Development of a simple, biocompatible and cost-effective Inulin-Diethylenetriamine based siRNA delivery system

Small interfering RNAs (siRNAs) have the potential to be of therapeutic value for many human diseases. So far, however, a serious obstacle to their therapeutic use is represented by the absence of appropriate delivery systems able to protect them from degradation and to allow an efficient cellular uptake. In this work we developed a siRNA delivery system based on inulin (Inu), an abundant and natural polysaccharide. Inu was functionalized via the conjugation with diethylenetriamine (DETA) residues to form the complex Inu-DETA. We studied the size, surface charge and the shape of the Inu-DETA/siRNA complexes; additionally, the cytotoxicity, the silencing efficacy and the cell uptake-mechanis…

Current strategies to improve the efficacy and the delivery of nucleic acid based drugs

Guar gum/borax hydrogel: Rheological, low field NMR and release characterizations

Guar gum (GG) and Guar gum/borax (GGb) hydrogels are studied by means of rheology, Low Field Nuclear Magnetic Resonance (LF NMR) and model drug release tests. These three approaches are used to estimate the mesh size (ζ) of the polymeric network. A comparison with similar Scleroglucan systems is carried out. In the case of GGb, the rheological and Low Field NMR estimations of ζ lead to comparable results, while the drug release approach seems to underestimate ζ. Such discrepancy is attributed to the viscous effect of some polymeric chains that, although bound to the network to one end, can freely fluctuate among meshes. The viscous drag exerted by these chains slows down drug…

Galactosylated polyaspartamide copolymers for siRNA targeted delivery to hepatocellular carcinoma cells

The limited efficacy of available treatments for hepatocellular carcinoma (HCC) requires the development of novel therapeutic approaches. We synthesized a novel cationic polymer based on α,β-poly-(N-2-hydroxyethyl)-D,L-aspartamide (PHEA) for drug delivery to HCC cells. The copolymer was synthesized by subsequent derivatization of PHEA with diethylene triamine (DETA) and with a polyethylene glycol (PEG) derivative bearing galactose (GAL) molecules, obtaining the cationic derivative PHEA-DETA-PEG-GAL. PHEA-DETA-PEG-GAL has suitable chemical-physical characteristics for a potential systemic use and can effectively deliver a siRNA (siE2F1) targeted against the transcription factor E2F1, a gen…

Novel Lipid and Polymeric Materials as Delivery Systems for Nucleic Acid Based Drugs

Nucleic acid based drugs (NADBs) are short DNA/RNA molecules that include among others, antisense oligonucleotides, aptamers, small interfering RNAs and micro-interfering RNAs. Despite the different mechanisms of actions, NABDs have the ability to combat the effects of pathological gene expression in many experimental systems. Thus, nowadays, NABDs are considered to have a great therapeutic potential, possibly superior to that of available drugs. Unfortunately, however, the lack of effective delivery systems limits the practical use of NABDs. Due to their hydrophilic nature, NABDs cannot efficiently cross cellular membrane; in addition, they are subjected to fast degradation by cellular and…

Characterization of PLLA scaffolds for biomedical applications

The porosity and pore size distribution of three-dimensional scaffolds have direct implications on their biomedical applications (tissue engineering, drug delivery, and wound dressing). Accordingly, in this paper, a fast, facile, and conservative method relying on low-field nuclear magnetic resonance (LF-NMR) for the evaluation of mean pore size and pore size distribution of polymeric scaffolds is reported. The applicability of the technique is demonstrated on poly-L-lactic acid scaffolds fabricated using the thermal induced phase separation. Results obtained through LF-NMR are successfully compared to scanning electron microscope and X-ray microcomputed tomography micrographs.

