In Vitro Transfection Mediated by Dendrigraft Poly(L‐lysines): The Effect of Structure and Molecule Size

Abstract

Dendritic poly(L-lysines) (DGL) constitute promising nanomaterials applicable as a nonviral gene-delivery vector. In this study, we evaluate the transfection abilities of four DGL generations with special emphasis on the systematic description of the relationship of how generation (i.e., molecule size) affects the transfection efficacy. Using Hep2 cells, we demonstrated that the capability of unmodified DGL to deliver plasmid is of a magnitude lower than that of jetPEI. On the other hand, employing the Hep2 cell line stably transduced with eGFP, we observed that DGL G5 delivers the siRNA oligonucleotide with the same efficiency as Lipofectamine 2000. In further experiments, it was shown that DGL affords excellent ability to bind DNA, protect it against DNase I attack, and internalize it into cells.

Digitizing Poly-l-lysine Dendrigrafts: From Experimental Data to Molecular Dynamics Simulations

“…In contrast to PAMAM and PEI, DGLs are biodegradable, 2 exhibit low cellular toxicities, 3 and turned out to be non-immunogenic. 4 As a consequence, DGLs have recently gained a huge interest in the biomedical field during the last quinquennium, with numerous applications in drug and gene delivery, 2,3,[5][6][7][8][9][10][11][12][13][14][15][16][17][18] biomaterials and tissue engineering, [19][20][21][22][23][24][25] bio-imaging [26][27][28][29][30][31] and biosensing. [32][33][34] Despite this growing use of DGLs, a deeper understanding of the molecular level properties of these macromolecules is missing.…”

Molecular interactions of DNA with transfectants: a study based on infrared spectroscopy and quantum chemistry as aids to fluorescence spectroscopy and dynamic light scattering analyses

“…In order to improve their effectiveness, a number of studies dealing with establishing structure-function relationships have been reported. [5][6][7][8] Detailed and extensive characterization of the physicochemical and biophysical features of polyplexes does basically rely on multiple techniques and methods, in rst place dynamic light scattering (DLS) and electron microscopy (EM) for size measurements, [9][10][11] Doppler velocimetry for surface charge measurements, while agarose gel retardation analysis and dyeexclusion assays for the determination of the binding strength between the polymers and the nucleic acids and the stability of polyplexes over time and in different environments. [12][13][14] Unfortunately, mechanistic insights into polyplex formation and disassembly have been hampered by a lack of atomic resolution, and structural information on polymer/DNA complexes.…”

Everything You Always Wanted to Know about Poly‐l‐lysine Dendrigrafts (But Were Afraid to Ask)

“…[35] Drug and gene delivery Severalo bstacles such as fast degradation,l ow penetration through membranes, or low selectivityf or target cells, often hinder the efficient delivery of bioactive molecules,i ncluding peptides or proteins. [31] Regardingt heir plasmidD NA (pDNA)a nd silencing RNA (siRNA) internalizingp roperties in Hep2 cells, DGLs can be ranked as follows: G3 > G4 ! [36] The protection against enzymaticd egradation as well as the retardationo fi nsulin release in simulated intestinal fluid, which were found to be positively related to the number of guanidine end-groups, open interesting perspectives in the delivery of insulin to systemic circulation through oral administration.…”

“…Indeed, it was demonstrated that DGLs are readily soluble in water at least up to 60 gL À1 in water, [12] are stable under sterilization conditions, [28] and are partially degradable under the action of endogenous peptidases. [29] In addition, these biomacromolecules are non-immunogenic [30] and carry low cytotoxicity (i.e., IC50 values on Hep2 cellso f1 6.8, 13.2, and 13.9 mgmL À1 from second-to fourth-generation DGL, respectively), [31] in comparison with other polycationic polymers (e.g.,a bout 90 %v iability for DGL G3 versusl ess than 50 %v iability for hyperbranched PEI at as ame concentration of 12.5 mgmL À1 on HeLa cells). [32] As aresult, DGLs have already attracted the interest of multiple researchg roups during the last fivey ears, and found numerous applications in the biomedical field, in which the ability of the arborescent polymers to display multiple functionalities has been exploited.…”