Graft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfection

The first chapter reviews the most important aspects of the polymeric vectors used in gene therapy. This type of therapy involves the introduction of genetic material into specific cells to treat certain diseases, using vectors that mediate the cellular internalization of nucleic acids and protect t...

Full description

Autores:
Díaz Ariza, Ivonne Lorena
Tipo de recurso:
Doctoral thesis
Fecha de publicación:
2020
Institución:
Universidad Nacional de Colombia
Repositorio:
Universidad Nacional de Colombia
Idioma:
eng
OAI Identifier:
oai:repositorio.unal.edu.co:unal/78367
Acceso en línea:
https://repositorio.unal.edu.co/handle/unal/78367
Palabra clave:
660 - Ingeniería química
547 - Química orgánica
PDMAEMA
PEI
DNA transfection
Gene therapy
Polymeric vectors
Cationic graft copolymers
PDMAEMA
PEI
Terapia génica
Vectores poliméricos
Copolímeros de injerto catiónicos
Transfección de ADN
Rights
openAccess
License
Atribución-SinDerivadas 4.0 Internacional
id UNACIONAL2_b11fc4e1c1005ddadb34945604a5c422
oai_identifier_str oai:repositorio.unal.edu.co:unal/78367
network_acronym_str UNACIONAL2
network_name_str Universidad Nacional de Colombia
repository_id_str
dc.title.spa.fl_str_mv Graft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfection
title Graft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfection
spellingShingle Graft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfection
660 - Ingeniería química
547 - Química orgánica
PDMAEMA
PEI
DNA transfection
Gene therapy
Polymeric vectors
Cationic graft copolymers
PDMAEMA
PEI
Terapia génica
Vectores poliméricos
Copolímeros de injerto catiónicos
Transfección de ADN
title_short Graft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfection
title_full Graft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfection
title_fullStr Graft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfection
title_full_unstemmed Graft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfection
title_sort Graft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfection
dc.creator.fl_str_mv Díaz Ariza, Ivonne Lorena
dc.contributor.advisor.spa.fl_str_mv Pérez Pérez, León Darío
dc.contributor.author.spa.fl_str_mv Díaz Ariza, Ivonne Lorena
dc.contributor.researchgroup.spa.fl_str_mv Macromoléculas
dc.subject.ddc.spa.fl_str_mv 660 - Ingeniería química
547 - Química orgánica
topic 660 - Ingeniería química
547 - Química orgánica
PDMAEMA
PEI
DNA transfection
Gene therapy
Polymeric vectors
Cationic graft copolymers
PDMAEMA
PEI
Terapia génica
Vectores poliméricos
Copolímeros de injerto catiónicos
Transfección de ADN
dc.subject.proposal.eng.fl_str_mv PDMAEMA
PEI
DNA transfection
Gene therapy
Polymeric vectors
Cationic graft copolymers
dc.subject.proposal.spa.fl_str_mv PDMAEMA
PEI
Terapia génica
Vectores poliméricos
Copolímeros de injerto catiónicos
Transfección de ADN
description The first chapter reviews the most important aspects of the polymeric vectors used in gene therapy. This type of therapy involves the introduction of genetic material into specific cells to treat certain diseases, using vectors that mediate the cellular internalization of nucleic acids and protect them from degradation. Synthetic cationic polymers have shown promising results as gene vectors, due to their advantages such as higher safety, simple fabrication process and modifiable structure. Poly(ethyleneimine) (PEI) and poly(2- (dimethylamino)ethyl methacrylate) (PDMAEMA) have been highlighted in this context due to their high transfection efficiency; however, the non-biodegradable character and associated cytotoxicity have limited their applications. To overcome these disadvantages, several researchers have studied the implementation of strategies such as PEGylation, copolymerization with hydrophobic segments and the use of branched architectures, which have demonstrated to significantly reduce the toxicity and increase the transfection efficiency. In addition, these structural modifications can be perform using synthetic methods, such as controlled polymerizations and selective coupling reactions, which allow the design of molecules with well-defined properties. The second chapter describes the synthesis and characterization of copolymers composed of methoxy poly(ethylene glycol) and a hydrophobic block of poly(ɛ-caprolactone-co-propargyl carbonate) grafted with low molecular weight PDMAEMA or linear PEI (lPEI), using a combination of ring opening polymerization, click chemistry and atom transfer radical polymerization. In this way, materials with different grafting densities and lengths of the hydrophobic segment were obtained. Following the proposed synthetic route, the synthesis of graft copolymers based on PDMAEMA with target structure and composition was achieved, while for copolymers composed of lPEI, lower grafting densities than the target ones were obtained. These materials could self-assembled in aqueous medium to form positively charged particles with average sizes between 150 and 380 nm. The third chapter covers the formation and characterization of the copolymer/DNA complexes with different nitrogen/phosphorus (N/P) ratios and complexation matrices. The study of the condensation ability showed that all PDMAEMA copolymers were able to complex the DNA molecules at N/P ratios ≥ 1, regardless of the used conditions, while lPEI copolymers required N/P ratios from 7 to 20, depending on their composition and the complexation solution. In the case of the materials composed of PDMAEMA, the measurements of particle size and zeta potential indicated the formation of positively charged complexes with sizes below 300 nm, which are suitable for cellular uptake. On the other hand, the lPEI based complexes exhibited negative or neutral surface charges with hydrodynamic diameters below 500 nm. These physicochemical properties could generate restrictions for the effective delivery of DNA. The fourth chapter describes the in vitro cytotoxicity of graft copolymers and the transfection efficiency of the resulting polyplexes, using 25 kDa lPEI and 20 kDa PDMAEMA as controls. The copolymers were less cytotoxic to L929 cells than the control homopolymers, exhibiting higher cell viability by reducing the length of the hydrophobic segment and the amount of cationic grafts. Polyplexes formed from PDMAEMA copolymers were more efficient in the delivery of DNA in HEK-293 cells than the standard polycations, without affecting the cell viability. On the other hand, the complexes based on lPEI copolymers exhibited a low transfection efficiency, even using high polymer concentrations and N/P ratios. In general, copolymers composed of long hydrophobic segments and higher grafting density showed a better performance. In particular, the material PP6D5 showed the most promising features for its application as DNA transfection agent, for both in vitro and in vivo assays. This thesis represents the starting point for future research related to the rational design of grafted structures based on cationic polymers that may have improved properties and performance for their application as gene delivery systems.
publishDate 2020
dc.date.accessioned.spa.fl_str_mv 2020-09-03T03:57:31Z
dc.date.available.spa.fl_str_mv 2020-09-03T03:57:31Z
dc.date.issued.spa.fl_str_mv 2020-02-14
dc.type.spa.fl_str_mv Trabajo de grado - Doctorado
dc.type.driver.spa.fl_str_mv info:eu-repo/semantics/doctoralThesis
dc.type.version.spa.fl_str_mv info:eu-repo/semantics/acceptedVersion
dc.type.coar.spa.fl_str_mv http://purl.org/coar/resource_type/c_db06
dc.type.content.spa.fl_str_mv Text
format http://purl.org/coar/resource_type/c_db06
status_str acceptedVersion
dc.identifier.uri.none.fl_str_mv https://repositorio.unal.edu.co/handle/unal/78367
url https://repositorio.unal.edu.co/handle/unal/78367
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.references.spa.fl_str_mv 1. Ospina, M. L.; Huertas, J. A.; Montaño, J. I.; Rivillas, J. C., Observatorio Nacional de Cáncer Colombia/The Colombian National Cancer Observatory/Observatório Nacional de Câncer Colômbia. Revista de la Facultad Nacional de Salud Pública 2015, 33, (2), 262.
2. Lozano, R.; Naghavi, M.; Foreman, K.; Lim, S.; Shibuya, K.; Aboyans, V.; Abraham, J.; Adair, T.; Aggarwal, R.; Ahn, S. Y., Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. The lancet 2012, 380, (9859), 2095-2128.
3. Mintzer, M. A.; Simanek, E. E., Nonviral vectors for gene delivery. Chemical reviews 2008, 109, (2), 259-302.
4. Naldini, L., Gene therapy returns to centre stage. Nature 2015, 526, (7573), 351-360.
5. Liu, X. Q.; Sun, C. Y.; Yang, X. Z.; Wang, J., Polymeric‐Micelle‐Based Nanomedicine for siRNA Delivery. Particle & Particle Systems Characterization 2013, 30, (3), 211-228.
6. Falamarzian, A.; Xiong, X.-B.; Uludag, H.; Lavasanifar, A., Polymeric micelles for siRNA delivery. Journal of Drug Delivery Science and Technology 2012, 22, (1), 43-54.
7. Ramamoorth, M.; Narvekar, A., Non viral vectors in gene therapy-an overview. Journal of clinical and diagnostic research: JCDR 2015, 9, (1), GE01.
8. Samal, S. K.; Dash, M.; Van Vlierberghe, S.; Kaplan, D. L.; Chiellini, E.; Van Blitterswijk, C.; Moroni, L.; Dubruel, P., Cationic polymers and their therapeutic potential. Chemical Society Reviews 2012, 41, (21), 7147-7194.
9. Samsonova, O.; Pfeiffer, C.; Hellmund, M.; Merkel, O. M.; Kissel, T., Low molecular weight pDMAEMA-block-pHEMA block-copolymers synthesized via RAFT-polymerization: potential non-viral gene delivery agents? Polymers 2011, 3, (2), 693-718.
10. Lin, D.; Huang, Y.; Jiang, Q.; Zhang, W.; Yue, X.; Guo, S.; Xiao, P.; Du, Q.; Xing, J.; Deng, L., Structural contributions of blocked or grafted poly (2-dimethylaminoethyl methacrylate) on PEGylated polycaprolactone nanoparticles in siRNA delivery. Biomaterials 2011, 32, (33), 8730-8742.
11. Guo, S.; Huang, Y.; Wei, T.; Zhang, W.; Wang, W.; Lin, D.; Zhang, X.; Kumar, A.; Du, Q.; Xing, J., Amphiphilic and biodegradable methoxy polyethylene glycol-block-(polycaprolactone-graft-poly (2-(dimethylamino) ethyl methacrylate)) as an effective gene carrier. Biomaterials 2011, 32, (3), 879-889.
12. Díaz-Ariza, I. L.; Sierra, C. A.; Pérez-Pérez, L. D., Desarrollo de vectores génicos basados en polímeros sintéticos: PEI y PDMAEMA. Revista Colombiana de Ciencias Químico-Farmacéuticas 2018, 47, (3), 350-374.
13. Zhou, Z.; Liu, X.; Zhu, D.; Wang, Y.; Zhang, Z.; Zhou, X.; Qiu, N.; Chen, X.; Shen, Y., Nonviral cancer gene therapy: delivery cascade and vector nanoproperty integration. Advanced drug delivery reviews 2017, 115, 115-154.
14. Nayerossadat, N.; Maedeh, T.; Ali, P. A., Viral and nonviral delivery systems for gene delivery. Advanced biomedical research 2012, 1.
15. Sun, X.; Zhang, N., Cationic polymer optimization for efficient gene delivery. Mini reviews in medicinal chemistry 2010, 10, (2), 108-125.
16. Caffery, B.; Lee, J. S.; Alexander-Bryant, A. A., Vectors for glioblastoma gene therapy: viral & non-viral delivery strategies. Nanomaterials 2019, 9, (1), 105.
17. Levine, R. M.; Scott, C. M.; Kokkoli, E., Peptide functionalized nanoparticles for nonviral gene delivery. Soft Matter 2013, 9, (4), 985-1004.
18. Junquera, E.; Aicart, E., Cationic lipids as transfecting agents of DNA in gene therapy. Current topics in medicinal chemistry 2014, 14, (5), 649-663.
19. Zhi, D.; Bai, Y.; Yang, J.; Cui, S.; Zhao, Y.; Chen, H.; Zhang, S., A review on cationic lipids with different linkers for gene delivery. Advances in Colloid and Interface Science 2017.
20. Pahle, J.; Walther, W., Vectors and strategies for nonviral cancer gene therapy. Expert opinion on biological therapy 2016, 16, (4), 443-461.
21. Pushpendra, S.; Arvind, P.; Anil, B., Nucleic acids as therapeutics. In From Nucleic Acids Sequences to Molecular Medicine, Springer: 2012; pp 19-45.
22. Sridharan, K.; Gogtay, N. J., Therapeutic nucleic acids: current clinical status. British journal of clinical pharmacology 2016, 82, (3), 659-672.
23. Schultz, N.; Marenstein, D. R.; De Angelis, D. A.; Wang, W.-Q.; Nelander, S.; Jacobsen, A.; Marks, D. S.; Massagué, J.; Sander, C., Off-target effects dominate a large-scale RNAi screen for modulators of the TGF-β pathway and reveal microRNA regulation of TGFBR2. Silence 2011, 2, (1), 3.
24. Foldvari, M.; Chen, D. W.; Nafissi, N.; Calderon, D.; Narsineni, L.; Rafiee, A., Non-viral gene therapy: Gains and challenges of non-invasive administration methods. Journal of Controlled Release 2016, 240, 165-190.
25. Uludag, H.; Ubeda, A.; Ansari, A., At the Intersection of Biomaterials and Gene Therapy: Progress in Non-viral Delivery of Nucleic Acids. Frontiers in Bioengineering and Biotechnology 2019, 7.
26. Walther, W.; Petkov, S.; Kuvardina, O.; Aumann, J.; Kobelt, D.; Fichtner, I.; Lemm, M.; Piontek, J.; Blasig, I.; Stein, U., Novel Clostridium perfringens enterotoxin suicide gene therapy for selective treatment of claudin-3-and-4-overexpressing tumors. Gene therapy 2012, 19, (5), 494.
27. Nakhlband, A.; Barar, J.; Bidmeshkipour, A.; Heidari, H. R.; Omidi, Y., Bioimpacts of anti epidermal growth receptor antisense complexed with polyamidoamine dendrimers in human lung epithelial adenocarcinoma cells. Journal of biomedical nanotechnology 2010, 6, (4), 360-369.
28. Chen, X.-A.; Zhang, L.-J.; He, Z.-J.; Wang, W.-W.; Xu, B.; Zhong, Q.; Shuai, X.-T.; Yang, L.-Q.; Deng, Y.-B., Plasmid-encapsulated polyethylene glycol-grafted polyethylenimine nanoparticles for gene delivery into rat mesenchymal stem cells. International journal of nanomedicine 2011, 6, 843.
29. Senovilla, L.; Vacchelli, E.; Garcia, P.; Eggermont, A.; Fridman, W. H.; Galon, J.; Zitvogel, L.; Kroemer, G.; Galluzzi, L., Trial watch: DNA vaccines for cancer therapy. Oncoimmunology 2013, 2, (4), e23803.
30. Lächelt, U.; Wagner, E., Nucleic acid therapeutics using polyplexes: a journey of 50 years (and beyond). Chemical reviews 2015, 115, (19), 11043-11078.
31. Yin, H.; Kanasty, R. L.; Eltoukhy, A. A.; Vegas, A. J.; Dorkin, J. R.; Anderson, D. G., Non-viral vectors for gene-based therapy. Nature Reviews Genetics 2014, 15, (8), 541.
32. Grandinetti, G.; Reineke, T. M., Exploring the mechanism of plasmid DNA nuclear internalization with polymer-based vehicles. Molecular pharmaceutics 2012, 9, (8), 2256-2267.
33. Kendrick, J.; Matthews, K.; Straughn Jr, J.; Barnes, M.; Fewell, J.; Anwer, K.; Alvarez, R. In A phase I trial of intraperitoneal EGEN-001, a novel IL-12 gene therapeutic, administered alone or in combination with chemotherapy in patients with recurrent ovarian cancer, ASCO Annual Meeting Proceedings, 2008; 2008; p 5572.
34. Sinn, P. L.; Anthony, R. M.; McCray Jr, P. B., Genetic therapies for cystic fibrosis lung disease. Human molecular genetics 2011, 20, (R1), R79-R86.
