Electrostatic immobilization of DNA polyplexes on small intestinal submucosa for tissue substrate-mediated transfection

Tseng, S-Ja; Chuang, Chern; Tang, Shiue–Cheng

doi:10.1016/j.actbio.2008.01.018

Cited by 15 publications

(11 citation statements)

References 31 publications

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“…Similar to forward transfection, increasing the amount of DNA adsorbed to a surface during reverse transfection increases expression levels, up to a limit. PEI polyplexes with increasingly positive zeta potentials adhere in greater numbers to acellular intestinal submucosa (rich in negatively charged glycosaminoglycans), leading to increased transgene expression in fibroblasts seeded in direct contact with the adsorbed complexes [74]. The size of printed lipoplex spots depends on the hydrophobicity of the substrate; more hydrophobic substrates produce smaller spot sizes, higher local DNA concentrations, and elevated expression levels [75].…”

Section: Relationship Between Substrate Surface Nanotechnology Andmentioning

confidence: 99%

Emerging links between surface nanotechnology and endocytosis: Impact on nonviral gene delivery

2010

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Significant effort continues to be exerted toward the improvement of transfection mediated by nonviral vectors. These endeavors are often focused on the design of particulate carriers with properties that encourage efficient accumulation at the membrane surface, particle uptake, and endosomal escape. Despite its demonstrated importance in successful nonviral transfection, relatively little investigation has been done to understand the pressures driving internalized vectors into favorable nondegradative endocytic pathways. Improvements in transfection efficiency have been noted for complexes delivered with a substrate-mediated approach, but the reasons behind such enhancements remain unclear. The phenotypic changes exhibited by cells interacting with nano-and micro-featured substrates offer hints that may explain these effects. This review describes nanoscale particulate and substrate parameters that influence both the uptake of nonviral gene carriers and the endocytic phenotype of interacting cells, and explores the molecular links that may mediate these interactions. Substrate-mediated control of endocytosis represents an exciting new design parameter that will guide the creation of efficient transgene carriers.

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Section: Relationship Between Substrate Surface Nanotechnology Andmentioning

confidence: 99%

Emerging links between surface nanotechnology and endocytosis: Impact on nonviral gene delivery

2010

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“…The PEI/DNA mixture was incubated at room temperature for 30 min to allow for self-assembly of polyplexes. The hydrodynamic size and the surface charge of the polyplexes (such as the PEI/pCMV-EGFP) were approximately at 100 nm and ?20 mV, respectively (Tseng et al 2008). …”

Section: Preparation Of the Bfgf Expression Plasmidmentioning

confidence: 97%

“…We first measured the surface zeta potential of H-CM using a streaming potential method (Tseng et al 2008;Fievet et al 2003;Hunter 1981) and obtained a negative surface charge at -17 mV at neutral pH. We then applied the cationic poly-ethylenimine (PEI)/DNA polyplexes (zeta potential at ?20 mV) as the genetransfer vectors and studied their adsorption onto H-CM.…”

Section: Electrostatic Adsorption Of Polyplexes Onto H-cmmentioning

confidence: 99%

“…Samples were withdrawn from the solution at various time points for measurement of DNA concentration. A PicoGreen dye (Molecular Probes, Eugene, OR, USA) assay was used to measure the DNA amount as previously described Cellulose (2011) 18:95-104 97 (Tseng et al 2008). The amount of DNA adsorbed onto the membrane was determined from the difference between the feed DNA and the residual DNA in solution.…”

Section: Preparation Of the Bfgf Expression Plasmidmentioning

confidence: 99%

“…Second, growth factors were produced from the genetically modified cells on the scaffolds. In this approach, viral (Ito et al 2005;Yue et al 2007) or non-viral (Pannier and Shea 2004;Tseng et al 2008;Lim et al 2006) vectors carrying the growth factor-encoding gene were immobilized on the tissue substrate, where, upon adhesion/infiltration, cells were transduced or transfected to produce growth factors. However, to the best of authors' knowledge, neither of the two approaches has been applied simultaneously to stimulate cellular proliferation on the cellulose scaffold.…”

Section: Introductionmentioning

confidence: 99%

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Application of heparinized cellulose matrices for substrate-mediated bFGF peptide and transgene delivery to stimulate cellular proliferation

2010

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We investigated the use of heparinized cellulose matrices (H-CM) as affinity substrates for binding of basic fibroblast growth factor (bFGF), a heparin-binding peptide, to facilitate cellular proliferation and substrate-mediated transgene delivery. Using human HT-1080 fibroblasts and Saos-2 osteoblasts as cellular models, we showed that H-CM was a friendly substrate for cellular adhesion. Once adhered, cells received stimulation from the bound bFGF, leading to enhanced proliferation. Furthermore, taking advantage of the negative zeta potential of H-CM, we applied electrostatic adsorption to immobilize cationic poly-ethylenimine/DNA polyplexes onto the surface for transgene delivery upon cellular adhesion. Because bFGF stimulated cellular proliferation, we observed a significant increase in transfection efficiency in comparison to transfection on H-CM without the bFGF binding. We showed that H-CM was capable of mediating both bFGF peptide and bFGF transgene delivery to induce a synergistic stimulation of cellular proliferation, thus offering a useful device for fabrication of tissue scaffolds.

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Nonviral Gene Delivery for Applications in Regenerative Medicine

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Sullivan

2011

Engineering Biomaterials for Regenerative Medicine

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Electrostatic immobilization of DNA polyplexes on small intestinal submucosa for tissue substrate-mediated transfection

Cited by 15 publications

References 31 publications

Emerging links between surface nanotechnology and endocytosis: Impact on nonviral gene delivery

Emerging links between surface nanotechnology and endocytosis: Impact on nonviral gene delivery

Application of heparinized cellulose matrices for substrate-mediated bFGF peptide and transgene delivery to stimulate cellular proliferation

Nonviral Gene Delivery for Applications in Regenerative Medicine

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