Delivery of Antisense Oligonucleotides Mediated by a Hydrogel System: In Vitro and In Vivo Application in the Context of Spinal Cord Injury

  • Pedro M. D. Moreno
  • Teresa Rodrigues
  • Marília Torrado
  • Isabel F. Amaral
  • Ana P. PêgoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2036)


Biomaterials-based hydrogels are attractive drug-eluting vehicles in the context of RNA therapeutics, such as those utilizing antisense oligonucleotide or RNA interference based drugs, as they can potentially reduce systemic toxicity and enhance in vivo efficacy by increasing in situ concentrations. Here we describe the preparation of antisense oligonucleotide-loaded fibrin hydrogels exploring their applications in the context of the nervous system utilizing an organotypic dorsal root ganglion explant in vitro system and an in vivo model of spinal cord injury.

Key words

Antisense oligonucleotides Hydrogel Fibrin Gene silencing Central nervous system Spinal cord injury Dorsal root ganglion 



This work was supported by Fundação para a Ciência e a Tecnologia (FCT, Portugal) in the framework of the Harvard-Portugal Medical School Program [HMSP-ICT/0020/2010]; Project NORTE-01-0145-FEDER-000008, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020—Operacional Program for Competitiveness and Internationalization (POCI), Portugal 2020; by Portuguese funds through FCT/Ministério da Ciência, Tecnologia e Ensino Superior in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274); Santa Casa da Misericordia de Lisboa—Prémio Neurociências Mello e Castro (MC-1068-2015) and the fellowships SFRH/BPD/108738/2015 (FCT) to P.M.D.M and Infarmed (FIS-FIS-2015-01_CCV_20150630-88) to M.T.


  1. 1.
    Chai Q, Jiao Y, Yu X (2017) Hydrogels for biomedical applications: their characteristics and the mechanisms behind them. Gels 3:6–15. Scholar
  2. 2.
    Pêgo AP, Kubinova S, Cizkova D et al (2012) Regenerative medicine for the treatment of spinal cord injury: more than just promises? J Cell Mol Med 16:2564–2582. Scholar
  3. 3.
    Carballo-Molina OA, Velasco I (2015) Hydrogels as scaffolds and delivery systems to enhance axonal regeneration after injuries. Front Cell Neurosci 9:13. Scholar
  4. 4.
    Stein CA, Hansen JB, Lai J et al (2010) Efficient gene silencing by delivery of locked nucleic acid antisense oligonucleotides, unassisted by transfection reagents. Nucleic Acids Res 38:e3. Scholar
  5. 5.
    Straarup EM, Fisker N, Hedtjärn M et al (2010) Short locked nucleic acid antisense oligonucleotides potently reduce apolipoprotein B mRNA and serum cholesterol in mice and non-human primates. Nucleic Acids Res 38:7100–7111. Scholar
  6. 6.
    Crooke ST, Wang S, Vickers TA et al (2017) Cellular uptake and trafficking of antisense oligonucleotides. Nat Biotechnol 35:230–237. Scholar
  7. 7.
    Passini MA, Bu J, Richards AM et al (2011) Antisense oligonucleotides delivered to the mouse CNS ameliorate symptoms of severe spinal muscular atrophy. Sci Transl Med 3:72ra18. Scholar
  8. 8.
    Kordasiewicz HB, Stanek LM, Wancewicz EV et al (2012) Sustained therapeutic reversal of Huntington's disease by transient repression of Huntingtin synthesis. Neuron 74:1031–1044. Scholar
  9. 9.
    Khorkova O, Wahlestedt C (2017) Oligonucleotide therapies for disorders of the nervous system. Nat Biotechnol 35:249–263. Scholar
  10. 10.
    Smith CIE, Zain R (2018) Therapeutic oligonucleotides: state of the art. Annu Rev Pharmacol Toxicol. Scholar
  11. 11.
    Johnson PJ, Parker SR, Sakiyama-Elbert SE (2009) Controlled release of neurotrophin-3 from fibrin-based tissue engineering scaffolds enhances neural fiber sprouting following subacute spinal cord injury. Biotechnol Bioeng 104:1207–1214. Scholar
  12. 12.
    King VR, Alovskaya A, Wei DYT et al (2010) The use of injectable forms of fibrin and fibronectin to support axonal ingrowth after spinal cord injury. Biomaterials 31:4447–4456. Scholar
  13. 13.
    Sharp KG, Yee KM, Steward O (2014) A re-assessment of long distance growth and connectivity of neural stem cells after severe spinal cord injury. Exp Neurol 257:186–204. Scholar
  14. 14.
    Moreno PMD, Ferreira AR, Salvador D et al (2018) Hydrogel-assisted antisense LNA Gapmer delivery for in situ gene silencing in spinal cord injury. Mol Ther Nucleic Acids 11:393–406. Scholar
  15. 15.
    Pires LR, Rocha DN, Ambrosio L, Pêgo AP (2015) The role of the surface on microglia function: implications for central nervous system tissue engineering. J R Soc Interface 12:20141224–20141224. Scholar
  16. 16.
    Pires LR, Lopes CDF, Salvador D et al (2017) Ibuprofen-loaded fibrous patches-taming inhibition at the spinal cord injury site. J Mater Sci Mater Med 28:157. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Pedro M. D. Moreno
    • 1
    • 2
  • Teresa Rodrigues
    • 1
    • 2
  • Marília Torrado
    • 1
    • 2
  • Isabel F. Amaral
    • 1
    • 2
  • Ana P. Pêgo
    • 1
    • 2
    • 3
    • 4
    Email author
  1. 1.i3S—Instituto de Investigação e Inovação em SaúdeUniversidade do PortoPortoPortugal
  2. 2.INEB—Instituto de Engenharia BiomédicaUniversidade do PortoPortoPortugal
  3. 3.Faculdade de Engenharia da Universidade do PortoPortoPortugal
  4. 4.Instituto de Ciências Biomédicas Abel Salazar (ICBAS)Universidade do PortoPortoPortugal

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