Layer-by-layer nanoparticles as an efficient siRNA delivery vehicle for SPARC silencing
Efficient and safe delivery systems for siRNA therapeutics remain a challenge. Elevated secreted protein, acidic, and rich in cysteine (SPARC) protein expression is associated with tissue scarring and fibrosis. Here we investigate the feasibility of encapsulating SPARC-siRNA in the bilayers of layer...
| Main Authors: | , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
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2014
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/103586 http://hdl.handle.net/10220/19326 |
| _version_ | 1826127841551450112 |
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| author | Tan, Yang Fei Chen, Min Hui Averil Lessig, Jacqueline Neu, Björn Mundargi, Raghavendra C. Venkatraman, Subbu S. Wong, Tina T. |
| author2 | School of Materials Science & Engineering |
| author_facet | School of Materials Science & Engineering Tan, Yang Fei Chen, Min Hui Averil Lessig, Jacqueline Neu, Björn Mundargi, Raghavendra C. Venkatraman, Subbu S. Wong, Tina T. |
| author_sort | Tan, Yang Fei |
| collection | NTU |
| description | Efficient and safe delivery systems for siRNA therapeutics remain a challenge. Elevated secreted protein, acidic, and rich in cysteine (SPARC) protein expression is associated with tissue scarring and fibrosis. Here we investigate the feasibility of encapsulating SPARC-siRNA in the bilayers of layer-by-layer (LbL) nanoparticles (NPs) with poly(L-arginine) (ARG) and dextran (DXS) as polyelectrolytes. Cellular binding and uptake of LbL NPs as well as siRNA delivery were studied in FibroGRO cells. siGLO-siRNA and SPARC-siRNA were efficiently coated onto hydroxyapatite nanoparticles. The multilayered NPs were characterized with regard to particle size, zeta potential and surface morphology using dynamic light scattering and transmission electron microscopy. The SPARC-gene silencing and mRNA levels were analyzed using ChemiDOC western blot technique and RT-PCR. The multilayer SPARC-siRNA incorporated nanoparticles are about 200 nm in diameter and are efficiently internalized into FibroGRO cells. Their intracellular fate was also followed by tagging with suitable reporter siRNA as well as with lysotracker dye; confocal microscopy clearly indicates endosomal escape of the particles. Significant (60%) SPARC-gene knock down was achieved by using 0.4 pmole siRNA/μg of LbL NPs in FibroGRO cells and the relative expression of SPARC mRNA reduced significantly (60%) against untreated cells. The cytotoxicity as evaluated by xCelligence real-time cell proliferation and MTT cell assay, indicated that the SPARC-siRNA-loaded LbL NPs are non-toxic. In conclusion, the LbL NP system described provides a promising, safe and efficient delivery platform as a non-viral vector for siRNA delivery that uses biopolymers to enhance the gene knock down efficiency for the development of siRNA therapeutics. |
| first_indexed | 2024-10-01T07:15:24Z |
| format | Journal Article |
| id | ntu-10356/103586 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:15:24Z |
| publishDate | 2014 |
| record_format | dspace |
| spelling | ntu-10356/1035862020-06-01T10:13:51Z Layer-by-layer nanoparticles as an efficient siRNA delivery vehicle for SPARC silencing Tan, Yang Fei Chen, Min Hui Averil Lessig, Jacqueline Neu, Björn Mundargi, Raghavendra C. Venkatraman, Subbu S. Wong, Tina T. School of Materials Science & Engineering DRNTU::Engineering::Materials::Nanostructured materials Efficient and safe delivery systems for siRNA therapeutics remain a challenge. Elevated secreted protein, acidic, and rich in cysteine (SPARC) protein expression is associated with tissue scarring and fibrosis. Here we investigate the feasibility of encapsulating SPARC-siRNA in the bilayers of layer-by-layer (LbL) nanoparticles (NPs) with poly(L-arginine) (ARG) and dextran (DXS) as polyelectrolytes. Cellular binding and uptake of LbL NPs as well as siRNA delivery were studied in FibroGRO cells. siGLO-siRNA and SPARC-siRNA were efficiently coated onto hydroxyapatite nanoparticles. The multilayered NPs were characterized with regard to particle size, zeta potential and surface morphology using dynamic light scattering and transmission electron microscopy. The SPARC-gene silencing and mRNA levels were analyzed using ChemiDOC western blot technique and RT-PCR. The multilayer SPARC-siRNA incorporated nanoparticles are about 200 nm in diameter and are efficiently internalized into FibroGRO cells. Their intracellular fate was also followed by tagging with suitable reporter siRNA as well as with lysotracker dye; confocal microscopy clearly indicates endosomal escape of the particles. Significant (60%) SPARC-gene knock down was achieved by using 0.4 pmole siRNA/μg of LbL NPs in FibroGRO cells and the relative expression of SPARC mRNA reduced significantly (60%) against untreated cells. The cytotoxicity as evaluated by xCelligence real-time cell proliferation and MTT cell assay, indicated that the SPARC-siRNA-loaded LbL NPs are non-toxic. In conclusion, the LbL NP system described provides a promising, safe and efficient delivery platform as a non-viral vector for siRNA delivery that uses biopolymers to enhance the gene knock down efficiency for the development of siRNA therapeutics. 2014-05-15T01:14:54Z 2019-12-06T21:16:00Z 2014-05-15T01:14:54Z 2019-12-06T21:16:00Z 2014 2014 Journal Article Tan, Y. F., Mundargi, R. C., Chen, M. H. A., Lessig, J., Neu, B., Venkatraman, S. S., & Wong, T. T. (2014). Layer-by-Layer Nanoparticles as an Efficient siRNA Delivery Vehicle for SPARC Silencing . Small, 10(9), 1790-1798. 1613-6810 https://hdl.handle.net/10356/103586 http://hdl.handle.net/10220/19326 10.1002/smll.201303201 178779 en Small © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. |
| spellingShingle | DRNTU::Engineering::Materials::Nanostructured materials Tan, Yang Fei Chen, Min Hui Averil Lessig, Jacqueline Neu, Björn Mundargi, Raghavendra C. Venkatraman, Subbu S. Wong, Tina T. Layer-by-layer nanoparticles as an efficient siRNA delivery vehicle for SPARC silencing |
| title | Layer-by-layer nanoparticles as an efficient siRNA delivery vehicle for SPARC silencing |
| title_full | Layer-by-layer nanoparticles as an efficient siRNA delivery vehicle for SPARC silencing |
| title_fullStr | Layer-by-layer nanoparticles as an efficient siRNA delivery vehicle for SPARC silencing |
| title_full_unstemmed | Layer-by-layer nanoparticles as an efficient siRNA delivery vehicle for SPARC silencing |
| title_short | Layer-by-layer nanoparticles as an efficient siRNA delivery vehicle for SPARC silencing |
| title_sort | layer by layer nanoparticles as an efficient sirna delivery vehicle for sparc silencing |
| topic | DRNTU::Engineering::Materials::Nanostructured materials |
| url | https://hdl.handle.net/10356/103586 http://hdl.handle.net/10220/19326 |
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