35. Nguyen, D. N.; Green, J. J.; Chan, J. M.; Langer, R.; Anderson, D. G., Polymeric materials for gene delivery and DNA vaccination. Advanced Materials 2009, 21, (8), 847-867.
36. Prabu, S. L.; Ruckmani, K., Biopolymer in Gene Delivery. Advanced Technology for Delivering Therapeutics 2017, 137.
37. Oskolkov, N. N.; Potemkin, I. I., Complexation in asymmetric solutions of oppositely charged polyelectrolytes: Phase diagram. Macromolecules 2007, 40, (23), 8423-8429.
38. Ou, Z.; Muthukumar, M., Entropy and enthalpy of polyelectrolyte complexation: Langevin dynamics simulations. The Journal of chemical physics 2006, 124, (15), 154902.
39. Zhang, P.; Wagner, E., History of polymeric gene delivery systems. Topics in Current Chemistry 2017, 375, (2), 26.
40. Vader, P.; van der Aa, L. J.; Engbersen, J. F.; Storm, G.; Schiffelers, R. M., Disulfide-based poly (amido amine) s for siRNA delivery: effects of structure on siRNA complexation, cellular uptake, gene silencing and toxicity. Pharmaceutical research 2011, 28, (5), 1013-1022.
41. Bishop, C. J.; Ketola, T.-M.; Tzeng, S. Y.; Sunshine, J. C.; Urtti, A.; Lemmetyinen, H.; Vuorimaa-Laukkanen, E.; Yliperttula, M.; Green, J. J., The Effect and Role of Carbon Atoms in Poly (β-amino ester) s for DNA Binding and Gene Delivery. Journal of the American Chemical Society 2013, 135, (18), 6951-6957.
42. Mathew, A.; Cao, H.; Collin, E.; Wang, W.; Pandit, A., Hyperbranched PEGmethacrylate linear pDMAEMA block copolymer as an efficient non-viral gene delivery vector. International journal of pharmaceutics 2012, 434, (1-2), 99-105.
43. Verbaan, F. J.; Klouwenberg, P. K.; van Steenis, J. H.; Snel, C. J.; Boerman, O.; Hennink, W. E.; Storm, G., Application of poly (2-(dimethylamino) ethyl methacrylate)-based polyplexes for gene transfer into human ovarian carcinoma cells. International journal of pharmaceutics 2005, 304, (1), 185-192.
44. Agarwal, S.; Zhang, Y.; Maji, S.; Greiner, A., PDMAEMA based gene delivery materials. Materials Today 2012, 15, (9), 388-393.
45. Synatschke, C. V.; Schallon, A.; Jérôme, V. r.; Freitag, R.; Müller, A. H., Influence of polymer architecture and molecular weight of poly (2-(dimethylamino) ethyl methacrylate) polycations on transfection efficiency and cell viability in gene delivery. Biomacromolecules 2011, 12, (12), 4247-4255.
46. Zhao, G.; Long, L.; Zhang, L.; Peng, M.; Cui, T.; Wen, X.; Zhou, X.; Sun, L.; Che, L., Smart pH-sensitive nanoassemblies with cleavable PEGylation for tumor targeted drug delivery. Scientific reports 2017, 7, (1), 3383.
47. Qiao, Y.; Huang, Y.; Qiu, C.; Yue, X.; Deng, L.; Wan, Y.; Xing, J.; Zhang, C.; Yuan, S.; Dong, A., The use of PEGylated poly [2-(N, N-dimethylamino) ethyl methacrylate] as a mucosal DNA delivery vector and the activation of innate immunity and improvement of HIV-1-specific immune responses. Biomaterials 2010, 31, (1), 115-123.
48. Jiang, X.; Lok, M. C.; Hennink, W. E., Degradable-brushed pHEMA–pDMAEMA synthesized via ATRP and click chemistry for gene delivery. Bioconjugate chemistry 2007, 18, (6), 2077-2084.
49. Song, Y.; Zhang, T.; Song, X.; Zhang, L.; Zhang, C.; Xing, J.; Liang, X.-J., Polycations with excellent gene transfection ability based on PVP-g-PDMAEMA with random coil and micelle structures as non-viral gene vectors. Journal of Materials Chemistry B 2015, 3, (5), 911-918.
50. Xu, F.; Ping, Y.; Ma, J.; Tang, G.; Yang, W.; Li, J.; Kang, E.; Neoh, K., Comb-shaped copolymers composed of hydroxypropyl cellulose backbones and cationic poly ((2-dimethyl amino) ethyl methacrylate) side chains for gene delivery. Bioconjugate chemistry 2009, 20, (8), 1449-1458.
51. Malikmammadov, E.; Tanir, T. E.; Kiziltay, A.; Hasirci, V.; Hasirci, N., PCL and PCL-based materials in biomedical applications. Journal of Biomaterials Science, Polymer Edition 2018, 29, (7-9), 863-893.
52. Ulery, B. D.; Nair, L. S.; Laurencin, C. T., Biomedical applications of biodegradable polymers. Journal of polymer science Part B: polymer physics 2011, 49, (12), 832-864.
53. Guo, S.; Qiao, Y.; Wang, W.; He, H.; Deng, L.; Xing, J.; Xu, J.; Liang, X.-J.; Dong, A., Poly (ε-caprolactone)-graft-poly (2-(N, N-dimethylamino) ethyl methacrylate) nanoparticles: pH dependent thermo-sensitive multifunctional carriers for gene and drug delivery. Journal of Materials Chemistry 2010, 20, (33), 6935-6941.
54. Huang, Y.; Lin, D.; Jiang, Q.; Zhang, W.; Guo, S.; Xiao, P.; Zheng, S.; Wang, X.; Chen, H.; Zhang, H.-Y., Binary and ternary complexes based on polycaprolactone-graft-poly (N, N-dimethylaminoethyl methacrylate) for targeted siRNA delivery. Biomaterials 2012, 33, (18), 4653-4664.
55. Zhu, C.; Jung, S.; Luo, S.; Meng, F.; Zhu, X.; Park, T. G.; Zhong, Z., Co-delivery of siRNA and paclitaxel into cancer cells by biodegradable cationic micelles based on PDMAEMA–PCL–PDMAEMA triblock copolymers. Biomaterials 2010, 31, (8), 2408-2416.
56. Yue, X.; Qiao, Y.; Qiao, N.; Guo, S.; Xing, J.; Deng, L.; Xu, J.; Dong, A., Amphiphilic Methoxy Poly (ethylene glycol)-b-poly (ε-caprolactone)-b-poly (2-dimethylaminoethyl methacrylate) Cationic Copolymer Nanoparticles as a Vector for Gene and Drug Delivery. Biomacromolecules 2010, 11, (9), 2306-2312.
57. Qian, X.; Long, L.; Shi, Z.; Liu, C.; Qiu, M.; Sheng, J.; Pu, P.; Yuan, X.; Ren, Y.; Kang, C., Star-branched amphiphilic PLA-b-PDMAEMA copolymers for co-delivery of miR-21 inhibitor and doxorubicin to treat glioma. Biomaterials 2014, 35, (7), 2322-2335.
58. Ziebarth, J. D.; Wang, Y., Understanding the protonation behavior of linear polyethylenimine in solutions through Monte Carlo simulations. Biomacromolecules 2009, 11, (1), 29-38.
59. Yu, Q.-Y.; Zhan, Y.-R.; Zhang, J.; Luan, C.-R.; Wang, B.; Yu, X.-Q., Aromatic Modification of Low Molecular Weight PEI for Enhanced Gene Delivery. Polymers 2017, 9, (8), 362.
60. Hu, J.; Zhao, W.; Liu, K.; Yu, Q.; Mao, Y.; Lu, Z.; Zhang, Y.; Zhu, M., Low-molecular weight polyethylenimine modified with pluronic 123 and rgd-or chimeric rgd-nls peptide: Characteristics and transfection efficacy of their complexes with plasmid DNA. Molecules 2016, 21, (5), 655.
61. Merdan, T.; Kunath, K.; Petersen, H.; Bakowsky, U.; Voigt, K. H.; Kopecek, J.; Kissel, T., PEGylation of poly (ethylene imine) affects stability of complexes with plasmid DNA under in vivo conditions in a dose-dependent manner after intravenous injection into mice. Bioconjugate chemistry 2005, 16, (4), 785-792.
62. Mao, S.; Neu, M.; Germershaus, O.; Merkel, O.; Sitterberg, J.; Bakowsky, U.; Kissel, T., Influence of polyethylene glycol chain length on the physicochemical and biological properties of poly (ethylene imine)-graft-poly (ethylene glycol) block copolymer/SiRNA polyplexes. Bioconjugate chemistry 2006, 17, (5), 1209-1218.
63. Ochrimenko, S.; Vollrath, A.; Tauhardt, L.; Kempe, K.; Schubert, S.; Schubert, U. S.; Fischer, D., Dextran-graft-linear poly (ethylene imine) s for gene delivery: importance of the linking strategy. Carbohydrate polymers 2014, 113, 597-606.
64. Min, S.-H.; Park, K. C.; Yeom, Y. I., Chitosan-mediated non-viral gene delivery with improved serum stability and reduced cytotoxicity. Biotechnology and bioprocess engineering 2014, 19, (6), 1077-1082.
65. Cao, N.; Cheng, D.; Zou, S.; Ai, H.; Gao, J.; Shuai, X., The synergistic effect of hierarchical assemblies of siRNA and chemotherapeutic drugs co-delivered into hepatic cancer cells. Biomaterials 2011, 32, (8), 2222-2232.
66. Qiu, L. Y.; Bae, Y. H., Self-assembled polyethylenimine-graft-poly (ε-caprolactone) micelles as potential dual carriers of genes and anticancer drugs. Biomaterials 2007, 28, (28), 4132-4142.
67. Endres, T.; Zheng, M.; Kılıç, A. e.; Turowska, A.; Beck-Broichsitter, M.; Renz, H.; Merkel, O. M.; Kissel, T., Amphiphilic biodegradable PEG-PCL-PEI triblock copolymers for FRET-capable in vitro and in vivo delivery of siRNA and quantum dots. Molecular pharmaceutics 2014, 11, (4), 1273-1281.
68. Zheng, M.; Liu, Y.; Samsonova, O.; Endres, T.; Merkel, O.; Kissel, T., Amphiphilic and biodegradable hy-PEI-g-PCL-b-PEG copolymers efficiently mediate transgene expression depending on their graft density. International journal of pharmaceutics 2012, 427, (1), 80-87.
69. Liu, Y.; Samsonova, O.; Sproat, B.; Merkel, O.; Kissel, T., Biophysical characterization of hyper-branched polyethylenimine-graft-polycaprolactone-block-mono-methoxyl-poly (ethylene glycol) copolymers (hy-PEI-PCL-mPEG) for siRNA delivery. Journal of controlled release 2011, 153, (3), 262-268.
70. Zheng, M.; Librizzi, D.; Kılıç, A.; Liu, Y.; Renz, H.; Merkel, O. M.; Kissel, T., Enhancing in vivo circulation and siRNA delivery with biodegradable polyethylenimine-graft-polycaprolactone-block-poly (ethylene glycol) copolymers. Biomaterials 2012, 33, (27), 6551-6558.
71. Wu, Y.; Zhang, Y.; Zhang, W.; Sun, C.; Wu, J.; Tang, J., Reversing of multidrug resistance breast cancer by co-delivery of P-gp siRNA and doxorubicin via folic acid-modified core-shell nanomicelles. Colloids and Surfaces B: Biointerfaces 2016, 138, 60-69.
72. Coulembier, O.; Moins, S.; Maji, S.; Zhang, Z.; De Geest, B. G.; Dubois, P.; Hoogenboom, R., Linear polyethylenimine as (multi) functional initiator for organocatalytic l-lactide polymerization. Journal of Materials Chemistry B 2015, 3, (4), 612-619.
73. Gaspar, V. M.; Baril, P.; Costa, E. C.; de Melo-Diogo, D.; Foucher, F.; Queiroz, J. A.; Sousa, F.; Pichon, C.; Correia, I. J., Bioreducible poly (2-ethyl-2-oxazoline)–PLA–PEI-SS triblock copolymer micelles for co-delivery of DNA minicircles and Doxorubicin. Journal of Controlled Release 2015, 213, 175-191.
74. Lauter, V.; Lauter, H.; Glavic, A.; Toperverg, B., Reference module in materials science and materials engineering. In Elsevier: 2016.
75. Pitsikalis, M., Ionic polymerization. 2013.
76. Parker, G., Encyclopedia of materials: science and technology. 2001.
77. Uchida, S., Graft Copolymer Synthesis. Encyclopedia of Polymeric Nanomaterials 2015, 867-870.
78. Nuyken, O.; Pask, S., Ring-opening polymerization—An introductory review. Polymers 2013, 5, (2), 361-403.
79. Matyjaszewski, K.; Xia, J., Atom transfer radical polymerization. Chemical reviews 2001, 101, (9), 2921-2990.
80. Parrish, B.; Breitenkamp, R. B.; Emrick, T., PEG-and peptide-grafted aliphatic polyesters by click chemistry. Journal of the American Chemical Society 2005, 127, (20), 7404-7410.
81. Odian, G., Principles of polymerization. John Wiley & Sons: 2004.
82. Brunelle, D. J., Ring-Opening Polymerization. Mechanisms, Catalysis, Structure, Utility. Hanser Publishers, 1993 1993, 361.
83. Albertsson, A.-C.; Varma, I. K., Recent developments in ring opening polymerization of lactones for biomedical applications. Biomacromolecules 2003, 4, (6), 1466-1486.
84. Degée, P.; Dubois, P.; Jérǒme, R.; Jacobsen, S.; Fritz, H. G. In New catalysis for fast bulk ring‐opening polymerization of lactide monomers, Macromolecular Symposia, 1999; Wiley Online Library: 1999; pp 289-302.
85. Storey, R. F.; Sherman, J. W., Kinetics and mechanism of the stannous octoate-catalyzed bulk polymerization of ε-caprolactone. Macromolecules 2002, 35, (5), 1504-1512.
86. Braunecker, W. A.; Matyjaszewski, K., Controlled/living radical polymerization: Features, developments, and perspectives. Progress in Polymer Science 2007, 32, (1), 93-146.
87. Dong, H.; Matyjaszewski, K., ARGET ATRP of 2-(dimethylamino) ethyl methacrylate as an intrinsic reducing agent. Macromolecules 2008, 41, (19), 6868-6870.
88. Kolb, H. C.; Finn, M.; Sharpless, K. B., Click chemistry: diverse chemical function from a few good reactions. Angewandte Chemie International Edition 2001, 40, (11), 2004-2021.
89. Lecomte, P.; Riva, R.; Jérôme, C.; Jérôme, R., Macromolecular Engineering of Biodegradable Polyesters by Ring‐Opening Polymerization and ‘Click’Chemistry. Macromolecular Rapid Communications 2008, 29, (12‐13), 982-997.
90. Binder, W. H.; Sachsenhofer, R., ‘Click’chemistry in polymer and materials science. Macromolecular Rapid Communications 2007, 28, (1), 15-54.
91. Bock, V. D.; Hiemstra, H.; Van Maarseveen, J. H., CuI‐catalyzed alkyne–azide “click” cycloadditions from a mechanistic and synthetic perspective. European Journal of Organic Chemistry 2006, 2006, (1), 51-68.
92. Mespouille, L.; Vachaudez, M.; Suriano, F.; Gerbaux, P.; Coulembier, O.; Degée, P.; Flammang, R.; Dubois, P., One‐Pot Synthesis of Well‐Defined Amphiphilic and Adaptative Block Copolymers via Versatile Combination of “Click” Chemistry and ATRP. Macromolecular Rapid Communications 2007, 28, (22), 2151-2158.