Recent advances in smart biotechnology: Hydrogels and nanocarriers for tailored bioactive molecules depot

Over the past ten years, the global biopharmaceutical market has remarkably grown, with ten over the top twenty worldwide high performance medical treatment sales being biologics. Thus, biotech R&D (research and development) sector is becoming a key leading branch, with expanding revenues. Biotechnology offers considerable advantages compared to traditional therapeutic approaches, such as reducing side effects, specific treatments, higher patient compliance and therefore more effective treatments leading to lower healthcare costs. Within this sector, smart nanotechnology and colloidal self-assembling systems represent pivotal tools able to modulate the delivery of therapeutics. A comprehens…

Modulating carbohydrate-based hydrogels as viscoelastic lubricant substitute for articular cartilages

Viscosupplementation is a therapeutic approach for osteoarthritis treatment, where the synovial fluid, the natural lubricant of the joints, is replaced by viscoelastic solutions with rheological properties comparable or better than the starting material. This study presents the development of an innovative platform for viscosupplementation, based on the optimization of polysaccharide-based colloidal hydrogel, aiming to reduce on-site enzyme degradation and enhance the possibility of hyaluronic acid substitution with alternative biomaterials. Catanionic vesicles are proposed as physical crosslinker that can guarantee the formation of a 'soft', tunable network, offering a dual-therapeutic app…

Physical characterization of alginate-Pluronic F127 gel for endoluminal NABDs delivery

Here we focus the attention on the physical characteristics of a highly biocompatible hydrogel made up of crosslinked alginate and Pluronic F127 (PF127). This is a composite polymeric blend we propose for artery endoluminal delivery of an emerging class of molecules named nucleic acid based drugs (NABDs). The physical characterization of our composite gel, i.e. mesh size distribution and PF127-alginate mutual organization after crosslinking, can significantly determine the NABDs release kinetics. Thus, to explore these aspects, different technical approaches, i.e. rheology, low/high field NMR and TEM, were used. While rheology provided information at the macroscopic and nano-level, the othe…

Synthesis and characterization of polyaspartamide copolymers obtained by ATRP for nucleic acid delivery

Abstract Nucleic acid molecules such as small interfering RNAs (siRNAs) and plasmidic DNAs (pDNAs) have been shown to have the potential to be of therapeutic value in different human diseases. Their practical use is however compromised by the lack of appropriate release systems. Delivered as naked molecules, siRNAs/pDNAs are rapidly degraded by extracellular nucleases thus considerably reducing the amount of molecule which can reach the target cells. Additionally, the anionic charge of the phosphate groups present on the siRNAs/pDNAs backbone, disfavors the interaction with the negatively charged surface of the cell membrane. In this paper we describe the generation of a novel polymer able …

Effect of chest physiotherapy on cystic fibrosis sputum nanostructure: an experimental and theoretical approach.

AbstractCystic fibrosis (CF) is a disease characterized by the production of viscous mucoid secretions in multiple organs, particularly the airways. The pathological increase of proteins, mucin and biological polymers determines their arrangement into a three-dimensional polymeric network, affecting the whole mucus and impairing the muco-ciliary clearance which promotes inflammation and bacterial infection. Thus, to improve the efficacy of the drugs usually applied in CF therapy (e.g., mucolytics, anti-inflammatory and antibiotics), an in-depth understanding of the mucus nanostructure is of utmost importance. Drug diffusivity inside a gel-like system depends on the ratio between the diffusi…

Targeted delivery of siRNAs against hepatocellular carcinoma-related genes by a galactosylated polyaspartamide copolymer

Given the lack of effective treatments for Hepatocellular carcinoma (HCC), the development of novel therapeutic approaches is very urgent. Here, siRNAs were delivered to HCC cells by a synthetic polymer containing α,β-poly-(N-2-hydroxyethyl)-D,L-aspartamide-(PHEA) derivatized with diethylene triamine (DETA) and bearing in the side chain galactose (GAL) linked via a polyethylene glycol (PEG) to obtain (PHEA-DETA-PEG-GAL, PDPG). The GAL residue allows the targeting to the asialo-glycoprotein receptor (ASGPR), overexpressed in HCC cells compared to normal hepatocytes. Uptake studies performed using a model siRNA or a siRNA targeted against the enhanced green fluorescence protein, demonstrated …

Characterization of PLLA scaffolds for biomedical applications

The porosity and pore size distribution of three-dimensional scaffolds have direct implications on their biomedical applications (tissue engineering, drug delivery, and wound dressing). Accordingly, in this paper, a fast, facile, and conservative method relying on low-field nuclear magnetic resonance (LF-NMR) for the evaluation of mean pore size and pore size distribution of polymeric scaffolds is reported. The applicability of the technique is demonstrated on poly-L-lactic acid scaffolds fabricated using the thermal induced phase separation. Results obtained through LF-NMR are successfully compared to scanning electron microscope and X-ray microcomputed tomography micrographs.