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Biomedical subjects

Tiffany L Houchin

Publications and source records attributed to Tiffany L Houchin.

4 recordsLinked to original sources

Surface adsorption of DNA to tissue engineering scaffolds for efficient gene delivery.

Gene delivery from tissue engineering scaffolds has potential to promote localized transgene expression that can induce the formation of functional tissues. Substrate-mediated delivery, an alternative delivery strategy to sustained release, is based on immobilization of DNA complexes to the polymer surface for subsequent delivery to cells cultured on the substrate. We investigate polyethylenimine (PEI)/DNA complex immobilization and subsequent cellular transfection on tissue engineering scaffolds fabricated from poly(lactide-co-glycolide) (PLG). The properties of the substrate and the complex affect both immobilization and cellular transfection. PLG promotes binding of PEI/DNA complexes, with percent bound independent of the N/P ratio or the DNA dosage. The levels of transgene expression are similar to or greater than control studies based on bolus DNA delivery, with orders of magnitude less DNA. Immobilization also homogeneously distributes the DNA throughout the scaffold, resulting in large numbers of transfected cells (>60%) at low surface quantities (<50 ng). Importantly, this approach can be employed to transfect cells throughout a three-dimensional scaffold. Tissue engineering scaffolds that are prefabricated into various shapes from a range of materials could potentially employ this strategy for numerous applications.

Adsorption↗

Modular design of non-viral vectors with bioactive components.

Inefficient gene delivery continues to limit gene therapy applications to both basic and applied sciences. Approaches for engineering vectors increasingly include bioactive components that bind cellular receptors, disrupt membranes, or enhance nuclear transport. Recently, a novel cationic lipid was developed by modifying the glucocorticoid dexamethasone. This cationic corticosteroid condenses plasmids for gene delivery, while also modulating inflammation. Modular vectors containing bioactive components that target various cellular processes can overcome the barriers limiting gene transfer.

Active Transport, Cell Nucleus↗

Measurement of rheology of distiller's grain slurries using a helical impeller viscometer.

Current research is focused on developing a process to convert the cellulose and hemicellulose in distiller's grains into fermentable sugars, increasing both ethanol yield and the amount of protein in the remaining solid product. The rheologic properties of distiller's grain slurries were determined for concentrations of 21, 23, and 25%. Distiller's grain slurries are non-Newtonian, heterogeneous fluids subject to particle settling. Traditional methods of viscosity measurement, such as cone-and-plate and concentric cylinder viscometers, are not adequate for these fluids. A helical impeller viscometer was employed to measure impeller torque over a range of rotational speeds. Newtonian and non-Newtonian calibration fluids were utilized to obtain constants that relate shear stresses and shear rates to the experimental data. The Newtonian impeller constant, c, was 151; the non-Newtonian shear rate constant, k, was 10.30. Regression analysis of experimental data was utilized for comparison to power law, Herschel-Bulkley, and Casson viscosity models with regression coefficients exceeding 0.99 in all cases.

Biotechnology↗

Gene delivery from polymer scaffolds for tissue engineering.

The combination of gene therapy with tissue engineering offers the potential to direct progenitor cell proliferation and differentiation into functional tissue replacements. Many approaches to engineering tissue replacements feature a polymer scaffold to create and maintain a space, support cell adhesion, and organize tissue formation. Polymer scaffolds, either natural, synthetic, or a combination of the two, have also been adapted to serve as delivery vehicles for viral and nonviral vectors, which can induce the expression of tissue inductive factors. Gene delivery is a versatile approach, capable of targeting any cellular process through localized expression of tissue inductive factors. The design and application of tissue engineering scaffolds for localized gene transfer are reviewed. Scaffolds are designed either to release the vector into the local tissue environment or maintain the vector at the polymer surface, which is regulated by the effective affinity of the vector for the polymer. Polymeric delivery can enhance gene transfer locally, promote and extend transgene expression, avoid vector distribution to distant tissues, and reduce the immune response to the vector. Scaffolds capable of controlled DNA delivery can provide a fundamental tool for directing progenitor cell function, which has applications with the engineering of numerous types of tissue. The utility of this approach will increase with the development of design parameters that correlate release and transgene expression, and with continued research into the biology of tissue formation.

Cell Culture Techniques↗