Impact of lipid substitution on assembly and delivery of siRNA by cationic polymers

Characterization of a polymer library engineered to enhance their ability to protect and deliver their nucleotide cargo to the cells is reported. The ζ-potential continuously increased with higher polymer:siRNA weight ratio, and the ζ-potential of lipid-modified polymers:siRNA complexes were higher than PEI2 at all ratios. At polymer:siRNA ratio of 1:1, all lipid-substituted polymers showed complete protection against degradation. Lipid-modified polymers significantly increased the cellular uptake of siRNA complexes and down-regulation of GAPDH and P-gp (max. 66% and 67%, respectively). The results indicate that hydrophobic modification of low molecular PEI could render this otherwise ineffective polymer to a safe effective delivery system for intracellular siRNA delivery and protein silencing.     

In vitro biocompatibility of electrospun hexanoyl chitosan fibrous scaffolds towards human keratinocytes and fibroblasts

With the ability to form a submicron-sized fibrous structure with interconnected pores mimicking the extracellular matrix (ECM) for tissue formation, electrospinning was used to fabricate ultra-fine fiber mats of hexanoyl chitosan (H-chitosan) for potential use as skin tissue scaffolds. In the present communication, the in vitro biocompatibility of the electrospun fiber mats was evaluated. Indirect cytotoxicity evaluation of the fiber mats with mouse fibroblasts (L929) revealed that the materials were non-toxic and did not release substances harmful to living cells. The potential for use of the fiber mats as skin tissue scaffolds was further assessed in terms of the attachment and the proliferation of human keratinocytes (HaCaT) and human foreskin fibroblasts (HFF) that were seeded or cultured on the scaffolds at different times. The results showed that the electrospun fibrous scaffolds could support the attachment and the proliferation of both types of cells, especially for HaCaT. In addition, the cells cultured on the fibrous scaffolds exhibited normal cell shapes and integrated well with surrounding fibers. The obtained results confirmed the potential for use of the electrospun H-chitosan fiber mats as scaffolds for skin tissue engineering.     

In vitro biocompatibility of electrospun and solvent-cast chitosan substrata towards Schwann, osteoblast, keratinocyte and fibroblast cells

Chitosan or poly(N-acetyl-d-glucosamine-co-d-glucosamine) with a degree of deacetylation of about 85% was fabricated into nanofibrous membranes by electrospinning from 7% w/v chitosan solution in 70:30 v/v trifluoroacetic acid/dichloromethane solvent system. The obtained fibers were smooth without the presence of beads. The diameters of the individual fiber segments were 126 ± 20 nm. The potential for use of the electrospun chitosan nanofibrous membranes as substrates for cell/tissue culture was evaluated with four different cell types, i.e., Schwann cells, osteoblast-like cells, keratinocytes and fibroblasts, in terms of the attachment and the proliferation of the cells as well as the morphology of the seeded and the cultured cells. The results were compared with the corresponding solvent-cast films. Both types of the chitosan substrates supported the attachment and, at the same time, promoted the largest increase in the viability of the cultured keratinocytes. The viability of Schwann cells cultured on these substrates increased marginally well, but the attachment of the cells on the surfaces was relatively poor. Finally, both types of the chitosan substrates showed cytostatic property towards both osteoblast-like cells and fibroblasts, despite the convincingly good attachment of osteoblast-like cells on the surfaces.