Nerve growth factor expression by PLG-mediated lipofection

Whittlesey, Kevin J.; Shea, Lonnie D.

doi:10.1016/j.biomaterials.2005.11.016

Cited by 35 publications

(30 citation statements)

References 46 publications

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“…These non-viral DNA complexes were immobilized to the PLG surface and both transfection efficiency and transgene expression were investigated as a function of channel width and DNA amount. DNA encoding NGF was subsequently non-specifically immobilized to the PLG microchannels and an in vitro co-culture model was employed to assess neuron survival and neurite extension [19]. Additionally, the ability to localize gene transfer to the channel in order to tailor channels for specific axonal populations was investigated.…”

Section: Introductionmentioning

confidence: 99%

Patterned PLG substrates for localized DNA delivery and directed neurite extension

et al. 2007

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Tissue engineering strategies that enable nerve regeneration will require methods that can promote and direct neurite extension across the lesion. In this report, we investigate an in vitro combinatorial approach to directed neurite outgrowth using gene delivery from topographically patterned substrates, which can induce expression of neurotrophic factors to promote neurite extension and direct the extending neurites. Poly(lactide-co-glycolide) (PLG), which has been used to fabricate conduits or bridges for regeneration, was compression molded to create channels with 100, 150, and 250 microm widths. DNA complexes were immobilized to the PLG, and cells cultured on the substrate were transfected with efficiencies dependent on channel width and DNA amount. A co-culture model consisting of primary neurons and accessory cells was employed to investigate neurite outgrowth within the channels. Localized secretion of nerve growth factor (NGF) by the accessory cells promoted neuron survival and neurite extension. Neurons cultured in channels with NGF expression exhibited longer primary neurites than in the absence of channels. Neurons cultured in smaller width PLG microchannels exhibited a greater degree of directionality and less secondary sprouting than larger channels. Finally, surface immobilization allowed for the delivery of distinct plasmids from each channel, which may enable channels to be tailored for specific nerve tracts. This approach demonstrates the ability to combine gene delivery with physical guidance, and can be tailored to target specific axonal populations with varying neurotrophic factor requirements.

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Section: Introductionmentioning

confidence: 99%

Patterned PLG substrates for localized DNA delivery and directed neurite extension

et al. 2007

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“…Traditional means of transfection, protein transduction, and macromolecular delivery have proven difficult in neurons due to their terminally differentiated state (6). A number of commercially available methods rely on use of liposomes or other charged lipid formulations, which have limited complex stability in serum and often high toxicity over time (7). Electroporation-based techniques often yield higher rates of transfection but are only effective when performed during a specific window of development in young, healthy, and undifferentiated cells, with eventual loss of expression or bioactivity over time (8).…”

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Functional Protein Delivery into Neurons Using Polymeric Nanoparticles

Hasadsri

Kreuter

Hashimoto³

et al. 2009

Journal of Biological Chemistry

118

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An efficient route for delivering specific proteins and peptides into neurons could greatly accelerate the development of therapies for various diseases, especially those involving intracellular defects such as Parkinson disease. Here we report the novel use of polybutylcyanoacrylate nanoparticles for delivery of intact, functional proteins into neurons and neuronal cell lines. Uptake of these particles is primarily dependent on endocytosis via the low density lipoprotein receptor. The nanoparticles are rapidly turned over and display minimal toxicity to cultured neurons. Delivery of three different functional cargo proteins is demonstrated. When primary neuronal cultures are treated with recombinant Escherichia coli ␤-galactosidase as nanoparticle cargo, persistent enzyme activity is measured beyond the period of nanoparticle degradation. Delivery of the small GTPase rhoG induces neurite outgrowth and differentiation in PC12 cells. Finally, a monoclonal antibody directed against synuclein is capable of interacting with endogenous ␣-synuclein in cultured neurons following delivery via nanoparticles. Polybutylcyanoacrylate nanoparticles are thus useful for intracellular protein delivery in vitro and have potential as carriers of therapeutic proteins for treatment of neuronal disorders in vivo.

