Skip to main content

Advertisement

SpringerLink
Log in

Comparison of Polymeric siRNA Nanocarriers in a Murine LPS-Activated Macrophage Cell Line: Gene Silencing, Toxicity and Off-Target Gene Expression

  • Research Paper
  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

ABSTRACT

Purpose

Tumor necrosis factor α (TNF-α) plays a key role in the progression of rheumatoid arthritis and is an important target for anti-rheumatic therapies. TNF-α expression can be silenced with small interfering RNA (siRNA), but efficacy is dependent on efficient and safe siRNA delivery vehicles. We aimed to identify polymeric nanocarriers for anti-TNF-α siRNA with optimal efficacy and minimal off-target effects in vitro.

Methods

TNF-α silencing with polymeric siRNA nanocarriers was compared in lipopolysaccharide-activated RAW 264.7 macrophages by real-time reverse transcription (RT)-PCR. Expression of non-target genes involved in inflammation, apoptosis, and cell cycle progression was determined by RT-PCR, toxicity evaluated by propidium iodide and annexin V staining.

Results

PAMAM dendrimers (G4 and G7) and dextran nanogels mediated remarkably high concentration-dependent gene silencing and low toxicity; dioleoyltrimethylammoniumpropane-modified poly(DL-lactide-co-glycolide acid) nanoparticles, thiolated, trimethylated chitosan and poly[(2-hydroxypropyl)methacrylamide 1-methyl-2-piperidine methanol] polyplexes were less efficient transfectants. There were minor changes in the regulation of off-target genes, mainly dependent on nanocarrier and siRNA concentration.

Conclusions

Dextran nanogels and PAMAM dendrimers mediated high gene silencing with minor toxicity and off-target transcriptional changes and are therefore expected to be suitable siRNA delivery systems in vivo.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

Act:

β-actin

AEMA:

2-aminoethyl methacrylate hydrochloride

ANOVA:

analysis of variance

Ccna2:

cyclin a2

Cdk7:

cyclin-dependent kinase 7

CP:

crossing point

Cse1L:

cellular apoptosis susceptibility protein 1L

DAB:

diaminobutane dendrimers

dex-HEMA:

dextran hydroxyethyl methacrylate

dex-MA:

dextran methacrylate

DMEM:

Dulbecco´s Modified Eagle´s Medium

DOTAP:

1,2-dioleoyloxy-3-trimethylammoniumpropane

EE:

encapsulation efficiency

EGFP:

enhanced green fluorescent protein

FBS:

fetal bovine serum

FITC:

fluorescein isothiocyanate

G:

generation

Gus:

β-glucuronidase

HEPES:

4-(2-hydroxyethyl)-piperazine-1-ethanesulfonic acid

IL:

interleukin

INF:

interferon

LPS:

lipopolysaccharide

LUC:

luciferase

NHS-PEG:

N-hydroxysuccinimidyl-activated methoxypolyethylene glycol 5000 propionic acid

N/P:

amine-to-phosphate ratio

OAS1d:

oligoadenylate Synthetase-Like Protein 1d

OF:

oligofectamine

PAMAM:

poly(amidoamine)

PCI:

photochemical internalization

PDI:

polydispersity index

PEG:

polyethylene glycol

PEI:

polyethylenimine

PF:

polyfect (based on PAMAM dendrimers)

pHPMA-MPPM:

poly((2-hydroxypropyl)methacrylamide 1-methyl-2-piperidine methanol)

PI:

propidium iodide

PLGA:

poly(DL-lactide-co-glycolide acid)

PVA:

polyvinylalcohol

RA:

rheumatoid arthritis

RNAi:

RNA interference

RT:

reverse transcription

siRNA:

small interfering RNA

TLR:

toll-like receptor

TMAEMA:

[2-(methacryloyloxy)ethyl]-trimethylammonium chloride

TMC-SH:

thiolated N,N,N-trimethylated chitosan

TNF-α:

tumor necrosis factor α

REFERENCES

  1. Doan T, Massarotti E. Rheumatoid arthritis: An overview of new and emerging therapies. J Clin Pharmacol. 2005;45(7):751–62.

    Article  PubMed  CAS  Google Scholar 

  2. Kinne RW, Brauer R, Stuhlmuller B, Palombo-Kinne E, Burmester GR. Macrophages in rheumatoid arthritis. Arthritis Res. 2000;2(3):189–202.

