Magdalena Kita

Skeletal muscle cell proliferation and differentiation on polypyrrole substrates doped with extracellular matrix components

Conducting polymers have been developed as substrates for in vitro studies with a range of cell types including electrically-excitable cells such as nerve and smooth muscle. The goal of this study was to optimise and characterise a range of polypyrrole materials to act as substrates for electrical stimulation of differentiating skeletal myoblasts. Although all of the polymer materials provided suitable substrates for myoblast adhesion and proliferation, significant differences became apparent under the low-serum conditions used for differentiation of primary myoblasts. The significance of the work lies in the design and control of polymer materials to facilitate different stages of skeletal muscle cell proliferation and/or differentiation, opening up opportunities for engineering of this tissue. This paper therefore constitutes not just a biocompatibility assessment but a comprehensive study of how synthesis conditions affect the final outcome in terms of cell response.

A conducting-polymer platform with biodegradable fibers for stimulation and guidance of axonal growth

A biosynthetic platform composed of a conducting polypyrrole sheet embedded with unidirectional biodegradable polymer fibers is described (see image; scale bar = 50 µm). Such hybrid systems can promote rapid directional nerve growth for neuro-regenerative scaffolds and act as interfaces between the electronic circuitry of medical bionic devices and the nervous system.

Electrical Stimulation of Myoblast Proliferation and Differentiation on Aligned Nanostructured Conductive Polymer Platforms

In this study, nanostructured conductive platforms synthesized from aligned multiwalled carbon nanotubes and polypyrrole are investigated as myo-regenerative scaffolds. Myotube formation follows a linear path on the platforms coinciding with extent of nanotopography. In addition, electrical stimulation enhances myo-nuclear number and differentiation. These studies demonstrate that conductive polymer platforms can be used to influence muscle cell behaviour through nanostructure and electrical stimulation.

Creating Conductive Structures for Cell Growth: Growth and alignment of Myogenic cell types on polythiophenes.

Conducting polymers provide suitable substrates for the in vitro study of excitable cells, including skeletal muscle cells, due to their inherent conductivity and electroactivity. The thiophene family of conducting polymers offers unique flexibility for tailoring of polymer properties as a result of the ease of functionalization of the parent monomer. This article describes the preparation of films and electrospun fibers from an ester-functionalized organic solvent-soluble polythiophene (poly-octanoic acid 2-thiophen-3-yl-ethyl ester) and details the changes in properties that result from post-polymerization hydrolysis of the ester linkage. The polymer films supported the proliferation and differentiation of both primary and transformed skeletal muscle myoblasts. In addition, aligned electrospun fibers formed from the polymers provided scaffolds for the guided differentiation of linearly aligned primary myotubes, suggesting their suitability as three-dimensional substrates for the in vitro engineering of skeletal muscle tissue.

DNA electroporation in vivo targets mature fibres in dystrophic mdx muscle

Non-viral gene transfer into skeletal muscle is enhanced by electroporation and myotoxin preconditioning of muscle following plasmid injection. We investigated in vivo delivery of naked DNA to mdx mouse muscle, utilising enhanced green fluorescent protein reporter vector (pEGFP) and a corrective nucleic acid to promote targeted corrective gene conversion at the mutant mdx mouse dystrophin (DMDmdx) locus. Electroporation, myoablation with bupivacaine and a combined protocol, were applied to mdx muscle. We report up to 90% EGFP expression in electroporated mdx tibialis anterior muscle. Muscles preconditioned with bupivacaine showed low transgene expression with or without EP. Single EGFP+ve muscle fibre explants showed EGFP expression in mature fibres in preference to satellite cells. We observed a two-fold increase (P<0.005; t) in dystrophin protein, accompanied by wild-type (wt) DMD transcript in muscles injected with corrective nucleic acid over contralateral saline-injected TAs. By targeting the muscle fibres in preference to the satellite cells, plasmid-bourne transgenes delivered to dystrophic muscle will not penetrate the regenerative component of muscle. Whether in the context of targeted corrective gene conversion or therapeutic non-viral transgenes, under these conditions periodic re-administration will be required to promote phenotypic benefits in dystrophic muscle.

Genetic analysis of stress responsiveness in a mouse model

The purpose of the present paper was to look for genes that might be involved in anxiety-related behaviours by undertaking a genetic analysis of a simple mouse model of stress responsiveness. Two inbred mouse strains have been identified that show either high or low stress responsiveness. These strains were crossed to generate F1 progeny, which were then crossed to generate F2 progeny, and in which there is segregation of genotype within individual animals. DNA was isolated from these animals and a genome scan was conducted in order to find regions on the genome that correlate with the stress responsiveness. Several regions on the mouse genome show significant linkage with the stress phenotype. One region in particular, on chromosome 12, was further characterised and the most significant linkage was found between 32.8 and 44.8 cM. These chromosomal regions may contain genes encoding proteins that are involved in the underlying neural circuitry involved in stress responsiveness.

Nerve Repair: A Conducting‐Polymer Platform with Biodegradable Fibers for Stimulation and Guidance of Axonal Growth (Adv. Mater. 43/2009)

Effective functional innervation of medical bionic devices, as well as re-innervation of target tissue in nerve and spinal cord injuries, requires a platform that can stimulate and orientate neural growth. Gordon Wallace and co-workers report on p. 4393 that conducting and nonconducting biodegradable polymers show excellent potential as suitable hybrid substrata for neural regeneration and may form the basis of electrically active conduits designed to accelerate nerve repair.

Peptide nucleic acids for selective inhibition of cells containing the Mdx dystrophin locus

No abstract is available for this article.

Bionic materials for neuromuscular restoration and maintenance

No abstract is available for this article.

Nanostructured conducting polymer scaffolds for skeletal muscle growth

S THE JOURNAL OF GENE MEDICINE J Gene Med 2011; 13: 410 446 Published online in Wiley Online Library (wileyonlinelibrary.com) DOI: 10.1002/jgm.1582 7 th AUSTRALASIAN GENE THERAPY SOCIETY MEETING