93. Darcos, V.; El Habnouni, S.; Nottelet, B.; El Ghzaoui, A.; Coudane, J., Well-defined PCL-graft-PDMAEMA prepared by ring-opening polymerisation and click chemistry. Polymer Chemistry 2010, 1, (3), 280-282.
94. Mespouille, L.; Coulembier, O.; Paneva, D.; Degée, P.; Rashkov, I.; Dubois, P., Synthesis of adaptative and amphiphilic polymer model conetworks by versatile combination of ATRP, ROP, and “Click chemistry”. Journal of Polymer Science Part A: Polymer Chemistry 2008, 46, (15), 4997-5013.
95. Liu, J.; Xu, L.; Jiang, X.; Hennink, W. E.; Wang, X.; Zhuo, R., Disulfide-containing cross-linked PEI derivative synthesized by click chemistry for non-viral gene delivery. Journal of Controlled Release 2011, 152, e157-e159.
96. Zhang, X.; Li, Y.; Chen, Y. E.; Chen, J.; Ma, P. X., Cell-free 3D scaffold with two-stage delivery of miRNA-26a to regenerate critical-sized bone defects. Nature communications 2016, 7, 10376.
97. Farshbaf, M.; Davaran, S.; Zarebkohan, A.; Annabi, N.; Akbarzadeh, A.; Salehi, R., Significant role of cationic polymers in drug delivery systems. Artificial cells, nanomedicine, and biotechnology 2018, 46, (8), 1872-1891.
98. Bahadur, K. R.; Uludağ, H., PEI and its derivatives for gene therapy. In Polymers and Nanomaterials for Gene Therapy, Elsevier: 2016; pp 29-54.
99. Lee, S. B.; Russell, A. J.; Matyjaszewski, K., ATRP synthesis of amphiphilic random, gradient, and block copolymers of 2-(dimethylamino) ethyl methacrylate and n-butyl methacrylate in aqueous media. Biomacromolecules 2003, 4, (5), 1386-1393.
100. Lambermont-Thijs, H. M.; van der Woerdt, F. S.; Baumgaertel, A.; Bonami, L.; Du Prez, F. E.; Schubert, U. S.; Hoogenboom, R., Linear poly (ethylene imine) s by acidic hydrolysis of poly (2-oxazoline) s: kinetic screening, thermal properties, and temperature-induced solubility transitions. Macromolecules 2009, 43, (2), 927-933.
101. Neu, M.; Fischer, D.; Kissel, T., Recent advances in rational gene transfer vector design based on poly (ethylene imine) and its derivatives. The Journal of Gene Medicine: A cross‐disciplinary journal for research on the science of gene transfer and its clinical applications 2005, 7, (8), 992-1009.
102. Zhang, W.; He, J.; Liu, Z.; Ni, P.; Zhu, X., Biocompatible and pH‐responsive triblock copolymer mPEG‐b‐PCL‐b‐PDMAEMA: synthesis, self‐assembly, and application. Journal of Polymer Science Part A: Polymer Chemistry 2010, 48, (5), 1079-1091.
103. Brannon-Peppas, L., Poly (ethylene glycol): Chemistry and Biological Applications-JM Harris and S. Zalipsky, editors, American Chemical Society, Washington DC, 1997, 489 pp. Journal of Controlled Release 2000, 2, (66), 321.
104. Chen, W.; Yang, H.; Wang, R.; Cheng, R.; Meng, F.; Wei, W.; Zhong, Z., Versatile synthesis of functional biodegradable polymers by combining ring-opening polymerization and postpolymerization modification via Michael-type addition reaction. Macromolecules 2009, 43, (1), 201-207.
105. Mullen, B. D.; Tang, C. N.; Storey, R. F., New aliphatic poly (ester‐carbonates) based on 5‐methyl‐5‐allyloxycarbonyl‐1, 3‐dioxan‐2‐one. Journal of Polymer Science Part A: Polymer Chemistry 2003, 41, (13), 1978-1991.
106. Chen, W.; Zou, Y.; Jia, J.; Meng, F.; Cheng, R.; Deng, C.; Feijen, J.; Zhong, Z., Functional poly (ε-caprolactone) s via copolymerization of ε-caprolactone and pyridyl disulfide-containing cyclic carbonate: controlled synthesis and facile access to reduction-sensitive biodegradable graft copolymer micelles. Macromolecules 2013, 46, (3), 699-707.
107. Shi, Q.; Chen, X.; Lu, T.; Jing, X., The immobilization of proteins on biodegradable polymer fibers via click chemistry. Biomaterials 2008, 29, (8), 1118-1126.
108. Xu, J.; Prifti, F.; Song, J., A Versatile Monomer for Preparing Well-Defined Functional Polycarbonates and Poly (ester− carbonates). Macromolecules 2011, 44, (8), 2660-2667.
109. Mespouille, L.; Vachaudez, M.; Suriano, F.; Gerbaux, P.; Van Camp, W.; Coulembier, O.; Degée, P.; Flammang, R.; Du Prez, F.; Dubois, P., Controlled synthesis of amphiphilic block copolymers based on polyester and poly (amino methacrylate): Comprehensive study of reaction mechanisms. Reactive and Functional Polymers 2008, 68, (5), 990-1003.
110. Lu, C.; Shi, Q.; Chen, X.; Lu, T.; Xie, Z.; Hu, X.; Ma, J.; Jing, X., Sugars‐grafted aliphatic biodegradable poly (L‐lactide‐co‐carbonate) s by click reaction and their specific interaction with lectin molecules. Journal of Polymer Science Part A: Polymer Chemistry 2007, 45, (15), 3204-3217.
111. Li, C.; Liu, X.; He, S.; Huang, Y.; Cui, D., Synthesis and AIE properties of PEG–PLA–PMPC based triblock amphiphilic biodegradable polymers. Polymer Chemistry 2016, 7, (5), 1121-1128.
112. Cordeiro, R. A.; Farinha, D.; Rocha, N.; Serra, A. C.; Faneca, H.; Coelho, J. F., Novel Cationic Triblock Copolymer of Poly [2‐(dimethylamino) ethyl methacrylate]‐block‐poly (β‐amino ester)‐block‐poly [2‐(dimethylamino) ethyl methacrylate]: A Promising Non‐Viral Gene Delivery System. Macromolecular bioscience 2015, 15, (2), 215-228.
113. Tauhardt, L.; Kempe, K.; Knop, K.; Altuntaş, E.; Jäger, M.; Schubert, S.; Fischer, D.; Schubert, U. S., Linear polyethyleneimine: optimized synthesis and characterization–on the way to “pharmagrade” batches. Macromolecular Chemistry and Physics 2011, 212, (17), 1918-1924.
114. Tempelaar, S.; Barker, I. A.; Truong, V. X.; Hall, D. J.; Mespouille, L.; Dubois, P.; Dove, A. P., Organocatalytic synthesis and post-polymerization functionalization of propargyl-functional poly (carbonate) s. Polymer Chemistry 2013, 4, (1), 174-183.
115. Albertsson, A. C.; Eklund, M., Synthesis of copolymers of 1, 3‐dioxan‐2‐one and oxepan‐2‐one using coordination catalysts. Journal of Polymer Science Part A: Polymer Chemistry 1994, 32, (2), 265-279.
116. Wei, Z.; Liu, L.; Qi, M., Kinetics and mechanism of the ring opening polymerization of (R, S)-β-butyrolactone initiated with dibutylmagnesium. European polymer journal 2007, 43, (4), 1210-1218.
117. Tempelaar, S. Synthesis and Post-polymerisation Functionalisation of Aliphatic Poly (carbonate) s. University of Warwick, 2012.
118. Diaz, I. L.; Perez, L. D., Synthesis and micellization properties of triblock copolymers PDMAEMA-b-PCL-b-PDMAEMA and their applications in the fabrication of amphotericin B-loaded nanocontainers. Colloid and Polymer Science 2015, 293, (3), 913-923.
119. Muñoz-Bonilla, A.; Fernández-García, M.; Haddleton, D. M., Synthesis and aqueous solution properties of stimuli-responsive triblock copolymers. Soft Matter 2007, 3, (6), 725-731.
120. Song, J.; Jung, Y.; Lee, I.; Jang, J., Fabrication of pDMAEMA-coated silica nanoparticles and their enhanced antibacterial activity. Journal of colloid and interface science 2013, 407, 205-209.
121. Yin, J. J.; Wahid, F.; Zhang, Q.; Tao, Y. C.; Zhong, C.; Chu, L. Q., Facile Incorporation of Silver Nanoparticles into Quaternized Poly (2‐(Dimethylamino) Ethyl Methacrylate) Brushes as Bifunctional Antibacterial Coatings. Macromolecular Materials and Engineering 2017, 302, (6), 1700069.
122. Bonami, L.; Van Camp, W.; Van Rijckegem, D.; Du Prez, F. E., Facile Access to an Efficient Solid‐Supported Click Catalyst System Based on Poly (ethyleneimine). Macromolecular rapid communications 2009, 30, (1), 34-38.
123. Zhang, G.; Liu, J.; Yang, Q.; Zhuo, R.; Jiang, X., Disulfide-containing brushed polyethylenimine derivative synthesized by click chemistry for nonviral gene delivery. Bioconjugate chemistry 2012, 23, (6), 1290-1299.
124. Müller, A. J.; Michell, R. M., Differential scanning calorimetry of polymers. Polym. Morphol. Princ. Charact. Process 2016, 72-99.
125. Du, Z. X.; Xu, J. T.; Yang, Y.; Fan, Z. Q., Synthesis and characterization of poly (ϵ‐caprolactone)‐b‐poly (ethylene glycol) block copolymers prepared by a salicylaldimine‐aluminum complex. Journal of applied polymer science 2007, 105, (2), 771-776.
126. Tran, T.-Q.-M.; Hsieh, M.-F.; Chang, K.-L.; Pho, Q.-H.; Nguyen, V.-C.; Cheng, C.-Y.; Huang, C.-M., Bactericidal effect of lauric acid-loaded PCL-PEG-PCL nano-sized micelles on skin commensal Propionibacterium acnes. Polymers 2016, 8, (9), 321.
127. Nojima, S.; Ono, M.; Ashida, T., Crystallization of block copolymers II. Morphological study of poly (ethylene glycol)-poly (ε-caprolactone) block copolymers. Polymer journal 1992, 24, (11), 1271.
128. Hu, X.; Chen, X.; Xie, Z.; Cheng, H.; Jing, X., Aliphatic poly (ester‐carbonate) s bearing amino groups and its RGD peptide grafting. Journal of Polymer Science Part A: Polymer Chemistry 2008, 46, (21), 7022-7032.
129. Bruce, C.; Javakhishvili, I.; Fogelström, L.; Carlmark, A.; Hvilsted, S.; Malmström, E., Well-defined ABA-and BAB-type block copolymers of PDMAEMA and PCL. Rsc Advances 2014, 4, (49), 25809-25818.
130. Weyts, K. F.; Goethals, E. J., New synthesis of linear polyethyleneimine. Polymer bulletin 1988, 19, (1), 13-19.
131. Tanaka, R.; Ueoka, I.; Takaki, Y.; Kataoka, K.; Saito, S., High molecular weight linear polyethylenimine and poly (N-methylethylenimine). Macromolecules 1983, 16, (6), 849-853.
132. Lambermont-Thijs, H. M.; Bonami, L.; Du Prez, F. E.; Hoogenboom, R., Linear poly (alkyl ethylene imine) with varying side chain length: synthesis and physical properties. Polymer Chemistry 2010, 1, (5), 747-754.
133. Shuai, X.; Merdan, T.; Unger, F.; Wittmar, M.; Kissel, T., Novel biodegradable ternary copolymers hy-PEI-g-PCL-b-PEG: synthesis, characterization, and potential as efficient nonviral gene delivery vectors. Macromolecules 2003, 36, (15), 5751-5759.
134. Cao, P. F.; Felipe, M. J.; Advincula, R. C., On the Formation and Electropolymerization of a Star Copolymer With Peripheral Carbazoles. Macromolecular Chemistry and Physics 2013, 214, (3), 386-395.
135. Riess, G., Micellization of block copolymers. Progress in Polymer Science 2003, 28, (7), 1107-1170.
136. Mai, Y.; Eisenberg, A., Self-assembly of block copolymers. Chemical Society Reviews 2012, 41, (18), 5969-5985.
137. Munk, P., Equilibrium and nonequilibrium polymer micelles. In Solvents and Self-organization of Polymers, Springer: 1996; pp 19-32.
138. Loh, X. J.; Wu, Y.-L.; Seow, W. T. J.; Norimzan, M. N. I.; Zhang, Z.-X.; Xu, F.-J.; Kang, E.-T.; Neoh, K.-G.; Li, J., Micellization and phase transition behavior of thermosensitive poly (N-isopropylacrylamide)–poly (ɛ-caprolactone)–poly (N-isopropylacrylamide) triblock copolymers. Polymer 2008, 49, (23), 5084-5094.
139. Zhang, X.; Zhu, X.; Ke, F.; Ye, L.; Chen, E.-q.; Zhang, A.-y.; Feng, Z.-g., Preparation and self-assembly of amphiphilic triblock copolymers with polyrotaxane as a middle block and their application as carrier for the controlled release of Amphotericin B. Polymer 2009, 50, (18), 4343-4351.
140. Ginn, S. L.; Alexander, I. E.; Edelstein, M. L.; Abedi, M. R.; Wixon, J., Gene therapy clinical trials worldwide to 2012–an update. The journal of gene medicine 2013, 15, (2), 65-77.
141. Yin, H.; Kanasty, R. L.; Eltoukhy, A. A.; Vegas, A. J.; Dorkin, J. R.; Anderson, D. G., Non-viral vectors for gene-based therapy. Nature Reviews Genetics 2014, 15, (8), 541-555.
142. Pack, D. W.; Hoffman, A. S.; Pun, S.; Stayton, P. S., Design and development of polymers for gene delivery. Nature reviews Drug discovery 2005, 4, (7), 581.
143. Endres, T. Biodegradable amphiphilic PEG-PCL-PEI triblock copolymers designed for the self-assembly of multifunctional gene carriers. Philipps-Universität Marburg, 2012.
144. Van der Gucht, J.; Spruijt, E.; Lemmers, M.; Stuart, M. A. C., Polyelectrolyte complexes: bulk phases and colloidal systems. Journal of colloid and interface science 2011, 361, (2), 407-422.
145. Bucur, C. B.; Sui, Z.; Schlenoff, J. B., Ideal mixing in polyelectrolyte complexes and multilayers: Entropy driven assembly. Journal of the American Chemical Society 2006, 128, (42), 13690-13691.
146. Che, J.; Tao, A.; Chen, S.; Li, X.; Zhao, Y.; Yuan, W., Biologically responsive carrier-mediated anti-angiogenesis shRNA delivery for tumor treatment. Scientific reports 2016, 6, 35661.
147. Kunath, K.; von Harpe, A.; Fischer, D.; Petersen, H.; Bickel, U.; Voigt, K.; Kissel, T., Low-molecular-weight polyethylenimine as a non-viral vector for DNA delivery: comparison of physicochemical properties, transfection efficiency and in vivo distribution with high-molecular-weight polyethylenimine. Journal of Controlled Release 2003, 89, (1), 113-125.
148. Mosqueira, V. C. F.; Legrand, P.; Gref, R.; Heurtault, B.; Appel, M.; Barratt, G., Interactions between a macrophage cell line (J774A1) and surface-modified poly (D, L-lactide) nanocapsules bearing poly (ethylene glycol). Journal of drug targeting 1999, 7, (1), 65-78.
149. Shen, Z.-l.; Xia, Y.-q.; Yang, Q.-s.; Chen, K.; Ma, Y.-q., Polymer–nucleic acid interactions. In Polymeric Gene Delivery Systems, Springer: 2017; pp 41-64.