    Article  PubMed  CAS  Google Scholar 

  3. Kinne RW, Stuhlmuller B, Burmester GR. Cells of the synovium in rheumatoid arthritis—Macrophages. Arthritis Res Ther. 2007;9(6).

  4. Scott DL, Kingsley GH. Tumor necrosis factor inhibitors for rheumatoid arthritis. N Engl J Med. 2006;355(7):704–12.

    Article  PubMed  CAS  Google Scholar 

  5. van Vollenhoven RF. Treatment of rheumatoid arthritis: state of the art 2009. Nat Rev Rheumatol. 2009;5(10):531–41.

    Article  PubMed  Google Scholar 

  6. Huber LC, Distler O, Gay RE, Gay S. Antisense strategies in degenerative joint diseases: Sense or nonsense? Adv Drug Deliv Rev. 2006;58(2):285–99.

    Article  PubMed  CAS  Google Scholar 

  7. Khoury M, Jorgensen C, Apparailly F. RNAi in arthritis: Prospects of a future antisense therapy in inflammation. Curr Opin Mol Ther. 2007;9(5):483–9.

    PubMed  CAS  Google Scholar 

  8. Castanotto D, Rossi JJ. The promises and pitfalls of RNA-interference-based therapeutics. Nature. 2009;457(7228):426–33.

    Article  PubMed  CAS  Google Scholar 

  9. Schiffelers RM, Xu J, Storm G, Woodle MC, Scaria PV. Effects of treatment with small interfering RNA on joint inflammation in mice with collagen-induced arthritis. Arthritis Rheum. 2005;52(4):1314–8.

    Article  PubMed  CAS  Google Scholar 

  10. Howard KA, Paludan SR, Behlke MA, Besenbacher F, Deleuran B, Kjems J. Chitosan/siRNA nanoparticle-mediated TNF-alpha knockdown in peritoneal macrophages for anti-inflammatory treatment in a murine arthritis model. Mol Ther. 2009;17(1):162–8.

    Article  PubMed  CAS  Google Scholar 

  11. Khoury M, Louis-Plence P, Escriou V, Noel D, Largeau C, Cantos C, et al. Efficient new cationic liposome formulation for systemic delivery of small interfering RNA silencing tumor necrosis factor alpha in experimental arthritis. Arthritis Rheum. 2006;54(6):1867–77.

    Article  PubMed  CAS  Google Scholar 

  12. Khoury M, Courties G, Fabre S, Bouffi C, Seemayer CA, Vervoordeldonk MJ, et al. Adeno-associated virus type 5-mediated intraarticular administration of tumor necrosis factor small interfering RNA improves collagen-induced arthritis. Arthritis Rheum. 2010;62(3):765–70.

    Article  PubMed  CAS  Google Scholar 

  13. Scaggiante B, Dapas B, Farra R, Grassi M, Pozzato G, Giansante C, et al. Improving siRNA bio-distribution and minimizing side effects. Curr Drug Metab. 2011;12(1):11–23.

    Article  PubMed  CAS  Google Scholar 

  14. Whitehead KA, Langer R, Anderson DG. Knocking down barriers: advances in siRNA delivery. Nat Rev Drug Discov. 2009;8(2):129–38.

    Article  PubMed  CAS  Google Scholar 

  15. Cun D, Jensen LB, Nielsen HM, Moghimi M, Foged C. Polymeric nanocarriers for siRNA delivery: Challenges and future prospects. J Biomed Nanotechnol. 2008;4(3):258–75.

    Article  CAS  Google Scholar 

  16. Mislick KA, Baldeschwieler JD. Evidence for the role of proteoglycans in cation-mediated gene transfer. Proc Natl Acad Sci USA. 1996;93(22):12349–54.

    Article  PubMed  CAS  Google Scholar 

  17. Urban-Klein B, Werth S, Abuharbeid S, Czubayko F, Aigner A. RNAi-mediated gene-targeting through systemic application of polyethylenimine (PEI)-complexed siRNA in vivo. Gene Ther. 2005;12(5):461–6.

