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Vapor nanobubble is the more reliable photothermal mechanism for inducing endosomal escape of siRNA without disturbing cell homeostasis

Juan Fraire (UGent) , Gaëlle Houthaeve (UGent) , Jing Liu (UGent) , Laurens Raes (UGent) , Lotte Vermeulen (UGent) , Stephan Stremersch (UGent) , Toon Brans (UGent) , Gerardo Garcia-Diaz Barriga, Sarah De Keulenaer (UGent) , Filip Van Nieuwerburgh (UGent) , et al.
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Abstract
Strategies for controlled delivery of therapeutic siRNA into living cells are in high demand as endosomal escape remains the most prominent bottleneck at the intracellular level. Photothermal properties of gold nanoparticles (AuNP) can be used to overcome the endosomal membrane barrier upon laser irradiation by two mechanisms: endosomal rupture by mechanical energy from water vapor nanobubbles (VNBs), or permeabilization of the endosomal membrane by heat diffusion. Here we evaluated how both mechanisms influence cargo release, transfection efficiency, acute cytotoxicity and cell homeostasis. Using a siRNA/AuNP drug delivery system we found that the in vitro release of siRNA from the AuNP carrier occurs equally efficiently by VNB formation or heat generation. Heat-mediated endosomal escape happened more efficiently in cells that had more particles per endosome, resulting in variable siRNA-induced downregulation (20-50%). VNB-mediated endosomal escape did not dependent on the number of AuNP per endosome, yielding high downregulations (50-60%) independent of the cell type. Effects on cell homeostasis by whole transcriptome analysis, showed a quick recover after 24 h or 48 h for either of both photothermal mechanisms. We conclude that VNBs are more consistent to induce efficient endosomal escape and gene silencing independent of the cell type without long lasting effects on cell homeostasis.
Keywords
Gold nanoparticles, Endosomal escape, Vapor nanobubbles, Intracellular delivery, siRNA, Transfection, Photoporation, Transcriptomics, INTRACELLULAR DELIVERY, GENERATION, BARRIERS

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MLA
Fraire, Juan, et al. “Vapor Nanobubble Is the More Reliable Photothermal Mechanism for Inducing Endosomal Escape of SiRNA without Disturbing Cell Homeostasis.” JOURNAL OF CONTROLLED RELEASE, vol. 319, 2020, pp. 262–75, doi:10.1016/j.jconrel.2019.12.050.
APA
Fraire, J., Houthaeve, G., Liu, J., Raes, L., Vermeulen, L., Stremersch, S., … Braeckmans, K. (2020). Vapor nanobubble is the more reliable photothermal mechanism for inducing endosomal escape of siRNA without disturbing cell homeostasis. JOURNAL OF CONTROLLED RELEASE, 319, 262–275. https://doi.org/10.1016/j.jconrel.2019.12.050
Chicago author-date
Fraire, Juan, Gaëlle Houthaeve, Jing Liu, Laurens Raes, Lotte Vermeulen, Stephan Stremersch, Toon Brans, et al. 2020. “Vapor Nanobubble Is the More Reliable Photothermal Mechanism for Inducing Endosomal Escape of SiRNA without Disturbing Cell Homeostasis.” JOURNAL OF CONTROLLED RELEASE 319: 262–75. https://doi.org/10.1016/j.jconrel.2019.12.050.
Chicago author-date (all authors)
Fraire, Juan, Gaëlle Houthaeve, Jing Liu, Laurens Raes, Lotte Vermeulen, Stephan Stremersch, Toon Brans, Gerardo Garcia-Diaz Barriga, Sarah De Keulenaer, Filip Van Nieuwerburgh, Riet De Rycke, Jo Vandesompele, Pieter Mestdagh, Koen Raemdonck, Winnok H. De Vos, Stefaan De Smedt, and Kevin Braeckmans. 2020. “Vapor Nanobubble Is the More Reliable Photothermal Mechanism for Inducing Endosomal Escape of SiRNA without Disturbing Cell Homeostasis.” JOURNAL OF CONTROLLED RELEASE 319: 262–275. doi:10.1016/j.jconrel.2019.12.050.
Vancouver
1.
Fraire J, Houthaeve G, Liu J, Raes L, Vermeulen L, Stremersch S, et al. Vapor nanobubble is the more reliable photothermal mechanism for inducing endosomal escape of siRNA without disturbing cell homeostasis. JOURNAL OF CONTROLLED RELEASE. 2020;319:262–75.
IEEE
[1]
J. Fraire et al., “Vapor nanobubble is the more reliable photothermal mechanism for inducing endosomal escape of siRNA without disturbing cell homeostasis,” JOURNAL OF CONTROLLED RELEASE, vol. 319, pp. 262–275, 2020.
@article{8659985,
  abstract     = {Strategies for controlled delivery of therapeutic siRNA into living cells are in high demand as endosomal escape remains the most prominent bottleneck at the intracellular level. Photothermal properties of gold nanoparticles (AuNP) can be used to overcome the endosomal membrane barrier upon laser irradiation by two mechanisms: endosomal rupture by mechanical energy from water vapor nanobubbles (VNBs), or permeabilization of the endosomal membrane by heat diffusion. Here we evaluated how both mechanisms influence cargo release, transfection efficiency, acute cytotoxicity and cell homeostasis. Using a siRNA/AuNP drug delivery system we found that the in vitro release of siRNA from the AuNP carrier occurs equally efficiently by VNB formation or heat generation. Heat-mediated endosomal escape happened more efficiently in cells that had more particles per endosome, resulting in variable siRNA-induced downregulation (20-50%). VNB-mediated endosomal escape did not dependent on the number of AuNP per endosome, yielding high downregulations (50-60%) independent of the cell type. Effects on cell homeostasis by whole transcriptome analysis, showed a quick recover after 24 h or 48 h for either of both photothermal mechanisms. We conclude that VNBs are more consistent to induce efficient endosomal escape and gene silencing independent of the cell type without long lasting effects on cell homeostasis.},
  author       = {Fraire, Juan and Houthaeve, Gaëlle and Liu, Jing and Raes, Laurens and Vermeulen, Lotte and Stremersch, Stephan and Brans, Toon and Barriga, Gerardo Garcia-Diaz and De Keulenaer, Sarah and Van Nieuwerburgh, Filip and De Rycke, Riet and Vandesompele, Jo and Mestdagh, Pieter and Raemdonck, Koen and De Vos, Winnok H. and De Smedt, Stefaan and Braeckmans, Kevin},
  issn         = {0168-3659},
  journal      = {JOURNAL OF CONTROLLED RELEASE},
  keywords     = {Gold nanoparticles,Endosomal escape,Vapor nanobubbles,Intracellular delivery,siRNA,Transfection,Photoporation,Transcriptomics,INTRACELLULAR DELIVERY,GENERATION,BARRIERS},
  language     = {eng},
  pages        = {262--275},
  title        = {Vapor nanobubble is the more reliable photothermal mechanism for inducing endosomal escape of siRNA without disturbing cell homeostasis},
  url          = {http://dx.doi.org/10.1016/j.jconrel.2019.12.050},
  volume       = {319},
  year         = {2020},
}

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