150. Stahlschmidt, U.; Jérôme, V.; Majewski, A.; Müller, A.; Freitag, R., Systematic Study of a Library of PDMAEMA-Based, Superparamagnetic Nano-Stars for the Transfection of CHO-K1 Cells. Polymers 2017, 9, (5), 156.
151. Guzaev, M.; Li, X.; Park, C.; Leung, W.-Y.; Roberts, L., Comparison of nucleic acid gel stains cell permeability, safety, and sensitivity of ethidium bromide alternatives. Online at https://biotium. com/wp-content/uploads/2017/02/Gel-Stains-Comparison. pdf 2017.
152. Raup, A.; Wang, H.; Synatschke, C. V.; Jérôme, V. r.; Agarwal, S.; Pergushov, D. V.; Müller, A. H.; Freitag, R., Compaction and transmembrane delivery of pdna: Differences between l-PEI and two types of amphiphilic block copolymers. Biomacromolecules 2017, 18, (3), 808-818.
153. Visakh, P., Polyelectrolyte: Thermodynamics and Rheology. In Polyelectrolytes, Springer: 2014; pp 1-17.
154. Riley, T.; Govender, T.; Stolnik, S.; Xiong, C.; Garnett, M.; Illum, L.; Davis, S., Colloidal stability and drug incorporation aspects of micellar-like PLA–PEG nanoparticles. Colloids and surfaces B: Biointerfaces 1999, 16, (1-4), 147-159.
155. Laugel, N.; Betscha, C.; Winterhalter, M.; Voegel, J.-C.; Schaaf, P.; Ball, V., Relationship between the growth regime of polyelectrolyte multilayers and the polyanion/polycation complexation enthalpy. The journal of physical chemistry B 2006, 110, (39), 19443-19449.
156. Khalil, I. A.; Kogure, K.; Akita, H.; Harashima, H., Uptake pathways and subsequent intracellular trafficking in nonviral gene delivery. Pharmacological reviews 2006, 58, (1), 32-45.
157. Aied, A.; Greiser, U.; Pandit, A.; Wang, W., Polymer gene delivery: overcoming the obstacles. Drug discovery today 2013, 18, (21-22), 1090-1098.
158. Liu, S.; Gao, Y.; Zhou, D.; Zeng, M.; Alshehri, F.; Newland, B.; Lyu, J.; O’Keeffe-Ahern, J.; Greiser, U.; Guo, T., Highly branched poly (β-amino ester) delivery of minicircle DNA for transfection of neurodegenerative disease related cells. Nature communications 2019, 10, (1), 1-14.
159. Li, X.; Guo, X.; Cheng, Y.; Zhao, X.; Fang, Z.; Luo, Y.; Xia, S.; Feng, Y.; Chen, J.; Yuan, W.-E., pH-responsive cross-linked low molecular weight polyethylenimine as an efficient gene vector for delivery of plasmid DNA encoding anti-VEGF-shRNA for tumor treatment. Frontiers in oncology 2018, 8.
160. Faneca, H.; Simoes, S.; De Lima, M. P., Association of albumin or protamine to lipoplexes: enhancement of transfection and resistance to serum. The journal of gene medicine 2004, 6, (6), 681-692.
161. Cordeiro, R. A.; Santo, D.; Farinha, D.; Serra, A.; Faneca, H.; Coelho, J. F., High transfection efficiency promoted by tailor-made cationic tri-block copolymer-based nanoparticles. Acta biomaterialia 2017, 47, 113-123.
162. Jones, C. H.; Chen, C.-K.; Ravikrishnan, A.; Rane, S.; Pfeifer, B. A., Overcoming nonviral gene delivery barriers: perspective and future. Molecular pharmaceutics 2013, 10, (11), 4082-4098.
163. Sun, H.; Zhou, L.; Chen, X.; Han, X.; Wang, R.; Liu, H., Microscopic insight into the DNA condensation process of a zwitterion‐functionalized polycation. Biopolymers 2016, 105, (11), 802-810.
164. Xiao, K.; Li, Y.; Luo, J.; Lee, J. S.; Xiao, W.; Gonik, A. M.; Agarwal, R. G.; Lam, K. S., The effect of surface charge on in vivo biodistribution of PEG-oligocholic acid based micellar nanoparticles. Biomaterials 2011, 32, (13), 3435-3446.
165. Pezzoli, D.; Giupponi, E.; Mantovani, D.; Candiani, G., Size matters for in vitro gene delivery: investigating the relationships among complexation protocol, transfection medium, size and sedimentation. Scientific reports 2017, 7, 44134.
166. Dash, P.; Read, M.; Barrett, L.; Wolfert, M.; Seymour, L., Factors affecting blood clearance and in vivo distribution of polyelectrolyte complexes for gene delivery. Gene therapy 1999, 6, (4), 643.
167. Santo, D.; Cordeiro, R. A.; Sousa, A.; Serra, A. n.; Coelho, J. F.; Faneca, H., Combination of Poly [(2-dimethylamino) ethyl methacrylate] and Poly (β-amino ester) Results in a Strong and Synergistic Transfection Activity. Biomacromolecules 2017, 18, (10), 3331-3342.
168. Venkataraman, S.; Hedrick, J. L.; Ong, Z. Y.; Yang, C.; Ee, P. L. R.; Hammond, P. T.; Yang, Y. Y., The effects of polymeric nanostructure shape on drug delivery. Advanced drug delivery reviews 2011, 63, (14-15), 1228-1246.
169. Gary, D. J.; Lee, H.; Sharma, R.; Lee, J.-S.; Kim, Y.; Cui, Z. Y.; Jia, D.; Bowman, V. D.; Chipman, P. R.; Wan, L., Influence of nano-carrier architecture on in vitro siRNA delivery performance and in vivo biodistribution: polyplexes vs micelleplexes. ACS nano 2011, 5, (5), 3493-3505.
170. Zhang, J.; Li, X.; Lou, L.; Li, X.; Jia, Y.; Jin, Z.; Zhu, Y., Non-viral gene therapy. In Intracellular Delivery, Springer: 2011; pp 599-699.
171. Majewski, A. P.; Stahlschmidt, U.; Jérôme, V. r.; Freitag, R.; Müller, A. H.; Schmalz, H., PDMAEMA-grafted core–shell–corona particles for nonviral gene delivery and magnetic cell separation. Biomacromolecules 2013, 14, (9), 3081-3090.
172. Endres, T. K.; Beck-Broichsitter, M.; Samsonova, O.; Renette, T.; Kissel, T. H., Self-assembled biodegradable amphiphilic PEG–PCL–lPEI triblock copolymers at the borderline between micelles and nanoparticles designed for drug and gene delivery. Biomaterials 2011, 32, (30), 7721-7731.
173. Lu, X.; Liu, L., Asymmetric polyplex-nanocapsules loaded with photosentisizer for light-assisted gene transfer. Journal of Photochemistry and Photobiology B: Biology 2017, 174, 269-275.
174. Dai, J.; Zou, S.; Pei, Y.; Cheng, D.; Ai, H.; Shuai, X., Polyethylenimine-grafted copolymer of poly (l-lysine) and poly (ethylene glycol) for gene delivery. Biomaterials 2011, 32, (6), 1694-1705.
175. Abebe, D. G.; Kandil, R.; Kraus, T.; Elsayed, M.; Merkel, O. M.; Fujiwara, T., Three‐Layered Biodegradable Micelles Prepared by Two‐Step Self‐Assembly of PLA‐PEI‐PLA and PLA‐PEG‐PLA Triblock Copolymers as Efficient Gene Delivery System. Macromolecular bioscience 2015, 15, (5), 698-711.
176. Zakeri, A.; Kouhbanani, M. A. J.; Beheshtkhoo, N.; Beigi, V.; Mousavi, S. M.; Hashemi, S. A. R.; Karimi Zade, A.; Amani, A. M.; Savardashtaki, A.; Mirzaei, E., Polyethylenimine-based nanocarriers in co-delivery of drug and gene: a developing horizon. Nano reviews & experiments 2018, 9, (1), 1488497.
177. Schallon, A.; Jérôme, V.; Walther, A.; Synatschke, C. V.; Müller, A. H.; Freitag, R., Performance of three PDMAEMA-based polycation architectures as gene delivery agents in comparison to linear and branched PEI. Reactive and Functional Polymers 2010, 70, (1), 1-10.
178. Saqafi, B.; Rahbarizadeh, F., Effect of PEI surface modification with PEG on cytotoxicity and transfection efficiency. Micro & Nano Letters 2018, 13, (8), 1090-1095.
179. Xing, H.; Lu, M.; Yang, T.; Liu, H.; Sun, Y.; Zhao, X.; Xu, H.; Yang, L.; Ding, P., Structure-function relationships of nonviral gene vectors: Lessons from antimicrobial polymers. Acta biomaterialia 2018.
180. Schallon, A.; Synatschke, C. V.; Jérôme, V. r.; Müller, A. H.; Freitag, R., Nanoparticulate nonviral agent for the effective delivery of pDNA and siRNA to differentiated cells and primary human T lymphocytes. Biomacromolecules 2012, 13, (11), 3463-3474.
181. Standardization, I. O. f., ISO 10993‐5: Biological Evaluation of Medical Devices. Part 5: Tests For In Vitro Cytotoxicity. In ISO Geneva, Switzerland: 2009.
182. Fischer, D.; Dautzenberg, H.; Kunath, K.; Kissel, T., Poly (diallyldimethylammonium chlorides) and their N-methyl-N-vinylacetamide copolymer-based DNA-polyplexes: role of molecular weight and charge density in complex formation, stability, and in vitro activity. International journal of pharmaceutics 2004, 280, (1-2), 253-269.
183. Fischer, D.; Li, Y.; Ahlemeyer, B.; Krieglstein, J.; Kissel, T., In vitro cytotoxicity testing of polycations: influence of polymer structure on cell viability and hemolysis. Biomaterials 2003, 24, (7), 1121-1131.
184. Ivanova, E. P.; Bazaka, K.; Crawford, R. J., New functional biomaterials for medicine and healthcare. Woodhead publishing New Delhi, India:: 2014; Vol. 67.
185. van de Wetering, P.; Cherng, J.-Y.; Talsma, H.; Hennink, W. E., Relation between transfection efficiency and cytotoxicity of poly (2-(dimethylamino) ethyl methacrylate)/plasmid complexes. Journal of Controlled Release 1997, 49, (1), 59-69.
186. Beyerle, A.; Merkel, O.; Stoeger, T.; Kissel, T., PEGylation affects cytotoxicity and cell-compatibility of poly (ethylene imine) for lung application: structure–function relationships. Toxicology and applied pharmacology 2010, 242, (2), 146-154.
187. Zhang, H.; Chen, Z.; Du, M.; Li, Y.; Chen, Y., Enhanced gene transfection efficiency by low-dose 25 kDa polyethylenimine by the assistance of 1.8 kDa polyethylenimine. Drug delivery 2018, 25, (1), 1740-1745.
188. Forcato, D.; Fili, A.; Alustiza, F.; Martínez, J. L.; Abel, S. B.; Nicotra, M. O.; Alessio, A.; Rodríguez, N.; Barbero, C.; Bosch, P., Transfection of bovine fetal fibroblast with polyethylenimine (PEI) nanoparticles: effect of particle size and presence of fetal bovine serum on transgene delivery and cytotoxicity. Cytotechnology 2017, 69, (4), 655-665.
189. Wahlfors, J.; Loimas, S.; Pasanen, T.; Hakkarainen, T., Green fluorescent protein (GFP) fusion constructs in gene therapy research. Histochemistry and cell biology 2001, 115, (1), 59-65.
190. Liu, H. C.; Zhao, H.; Chen, J.; Wu, W. L.; Wang, H. L.; Jiao, G. J.; Chen, Y. Z., Role of recombinant plasmid pEGFP-N1-IGF-1 transfection in alleviating osteoporosis in ovariectomized rats. Journal of molecular histology 2013, 44, (5), 535-544.
191. Riedl, S.; Kaiser, P.; Raup, A.; Synatschke, C.; Jérôme, V.; Freitag, R., Non-Viral Transfection of Human T Lymphocytes. Processes 2018, 6, (10), 188.
192. Raup, A.; Stahlschmidt, U.; Jérôme, V.; Synatschke, C. V.; Müller, A. H.; Freitag, R., Influence of polyplex formation on the performance of star-shaped polycationic transfection agents for mammalian cells. Polymers 2016, 8, (6), 224.
193. Yue, Y.; Jin, F.; Deng, R.; Cai, J.; Dai, Z.; Lin, M. C.; Kung, H.-F.; Mattebjerg, M. A.; Andresen, T. L.; Wu, C., Revisit complexation between DNA and polyethylenimine—Effect of length of free polycationic chains on gene transfection. Journal of controlled release 2011, 152, (1), 143-151.
194. Wen, Y.; Pan, S.; Luo, X.; Zhang, W.; Shen, Y.; Feng, M., PEG-and PDMAEG-graft-modified branched PEI as novel gene vector: synthesis, characterization and gene transfection. Journal of Biomaterials Science, Polymer Edition 2010, 21, (8-9), 1103-1126.
195. Chen, B.; Synatschke, C. V.; Jérôme, V.; Müller, A. H.; Freitag, R.; Wu, C., Co-transfection of star-shaped PDMAEMAs enhance transfection efficiency of protamine/pDNA complexes in the presence of serum. European Polymer Journal 2018, 103, 362-369.
196. Cai, J.; Yue, Y.; Wang, Y.; Jin, Z.; Jin, F.; Wu, C., Quantitative study of effects of free cationic chains on gene transfection in different intracellular stages. Journal of Controlled Release 2016, 238, 71-79.
197. Varga, C.; Tedford, N.; Thomas, M.; Klibanov, A.; Griffith, L.; Lauffenburger, D., Quantitative comparison of polyethylenimine formulations and adenoviral vectors in terms of intracellular gene delivery processes. Gene therapy 2005, 12, (13), 1023.