    Article  PubMed  CAS  Google Scholar 

  18. Werth S, Urban-Klein B, Dai L, Hobel S, Grzelinski M, Bakowsky U, et al. A low molecular weight fraction of polyethylenimine (PEI) displays increased transfection efficiency of DNA and siRNA in fresh or lyophilized complexes. J Control Release. 2006;112(2):257–70.

    Article  PubMed  CAS  Google Scholar 

  19. Perez AP, Romero EL, Morilla MJ. Ethylendiamine core PAMAM dendrimers/siRNA complexes as in vitro silencing agents. Int J Pharm. 2009;380(1–2):189–200.

    Article  PubMed  CAS  Google Scholar 

  20. Zhou JH, Wu JY, Hafdi N, Behr JP, Erbacher P, Peng L. PAMAM dendrimers for efficient siRNA delivery and potent gene silencing. Chem Comm. 2006;22:2362–4.

    Article  PubMed  Google Scholar 

  21. Duncan R, Izzo L. Dendrimer biocompatibility and toxicity. Adv Drug Deliv Rev. 2005;57(15):2215–37.

    Article  PubMed  CAS  Google Scholar 

  22. Lv H, Zhang S, Wang B, Cui S, Yan J. Toxicity of cationic lipids and cationic polymers in gene delivery. J Control Release. 2006;114(1):100–9.

    Article  PubMed  CAS  Google Scholar 

  23. Luten J, van Nostruin CF, De Smedt SC, Hennink WE. Biodegradable polymers as non-viral carriers for plasmid DNA delivery. J Control Release. 2008;126(2):97–110.

    Article  PubMed  CAS  Google Scholar 

  24. Kim TH, Jiang HL, Jere D, Park IK, Cho MH, Nah JW, et al. Chemical modification of chitosan as a gene carrier in vitro and in vivo. Progr Polymer Sci. 2007;32(7):726–53.

    Article  CAS  Google Scholar 

  25. Lee KY. Chitosan and its derivatives for gene delivery. Macromol Res. 2007;15(3):195–201.

    Article  CAS  Google Scholar 

  26. Raemdonck K, Demeester J, De Smedt S. Advanced nanogel engineering for drug delivery. Soft Matter. 2009;5(4):707–15.

    Article  CAS  Google Scholar 

  27. Raemdonck K, Naeye B, Buyens K, Vandenbroucke RE, Hogset A, Demeester J, et al. Biodegradable dextran nanogels for RNA interference: Focusing on endosomal escape and intracellular siRNA delivery. Adv Funct Mater. 2009;19(9):1406–15.

    Article  CAS  Google Scholar 

  28. Cun D, Jensen DK, Maltesen MJ, Bunker M, Whiteside P, Scurr D, et al. High loading efficiency and sustained release of siRNA encapsulated in PLGA nanoparticles: quality by design optimization and characterization. Eur J Pharm Biopharm. 2011;77(1):26–35.

    Article  PubMed  CAS  Google Scholar 

  29. Jackson AL, Linsley PS. Recognizing and avoiding siRNA off-target effects for target identification and therapeutic application. Nat Rev Drug Discov. 2010;9(1):57–67.

    Article  PubMed  CAS  Google Scholar 

  30. Robbins M, Judge A, MacLachlan I. siRNA and innate immunity. Oligonucleotides. 2009;19(2):89–102.

    Article  PubMed  CAS  Google Scholar 

  31. Judge A, MacLachlan I. Overcoming the innate immune response to small interfering RNA. Hum Gene Ther. 2008;19(2):111–24.

    Article  PubMed  CAS  Google Scholar 

  32. Omidi Y, Hollins AJ, Benboubetra M, Drayton R, Benter IF, Akhtar S. Toxicogenomics of non-viral vectors for gene therapy: A microarray study of lipofectin- and oligofectamine-induced gene expression changes in human epithelial cells. J Drug Target. 2003;11(6):311–23.

    Article  PubMed  CAS  Google Scholar 

  33. Akhtar S. Cationic nanosystems for the delivery of small interfering ribonucleic acid therapeutics: a focus on toxicogenomics. Expert Opin Drug Metab Toxicol. 2010;6(11):1347–62.

    Article  PubMed  CAS  Google Scholar 

  34. Akhtar S, Benter I. Toxicogenomics of non-viral drug delivery systems for RNAi: Potential impact on siRNA-mediated gene silencing activity and specificity. Adv Drug Deliv Rev. 2007;59(2–3):164–82.

