Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro
New amphiphilic 1,4-DHP derivative C12-Man-Q with remoted cationic moieties at positions 2 and 6 was synthesised to study DNA delivery activity. The results were compared with data obtained for cationic 1,4-DHP derivative D19, which is known to be the most efficient one among the previously tested 1...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2018-06-01
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| Series: | Molecules |
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| Online Access: | http://www.mdpi.com/1420-3049/23/7/1540 |
| _version_ | 1818153175292575744 |
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| author | Gunita Apsite Irena Timofejeva Aleksandra Vezane Brigita Vigante Martins Rucins Arkadij Sobolev Mara Plotniece Karlis Pajuste Tatjana Kozlovska Aiva Plotniece |
| author_facet | Gunita Apsite Irena Timofejeva Aleksandra Vezane Brigita Vigante Martins Rucins Arkadij Sobolev Mara Plotniece Karlis Pajuste Tatjana Kozlovska Aiva Plotniece |
| author_sort | Gunita Apsite |
| collection | DOAJ |
| description | New amphiphilic 1,4-DHP derivative C12-Man-Q with remoted cationic moieties at positions 2 and 6 was synthesised to study DNA delivery activity. The results were compared with data obtained for cationic 1,4-DHP derivative D19, which is known to be the most efficient one among the previously tested 1,4-DHP amphiphiles. We analysed the effects of C12-Man-Q concentration, complexation media, and complex/cell contact time on the gene delivery effectiveness and cell viability. Transmission electron microscopy data confirms that lipoplexes formed by the compound C12-Man-Q were quite uniform, vesicular-like structures with sizes of about 50 nm, and lipoplexes produced by compound D19 were of irregular shapes, varied in size in the range of 25–80 nm. Additionally, confocal microscopy results revealed that both amphiphiles effectively delivered green fluorescent protein expression plasmid into BHK-21 cells and produced a fluorescent signal with satisfactory efficiency, although compound C12-Man-Q was more cytotoxic to the BHK-21 cells with an increase of concentration. It can be concluded that optimal conditions for C12-Man-Q lipoplexes delivery in BHK-21 cells were the serum free media without 0.15 M NaCl, at an N/P ratio of 0.9. Compound D19 showed higher transfection efficiency to transfect BHK-21 and Cos-7 cell lines, when transfecting active proliferating cells. Although D19 was not able to transfect all studied cell lines we propose that it could be cell type specific. The compound C12-Man-Q showed modest delivery activity in all used cell lines, and higher activity was obtained in the case of H2-35 and B16 cells. The transfection efficiency in cell lines MCF-7, HeLa, and Huh-7 appears to be comparable to the reference compound D19 and minimal in the HepG2 cell line. |
| first_indexed | 2024-12-11T14:06:26Z |
| format | Article |
| id | doaj.art-611c777d0f414362a47fcbfe447e4b6d |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-11T14:06:26Z |
| publishDate | 2018-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-611c777d0f414362a47fcbfe447e4b6d2022-12-22T01:03:39ZengMDPI AGMolecules1420-30492018-06-01237154010.3390/molecules23071540molecules23071540Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In VitroGunita Apsite0Irena Timofejeva1Aleksandra Vezane2Brigita Vigante3Martins Rucins4Arkadij Sobolev5Mara Plotniece6Karlis Pajuste7Tatjana Kozlovska8Aiva Plotniece9Latvian Biomedical Research and Study Centre, Ratsupites iela 1, LV-1067 Riga, LatviaLatvian Biomedical Research and Study Centre, Ratsupites iela 1, LV-1067 Riga, LatviaLatvian Biomedical Research and Study Centre, Ratsupites iela 1, LV-1067 Riga, LatviaLatvian Institute of Organic Synthesis, Aizkraukles iela 21, LV-1006 Riga, LatviaLatvian Institute of Organic Synthesis, Aizkraukles iela 21, LV-1006 Riga, LatviaLatvian Institute of Organic Synthesis, Aizkraukles iela 21, LV-1006 Riga, LatviaLatvian Institute of Organic Synthesis, Aizkraukles iela 21, LV-1006 Riga, LatviaLatvian Institute of Organic Synthesis, Aizkraukles iela 21, LV-1006 Riga, LatviaLatvian Biomedical Research and Study Centre, Ratsupites iela 1, LV-1067 Riga, LatviaLatvian Institute of Organic Synthesis, Aizkraukles iela 21, LV-1006 Riga, LatviaNew amphiphilic 1,4-DHP derivative C12-Man-Q with remoted cationic moieties at positions 2 and 6 was synthesised to study DNA delivery activity. The results were compared with data obtained for cationic 1,4-DHP derivative D19, which is known to be the most efficient one among the previously tested 1,4-DHP amphiphiles. We analysed the effects of C12-Man-Q concentration, complexation media, and complex/cell contact time on the gene delivery effectiveness and cell viability. Transmission electron microscopy data confirms that lipoplexes formed by the compound C12-Man-Q were quite uniform, vesicular-like structures with sizes of about 50 nm, and lipoplexes produced by compound D19 were of irregular shapes, varied in size in the range of 25–80 nm. Additionally, confocal microscopy results revealed that both amphiphiles effectively delivered green fluorescent protein expression plasmid into BHK-21 cells and produced a fluorescent signal with satisfactory efficiency, although compound C12-Man-Q was more cytotoxic to the BHK-21 cells with an increase of concentration. It can be concluded that optimal conditions for C12-Man-Q lipoplexes delivery in BHK-21 cells were the serum free media without 0.15 M NaCl, at an N/P ratio of 0.9. Compound D19 showed higher transfection efficiency to transfect BHK-21 and Cos-7 cell lines, when transfecting active proliferating cells. Although D19 was not able to transfect all studied cell lines we propose that it could be cell type specific. The compound C12-Man-Q showed modest delivery activity in all used cell lines, and higher activity was obtained in the case of H2-35 and B16 cells. The transfection efficiency in cell lines MCF-7, HeLa, and Huh-7 appears to be comparable to the reference compound D19 and minimal in the HepG2 cell line.http://www.mdpi.com/1420-3049/23/7/1540cationic amphiphiles1,4-dihydropyridineslipoplexesDNA transfectiongene deliverycytotoxicity |
| spellingShingle | Gunita Apsite Irena Timofejeva Aleksandra Vezane Brigita Vigante Martins Rucins Arkadij Sobolev Mara Plotniece Karlis Pajuste Tatjana Kozlovska Aiva Plotniece Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro Molecules cationic amphiphiles 1,4-dihydropyridines lipoplexes DNA transfection gene delivery cytotoxicity |
| title | Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro |
| title_full | Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro |
| title_fullStr | Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro |
| title_full_unstemmed | Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro |
| title_short | Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro |
| title_sort | synthesis and comparative evaluation of novel cationic amphiphile c12 man q as an efficient dna delivery agent in vitro |
| topic | cationic amphiphiles 1,4-dihydropyridines lipoplexes DNA transfection gene delivery cytotoxicity |
| url | http://www.mdpi.com/1420-3049/23/7/1540 |
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