dc.rights.spa.fl_str_mv Derechos reservados - Universidad Nacional de Colombia
dc.rights.coar.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.rights.license.spa.fl_str_mv Atribución-SinDerivadas 4.0 Internacional
dc.rights.spa.spa.fl_str_mv Acceso abierto
dc.rights.uri.spa.fl_str_mv http://creativecommons.org/licenses/by-nd/4.0/
dc.rights.accessrights.spa.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv Atribución-SinDerivadas 4.0 Internacional
Derechos reservados - Universidad Nacional de Colombia
Acceso abierto
http://creativecommons.org/licenses/by-nd/4.0/
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.extent.spa.fl_str_mv 155
dc.format.mimetype.spa.fl_str_mv application/pdf
dc.publisher.program.spa.fl_str_mv Bogotá - Ciencias - Doctorado en Ciencias - Química
dc.publisher.department.spa.fl_str_mv Departamento de Química
dc.publisher.branch.spa.fl_str_mv Universidad Nacional de Colombia - Sede Bogotá
institution Universidad Nacional de Colombia
bitstream.url.fl_str_mv https://repositorio.unal.edu.co/bitstream/unal/78367/1/1015402910.2020.pdf
https://repositorio.unal.edu.co/bitstream/unal/78367/3/license_rdf
https://repositorio.unal.edu.co/bitstream/unal/78367/4/license.txt
https://repositorio.unal.edu.co/bitstream/unal/78367/5/1015402910.2020.pdf.jpg
bitstream.checksum.fl_str_mv a089d4843b1fe1441b37c87492c36880
dab767be7a093b539031785b3bf95490
e2f63a891b6ceb28c3078128251851bf
82682bcb555abae99f118ef4bfcad81a
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
MD5
repository.name.fl_str_mv Repositorio Institucional Universidad Nacional de Colombia
repository.mail.fl_str_mv repositorio_nal@unal.edu.co
_version_ 1814089847661395968
spelling Atribución-SinDerivadas 4.0 InternacionalDerechos reservados - Universidad Nacional de ColombiaAcceso abiertohttp://creativecommons.org/licenses/by-nd/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Pérez Pérez, León Darío26c85caa-172a-4f24-8364-71a25423a2a9Díaz Ariza, Ivonne Lorenaed5d9758-e97b-4f16-a9e3-ec7596a1c58aMacromoléculas2020-09-03T03:57:31Z2020-09-03T03:57:31Z2020-02-14https://repositorio.unal.edu.co/handle/unal/78367The first chapter reviews the most important aspects of the polymeric vectors used in gene therapy. This type of therapy involves the introduction of genetic material into specific cells to treat certain diseases, using vectors that mediate the cellular internalization of nucleic acids and protect them from degradation. Synthetic cationic polymers have shown promising results as gene vectors, due to their advantages such as higher safety, simple fabrication process and modifiable structure. Poly(ethyleneimine) (PEI) and poly(2- (dimethylamino)ethyl methacrylate) (PDMAEMA) have been highlighted in this context due to their high transfection efficiency; however, the non-biodegradable character and associated cytotoxicity have limited their applications. To overcome these disadvantages, several researchers have studied the implementation of strategies such as PEGylation, copolymerization with hydrophobic segments and the use of branched architectures, which have demonstrated to significantly reduce the toxicity and increase the transfection efficiency. In addition, these structural modifications can be perform using synthetic methods, such as controlled polymerizations and selective coupling reactions, which allow the design of molecules with well-defined properties. The second chapter describes the synthesis and characterization of copolymers composed of methoxy poly(ethylene glycol) and a hydrophobic block of poly(ɛ-caprolactone-co-propargyl carbonate) grafted with low molecular weight PDMAEMA or linear PEI (lPEI), using a combination of ring opening polymerization, click chemistry and atom transfer radical polymerization. In this way, materials with different grafting densities and lengths of the hydrophobic segment were obtained. Following the proposed synthetic route, the synthesis of graft copolymers based on PDMAEMA with target structure and composition was achieved, while for copolymers composed of lPEI, lower grafting densities than the target ones were obtained. These materials could self-assembled in aqueous medium to form positively charged particles with average sizes between 150 and 380 nm. The third chapter covers the formation and characterization of the copolymer/DNA complexes with different nitrogen/phosphorus (N/P) ratios and complexation matrices. The study of the condensation ability showed that all PDMAEMA copolymers were able to complex the DNA molecules at N/P ratios ≥ 1, regardless of the used conditions, while lPEI copolymers required N/P ratios from 7 to 20, depending on their composition and the complexation solution. In the case of the materials composed of PDMAEMA, the measurements of particle size and zeta potential indicated the formation of positively charged complexes with sizes below 300 nm, which are suitable for cellular uptake. On the other hand, the lPEI based complexes exhibited negative or neutral surface charges with hydrodynamic diameters below 500 nm. These physicochemical properties could generate restrictions for the effective delivery of DNA. The fourth chapter describes the in vitro cytotoxicity of graft copolymers and the transfection efficiency of the resulting polyplexes, using 25 kDa lPEI and 20 kDa PDMAEMA as controls. The copolymers were less cytotoxic to L929 cells than the control homopolymers, exhibiting higher cell viability by reducing the length of the hydrophobic segment and the amount of cationic grafts. Polyplexes formed from PDMAEMA copolymers were more efficient in the delivery of DNA in HEK-293 cells than the standard polycations, without affecting the cell viability. On the other hand, the complexes based on lPEI copolymers exhibited a low transfection efficiency, even using high polymer concentrations and N/P ratios. In general, copolymers composed of long hydrophobic segments and higher grafting density showed a better performance. In particular, the material PP6D5 showed the most promising features for its application as DNA transfection agent, for both in vitro and in vivo assays. This thesis represents the starting point for future research related to the rational design of grafted structures based on cationic polymers that may have improved properties and performance for their application as gene delivery systems.El primer capítulo revisa los aspectos más importantes de los vectores poliméricos usados en terapia génica. Este tipo de terapia envuelve la introducción de material genético en células específicas para el tratamiento de ciertas enfermedades, usando vectores que median la internalización celular de los ácidos nucleicos y los protege de la degradación. Dentro de los vectores que han mostrado resultados prometedores se encuentran los polímeros catiónicos sintéticos, los cuales presentan ventajas como mayor seguridad, procesos sencillos de fabricación y posibilidad de modificar su estructura. La poli(etilenimina) (PEI) y el poli(metacrilato de 2-(dimetilamino) etilo) (PDMAEMA) se han destacado en este contexto debido a su alta eficiencia de transfección; sin embargo, su carácter no biodegradable y citotoxicidad asociada han limitado sus aplicaciones. Para superar estas desventajas, varios investigadores han estudiado la implementación de estrategias como la PEGilación, la copolimerización con segmentos hidrófobos y el uso de arquitecturas ramificadas, las cuales han mostrado reducir significativamente la toxicidad y aumentar la eficiencia de transfección. Además, estas modificaciones estructurales pueden realizarse empleando métodos sintéticos, como polimerizaciones controladas y acoplamientos selectivos, que permiten diseñar moléculas con propiedades bien definidas. El segundo capítulo describe la síntesis y caracterización de una serie de copolímeros compuestos por metoxi-polietilenglicol y un bloque hidrófobo de poli(caprolactona-co-carbonato de propargilo) injertado con segmentos de PDMAEMA o PEI lineal (lPEI) de bajo peso molecular, empleando una combinación de polimerización por apertura de anillo, química click y polimerización radicalaria por transferencia de átomo. De esta manera se obtuvieron materiales con diferentes densidades de injerto y longitudes del segmento hidrófobo. Siguiendo la ruta sintética planteada, se logró sintetizar copolímeros de injerto basados en PDMAEMA con una estructura y composición target, mientras que para los copolímeros compuestos por lPEI, se obtuvieron densidades de injerto menores a las esperadas. Estos materiales se auto-ensamblaron en medio acuoso para formar partículas cargadas positivamente con tamaños promedio entre 150 y 380 nm. El tercer capítulo cubre la formación y caracterización de los complejos copolímero/ADN, variando parámetros como la relación nitrógeno/fósforo (N/P) y la matriz de complejación. Los estudios de habilidad de condensación mostraron que todos los copolímeros de PDMAEMA pudieron complejar las moléculas de ADN a relaciones N/P ≥ 1, sin importar las condiciones empleadas, mientras que los copolímeros de lPEI, necesitaron relaciones N/P de 7 a 20, dependiendo de su composición y la solución de complejación. En el caso de los materiales compuestos por PDMAEMA, las mediciones de tamaño de partícula y potencial zeta indicaron la formación de complejos cargados positivamente con tamaños inferiores a 300 nm, los cuales son adecuados para la internalización celular. Por su parte, los complejos basados en lPEI exhibieron cargas superficiales negativas o neutras con diámetros hidrodinámicos por debajo de 500 nm. Estas propiedades podrían generan restricciones para la entrega efectiva del ADN. El cuarto capítulo describe la citotoxicidad in vitro de los copolímeros de injerto y la eficiencia de transfección de los poliplejos resultantes, empleando como controles lPEI 25 kDa y PDMAEMA 20 kDa. Los copolímeros fueron menos citotóxicos que los homopolímeros control en células L929, exhibiendo una mayor viabilidad celular al reducir la longitud del segmento hidrófobo y la cantidad de injertos catiónicos. Los poliplejos formados a partir de los copolímeros de PDMAEMA fueron más eficientes en la entrega de ADN que los policationes estándar en células HEK-293, sin afectar su viabilidad. Por su parte, los complejos basados en los copolímeros de lPEI exhibieron una baja eficiencia de transfección, incluso usando altas concentraciones de polímero y relaciones N/P. En general, los copolímeros compuestos por segmentos hidrófobos largos y mayor densidad de injerto presentaron un mejor desempeño. En especial, el material denominado como PP6D5 mostró las características más prometedoras para su aplicación como agente de transfección de ADN, tanto en ensayos in vitro como in vivo. Esta tesis representa el punto de partida para futuras investigaciones relacionadas con el diseño racional de estructuras injertadas basadas en polímeros catiónicos que puedan presentar mejores propiedades y desempeño para su aplicación como sistemas de entrega de genes.Doctorado155application/pdfeng660 - Ingeniería química547 - Química orgánicaPDMAEMAPEIDNA transfectionGene therapyPolymeric vectorsCationic graft copolymersPDMAEMAPEITerapia génicaVectores poliméricosCopolímeros de injerto catiónicosTransfección de ADNGraft copolymers based on Poly(2-(dimethylamino)ethyl methacrylate) and Poly(ethyleneimine): synthesis and evaluation as vectors for DNA transfectionTrabajo de grado - Doctoradoinfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/acceptedVersionhttp://purl.org/coar/resource_type/c_db06TextBogotá - Ciencias - Doctorado en Ciencias - QuímicaDepartamento de QuímicaUniversidad Nacional de Colombia - Sede Bogotá1. Ospina, M. L.; Huertas, J. A.; Montaño, J. I.; Rivillas, J. C., Observatorio Nacional de Cáncer Colombia/The Colombian National Cancer Observatory/Observatório Nacional de Câncer Colômbia. Revista de la Facultad Nacional de Salud Pública 2015, 33, (2), 262.2. Lozano, R.; Naghavi, M.; Foreman, K.; Lim, S.; Shibuya, K.; Aboyans, V.; Abraham, J.; Adair, T.; Aggarwal, R.; Ahn, S. Y., Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. The lancet 2012, 380, (9859), 2095-2128.3. Mintzer, M. A.; Simanek, E. E., Nonviral vectors for gene delivery. Chemical reviews 2008, 109, (2), 259-302.4. Naldini, L., Gene therapy returns to centre stage. Nature 2015, 526, (7573), 351-360.5. Liu, X. Q.; Sun, C. Y.; Yang, X. Z.; Wang, J., Polymeric‐Micelle‐Based Nanomedicine for siRNA Delivery. Particle & Particle Systems Characterization 2013, 30, (3), 211-228.6. Falamarzian, A.; Xiong, X.