    Article  PubMed  CAS  Google Scholar 

  35. Hollins AJ, Omidi Y, Benter IF, Akhtar S. Toxicogenomics of drug delivery systems: Exploiting delivery system-induced changes in target gene expression to enhance siRNA activity. J Drug Target. 2007;15(1):83–8.

    Article  PubMed  CAS  Google Scholar 

  36. Gras R, Almonacid L, Ortega P, Serramia MJ, Gomez R, de la Mata FJ, et al. Changes in gene expression pattern of human primary macrophages induced by carbosilane dendrimer 2G-NN16. Pharm Res. 2009;26(3):577–86.

    Article  PubMed  CAS  Google Scholar 

  37. Andersen MO, Howard KA, Paludan SR, Besenbacher F, Kjems J. Delivery of siRNA from lyophilized polymeric surfaces. Biomaterials. 2008;29(4):506–12.

    Article  PubMed  CAS  Google Scholar 

  38. Amarzguioui M, Lundberg P, Cantin E, Hagstrom J, Behlke MA, Rossi JJ. Rational design and in vitro and in vivo delivery of Dicer substrate siRNA. Nat Protoc. 2006;1(2):508–17.

    Article  PubMed  CAS  Google Scholar 

  39. Varkouhi AK, Lammers T, Schiffelers RM, van Steenbergen MJ, Hennink WE, Storm G. Gene silencing activity of siRNA polyplexes based on biodegradable polymers. Eur J Pharm Biopharm. 2011;77(3):450–7.

    Article  PubMed  CAS  Google Scholar 

  40. Verheul RJ, van der Wal S, Hennink WE. Tailorable thiolated trimethyl chitosans for covalently stabilized nanoparticles. Biomacromolecules. 2010;11(8):1965–71.

    Article  PubMed  CAS  Google Scholar 

  41. Luten J, Akeroyd N, Funhoff A, Lok MC, Talsma H, Hennink WE. Methacrylamide polymers with hydrolysis-sensitive cationic side groups as degradable gene carriers. Bioconjug Chem. 2006;17(4):1077–84.

    Article  PubMed  CAS  Google Scholar 

  42. Varkouhi AK, Verheul RJ, Schiffelers RM, Lammers T, Storm G, Hennink WE. Gene silencing activity of siRNA polyplexes based on thiolated N, N, N-trimethylated chitosan. Bioconjug Chem. 2010;21(12):2339–46.

    Article  PubMed  CAS  Google Scholar 

  43. Jensen LB, Mortensen K, Pavan GM, Kasimova MR, Jensen DK, Gadzhyeva V, et al. Molecular characterization of the interaction between siRNA and PAMAM G7 dendrimers by SAXS, ITC, and molecular dynamics simulations. Biomacromolecules. 2010;11(12):3571–7.

    Article  PubMed  CAS  Google Scholar 

  44. Naeye B, Raemdonck K, Remaut K, Sproat B, Demeester J, De Smedt SC. PEGylation of biodegradable dextran nanogels for siRNA delivery. Eur J Pharm Sci. 2010;40(4):342–51.

    Article  PubMed  CAS  Google Scholar 

  45. Jensen DK, Jensen LB, Koocheki S, et al. Design of an inhalable dry powder formulation of DOTAP-modified PLGA nanoparticles loaded with siRNA. J Control Release. 2011;In press.

  46. Timm M, Hansen EW, Moesby L, Christensen JD. Utilization of the human cell line HL-60 for chemiluminescence based detection of microorganisms and related substances. Eur J Pharm Sci. 2006;27(2–3):252–8.

    Article  PubMed  CAS  Google Scholar 

  47. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucl Acids Res. 2001;29(9).

  48. Chapekar MS, Zaremba TG, Kuester RK, Hitchins VM. Synergistic induction of tumor necrosis factor alpha by bacterial lipopolysaccharide and lipoteichoic acid in combination with polytetrafluoroethylene particles in a murine macrophage cell line RAW 264.7. J Biomed Mater Res. 1996;31(2):251–6.

    Article  PubMed  CAS  Google Scholar 

  49. Amarzguioui M. Improved siRNA-mediated silencing in refractory adherent cell lines by detachment and transfection in suspension. Biotechniques. 2004;36(5):766–8.

