiol functional cross-linker of pentaerythritol tetrakis (3-mercaptopropionate) (QT) via thiol-ene Michael addition reaction. Furthermore, a semi-interpenetrated polymer networks (semi-IPN) structure was developed by combining this polymer with hyaluronic acid (HA), leading to an in situ cross-linkable hydrogel with significantly increased porosity, enhanced swelling behavior and improved cell adhesion and viability both in 2D and 3D cell culture models.

Hyperbranched PEGmethacrylate linear pDMAEMA block copolymer as an efficient non-viral gene delivery vector

A unique hyperbranched polymeric system with a linear poly-2-dimethylaminoethyl methacrylate (pDMAEMA) block and a hyperbranched polyethylene glycol methyl ether methacrylate (PEGMEMA) and ethylene dimethacrylate (EGDMA) block was designed and synthesized via deactivation enhanced atom transfer radical polymerisation (DE-ATRP) for efficient gene delivery. Using this unique structure, with a linear pDMAEMA block, which efficiently binds to plasmid DNA (pDNA) and hyperbranched polyethylene glycol (PEG) based block as a protective shell, we were able to maintain high transfection levels without sacrificing cellular viability even at high doses. The transfection capability and cytotoxicity of the polymers over a range of pDNA concentration were analysed and the results were compared to commercially available transfection vectors such as polyethylene imine (branched PEI, 25 kDa), partially degraded poly(amido amine)dendrimer (dPAMAM; commercial name: SuperFect(®)) in fibroblasts and adipose tissue derived stem cells (ADSCs).

Nano‐Structured Polymer‐Silica Composite Derived from a Marine Diatom via Deactivation Enhanced Atom Transfer Radical Polymerization Grafting

Nature exhibits phenomenally intricate architecture in various organisms and it is through these structures that researchers in the fi eld of materials science derive inspiration in creating new biomaterials. [ 1 ] One such organism that has drawn attention due to its architecture is the diatom, a unicellular algae, which has a characteristic ornate siliceous cell wall (known as a “frustule”) that current engineering practices cannot fabricate. The proposed applications for diatoms in catalysis, [ 2 ] separation science, [ 3 ] and optics [ 4 ] are heavily dependent on the architecture of the diatom. There has been an interest in altering the chemistry of the frustule, and replication of the frustule in order to expand on possible applications frustule. [ 5 ] Inorganic replicas have predominated: (i) MgO, [ 5a ] TiO 2 , [ 5b ] and ZrO 2 [ 5c ] replicas via gas/solid displacement reactions, and (ii) gold replicas by electroless deposition. [ 5d ] There has been a paucity of reports describing the generation of organic replicas. [ 6 ]

A fluorescently labeled, hyperbranched polymer synthesized from DE-ATRP for the detection of DNA hybridization

The early detection of oligonucleotide biomarkers of disease, such as microRNAs, has been established as a fundamental factor in cancer diagnosis. As the levels of these small molecules (microRNAs) in blood have recently been found to be significantly affected in cancer patients, they offer a means of early stage detection of cancer. Towards the goal of creating a novel method of DNA hybridization detection, we report the detection of specific sequences of small oligonucleotides in a model experiment carried out in serum. The results shown here display the versatility of the DE-ATRP method in synthesizing a specific polymer structure capable of changing its physical properties in the presence of double stranded DNA. The polymer was labeled and used to detect single-stranded DNA in serum successfully.

An antibody fragment functionalized dendritic PEGylated poly(2-(dimethylamino)ethyl diacrylate) as a vehicle of exogenous microRNA

The translation of interfering RNA to the clinic requires more effective delivery agents to enable safe and efficient delivery. The aim of this work was to create a multi-functional delivery agent using deactivation enhanced ATRP synthesis of poly(dimethylamino)ethyl methacrylate (pDMAEMA)-co-PEGMEA/PEGDA (pD-b-P/DA) with linear pDMAEMA as a macro-initiator. The pD-b-P/DA was characterized for its potential to bind synthetic microRNA mimics to form structures and reacted with antibody-derived fragments (Fabs) using Michael-type addition. Conjugation of antibody fragments was verified using SDS–PAGE. Functional delivery of these interfering RNA complexes was proven using a dual luciferase reporter assay. Functional silencing of a reporter gene was improved by complexation of microRNA mimics with pD-b-P/DA alone and with Fab-decorated pD-b-P/DA. The improved silencing with Fab-decorated pD-b-P/DA was evident at 48xa0h but disappeared at 96xa0h. The resultant agent enables complexation of nucleic acid (microRNA mimic) and facile conjugation of antibody fragments via a Michael-type addition. In conclusion, this platform is effective at silencing in this reporter system and has potential as an effective delivery system of interfering RNA.

A Structural Study of Escherichia coli Cells Using an In Situ Liquid Chamber TEM Technology

Studying cell microstructures and their behaviors under living conditions has been a challenging subject in microbiology. In this work, in situ liquid chamber TEM was used to study structures of Escherichia coli cells in aqueous solutions at a nanometer-scale resolution. Most of the cells remained intact under electron beam irradiation, and nanoscale structures were observed during the TEM imaging. The analysis revealed structures of pili surrounding the E. coli cells; the movements of the pili in the liquid were also observed during the in situ tests. This technology also allowed the observation of features of the nucleoid in the E. coli cells. Overall, in situ TEM can be applied as a valuable tool to study real-time microscopic structures and processes in microbial cells residing in native aqueous solutions.

An acetal-based polymeric crosslinker with controlled pH-sensitivity

Even though collagen hydrogels have been used in many biomedical applications due to their unique characteristics, they lack in mechanical stability and resistance to enzymatic degradation. Aiming at the elimination of these drawbacks, collagen hydrogels have been crosslinked with a variety of synthetic crosslinkers all of which presented increased toxicity, which resulted in their limited use in biomedical devices. In order to overcome this limitation, a branched copolymer based on N-hydroxysuccinimide (NHS) activated poly(ethylene glycol) (PEG) groups was synthesized by in situ deactivation enhanced atom transfer radical polymerization (DE-ATRP) and used as crosslinker for collagen hydrogels. The NHS-PEG polymer was modified with labile acetal linkages (acetal-NHS) which made it responsive to pH-alterations and its responsiveness was studied at various pH. The activated acetal-NHS polymeric crosslinker demonstrated long-term stability at neutral pH but underwent hydrolysis at acidic pH, showing strongly pH-dependent degradation properties. By using the acetal-NHS polymeric crosslinker, collagen hydrogels with controlled degradation profiles were fabricated. Both the acetal-NHS crosslinker and the crosslinked collagen hydrogels exhibited excellent cytocompatibility.

PEG based hyperbranched polymeric hollow nanospheres

The synthesis of a new PEG based hyperbranched copolymer of poly(ethylene glycol) methyl ether methacrylate-co-ethylene glycol dimethacrylate (PEGMEMA-co-EGDMA) was achieved via a one-step in situ deactivation enhanced atom transfer radical polymerization (DE-ATRP). Then, hollow PEG based nanospheres were fabricated from this polymer using a solvent evaporation method and post-stabilisation strategy. Furthermore, the analysis using a cellular metabolic activity assay proved that the copolymer did not affect cellular metabolism, indicating that this PEG based polymeric nanosphere has potential for use in drug delivery applications.