Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chai Q, Jiao Y, Yu X (2017) Hydrogels for biomedical applications: their characteristics and the mechanisms behind them. Gels 3:6–15. https://doi.org/10.3390/gels3010006
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. https://doi.org/10.1111/j.1582-4934.2012.01603.x
Carballo-Molina OA, Velasco I (2015) Hydrogels as scaffolds and delivery systems to enhance axonal regeneration after injuries. Front Cell Neurosci 9:13. https://doi.org/10.3389/fncel.2015.00013
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. https://doi.org/10.1093/nar/gkp841
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. https://doi.org/10.1093/nar/gkq457
Crooke ST, Wang S, Vickers TA et al (2017) Cellular uptake and trafficking of antisense oligonucleotides. Nat Biotechnol 35:230–237. https://doi.org/10.1038/nbt.3779
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. https://doi.org/10.1126/scitranslmed.3001777
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. https://doi.org/10.1016/j.neuron.2012.05.009
Khorkova O, Wahlestedt C (2017) Oligonucleotide therapies for disorders of the nervous system. Nat Biotechnol 35:249–263. https://doi.org/10.1038/nbt.3784
Smith CIE, Zain R (2018) Therapeutic oligonucleotides: state of the art. Annu Rev Pharmacol Toxicol. https://doi.org/10.1146/annurev-pharmtox-010818-021050
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. https://doi.org/10.1002/bit.22476
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. https://doi.org/10.1016/j.biomaterials.2010.02.018
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. https://doi.org/10.1016/j.expneurol.2014.04.008
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. https://doi.org/10.1016/j.omtn.2018.03.009
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. https://doi.org/10.1098/rsif.2014.1224
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. https://doi.org/10.1007/s10856-017-5967-7
Acknowledgments
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Moreno, P.M.D., Rodrigues, T., Torrado, M., Amaral, I.F., Pêgo, A.P. (2019). Delivery of Antisense Oligonucleotides Mediated by a Hydrogel System: In Vitro and In Vivo Application in the Context of Spinal Cord Injury. In: Gissberg, O., Zain, R., Lundin, K. (eds) Oligonucleotide-Based Therapies. Methods in Molecular Biology, vol 2036. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9670-4_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9670-4_12
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9669-8
Online ISBN: 978-1-4939-9670-4
eBook Packages: Springer Protocols