    PubMed  CAS  Google Scholar 

  50. Barar J, Hamzeiy H, Tabatabaei SAM, Hashemi-Aghdam SE, Omidi Y. Genomic signature and toxicogenomics comparison of polycationic gene delivery nanosystems in human alveolar epithelial A549 cells. Daru. 2009;17(3):139–47.

    CAS  Google Scholar 

  51. Saito Y, Higuchi Y, Kawakami S, Yamashita F, Hashida M. Immunostimulatory characteristics induced by linear polyethyleneimine-plasmid DNA complexes in cultured macrophages. Hum Gene Ther. 2009;20(2):137–45.

    Article  PubMed  CAS  Google Scholar 

  52. Shell SA, Hesse C, Morris SM, Milcarek C. Elevated levels of the 64-kDa cleavage stimulatory factor (CstF-64) in lipopolysaccharide-stimulated macrophages influence gene expression and induce alternative poly(A) site selection. J Biol Chem. 2005;280(48):39950–61.

    Article  PubMed  CAS  Google Scholar 

  53. Yan W, Ma L, Stein P, Pangas SA, Burns KH, Bai Y, et al. Mice deficient in oocyte-specific oligoadenylate synthetase-like protein OAS1D display reduced fertility. Mol Cell Biol. 2005;25(11):4615–24.

    Article  PubMed  CAS  Google Scholar 

  54. Merkel OM, Librizzi D, Pfestroff A, Schurrat T, Buyens K, Sanders NN, et al. Stability of siRNA polyplexes from poly(ethylenimine) and poly(ethylenimine)-g-poly(ethylene glycol) under in vivo conditions: effects on pharmacokinetics and biodistribution measured by Fluorescence Fluctuation Spectroscopy and Single Photon Emission Computed Tomography (SPECT) imaging. J Control Release. 2009;138(2):148–59.

    Article  PubMed  CAS  Google Scholar 

  55. Buyens K, Meyer M, Wagner E, Demeester J, De Smedt SC, Sanders NN. Monitoring the disassembly of siRNA polyplexes in serum is crucial for predicting their biological efficacy. J Control Release. 2010;141(1):38–41.

    Article  PubMed  CAS  Google Scholar 

  56. Brunner T, Cohen S, Monsonego A. Silencing of proinflammatory genes targeted to peritoneal-residing macrophages using siRNA encapsulated in biodegradable microspheres. Biomaterials. 2010;31(9):2627–36.

    Article  PubMed  CAS  Google Scholar 

  57. Owens DE, Peppas NA. Opsonization, biodistribution, and pharmacokinetics of polymeric nanoparticles. Int J Pharm. 2006;307(1):93–102.

    Article  PubMed  CAS  Google Scholar 

  58. Millili PG, Selekman JA, Blocker KM, Johnson DA, Naik UP, Sullivan MO. Structural and functional consequences of poly(ethylene glycol) inclusion on DNA condensation for gene delivery. Microsc Res Tech. 2010;73(9):866–77.

    Article  PubMed  CAS  Google Scholar 

  59. Raemdonck K, Van Thienen TG, Vandenbroucke RE, Sanders NN, Demeester J, De Smedt SC. Dextran microgels for time-controlled delivery of siRNA. Adv Funct Mater. 2008;18(7):993–1001.

    Article  CAS  Google Scholar 

  60. Raemdonck K, Naeye B, Hogset A, Demeester J, De Smedt SC. Prolonged gene silencing by combining siRNA nanogels and photochemical internalization. J Control Release. 2010;145(3):281–8.

    Article  PubMed  CAS  Google Scholar 

  61. Armstrong ME, Gantier M, Li LL, Chung WY, McCann A, Baugh JA, et al. Small interfering RNAs induce macrophage migration inhibitory factor production and proliferation in breast cancer cells via a double-stranded RNA-dependent protein kinase-dependent mechanism. J Immunol. 2008;180(11):7125–33.

    PubMed  CAS  Google Scholar 

  62. Omidi Y, Barar J, Heidari HR, Ahmadian S, Yazdi HA, Akhtar S. Microarray analysis of the toxicogenomics and the genotoxic potential of a cationic lipid-based gene delivery nanosystem in human alveolar epithelial A549 cells. Toxicol Mech Method. 2008;18(4):369–78.