-B.; Uludag, H.; Lavasanifar, A., Polymeric micelles for siRNA delivery. Journal of Drug Delivery Science and Technology 2012, 22, (1), 43-54.7. Ramamoorth, M.; Narvekar, A., Non viral vectors in gene therapy-an overview. Journal of clinical and diagnostic research: JCDR 2015, 9, (1), GE01.8. Samal, S. K.; Dash, M.; Van Vlierberghe, S.; Kaplan, D. L.; Chiellini, E.; Van Blitterswijk, C.; Moroni, L.; Dubruel, P., Cationic polymers and their therapeutic potential. Chemical Society Reviews 2012, 41, (21), 7147-7194.9. Samsonova, O.; Pfeiffer, C.; Hellmund, M.; Merkel, O. M.; Kissel, T., Low molecular weight pDMAEMA-block-pHEMA block-copolymers synthesized via RAFT-polymerization: potential non-viral gene delivery agents? Polymers 2011, 3, (2), 693-718.10. Lin, D.; Huang, Y.; Jiang, Q.; Zhang, W.; Yue, X.; Guo, S.; Xiao, P.; Du, Q.; Xing, J.; Deng, L., Structural contributions of blocked or grafted poly (2-dimethylaminoethyl methacrylate) on PEGylated polycaprolactone nanoparticles in siRNA delivery. Biomaterials 2011, 32, (33), 8730-8742.11. Guo, S.; Huang, Y.; Wei, T.; Zhang, W.; Wang, W.; Lin, D.; Zhang, X.; Kumar, A.; Du, Q.; Xing, J., Amphiphilic and biodegradable methoxy polyethylene glycol-block-(polycaprolactone-graft-poly (2-(dimethylamino) ethyl methacrylate)) as an effective gene carrier. Biomaterials 2011, 32, (3), 879-889.12. Díaz-Ariza, I. L.; Sierra, C. A.; Pérez-Pérez, L. D., Desarrollo de vectores génicos basados en polímeros sintéticos: PEI y PDMAEMA. Revista Colombiana de Ciencias Químico-Farmacéuticas 2018, 47, (3), 350-374.13. Zhou, Z.; Liu, X.; Zhu, D.; Wang, Y.; Zhang, Z.; Zhou, X.; Qiu, N.; Chen, X.; Shen, Y., Nonviral cancer gene therapy: delivery cascade and vector nanoproperty integration. Advanced drug delivery reviews 2017, 115, 115-154.14. Nayerossadat, N.; Maedeh, T.; Ali, P. A., Viral and nonviral delivery systems for gene delivery. Advanced biomedical research 2012, 1.15. Sun, X.; Zhang, N., Cationic polymer optimization for efficient gene delivery. Mini reviews in medicinal chemistry 2010, 10, (2), 108-125.16. Caffery, B.; Lee, J. S.; Alexander-Bryant, A. A., Vectors for glioblastoma gene therapy: viral & non-viral delivery strategies. Nanomaterials 2019, 9, (1), 105.17. Levine, R. M.; Scott, C. M.; Kokkoli, E., Peptide functionalized nanoparticles for nonviral gene delivery. Soft Matter 2013, 9, (4), 985-1004.18. Junquera, E.; Aicart, E., Cationic lipids as transfecting agents of DNA in gene therapy. Current topics in medicinal chemistry 2014, 14, (5), 649-663.19. Zhi, D.; Bai, Y.; Yang, J.; Cui, S.; Zhao, Y.; Chen, H.; Zhang, S., A review on cationic lipids with different linkers for gene delivery. Advances in Colloid and Interface Science 2017.20. Pahle, J.; Walther, W., Vectors and strategies for nonviral cancer gene therapy. Expert opinion on biological therapy 2016, 16, (4), 443-461.21. Pushpendra, S.; Arvind, P.; Anil, B., Nucleic acids as therapeutics. In From Nucleic Acids Sequences to Molecular Medicine, Springer: 2012; pp 19-45.22. Sridharan, K.; Gogtay, N. J., Therapeutic nucleic acids: current clinical status. British journal of clinical pharmacology 2016, 82, (3), 659-672.23. Schultz, N.; Marenstein, D. R.; De Angelis, D. A.; Wang, W.-Q.; Nelander, S.; Jacobsen, A.; Marks, D. S.; Massagué, J.; Sander, C., Off-target effects dominate a large-scale RNAi screen for modulators of the TGF-β pathway and reveal microRNA regulation of TGFBR2. Silence 2011, 2, (1), 3.24. Foldvari, M.; Chen, D. W.; Nafissi, N.; Calderon, D.; Narsineni, L.; Rafiee, A., Non-viral gene therapy: Gains and challenges of non-invasive administration methods. Journal of Controlled Release 2016, 240, 165-190.25. Uludag, H.; Ubeda, A.; Ansari, A., At the Intersection of Biomaterials and Gene Therapy: Progress in Non-viral Delivery of Nucleic Acids. Frontiers in Bioengineering and Biotechnology 2019, 7.26. Walther, W.; Petkov, S.; Kuvardina, O.; Aumann, J.; Kobelt, D.; Fichtner, I.; Lemm, M.; Piontek, J.; Blasig, I.; Stein, U., Novel Clostridium perfringens enterotoxin suicide gene therapy for selective treatment of claudin-3-and-4-overexpressing tumors. Gene therapy 2012, 19, (5), 494.27. Nakhlband, A.; Barar, J.; Bidmeshkipour, A.; Heidari, H. R.; Omidi, Y., Bioimpacts of anti epidermal growth receptor antisense complexed with polyamidoamine dendrimers in human lung epithelial adenocarcinoma cells. Journal of biomedical nanotechnology 2010, 6, (4), 360-369.28. Chen, X.-A.; Zhang, L.-J.; He, Z.-J.; Wang, W.-W.; Xu, B.; Zhong, Q.; Shuai, X.-T.; Yang, L.-Q.; Deng, Y.-B., Plasmid-encapsulated polyethylene glycol-grafted polyethylenimine nanoparticles for gene delivery into rat mesenchymal stem cells. International journal of nanomedicine 2011, 6, 843.29. Senovilla, L.; Vacchelli, E.; Garcia, P.; Eggermont, A.; Fridman, W. H.; Galon, J.; Zitvogel, L.; Kroemer, G.; Galluzzi, L., Trial watch: DNA vaccines for cancer therapy. Oncoimmunology 2013, 2, (4), e23803.30. Lächelt, U.; Wagner, E., Nucleic acid therapeutics using polyplexes: a journey of 50 years (and beyond). Chemical reviews 2015, 115, (19), 11043-11078.31. Yin, H.; Kanasty, R. L.; Eltoukhy, A. A.; Vegas, A. J.; Dorkin, J. R.; Anderson, D. G., Non-viral vectors for gene-based therapy. Nature Reviews Genetics 2014, 15, (8), 541.32. Grandinetti, G.; Reineke, T. M., Exploring the mechanism of plasmid DNA nuclear internalization with polymer-based vehicles. Molecular pharmaceutics 2012, 9, (8), 2256-2267.33. Kendrick, J.; Matthews, K.; Straughn Jr, J.; Barnes, M.; Fewell, J.; Anwer, K.; Alvarez, R. In A phase I trial of intraperitoneal EGEN-001, a novel IL-12 gene therapeutic, administered alone or in combination with chemotherapy in patients with recurrent ovarian cancer, ASCO Annual Meeting Proceedings, 2008; 2008; p 5572.34. Sinn, P. L.; Anthony, R. M.; McCray Jr, P. B., Genetic therapies for cystic fibrosis lung disease. Human molecular genetics 2011, 20, (R1), R79-R86.35. Nguyen, D. N.; Green, J. J.; Chan, J. M.; Langer, R.; Anderson, D. G., Polymeric materials for gene delivery and DNA vaccination. Advanced Materials 2009, 21, (8), 847-867.36. Prabu, S. L.; Ruckmani, K., Biopolymer in Gene Delivery. Advanced Technology for Delivering Therapeutics 2017, 137.37. Oskolkov, N. N.; Potemkin, I. I., Complexation in asymmetric solutions of oppositely charged polyelectrolytes: Phase diagram. Macromolecules 2007, 40, (23), 8423-8429.38. Ou, Z.; Muthukumar, M., Entropy and enthalpy of polyelectrolyte complexation: Langevin dynamics simulations. The Journal of chemical physics 2006, 124, (15), 154902.39. Zhang, P.; Wagner, E., History of polymeric gene delivery systems. Topics in Current Chemistry 2017, 375, (2), 26.40. Vader, P.; van der Aa, L. J.; Engbersen, J. F.; Storm, G.; Schiffelers, R. M., Disulfide-based poly (amido amine) s for siRNA delivery: effects of structure on siRNA complexation, cellular uptake, gene silencing and toxicity. Pharmaceutical research 2011, 28, (5), 1013-1022.41. Bishop, C. J.; Ketola, T.-M.; Tzeng, S. Y.; Sunshine, J. C.; Urtti, A.; Lemmetyinen, H.; Vuorimaa-Laukkanen, E.; Yliperttula, M.; Green, J. J., The Effect and Role of Carbon Atoms in Poly (β-amino ester) s for DNA Binding and Gene Delivery. Journal of the American Chemical Society 2013, 135, (18), 6951-6957.42. Mathew, A.; Cao, H.; Collin, E.; Wang, W.; Pandit, A., Hyperbranched PEGmethacrylate linear pDMAEMA block copolymer as an efficient non-viral gene delivery vector. International journal of pharmaceutics 2012, 434, (1-2), 99-105.43. Verbaan, F. J.; Klouwenberg, P. K.; van Steenis, J. H.; Snel, C. J.; Boerman, O.; Hennink, W. E.; Storm, G., Application of poly (2-(dimethylamino) ethyl methacrylate)-based polyplexes for gene transfer into human ovarian carcinoma cells. International journal of pharmaceutics 2005, 304, (1), 185-192.44. Agarwal, S.; Zhang, Y.; Maji, S.; Greiner, A., PDMAEMA based gene delivery materials. Materials Today 2012, 15, (9), 388-393.45. Synatschke, C. V.; Schallon, A.; Jérôme, V. r.; Freitag, R.; Müller, A. H., Influence of polymer architecture and molecular weight of poly (2-(dimethylamino) ethyl methacrylate) polycations on transfection efficiency and cell viability in gene delivery. Biomacromolecules 2011, 12, (12), 4247-4255.46. Zhao, G.; Long, L.; Zhang, L.; Peng, M.; Cui, T.; Wen, X.; Zhou, X.; Sun, L.; Che, L., Smart pH-sensitive nanoassemblies with cleavable PEGylation for tumor targeted drug delivery. Scientific reports 2017, 7, (1), 3383.47. Qiao, Y.; Huang, Y.; Qiu, C.; Yue, X.; Deng, L.; Wan, Y.; Xing, J.; Zhang, C.; Yuan, S.; Dong, A., The use of PEGylated poly [2-(N, N-dimethylamino) ethyl methacrylate] as a mucosal DNA delivery vector and the activation of innate immunity and improvement of HIV-1-specific immune responses. Biomaterials 2010, 31, (1), 115-123.48. Jiang, X.; Lok, M. C.; Hennink, W. E., Degradable-brushed pHEMA–pDMAEMA synthesized via ATRP and click chemistry for gene delivery. Bioconjugate chemistry 2007, 18, (6), 2077-2084.49. Song, Y.; Zhang, T.; Song, X.; Zhang, L.; Zhang, C.; Xing, J.; Liang, X.-J., Polycations with excellent gene transfection ability based on PVP-g-PDMAEMA with random coil and micelle structures as non-viral gene vectors. Journal of Materials Chemistry B 2015, 3, (5), 911-918.50. Xu, F.; Ping, Y.; Ma, J.; Tang, G.; Yang, W.; Li, J.; Kang, E.; Neoh, K., Comb-shaped copolymers composed of hydroxypropyl cellulose backbones and cationic poly ((2-dimethyl amino) ethyl methacrylate) side chains for gene delivery. Bioconjugate chemistry 2009, 20, (8), 1449-1458.51. Malikmammadov, E.; Tanir, T. E.; Kiziltay, A.; Hasirci, V.; Hasirci, N., PCL and PCL-based materials in biomedical applications. Journal of Biomaterials Science, Polymer Edition 2018, 29, (7-9), 863-893.52. Ulery, B. D.; Nair, L. S.; Laurencin, C. T., Biomedical applications of biodegradable polymers. Journal of polymer science Part B: polymer physics 2011, 49, (12), 832-864.53. Guo, S.; Qiao, Y.; Wang, W.; He, H.; Deng, L.; Xing, J.; Xu, J.; Liang, X.-J.; Dong, A., Poly (ε-caprolactone)-graft-poly (2-(N, N-dimethylamino) ethyl methacrylate) nanoparticles: pH dependent thermo-sensitive multifunctional carriers for gene and drug delivery. Journal of Materials Chemistry 2010, 20, (33), 6935-6941.54. Huang, Y.; Lin, D.; Jiang, Q.; Zhang, W.; Guo, S.; Xiao, P.; Zheng, S.; Wang, X.; Chen, H.; Zhang, H.-Y., Binary and ternary complexes based on polycaprolactone-graft-poly (N, N-dimethylaminoethyl methacrylate) for targeted siRNA delivery. Biomaterials 2012, 33, (18), 4653-4664.55. Zhu, C.; Jung, S.; Luo, S.; Meng, F.; Zhu, X.; Park, T. G.; Zhong, Z., Co-delivery of siRNA and paclitaxel into cancer cells by biodegradable cationic micelles based on PDMAEMA–PCL–PDMAEMA triblock copolymers. Biomaterials 2010, 31, (8), 2408-2416.56. Yue, X.; Qiao, Y.; Qiao, N.; Guo, S.; Xing, J.; Deng, L.; Xu, J.; Dong, A., Amphiphilic Methoxy Poly (ethylene glycol)-b-poly (ε-caprolactone)-b-poly (2-dimethylaminoethyl methacrylate) Cationic Copolymer Nanoparticles as a Vector for Gene and Drug Delivery. Biomacromolecules 2010, 11, (9), 2306-2312.57. Qian, X.; Long, L.; Shi, Z.; Liu, C.; Qiu, M.; Sheng, J.; Pu, P.; Yuan, X.; Ren, Y.; Kang, C., Star-branched amphiphilic PLA-b-PDMAEMA copolymers for co-delivery of miR-21 inhibitor and doxorubicin to treat glioma. Biomaterials 2014, 35, (7), 2322-2335.58. Ziebarth, J. D.; Wang, Y., Understanding the protonation behavior of linear polyethylenimine in solutions through Monte Carlo simulations. Biomacromolecules 2009, 11, (1), 29-38.59. Yu, Q.-Y.; Zhan, Y.-R.; Zhang, J.; Luan, C.-R.; Wang, B.; Yu, X.-Q., Aromatic Modification of Low Molecular Weight PEI for Enhanced Gene Delivery. Polymers 2017, 9, (8), 362.60. Hu, J.; Zhao, W.; Liu, K.; Yu, Q.; Mao, Y.; Lu, Z.; Zhang, Y.; Zhu, M., Low-molecular weight polyethylenimine modified with pluronic 123 and rgd-or chimeric rgd-nls peptide: Characteristics and transfection efficacy of their complexes with plasmid DNA. Molecules 2016, 21, (5), 655.61. Merdan, T.; Kunath, K.; Petersen, H.; Bakowsky, U.; Voigt, K. H.; Kopecek, J.; Kissel, T., PEGylation of poly (ethylene imine) affects stability of complexes with plasmid DNA under in vivo conditions in a dose-dependent manner after intravenous injection into mice. Bioconjugate chemistry 2005, 16, (4), 785-792.62. Mao, S.; Neu, M.; Germershaus, O.; Merkel, O.; Sitterberg, J.; Bakowsky, U.; Kissel, T., Influence of polyethylene glycol chain length on the physicochemical and biological properties of poly (ethylene imine)-graft-poly (ethylene glycol) block copolymer/SiRNA polyplexes. Bioconjugate chemistry 2006, 17, (5), 1209-1218.63. Ochrimenko, S.; Vollrath, A.; Tauhardt, L.; Kempe, K.; Schubert, S.; Schubert, U. S.; Fischer, D., Dextran-graft-linear poly (ethylene imine) s for gene delivery: importance of the linking strategy. Carbohydrate polymers 2014, 113, 597-606.64. Min, S.-H.; Park, K. C.; Yeom, Y. I., Chitosan-mediated non-viral gene delivery with improved serum stability and reduced cytotoxicity. Biotechnology and bioprocess engineering 2014, 19, (6), 1077-1082.65. Cao, N.; Cheng, D.; Zou, S.; Ai, H.; Gao, J.; Shuai, X., The synergistic effect of hierarchical assemblies of siRNA and chemotherapeutic drugs co-delivered into hepatic cancer cells. Biomaterials 2011, 32, (8), 2222-2232.66. Qiu, L. Y.; Bae, Y. H., Self-assembled polyethylenimine-graft-poly (ε-caprolactone) micelles as potential dual carriers of genes and anticancer drugs. Biomaterials 2007, 28, (28), 4132-4142.67. Endres, T.; Zheng, M.; Kılıç, A. e.; Turowska, A.; Beck-Broichsitter, M.; Renz, H.; Merkel, O. M.; Kissel, T., Amphiphilic biodegradable PEG-PCL-PEI triblock copolymers for FRET-capable in vitro and in vivo delivery of siRNA and quantum dots. Molecular pharmaceutics 2014, 11, (4), 1273-1281.68. Zheng, M.; Liu, Y.; Samsonova, O.; Endres, T.; Merkel, O.; Kissel, T., Amphiphilic and biodegradable hy-PEI-g-PCL-b-PEG copolymers efficiently mediate transgene expression depending on their graft density. International journal of pharmaceutics 2012, 427, (1), 80-87.69. Liu, Y.; Samsonova, O.; Sproat, B.; Merkel, O.; Kissel, T., Biophysical characterization of hyper-branched polyethylenimine-graft-polycaprolactone-block-mono-methoxyl-poly (ethylene glycol) copolymers (hy-PEI-PCL-mPEG) for siRNA delivery. Journal of controlled release 2011, 153, (3), 262-268.70. Zheng, M.; Librizzi, D.; Kılıç, A.; Liu, Y.; Renz, H.; Merkel, O. M.; Kissel, T., Enhancing in vivo circulation and siRNA delivery with biodegradable polyethylenimine-graft-polycaprolactone-block-poly (ethylene glycol) copolymers. Biomaterials 2012, 33, (27), 6551-6558.71. Wu, Y.; Zhang, Y.; Zhang, W.; Sun, C.; Wu, J.; Tang, J., Reversing of multidrug resistance breast cancer by co-delivery of P-gp siRNA and doxorubicin via folic acid-modified core-shell nanomicelles. Colloids and Surfaces B: Biointerfaces 2016, 138, 60-69.72. Coulembier, O.; Moins, S.; Maji, S.; Zhang, Z.; De Geest, B. G.; Dubois, P.; Hoogenboom, R., Linear polyethylenimine as (multi) functional initiator for organocatalytic l-lactide polymerization. Journal of Materials Chemistry B 2015, 3, (4), 612-619.73. Gaspar, V. M.; Baril, P.; Costa, E. C.; de Melo-Diogo, D.; Foucher, F.; Queiroz, J. A.; Sousa, F.; Pichon, C.; Correia, I. J., Bioreducible poly (2-ethyl-2-oxazoline)–PLA–PEI-SS triblock copolymer micelles for co-delivery of DNA minicircles and Doxorubicin. Journal of Controlled Release 2015, 213, 175-191.74. Lauter, V.; Lauter, H.; Glavic, A.; Toperverg, B., Reference module in materials science and materials engineering. In Elsevier: 2016.75. Pitsikalis, M., Ionic polymerization. 