JNK2 silencing and caspase-9 activation by hyperosmotic polymer inhibits tumor progression.

c-Jun N-terminal kinase 2 (JNK2) is primarily responsible for the oncogenic transformation of the transcription factor c-Jun. Expression of the proto-oncogene c-Jun progresses the cell cycle from G1 to S phase, but when its expression becomes awry it leads to uncontrolled proliferation and angiogenesis. Delivering a JNK2 siRNA (siJNK2) in tumor tissue was anticipated to reverse the condition with subsequent onset of apoptosis which predominantly requires an efficient delivering system capable of penetrating through the compact tumor mass. In the present study, it was demonstrated that polymannitol-based vector (PMGT) with inherent hyperosmotic properties was able to penetrate through and deliver the siJNK2 in the subcutaneous tumor of xenograft mice. Hyperosmotic activity of polymannitol was shown to account for the enhanced therapeutic delivery both in vitro and in vivo because of the induction of cyclooxygenase-2 (COX-2) which stimulates caveolin-1 for caveolae-mediated endocytosis of the polyplexes. Further suppression of JNK2 and hence c-Jun expression led to the activation of caspase-9 to induce apoptosis and inhibition of tumor growth in xenograft mice model. The study exemplifies PMGT as an efficient vector for delivering therapeutic molecules in compact tumor tissue and suppression of JNK2 introduces a strategy to inhibit tumor progression.

Hierarchically Micro- and Nanopatterned Topographical Cues for Modulation of Cellular Structure and Function.

Living cells receive biochemical and physical information from the surrounding microenvironment and respond to this information. Multiscale hierarchical substrates with micro- and nanogrooves have been shown to mimic the native extracellular matrix (ECM) better than conventional nanopatterned substrates; therefore, substrates with hierarchical topographical cues are considered suitable for investigating the role of physical factors in tissue functions. In this study, precisely controllable, multiscale hierarchical substrates that could mimic the micro- and nanotopography of complex ECMs were fabricated and used to culture various cell types, including fibroblasts, endothelial cells, osteoblasts, and human mesenchymal stem cells. These substrates had both microscale wrinkles and nanoscale patterns and enhanced the alignment and elongation of all the cells tested. In particular, the nanotopography on the microscale wrinkles promoted not only the adhesion, but also the functions of the cells. These findings suggest that the hierarchical multiscale substrates effectively regulated cellular structure and functions and that they can be used as a platform for tissue engineering and regenerative medicine.

Enhanced chitosan-DNA interaction by 2-acrylamido-2-methylpropane coupling for an efficient transfection in cancer cells.

Gene therapy is the treatment of human disorders by the introduction of genetic material to specific target cells of a patient. Chitosan and its derivatives show excellent biological properties including biocompatibility, biodegradability and nonallergenicity. Primary amines of chitosan are responsible for its cationic nature and hence binding and protection of DNA for intracellular delivery. But the transfection efficiency of chitosan based gene transporters is severely hampered by its poor physical properties such as low water solubility and high viscosity. In this study, primary amines of low molecular weight (LMW) chitosan were coupled with 2-acrylamido-2-methylpropane sulphonic acid (AMP) making it water soluble for its application in gene delivery. AMP modified chitosan (CSAMP) showed an enhanced interaction with DNA and a higher buffering capacity due to AMP amines leading to a higher transfection efficiency in cancer cells (A549, HeLa and HepG2) compared to native chitosan and Lipofectamine®. In vivo studies in Balb/c through intravenous injection demonstrated a higher luciferase expression compared to LMW chitosan.