    Article  CAS  Google Scholar 

  63. Omidi Y, Hollins AJ, Drayton RM, Akhtar S. Polypropylenimine dendrimer-induced gene expression changes: The effect of complexation with DNA, dendrimer generation and cell type. J Drug Target. 2005;13(7):431–43.

    Article  PubMed  CAS  Google Scholar 

  64. Kuo JHS, Liou MJ, Chiu HC. Evaluating the gene-expression profiles of HeLa cancer cells treated with activated and nonactivated poly(amidoamine) dendrimers, and their DNA complexes. Mol Pharm. 2010;7(3):805–14.

    Article  PubMed  CAS  Google Scholar 

  65. Vannucci L, Fiserova A, Sadalapure K, Lindhorst TK, Kuldova M, Rossmann P, et al. Effects of N-acetyl-glucosamine-coated glycodendrimers as biological modulators in the B16F10 melanoma model in vivo. Int J Oncol. 2003;23(2):285–96.

    PubMed  CAS  Google Scholar 

  66. Tai CJ, Hsu CH, Shen SC, Lee WR, Jiang MC. Cellular apoptosis susceptibility (CSE1L/CAS) protein in cancer metastasis and chemotherapeutic drug-induced apoptosis. J Exp Clin Cancer Res. 2010;29.

Download references

ACKNOWLEDGMENTS & DISCLOSURES

We gratefully thank Kirsten Vikkelsø Madsen for valuable scientific discussions concerning real time RT-PCR, Lasse Bengtson for preparation of the DOTAP/PLGA nanoparticles, and Dr. Michael Timm for testing pyrogen levels of the delivery systems (all from the Faculty of Pharmaceutical Sciences, University of Copenhagen). This study has been carried out with financial support from the Commission of the European Communities, Priority 3 “Nanotechnologies and Nanosciences, Knowledge Based Multifunctional Materials, New Production Processes and Devices” of the Sixth Framework Programme for Research and Technological Development (Targeted Delivery of Nanomedicine: NMP4-CT-2006-026668). We are grateful to the Danish Agency for Science, Technology and Innovation, the Drug Research Academy and the Carlsberg Foundation for financial support for the Zetasizer Nano ZS, Nanodrop 2000 C Spectrophotometer and the LightCycler® 480 system, respectively. The funding sources had no involvement in the study design, in the collection, analysis and interpretation of data, just as they had no involvement in the writing of the report and the decision to submit the paper for publication.

Author information

Author notes

  1. Joscha Griger

    Present address: Max Delbrueck Centre for Molecular Medicine, Robert-Roessle-Str. 10, 13125, Berlin-Buch, Germany

Authors and Affiliations

  1. Department of Pharmaceutics and Analytical Chemistry Faculty of Pharmaceutical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen O, Denmark

    Linda B. Jensen, Joscha Griger, Hanne M. Nielsen & Camilla Foged

  2. Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium

    Broes Naeye, Koen Raemdonck & Stefaan C. de Smedt

  3. Department of Pharmaceutics, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA, Utrecht, The Netherlands

    Amir K. Varkouhi, Raymond Schiffelers, Twan Lammers & Gert Storm

  4. Department of Experimental Molecular Imaging, RWTH Aachen University, Pauwelsstrasse 30, 52074, Aachen, Germany

    Twan Lammers

  5. Chemconsilium GCV, Jaarmarktstraat 48, 2221, Booischot, Belgium

    Brian S. Sproat

Authors
  1. Linda B. Jensen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Joscha Griger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Broes Naeye
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Amir K. Varkouhi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Koen Raemdonck
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Raymond Schiffelers
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Twan Lammers
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Gert Storm
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Stefaan C. de Smedt
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Brian S. Sproat
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Hanne M. Nielsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Camilla Foged
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Camilla Foged.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jensen, L.B., Griger, J., Naeye, B. et al. Comparison of Polymeric siRNA Nanocarriers in a Murine LPS-Activated Macrophage Cell Line: Gene Silencing, Toxicity and Off-Target Gene Expression. Pharm Res 29, 669–682 (2012). https://doi.org/10.1007/s11095-011-0589-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11095-011-0589-0

KEY WORDS

Search

Navigation