2013.76. Parker, G., Encyclopedia of materials: science and technology. 2001.77. Uchida, S., Graft Copolymer Synthesis. Encyclopedia of Polymeric Nanomaterials 2015, 867-870.78. Nuyken, O.; Pask, S., Ring-opening polymerization—An introductory review. Polymers 2013, 5, (2), 361-403.79. Matyjaszewski, K.; Xia, J., Atom transfer radical polymerization. Chemical reviews 2001, 101, (9), 2921-2990.80. Parrish, B.; Breitenkamp, R. B.; Emrick, T., PEG-and peptide-grafted aliphatic polyesters by click chemistry. Journal of the American Chemical Society 2005, 127, (20), 7404-7410.81. Odian, G., Principles of polymerization. John Wiley & Sons: 2004.82. Brunelle, D. J., Ring-Opening Polymerization. Mechanisms, Catalysis, Structure, Utility. Hanser Publishers, 1993 1993, 361.83. Albertsson, A.-C.; Varma, I. K., Recent developments in ring opening polymerization of lactones for biomedical applications. Biomacromolecules 2003, 4, (6), 1466-1486.84. Degée, P.; Dubois, P.; Jérǒme, R.; Jacobsen, S.; Fritz, H. G. In New catalysis for fast bulk ring‐opening polymerization of lactide monomers, Macromolecular Symposia, 1999; Wiley Online Library: 1999; pp 289-302.85. Storey, R. F.; Sherman, J. W., Kinetics and mechanism of the stannous octoate-catalyzed bulk polymerization of ε-caprolactone. Macromolecules 2002, 35, (5), 1504-1512.86. Braunecker, W. A.; Matyjaszewski, K., Controlled/living radical polymerization: Features, developments, and perspectives. Progress in Polymer Science 2007, 32, (1), 93-146.87. Dong, H.; Matyjaszewski, K., ARGET ATRP of 2-(dimethylamino) ethyl methacrylate as an intrinsic reducing agent. Macromolecules 2008, 41, (19), 6868-6870.88. Kolb, H. C.; Finn, M.; Sharpless, K. B., Click chemistry: diverse chemical function from a few good reactions. Angewandte Chemie International Edition 2001, 40, (11), 2004-2021.89. Lecomte, P.; Riva, R.; Jérôme, C.; Jérôme, R., Macromolecular Engineering of Biodegradable Polyesters by Ring‐Opening Polymerization and ‘Click’Chemistry. Macromolecular Rapid Communications 2008, 29, (12‐13), 982-997.90. Binder, W. H.; Sachsenhofer, R., ‘Click’chemistry in polymer and materials science. Macromolecular Rapid Communications 2007, 28, (1), 15-54.91. Bock, V. D.; Hiemstra, H.; Van Maarseveen, J. H., CuI‐catalyzed alkyne–azide “click” cycloadditions from a mechanistic and synthetic perspective. European Journal of Organic Chemistry 2006, 2006, (1), 51-68.92. Mespouille, L.; Vachaudez, M.; Suriano, F.; Gerbaux, P.; Coulembier, O.; Degée, P.; Flammang, R.; Dubois, P., One‐Pot Synthesis of Well‐Defined Amphiphilic and Adaptative Block Copolymers via Versatile Combination of “Click” Chemistry and ATRP. Macromolecular Rapid Communications 2007, 28, (22), 2151-2158.93. Darcos, V.; El Habnouni, S.; Nottelet, B.; El Ghzaoui, A.; Coudane, J., Well-defined PCL-graft-PDMAEMA prepared by ring-opening polymerisation and click chemistry. Polymer Chemistry 2010, 1, (3), 280-282.94. Mespouille, L.; Coulembier, O.; Paneva, D.; Degée, P.; Rashkov, I.; Dubois, P., Synthesis of adaptative and amphiphilic polymer model conetworks by versatile combination of ATRP, ROP, and “Click chemistry”. Journal of Polymer Science Part A: Polymer Chemistry 2008, 46, (15), 4997-5013.95. Liu, J.; Xu, L.; Jiang, X.; Hennink, W. E.; Wang, X.; Zhuo, R., Disulfide-containing cross-linked PEI derivative synthesized by click chemistry for non-viral gene delivery. Journal of Controlled Release 2011, 152, e157-e159.96. Zhang, X.; Li, Y.; Chen, Y. E.; Chen, J.; Ma, P. X., Cell-free 3D scaffold with two-stage delivery of miRNA-26a to regenerate critical-sized bone defects. Nature communications 2016, 7, 10376.97. Farshbaf, M.; Davaran, S.; Zarebkohan, A.; Annabi, N.; Akbarzadeh, A.; Salehi, R., Significant role of cationic polymers in drug delivery systems. Artificial cells, nanomedicine, and biotechnology 2018, 46, (8), 1872-1891.98. Bahadur, K. R.; Uludağ, H., PEI and its derivatives for gene therapy. In Polymers and Nanomaterials for Gene Therapy, Elsevier: 2016; pp 29-54.99. Lee, S. B.; Russell, A. J.; Matyjaszewski, K., ATRP synthesis of amphiphilic random, gradient, and block copolymers of 2-(dimethylamino) ethyl methacrylate and n-butyl methacrylate in aqueous media. Biomacromolecules 2003, 4, (5), 1386-1393.100. Lambermont-Thijs, H. M.; van der Woerdt, F. S.; Baumgaertel, A.; Bonami, L.; Du Prez, F. E.; Schubert, U. S.; Hoogenboom, R., Linear poly (ethylene imine) s by acidic hydrolysis of poly (2-oxazoline) s: kinetic screening, thermal properties, and temperature-induced solubility transitions. Macromolecules 2009, 43, (2), 927-933.101. Neu, M.; Fischer, D.; Kissel, T., Recent advances in rational gene transfer vector design based on poly (ethylene imine) and its derivatives. The Journal of Gene Medicine: A cross‐disciplinary journal for research on the science of gene transfer and its clinical applications 2005, 7, (8), 992-1009.102. Zhang, W.; He, J.; Liu, Z.; Ni, P.; Zhu, X., Biocompatible and pH‐responsive triblock copolymer mPEG‐b‐PCL‐b‐PDMAEMA: synthesis, self‐assembly, and application. Journal of Polymer Science Part A: Polymer Chemistry 2010, 48, (5), 1079-1091.103. Brannon-Peppas, L., Poly (ethylene glycol): Chemistry and Biological Applications-JM Harris and S. Zalipsky, editors, American Chemical Society, Washington DC, 1997, 489 pp. Journal of Controlled Release 2000, 2, (66), 321.104. Chen, W.; Yang, H.; Wang, R.; Cheng, R.; Meng, F.; Wei, W.; Zhong, Z., Versatile synthesis of functional biodegradable polymers by combining ring-opening polymerization and postpolymerization modification via Michael-type addition reaction. Macromolecules 2009, 43, (1), 201-207.105. Mullen, B. D.; Tang, C. N.; Storey, R. F., New aliphatic poly (ester‐carbonates) based on 5‐methyl‐5‐allyloxycarbonyl‐1, 3‐dioxan‐2‐one. Journal of Polymer Science Part A: Polymer Chemistry 2003, 41, (13), 1978-1991.106. Chen, W.; Zou, Y.; Jia, J.; Meng, F.; Cheng, R.; Deng, C.; Feijen, J.; Zhong, Z., Functional poly (ε-caprolactone) s via copolymerization of ε-caprolactone and pyridyl disulfide-containing cyclic carbonate: controlled synthesis and facile access to reduction-sensitive biodegradable graft copolymer micelles. Macromolecules 2013, 46, (3), 699-707.107. Shi, Q.; Chen, X.; Lu, T.; Jing, X., The immobilization of proteins on biodegradable polymer fibers via click chemistry. Biomaterials 2008, 29, (8), 1118-1126.108. Xu, J.; Prifti, F.; Song, J., A Versatile Monomer for Preparing Well-Defined Functional Polycarbonates and Poly (ester− carbonates). Macromolecules 2011, 44, (8), 2660-2667.109. Mespouille, L.; Vachaudez, M.; Suriano, F.; Gerbaux, P.; Van Camp, W.; Coulembier, O.; Degée, P.; Flammang, R.; Du Prez, F.; Dubois, P., Controlled synthesis of amphiphilic block copolymers based on polyester and poly (amino methacrylate): Comprehensive study of reaction mechanisms. Reactive and Functional Polymers 2008, 68, (5), 990-1003.110. Lu, C.; Shi, Q.; Chen, X.; Lu, T.; Xie, Z.; Hu, X.; Ma, J.; Jing, X., Sugars‐grafted aliphatic biodegradable poly (L‐lactide‐co‐carbonate) s by click reaction and their specific interaction with lectin molecules. Journal of Polymer Science Part A: Polymer Chemistry 2007, 45, (15), 3204-3217.111. Li, C.; Liu, X.; He, S.; Huang, Y.; Cui, D., Synthesis and AIE properties of PEG–PLA–PMPC based triblock amphiphilic biodegradable polymers. Polymer Chemistry 2016, 7, (5), 1121-1128.112. Cordeiro, R. A.; Farinha, D.; Rocha, N.; Serra, A. C.; Faneca, H.; Coelho, J. F., Novel Cationic Triblock Copolymer of Poly [2‐(dimethylamino) ethyl methacrylate]‐block‐poly (β‐amino ester)‐block‐poly [2‐(dimethylamino) ethyl methacrylate]: A Promising Non‐Viral Gene Delivery System. Macromolecular bioscience 2015, 15, (2), 215-228.113. Tauhardt, L.; Kempe, K.; Knop, K.; Altuntaş, E.; Jäger, M.; Schubert, S.; Fischer, D.; Schubert, U. S., Linear polyethyleneimine: optimized synthesis and characterization–on the way to “pharmagrade” batches. Macromolecular Chemistry and Physics 2011, 212, (17), 1918-1924.114. Tempelaar, S.; Barker, I. A.; Truong, V. X.; Hall, D. J.; Mespouille, L.; Dubois, P.; Dove, A. P., Organocatalytic synthesis and post-polymerization functionalization of propargyl-functional poly (carbonate) s. Polymer Chemistry 2013, 4, (1), 174-183.115. Albertsson, A. C.; Eklund, M., Synthesis of copolymers of 1, 3‐dioxan‐2‐one and oxepan‐2‐one using coordination catalysts. Journal of Polymer Science Part A: Polymer Chemistry 1994, 32, (2), 265-279.116. Wei, Z.; Liu, L.; Qi, M., Kinetics and mechanism of the ring opening polymerization of (R, S)-β-butyrolactone initiated with dibutylmagnesium. European polymer journal 2007, 43, (4), 1210-1218.117. Tempelaar, S. Synthesis and Post-polymerisation Functionalisation of Aliphatic Poly (carbonate) s. University of Warwick, 2012.118. Diaz, I. L.; Perez, L. D., Synthesis and micellization properties of triblock copolymers PDMAEMA-b-PCL-b-PDMAEMA and their applications in the fabrication of amphotericin B-loaded nanocontainers. Colloid and Polymer Science 2015, 293, (3), 913-923.119. Muñoz-Bonilla, A.; Fernández-García, M.; Haddleton, D. M., Synthesis and aqueous solution properties of stimuli-responsive triblock copolymers. Soft Matter 2007, 3, (6), 725-731.120. Song, J.; Jung, Y.; Lee, I.; Jang, J., Fabrication of pDMAEMA-coated silica nanoparticles and their enhanced antibacterial activity. Journal of colloid and interface science 2013, 407, 205-209.121. Yin, J. J.; Wahid, F.; Zhang, Q.; Tao, Y. C.; Zhong, C.; Chu, L. Q., Facile Incorporation of Silver Nanoparticles into Quaternized Poly (2‐(Dimethylamino) Ethyl Methacrylate) Brushes as Bifunctional Antibacterial Coatings. Macromolecular Materials and Engineering 2017, 302, (6), 1700069.122. Bonami, L.; Van Camp, W.; Van Rijckegem, D.; Du Prez, F. E., Facile Access to an Efficient Solid‐Supported Click Catalyst System Based on Poly (ethyleneimine). Macromolecular rapid communications 2009, 30, (1), 34-38.123. Zhang, G.; Liu, J.; Yang, Q.; Zhuo, R.; Jiang, X., Disulfide-containing brushed polyethylenimine derivative synthesized by click chemistry for nonviral gene delivery. Bioconjugate chemistry 2012, 23, (6), 1290-1299.124. Müller, A. J.; Michell, R. M., Differential scanning calorimetry of polymers. Polym. Morphol. Princ. Charact. Process 2016, 72-99.125. Du, Z. X.; Xu, J. T.; Yang, Y.; Fan, Z. Q., Synthesis and characterization of poly (ϵ‐caprolactone)‐b‐poly (ethylene glycol) block copolymers prepared by a salicylaldimine‐aluminum complex. Journal of applied polymer science 2007, 105, (2), 771-776.126. Tran, T.-Q.-M.; Hsieh, M.-F.; Chang, K.-L.; Pho, Q.-H.; Nguyen, V.-C.; Cheng, C.-Y.; Huang, C.-M., Bactericidal effect of lauric acid-loaded PCL-PEG-PCL nano-sized micelles on skin commensal Propionibacterium acnes. Polymers 2016, 8, (9), 321.127. Nojima, S.; Ono, M.; Ashida, T., Crystallization of block copolymers II. Morphological study of poly (ethylene glycol)-poly (ε-caprolactone) block copolymers. Polymer journal 1992, 24, (11), 1271.128. Hu, X.; Chen, X.; Xie, Z.; Cheng, H.; Jing, X., Aliphatic poly (ester‐carbonate) s bearing amino groups and its RGD peptide grafting. Journal of Polymer Science Part A: Polymer Chemistry 2008, 46, (21), 7022-7032.129. Bruce, C.; Javakhishvili, I.; Fogelström, L.; Carlmark, A.; Hvilsted, S.; Malmström, E., Well-defined ABA-and BAB-type block copolymers of PDMAEMA and PCL. Rsc Advances 2014, 4, (49), 25809-25818.130. Weyts, K. F.; Goethals, E. J., New synthesis of linear polyethyleneimine. Polymer bulletin 1988, 19, (1), 13-19.131. Tanaka, R.; Ueoka, I.; Takaki, Y.; Kataoka, K.; Saito, S., High molecular weight linear polyethylenimine and poly (N-methylethylenimine). Macromolecules 1983, 16, (6), 849-853.132. Lambermont-Thijs, H. M.; Bonami, L.; Du Prez, F. E.; Hoogenboom, R., Linear poly (alkyl ethylene imine) with varying side chain length: synthesis and physical properties. Polymer Chemistry 2010, 1, (5), 747-754.133. Shuai, X.; Merdan, T.; Unger, F.; Wittmar, M.; Kissel, T., Novel biodegradable ternary copolymers hy-PEI-g-PCL-b-PEG: synthesis, characterization, and potential as efficient nonviral gene delivery vectors. Macromolecules 2003, 36, (15), 5751-5759.134. Cao, P. F.; Felipe, M. J.; Advincula, R. C., On the Formation and Electropolymerization of a Star Copolymer With Peripheral Carbazoles. Macromolecular Chemistry and Physics 2013, 214, (3), 386-395.135. Riess, G., Micellization of block copolymers. Progress in Polymer Science 2003, 28, (7), 1107-1170.136. Mai, Y.; Eisenberg, A., Self-assembly of block copolymers. Chemical Society Reviews 2012, 41, (18), 5969-5985.137. Munk, P., Equilibrium and nonequilibrium polymer micelles. In Solvents and Self-organization of Polymers, Springer: 1996; pp 19-32.138. Loh, X. J.; Wu, Y.-L.; Seow, W. T. J.; Norimzan, M. N. I.; Zhang, Z.-X.; Xu, F.-J.; Kang, E.-T.; Neoh, K.-G.; Li, J., Micellization and phase transition behavior of thermosensitive poly (N-isopropylacrylamide)–poly (ɛ-caprolactone)–poly (N-isopropylacrylamide) triblock copolymers. Polymer 2008, 49, (23), 5084-5094.139. Zhang, X.; Zhu, X.; Ke, F.; Ye, L.; Chen, E.-q.; Zhang, A.-y.; Feng, Z.