Deciphering the role of substrate stiffness in enhancing the internalization efficiency of plasmid DNA in stem cells using lipid-based nanocarriers

Modaresi, Saman; Pacelli, Settimio; Whitlow, Jonathan; Paul, Arghya

doi:10.1039/c8nr01516c

Cited by 16 publications

(19 citation statements)

References 35 publications

(34 reference statements)

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“…BMP-2 transgene expression was significantly increased on 670 kPa versus 10 kPa hydrogels (2-fold increase), corresponding with an observed 6-fold significant increase of polyplex internalization. In comparing soft versus stiff substrates, Modaresi et al [120] tested delivery of pDNA encoding for vascular endothelial growth factor (VEGF) complexed with LF2000, to hAMSCs cultured on 0.5 or 32 kPa poly(dimethylsiloxane) PDMS surfaces coated with 1% ( w / v ) gelatin. VEGF transgene expression was significantly higher (4.5-fold) in hAMSCs cultured on 32 kPa versus 0.5 kPa PDMS, which correlated to a 2.5-fold significant increase in lipoplex internalization, which was shown to be correlated with increased caveolae-mediated endocytosis.…”

Section: Recent Approaches To Improve Nonviral Gene Delivery To Mscsmentioning

confidence: 99%

Nucleic acid delivery to mesenchymal stem cells: a review of nonviral methods and applications

2019

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BackgroundMesenchymal stem cells (MSCs) are multipotent stem cells that can be isolated and expanded from many tissues, and are being investigated for use in cell therapies. Though MSC therapies have demonstrated some success, none have been FDA approved for clinical use. MSCs lose stemness ex vivo, decreasing therapeutic potential, and face additional barriers in vivo, decreasing therapeutic efficacy. Culture optimization and genetic modification of MSCs can overcome these barriers. Viral transduction is efficient, but limited by safety concerns related to mutagenicity of integrating viral vectors and potential immunogenicity of viral antigens. Nonviral delivery methods are safer, though limited by inefficiency and toxicity, and are flexible and scalable, making them attractive for engineering MSC therapies.Main textTransfection method and nucleic acid determine efficiency and expression profile in transfection of MSCs. Transfection methods include microinjection, electroporation, and nanocarrier delivery. Microinjection and electroporation are efficient, but are limited by throughput and toxicity. In contrast, a variety of nanocarriers have been demonstrated to transfer nucleic acids into cells, however nanocarrier delivery to MSCs has traditionally been inefficient. To improve efficiency, plasmid sequences can be optimized by choice of promoter, inclusion of DNA targeting sequences, and removal of bacterial elements. Instead of DNA, RNA can be delivered for rapid protein expression or regulation of endogenous gene expression. Beyond choice of nanocarrier and nucleic acid, transfection can be optimized by priming cells with media additives and cell culture surface modifications to modulate barriers of transfection. Media additives known to enhance MSC transfection include glucocorticoids and histone deacetylase inhibitors. Culture surface properties known to modulate MSC transfection include substrate stiffness and specific protein coating. If nonviral gene delivery to MSCs can be sufficiently improved, MSC therapies could be enhanced by transfection for guided differentiation and reprogramming, transplantation survival and directed homing, and secretion of therapeutics. We discuss utilized delivery methods and nucleic acids, and resulting efficiency and outcomes, in transfection of MSCs reported for such applications.ConclusionRecent developments in transfection methods, including nanocarrier and nucleic acid technologies, combined with chemical and physical priming of MSCs, may sufficiently improve transfection efficiency, enabling scalable genetic engineering of MSCs, potentially bringing effective MSC therapies to patients.

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Section: Recent Approaches To Improve Nonviral Gene Delivery To Mscsmentioning

confidence: 99%

Nucleic acid delivery to mesenchymal stem cells: a review of nonviral methods and applications

2019

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“…51,72,73 Besides providing localized delivery, scaffolds can also be tuned to manipulate the extent and even the location of uptake within cells. 74 This can be achieved by altering the physico-chemical properties (elasticity, topology, composition, etc.) of the substrate, which the cells are attached to, thereby regulating cytoskeletal remodelling 75 and subsequently internalization pathways 74 (Figure 2 and Table 1).…”

Section: Tissue-localized Deliverymentioning

confidence: 99%

RNA interference in glial cells for nerve injury treatment

Lin

Kim

et al. 2020

J Tissue Eng

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Drivers of RNA interference are potent for manipulating gene and protein levels, which enable the restoration of dysregulated mRNA expression that is commonly associated with injuries and diseases. This review summarizes the potential of targeting neuroglial cells, using RNA interference, to treat nerve injuries sustained in the central nervous system. In addition, the various methods of delivering these RNA interference effectors will be discussed.

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“…It was previously reported that the surface properties of biomaterial, including composition, 47 stiffness, 48 topography 49 and so on, could affect cytoskeletal rearrangement, thereby influencing its cellular endocytosis. 50 In order to investigate whether the photothermal properties of the composite coating could lead to cytoskeletal rearrangement, we have used the laser confocal microscope for this study.…”

Section: Effects Of Visible-light Illumination On Cellular Endocytosis Of Lf/gfpmentioning

confidence: 99%

Bioactive nanocomposite coatings under visible light illumination promoted surface-mediated gene delivery

et al. 2020

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Deciphering the role of substrate stiffness in enhancing the internalization efficiency of plasmid DNA in stem cells using lipid-based nanocarriers

Cited by 16 publications

References 35 publications

Nucleic acid delivery to mesenchymal stem cells: a review of nonviral methods and applications

Nucleic acid delivery to mesenchymal stem cells: a review of nonviral methods and applications

RNA interference in glial cells for nerve injury treatment

Bioactive nanocomposite coatings under visible light illumination promoted surface-mediated gene delivery

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