Search results for "gene delivery."

Low RNA translation activity limits the efficacy of hydrodynamic gene transfer to pig liver “in vivo”

2014

Background Hydrodynamic gene delivery has proved an efficient strategy for nonviral gene therapy in the murine liver but it has been less efficient in pigs. The reason for such inefficiency remains unclear. The present study used a surgical strategy to seal the whole pig liver in vivo. Methods A solution of enhanced green fluorescent protein (eGFP) DNA was injected under two different venous injection conditions (anterograde and retrograde), employing flow rates of 10 and 20 ml/s in each case, with the aim of identifying the best gene transfer conditions. The gene delivery and information decoding steps were evaluated by measuring the eGFP DNA, mRNA and protein copy number 24 h after transf…

EMPLOYMENT OF CATIONIC SOLID-LIPID NANOPARTICLES AS RNA CARRIERS

2007

Gene transfer represents an important advance in the treatment of both genetic and acquired diseases. In this article, the suitability of cationically modified solid-lipid nanoparticles (SLN) as a nonviral vector for gene delivery was investigated, in order to obtain stable materials able to condense RNA. Cationic SLN were produced by microemulsion using Compritol ATO 888 as matrix lipid, Pluronic F68 as tenside, and dimethyldioctadecylammonium bromide (DDAB) as cationic lipid. The resulting particles were approximately 100 nm in size and showed a highly positive surface charge (+41 mV) in water. Size and shape were further characterized by scanning electron microscopy (SEM) measurements. M…

A surgical model for isolating the pig liver in vivo for gene therapy.

2013

Several studies report results that suggest the need of vascularization blocking for efficient gene transfer to the liver, especially in nonviral gene therapy. In this study, we describe a surgical strategy for in vivo isolation of the pig liver, resulting in a vascular watertight organ that allows the evaluation of several gene injection conditions. The hepatic artery and portal, suprahepatic and infrahepatic cava veins were dissected. Then, liver vascularization was excluded for 5-7 min. In that time, we first injected 200 ml saline solution containing the p3c-eGFP plasmid (20 µg/ml) simultaneously through two different catheters placed in the portal and cava veins, respectively. Vital co…

Combining reactive triblock copolymers with functional cross-linkers: A versatile pathway to disulfide stabilized-polyplex libraries and their applic…

2017

Therapeutic nucleic acids such as pDNA hold great promise for the treatment of multiple diseases. These therapeutic interventions are, however, compromised by the lack of efficient and safe non-viral delivery systems, which guarantee stability during blood circulation together with high transfection efficiency. To provide these desired properties within one system, we propose the use of reactive triblock copolypept(o)ides, which include a stealth-like block for efficient shielding, a hydrophobic block based on reactive disulfides for cross-linking and a cationic block for complexation of pDNA. After the complexation step, bifunctional cross-linkers can be employed to bio-reversibly stabiliz…

Glial Promoter Selectivity following AAV-Delivery to the Immature Brain

2013

Recombinant adeno-associated virus (AAV) vectors are versatile tools for gene transfer to the central nervous system (CNS) and proof-of-concept studies in adult rodents have shown that the use of cell type-specific promoters is sufficient to target AAV-mediated transgene expression to glia. However, neurological disorders caused by glial pathology usually have an early onset. Therefore, modelling and treatment of these conditions require expanding the concept of targeted glial transgene expression by promoter selectivity for gene delivery to the immature CNS. Here, we have investigated the AAV-mediated green fluorescent protein (GFP) expression driven by the myelin basic protein (MBP) or gl…

Cell Culture Systems for Studying Biomaterial Interactions with Biological Barriers

2011

The human body has numerous physical barriers that prevent most harmful or foreign compounds from entering the body. These barriers are formed by unique cell types, which through their location-specific biological cell characteristics prevent compounds from passing between or through them or selectively allow only specific compounds to move across the barrier that they form. Multiple cell types are involved that together form the functioning barrier in a particular organ or tissue. In many cases, in vitro human multicellular culture systems have been developed. These in vitro cell culture models have been extremely valuable in determining the toxic effects of novel compounds on cellular fun…

PROPERTIES AND APPLICATIONS OF POLYAMINOCYCLODEXTRIN DERIVATIVES

Nel presente lavoro sono state prese in esame le potenziali applicazioni delle poliamminociclodestrine. In particolare, diversi derivati poliamminociclodestrinici (amCD) sono stati ottenuti utilizzando un approccio sintetico diretto, tramite reazione di sostituzione nucleofila tra la heptakis-(6-deossi)-(6-bromo)-βCD (BrβCD) e poliammine lineari. Una volta effettuata l’opportuna caratterizzazione NMR, ESI-MS e potenziometrica, sono state investigate diverse possibili utilizzazioni di tali materiali. Anzitutto, sono state valutate le capacità di dare complessi di inclusione host-guest con diversi derivati organici sia delle amCD libere che di materiali costituiti da nanospugne a base di amCD…

Sonoporation, a redefined ultrasound modality as therapeutic aid: a review.

2011

Traditionally a diagnostic modality, ultrasound is emerging as a promising tool for non-invasive therapy, drug delivery, and gene therapy. The ultrasound is a mechanical wave energy generated in a medium as oscillating pressure in space and time at frequencies above 20 kHz, beyond the audible range. The ultrasound exposure generates bioeffects resulting in tissue heating, shear stress, and cavitation, which have been exploited for therapeutic applications. Ultrasound cavitation, enhanced by injected micro bubbles, perturbs cell membrane structures to cause sonoporation and increases the permeability to bioactive materials. Ultrasound-mediated gene delivery has been applied to heart, blood v…

Pig liver gene therapy by noninvasive interventionist catheterism

2006

The efficacy of noninvasive interventionist catheterism in large animals as an alternative to the hydrodynamic procedure, described for small animals, is evaluated. Basically, gene transfer is performed by implantation and fixation of a balloon catheter within the suprahepatic vein of anesthetized pigs, through the femoral vein. The catheter tip is identified by fluoroscopy, injecting a contrast solution that marks large or small hepatic territories. Animals were injected with a 100 ml pTG7101 plasmid solution (40 microg/ml), which contains the human alpha-1 antitrypsin gene, perfused at a rate of 7.5 ml/s and efficacy and toxicity of the procedure were evaluated. The results show: (i) the …

Enhanced baculovirus-mediated transduction of human cancer cells by tumor-homing peptides.

2006

ABSTRACT Tumor cells and vasculature offer specific targets for the selective delivery of therapeutic genes. To achieve tumor-specific gene transfer, baculovirus tropism was manipulated by viral envelope modification using baculovirus display technology. LyP-1, F3, and CGKRK tumor-homing peptides, originally identified by in vivo screening of phage display libraries, were fused to the transmembrane anchor of vesicular stomatitis virus G protein and displayed on the baculoviral surface. The fusion proteins were successfully incorporated into budded virions, which showed two- to fivefold-improved binding to human breast carcinoma (MDA-MB-435) and hepatocarcinoma (HepG2) cells. The LyP-1 pepti…