-g., Preparation and self-assembly of amphiphilic triblock copolymers with polyrotaxane as a middle block and their application as carrier for the controlled release of Amphotericin B. Polymer 2009, 50, (18), 4343-4351.140. Ginn, S. L.; Alexander, I. E.; Edelstein, M. L.; Abedi, M. R.; Wixon, J., Gene therapy clinical trials worldwide to 2012–an update. The journal of gene medicine 2013, 15, (2), 65-77.141. Yin, H.; Kanasty, R. L.; Eltoukhy, A. A.; Vegas, A. J.; Dorkin, J. R.; Anderson, D. G., Non-viral vectors for gene-based therapy. Nature Reviews Genetics 2014, 15, (8), 541-555.142. Pack, D. W.; Hoffman, A. S.; Pun, S.; Stayton, P. S., Design and development of polymers for gene delivery. Nature reviews Drug discovery 2005, 4, (7), 581.143. Endres, T. Biodegradable amphiphilic PEG-PCL-PEI triblock copolymers designed for the self-assembly of multifunctional gene carriers. Philipps-Universität Marburg, 2012.144. Van der Gucht, J.; Spruijt, E.; Lemmers, M.; Stuart, M. A. C., Polyelectrolyte complexes: bulk phases and colloidal systems. Journal of colloid and interface science 2011, 361, (2), 407-422.145. Bucur, C. B.; Sui, Z.; Schlenoff, J. B., Ideal mixing in polyelectrolyte complexes and multilayers: Entropy driven assembly. Journal of the American Chemical Society 2006, 128, (42), 13690-13691.146. Che, J.; Tao, A.; Chen, S.; Li, X.; Zhao, Y.; Yuan, W., Biologically responsive carrier-mediated anti-angiogenesis shRNA delivery for tumor treatment. Scientific reports 2016, 6, 35661.147. Kunath, K.; von Harpe, A.; Fischer, D.; Petersen, H.; Bickel, U.; Voigt, K.; Kissel, T., Low-molecular-weight polyethylenimine as a non-viral vector for DNA delivery: comparison of physicochemical properties, transfection efficiency and in vivo distribution with high-molecular-weight polyethylenimine. Journal of Controlled Release 2003, 89, (1), 113-125.148. Mosqueira, V. C. F.; Legrand, P.; Gref, R.; Heurtault, B.; Appel, M.; Barratt, G., Interactions between a macrophage cell line (J774A1) and surface-modified poly (D, L-lactide) nanocapsules bearing poly (ethylene glycol). Journal of drug targeting 1999, 7, (1), 65-78.149. Shen, Z.-l.; Xia, Y.-q.; Yang, Q.-s.; Chen, K.; Ma, Y.-q., Polymer–nucleic acid interactions. In Polymeric Gene Delivery Systems, Springer: 2017; pp 41-64.150. Stahlschmidt, U.; Jérôme, V.; Majewski, A.; Müller, A.; Freitag, R., Systematic Study of a Library of PDMAEMA-Based, Superparamagnetic Nano-Stars for the Transfection of CHO-K1 Cells. Polymers 2017, 9, (5), 156.151. Guzaev, M.; Li, X.; Park, C.; Leung, W.-Y.; Roberts, L., Comparison of nucleic acid gel stains cell permeability, safety, and sensitivity of ethidium bromide alternatives. Online at https://biotium. com/wp-content/uploads/2017/02/Gel-Stains-Comparison. pdf 2017.152. Raup, A.; Wang, H.; Synatschke, C. V.; Jérôme, V. r.; Agarwal, S.; Pergushov, D. V.; Müller, A. H.; Freitag, R., Compaction and transmembrane delivery of pdna: Differences between l-PEI and two types of amphiphilic block copolymers. Biomacromolecules 2017, 18, (3), 808-818.153. Visakh, P., Polyelectrolyte: Thermodynamics and Rheology. In Polyelectrolytes, Springer: 2014; pp 1-17.154. Riley, T.; Govender, T.; Stolnik, S.; Xiong, C.; Garnett, M.; Illum, L.; Davis, S., Colloidal stability and drug incorporation aspects of micellar-like PLA–PEG nanoparticles. Colloids and surfaces B: Biointerfaces 1999, 16, (1-4), 147-159.155. Laugel, N.; Betscha, C.; Winterhalter, M.; Voegel, J.-C.; Schaaf, P.; Ball, V., Relationship between the growth regime of polyelectrolyte multilayers and the polyanion/polycation complexation enthalpy. The journal of physical chemistry B 2006, 110, (39), 19443-19449.156. Khalil, I. A.; Kogure, K.; Akita, H.; Harashima, H., Uptake pathways and subsequent intracellular trafficking in nonviral gene delivery. Pharmacological reviews 2006, 58, (1), 32-45.157. Aied, A.; Greiser, U.; Pandit, A.; Wang, W., Polymer gene delivery: overcoming the obstacles. Drug discovery today 2013, 18, (21-22), 1090-1098.158. Liu, S.; Gao, Y.; Zhou, D.; Zeng, M.; Alshehri, F.; Newland, B.; Lyu, J.; O’Keeffe-Ahern, J.; Greiser, U.; Guo, T., Highly branched poly (β-amino ester) delivery of minicircle DNA for transfection of neurodegenerative disease related cells. Nature communications 2019, 10, (1), 1-14.159. Li, X.; Guo, X.; Cheng, Y.; Zhao, X.; Fang, Z.; Luo, Y.; Xia, S.; Feng, Y.; Chen, J.; Yuan, W.-E., pH-responsive cross-linked low molecular weight polyethylenimine as an efficient gene vector for delivery of plasmid DNA encoding anti-VEGF-shRNA for tumor treatment. Frontiers in oncology 2018, 8.160. Faneca, H.; Simoes, S.; De Lima, M. P., Association of albumin or protamine to lipoplexes: enhancement of transfection and resistance to serum. The journal of gene medicine 2004, 6, (6), 681-692.161. Cordeiro, R. A.; Santo, D.; Farinha, D.; Serra, A.; Faneca, H.; Coelho, J. F., High transfection efficiency promoted by tailor-made cationic tri-block copolymer-based nanoparticles. Acta biomaterialia 2017, 47, 113-123.162. Jones, C. H.; Chen, C.-K.; Ravikrishnan, A.; Rane, S.; Pfeifer, B. A., Overcoming nonviral gene delivery barriers: perspective and future. Molecular pharmaceutics 2013, 10, (11), 4082-4098.163. Sun, H.; Zhou, L.; Chen, X.; Han, X.; Wang, R.; Liu, H., Microscopic insight into the DNA condensation process of a zwitterion‐functionalized polycation. Biopolymers 2016, 105, (11), 802-810.164. Xiao, K.; Li, Y.; Luo, J.; Lee, J. S.; Xiao, W.; Gonik, A. M.; Agarwal, R. G.; Lam, K. S., The effect of surface charge on in vivo biodistribution of PEG-oligocholic acid based micellar nanoparticles. Biomaterials 2011, 32, (13), 3435-3446.165. Pezzoli, D.; Giupponi, E.; Mantovani, D.; Candiani, G., Size matters for in vitro gene delivery: investigating the relationships among complexation protocol, transfection medium, size and sedimentation. Scientific reports 2017, 7, 44134.166. Dash, P.; Read, M.; Barrett, L.; Wolfert, M.; Seymour, L., Factors affecting blood clearance and in vivo distribution of polyelectrolyte complexes for gene delivery. Gene therapy 1999, 6, (4), 643.167. Santo, D.; Cordeiro, R. A.; Sousa, A.; Serra, A. n.; Coelho, J. F.; Faneca, H., Combination of Poly [(2-dimethylamino) ethyl methacrylate] and Poly (β-amino ester) Results in a Strong and Synergistic Transfection Activity. Biomacromolecules 2017, 18, (10), 3331-3342.168. Venkataraman, S.; Hedrick, J. L.; Ong, Z. Y.; Yang, C.; Ee, P. L. R.; Hammond, P. T.; Yang, Y. Y., The effects of polymeric nanostructure shape on drug delivery. Advanced drug delivery reviews 2011, 63, (14-15), 1228-1246.169. Gary, D. J.; Lee, H.; Sharma, R.; Lee, J.-S.; Kim, Y.; Cui, Z. Y.; Jia, D.; Bowman, V. D.; Chipman, P. R.; Wan, L., Influence of nano-carrier architecture on in vitro siRNA delivery performance and in vivo biodistribution: polyplexes vs micelleplexes. ACS nano 2011, 5, (5), 3493-3505.170. Zhang, J.; Li, X.; Lou, L.; Li, X.; Jia, Y.; Jin, Z.; Zhu, Y., Non-viral gene therapy. In Intracellular Delivery, Springer: 2011; pp 599-699.171. Majewski, A. P.; Stahlschmidt, U.; Jérôme, V. r.; Freitag, R.; Müller, A. H.; Schmalz, H., PDMAEMA-grafted core–shell–corona particles for nonviral gene delivery and magnetic cell separation. Biomacromolecules 2013, 14, (9), 3081-3090.172. Endres, T. K.; Beck-Broichsitter, M.; Samsonova, O.; Renette, T.; Kissel, T. H., Self-assembled biodegradable amphiphilic PEG–PCL–lPEI triblock copolymers at the borderline between micelles and nanoparticles designed for drug and gene delivery. Biomaterials 2011, 32, (30), 7721-7731.173. Lu, X.; Liu, L., Asymmetric polyplex-nanocapsules loaded with photosentisizer for light-assisted gene transfer. Journal of Photochemistry and Photobiology B: Biology 2017, 174, 269-275.174. Dai, J.; Zou, S.; Pei, Y.; Cheng, D.; Ai, H.; Shuai, X., Polyethylenimine-grafted copolymer of poly (l-lysine) and poly (ethylene glycol) for gene delivery. Biomaterials 2011, 32, (6), 1694-1705.175. Abebe, D. G.; Kandil, R.; Kraus, T.; Elsayed, M.; Merkel, O. M.; Fujiwara, T., Three‐Layered Biodegradable Micelles Prepared by Two‐Step Self‐Assembly of PLA‐PEI‐PLA and PLA‐PEG‐PLA Triblock Copolymers as Efficient Gene Delivery System. Macromolecular bioscience 2015, 15, (5), 698-711.176. Zakeri, A.; Kouhbanani, M. A. J.; Beheshtkhoo, N.; Beigi, V.; Mousavi, S. M.; Hashemi, S. A. R.; Karimi Zade, A.; Amani, A. M.; Savardashtaki, A.; Mirzaei, E., Polyethylenimine-based nanocarriers in co-delivery of drug and gene: a developing horizon. Nano reviews & experiments 2018, 9, (1), 1488497.177. Schallon, A.; Jérôme, V.; Walther, A.; Synatschke, C. V.; Müller, A. H.; Freitag, R., Performance of three PDMAEMA-based polycation architectures as gene delivery agents in comparison to linear and branched PEI. Reactive and Functional Polymers 2010, 70, (1), 1-10.178. Saqafi, B.; Rahbarizadeh, F., Effect of PEI surface modification with PEG on cytotoxicity and transfection efficiency. Micro & Nano Letters 2018, 13, (8), 1090-1095.179. Xing, H.; Lu, M.; Yang, T.; Liu, H.; Sun, Y.; Zhao, X.; Xu, H.; Yang, L.; Ding, P., Structure-function relationships of nonviral gene vectors: Lessons from antimicrobial polymers. Acta biomaterialia 2018.180. Schallon, A.; Synatschke, C. V.; Jérôme, V. r.; Müller, A. H.; Freitag, R., Nanoparticulate nonviral agent for the effective delivery of pDNA and siRNA to differentiated cells and primary human T lymphocytes. Biomacromolecules 2012, 13, (11), 3463-3474.181. Standardization, I. O. f., ISO 10993‐5: Biological Evaluation of Medical Devices. Part 5: Tests For In Vitro Cytotoxicity. In ISO Geneva, Switzerland: 2009.182. Fischer, D.; Dautzenberg, H.; Kunath, K.; Kissel, T., Poly (diallyldimethylammonium chlorides) and their N-methyl-N-vinylacetamide copolymer-based DNA-polyplexes: role of molecular weight and charge density in complex formation, stability, and in vitro activity. International journal of pharmaceutics 2004, 280, (1-2), 253-269.183. Fischer, D.; Li, Y.; Ahlemeyer, B.; Krieglstein, J.; Kissel, T., In vitro cytotoxicity testing of polycations: influence of polymer structure on cell viability and hemolysis. Biomaterials 2003, 24, (7), 1121-1131.184. Ivanova, E. P.; Bazaka, K.; Crawford, R. J., New functional biomaterials for medicine and healthcare. Woodhead publishing New Delhi, India:: 2014; Vol. 67.185. van de Wetering, P.; Cherng, J.-Y.; Talsma, H.; Hennink, W. E., Relation between transfection efficiency and cytotoxicity of poly (2-(dimethylamino) ethyl methacrylate)/plasmid complexes. Journal of Controlled Release 1997, 49, (1), 59-69.186. Beyerle, A.; Merkel, O.; Stoeger, T.; Kissel, T., PEGylation affects cytotoxicity and cell-compatibility of poly (ethylene imine) for lung application: structure–function relationships. Toxicology and applied pharmacology 2010, 242, (2), 146-154.187. Zhang, H.; Chen, Z.; Du, M.; Li, Y.; Chen, Y., Enhanced gene transfection efficiency by low-dose 25 kDa polyethylenimine by the assistance of 1.8 kDa polyethylenimine. Drug delivery 2018, 25, (1), 1740-1745.188. Forcato, D.; Fili, A.; Alustiza, F.; Martínez, J. L.; Abel, S. B.; Nicotra, M. O.; Alessio, A.; Rodríguez, N.; Barbero, C.; Bosch, P., Transfection of bovine fetal fibroblast with polyethylenimine (PEI) nanoparticles: effect of particle size and presence of fetal bovine serum on transgene delivery and cytotoxicity. Cytotechnology 2017, 69, (4), 655-665.189. Wahlfors, J.; Loimas, S.; Pasanen, T.; Hakkarainen, T., Green fluorescent protein (GFP) fusion constructs in gene therapy research. Histochemistry and cell biology 2001, 115, (1), 59-65.190. Liu, H. C.; Zhao, H.; Chen, J.; Wu, W. L.; Wang, H. L.; Jiao, G. J.; Chen, Y. Z., Role of recombinant plasmid pEGFP-N1-IGF-1 transfection in alleviating osteoporosis in ovariectomized rats. Journal of molecular histology 2013, 44, (5), 535-544.191. Riedl, S.; Kaiser, P.; Raup, A.; Synatschke, C.; Jérôme, V.; Freitag, R., Non-Viral Transfection of Human T Lymphocytes. Processes 2018, 6, (10), 188.192. Raup, A.; Stahlschmidt, U.; Jérôme, V.; Synatschke, C. V.; Müller, A. H.; Freitag, R., Influence of polyplex formation on the performance of star-shaped polycationic transfection agents for mammalian cells. Polymers 2016, 8, (6), 224.193. Yue, Y.; Jin, F.; Deng, R.; Cai, J.; Dai, Z.; Lin, M. C.; Kung, H.-F.; Mattebjerg, M. A.; Andresen, T. L.; Wu, C., Revisit complexation between DNA and polyethylenimine—Effect of length of free polycationic chains on gene transfection. Journal of controlled release 2011, 152, (1), 143-151.194. Wen, Y.; Pan, S.; Luo, X.; Zhang, W.; Shen, Y.; Feng, M., PEG-and PDMAEG-graft-modified branched PEI as novel gene vector: synthesis, characterization and gene transfection. Journal of Biomaterials Science, Polymer Edition 2010, 21, (8-9), 1103-1126.195. Chen, B.; Synatschke, C. V.; Jérôme, V.; Müller, A. H.; Freitag, R.; Wu, C., Co-transfection of star-shaped PDMAEMAs enhance transfection efficiency of protamine/pDNA complexes in the presence of serum. European Polymer Journal 2018, 103, 362-369.196. Cai, J.; Yue, Y.; Wang, Y.; Jin, Z.; Jin, F.; Wu, C., Quantitative study of effects of free cationic chains on gene transfection in different intracellular stages. Journal of Controlled Release 2016, 238, 71-79.197. Varga, C.; Tedford, N.; Thomas, M.; Klibanov, A.; Griffith, L.; Lauffenburger, D., Quantitative comparison of polyethylenimine formulations and adenoviral vectors in terms of intracellular gene delivery processes. Gene therapy 2005, 12, (13), 1023.ORIGINAL1015402910.2020.pdf1015402910.2020.pdfapplication/pdf6374789https://repositorio.unal.edu.co/bitstream/unal/78367/1/1015402910.2020.pdfa089d4843b1fe1441b37c87492c36880MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8805https://repositorio.unal.edu.co/bitstream/unal/78367/3/license_rdfdab767be7a093b539031785b3bf95490MD53LICENSElicense.txtlicense.txttext/plain; charset=utf-83895https://repositorio.unal.edu.co/bitstream/unal/78367/4/license.txte2f63a891b6ceb28c3078128251851bfMD54THUMBNAIL1015402910.2020.pdf.jpg1015402910.2020.pdf.jpgGenerated Thumbnailimage/jpeg5236https://repositorio.unal.edu.co/bitstream/unal/78367/5/1015402910.2020.pdf.jpg82682bcb555abae99f118ef4bfcad81aMD55unal/78367oai:repositorio.unal.edu.co:unal/783672023-07-19 23:04:18.933Repositorio Institucional Universidad Nacional de Colombiarepositorio_nal@unal.edu.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