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“…Nerve conduits used for nerve repair also provide a channel for growth factor diffusion [81]. A combination of liposomes and neurotrophic factor genes used in another approach effectively facilitated nerve injury repair [82,83]. Furthermore, the release of acidic products when poly(D,L-lactic-coglycolic acid) is used as microspheres for carrying growth factors may result in protein inactivation [84].…”

Section: Future Prospectsmentioning

confidence: 99%

Neural Stem Cells and Induced Neurons for Nerve Injury Repair

Hsu¹,

Chen²,

Hsu³

et al. 2015

J Stem Trans Bio

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Cell transplantation can relieve the symptoms of or even reverse neurodegenerative diseases and repair nerve injuries. Fibroblast growth factor 1 promotes neuronal survival and stimulates axonal growth. A combination of fibroblast growth factor 1 and cell-based therapy is promising for nerve repair. Developers of future cell-based treatment should consider several key concerns: (1) the source of cells should be autologous, (2) consistent methods and protocols for cell isolation should be used, (3) the treatment should be tested in suitable animal models, and (4) the microenvironment of cells implanted should be optimally characterized. In addition, developing high temporal and spatial resolution images for cell tracking is crucial for evaluating the efficacy of cell transplantation. In this paper, we summarize recent progress in cellular reprogramming, such as induced neural stem cells and induced neurons, and the development of future cell-based therapy for peripheral nerve and spinal cord injury that includes conduits and growth factors. Fibroblast Growth Factor 1 for Nerve Injury RepairIn total, 22 mammalian fibroblast growth factors (FGFs) exist, grouped into seven subfamilies on the basis of differences in sequence homology and phylogeny. Notably, FGF1 and FGF2 share sequential and structural similarities and belong to the FGF1 subfamily [1]. The FGF ligands execute diverse functions by binding and activating the FGF receptor (FGFR) family of tyrosine kinase receptors with heparan sulfate proteoglycans. In total, four FGFR genes (FGFR1-FGFR4) encode receptors consisting of three extracellular immunoglobulin domains (D1-D3), a single-pass transmembrane domain, and a cytoplasmic tyrosine kinase domain [2]. Several FGFR isoforms exist because of exon skipping that removes the D1 domain and/or the acid box in FGFR1-FGFR4 [3]. Alternative splicing in the second half of the D3 domain of FGFR1-FGFR3 yields b (FGFR1b-FGFR3b) and c (FGFR1c-FGFR3c) isoforms that have distinct FGFbinding specificities [4] and are predominantly present in epithelial and mesenchymal cells, respectively. Each FGF binds to epithelial or mesenchymal FGFRs, with the exception of FGF1, which activates both spliced isoforms [3].The involvement of FGF signaling in human diseases has been thoroughly documented. Deregulated FGF signaling can contribute to pathological conditions through gain-or loss-of-function mutations in the FGFRs. Because both Fgf1 −/− and Fgf1mice are fertile and apparently normal [5], the physiological roles of FGF1 and FGF2 remain to be explored. However, FGF1 and FGF2 likely play a physiological role in the maintenance of the vascular tone because FGF1 and FGF2 administration lowers the blood pressure in rats [6] and can restore the nitric oxide synthase activity in spontaneously hypertensive rats [7]. In addition, blood vessels isolated from Fgf2 −/− mice exhibit a reduced response to vasoconstrictors. Although Fgf2 −/− mice had hypotension caused by reduced smooth muscle contractility [8], their blood pressu...

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Nerve growth factor expression by PLG-mediated lipofection

Cited by 35 publications

References 46 publications

Patterned PLG substrates for localized DNA delivery and directed neurite extension

Patterned PLG substrates for localized DNA delivery and directed neurite extension

Functional Protein Delivery into Neurons Using Polymeric Nanoparticles

Neural Stem Cells and Induced Neurons for Nerve Injury Repair

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