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*vitro* one (Ding et al., 2008) and positively charged lipoplexes are significantly more effective than negatively charged ones. Other investigations revealed that the best TE *in vivo*  corresponds to a low N/P ratio, small size of lipoplexes, and negative ξ-potential (Hattori et al., 2007; Gao & Hui, 2001). It is noteworthy when considering polycationic lipids, that lipoplexes that have a total negative charge and small size (200–300 nm) are optimal for both

It was revealed that the efficient gene delivery by polycationic lipids (Cooper et al., 1998) *in vivo* requires the cationic liposome systems, which are able to bind DNA more tightly than for the *in vitro* delivery. In comparison to CDAN(**18d**)/DOPE and DC-Chol(**1a**)/DOPE liposomes, CTAP(**18g**)/DOPE was able to neutralize, condense and encapsulate nucleic acids into lipoplex particles with a high efficiency. SDS stimulated the DNA release from a lipoplex and revealed the structure-activity relationships between the TE and lipid shielding of DNA (Bajaj et al., 2008c). Low shielding could facilitate the release of DNA and its hydrolysis within the cells. Certainly, this characteristic could be expected to be useful *in vivo,* given the greater complexity of the extracellular environment *in vivo* as compared to *in vitro*. Caminiti and co-workers demonstrated that both unstable and lipoplexes that are too stable, result in a strong and poor DNA release respectively, and exhibit a low transfection

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**16** 

*Spain* 

**Polycation-Mediated Gene Delivery:** 

Manuel Alatorre-Meda1, Eustolia Rodríguez-Velázquez2

*1Grupo de Física de Coloides y Polímeros, Departamento de Física de la Materia* 

*2Master student. Master en Ciencia y Tecnología de Materiales, Facultad de Física* 

*3Grupo de Nanomateriales y Materia Blanda, Departamento de Física de la Materia* 

Gene therapy is the process by which a foreign, corrective (or missing) gene is inserted toward biological tissues or cells aiming to alleviate symptoms or prevent disorders. Several clinical trials have demonstrated gene therapy as a promising option to treat diseases. However, therapeutic biological limitations (such as adverse immune responses of the body to incoming gene delivery systems) coupled with a poor understanding of the physicochemical motifs involved in the DNA compaction and delivery (transfection) processes have resulted in non-100% effective protocols. Aiming to contribute to a better understanding of the different physicochemical aspects of gene therapy, our group has committed for some years now to the physical chemistry characterization of the DNA compaction and transfection mediated by different kinds of compacting agents (vectors). In this chapter, we present an overview of the results we have obtained during the last three years regarding the DNA compaction and transfection mediated by cationic-liposomes and polymers (polycations). Two families of polycations, Chitosan and Poly (diallyldimethylammonium chloride) (pDADMAC), and one cationic lipid formulation extensively used worldwide in transfection assays, Metafectene® Pro (MEP), are studied as DNA vectors and compared with other systems already published. In particular, by varying the solution pH and polycation characteristics (chemical composition and molecular weight), we assess the influence of polycation-charge density (i.e., the mole fraction of the ionized groups along the polymer chain) and -valence (i.e., the total charge per polymer chain) on different parameters of the complexes formed that are important for gene therapy. The studied parameters include i) the hydrodynamic radius, RH, ii) the stability with time, iii) the vector to DNA ratio at which complexation takes place iv) the ζ-potential, v) the

**1. Introduction** 

**1.1 Chapter objectives** 

*Condensada. Facultad de Física, Universidad de Santiago de Compostela* 

*Condensada. Facultad de Física, Universidad de Santiago de Compostela* 

**The Physicochemical Aspects** 

**Governing the Process** 

*Universidad de Santiago de Compostela* 

and Julio R. Rodríguez3

