Effect of Tumor Suppressor MiR-34a Loaded on ZSM-5 Nanozeolite in Hepatocellular Carcinoma: In Vitro and In Vivo Approach.

BACKGROUND: MicroRNA modulation therapy has shown great promise to treat hepatocellular carcinoma (HCC), however Efficient tissue-specific and safe delivery remains a major challenge. OBJECTIVE: We sought to develop an inorganic-organic hybrid vehicle for the systemic delivery of the tumor suppressor miR-34a, and to investigate the efficiency of the delivered miR-34a in the treatment of HCC in vitro and in vivo. METHODS: In the present study, pEGP-miR cloning and expression vector, expressing miR-34a, was electrostatically bound to polyethyleneimine (PEI), and then loaded onto ZSM-5 zeolite nanoparticles (ZNP). Qualitative and quantitative assessment of the transfection efficiency of miR-34a construct in HepG2 cells was applied by GFP screening and qRT-PCR, respectively. The expression of miR-34a target genes was investigated by qRT-PCR in vitro and in vivo. RESULTS: ZNP/PEI/miR-34a nano-formulation could efficiently deliver into HepG2 cells with low cytotoxicity, indicating good biocompatibility of generated nanozeolite. Furthermore, five injected doses of ZNP/PEI/miR-34a nano-formulation in HCC induced male Balb-c mice, significantly inhibited tumor growth, and demonstrated improved cell structure, in addition to a significant decrease in alphafetoprotein level and liver enzymes activities, as compared to the positive control group. Moreover, injected ZNP/PEI/miR-34a nano-formulation led to a noticeable decrease in the CD44 and c-Myc levels. Results also showed that ZNP/PEI/miR-34a nano-formulation inhibited several target oncogenes including AEG-1, and SOX-9, in vitro and in vivo. CONCLUSION: Our results suggested that miR-34a is a powerful candidate in HCC treatment and that AEG-1 and SOX-9 are novel oncotargets of miR-34a in HCC. Results also demonstrated that our nano-formulation may serve as a candidate approach for miR-34a restoration for HCC therapy, and generally for safe gene delivery.

Improvement of physico-chemical properties of dextran-chitosan composite scaffolds by addition of nano-hydroxyapatite.

Nano-hydroxyapatite was incorporated into polymer matrix of Dextran/Chitosan to achieve a novel composite scaffold by freeze drying technique. The synthesized composite scaffolds were recognized by different performances such as: X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR) and Scanning electron microscope (SEM). The results revealed the complex formation between dextran and chitosan with an excellent dispersion of nHA inside the polymer matrix. The SEM images showed the presence of interconnected pore structure inside the scaffolds. The porosity of the composites was found to decrease from 82% to 67% by adding nanohydroxyapatite to the polymer matrix of Dextran/Chitosan. The mechanical properties of the scaffolds were measured by compression test. The obtained results verified that the presence of nHA can noticeably enhance young's modulus and compressive strength of the composite scaffolds. All the obtained results essentially recommend that these composites can be a good candidate for bone tissue engineering applications.

Crosstalk between GSK-3, c-Fos, NFκB and TNF-α signaling pathways play an ambitious role in Chitosan Nanoparticles Cancer Therapy.

Nanotechnology is a promising era of medicine for developing targeted drug delivery system. Chitosan nanoparticles (CNPs) have attracted increasing attention for their wide applications as anticancer drugs. This article is concerned with the therapeutic index of chitosan nanoparticles against diethyl nitrosamine (DEN) induced hepatocellular carcinoma (HCC). HCC was induced in rats via repeated DEN administration in a dose of 200 mg/kg BW IP, 2 weeks later rats received (2 ml/kg BW) CCl4 orally for 2 months followed by daily treatment with chitosan nanoparticles in an oral dose of 12 mg/kg for 1 month. Then the gene expression of glycogen synthase kinase-3 (GSK-3), (c-FOS), nuclear factor kappa-B (NFκB) and tumor necrosis factor- α (TNF-α) were reported in rats sera and the correlation between GSK-3, C-Fos, NFƘB and TNF-α and liver tumorigenesis was investigated. The results elucidated that DEN significantly increased serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Marked increments in serum malondialdehyde (MDA) and nitric oxide (NOx) levels along with a slight reduction of glutathione (GSH) level were evidenced in HCC. Liver injury triggered an inflammatory response by enhancing the mRNA gene expression of NFκB and TNF-α. DEN effectively activated apoptotic markers GSK-3 and c-FOS. Oral administration of CNPs alleviated the oxidative, inflammatory and apoptotic hazards induced via DEN. The histopathological examination reinforced these results. The present study highlights the anti-inflammatory and anti-apoptotic potentials of CNPs against DEN-induced HCC.

Development and bioactivity evaluation of bioglasses with low Na2O content based on the system Na 2O-CaO-MgO-P 2O 5-SiO 2.

Osteoconductive bioglasses, free of K(2)O and Al(2)O(3) and with content of Na(2)O lower than 10 mol%, were designed based on the ratio (SiO(2) + MgO)/(P(2)O(5) + CaO + Na(2)O) in the system Na(2)O-CaO-MgO-P(2)O(5)-SiO(2). The developed glasses have shown a strong potential for the formation of hydroxycarbonated apatite (HCA) in vitro. The particles of HCA aggregates tend to be of finer size with increasing the ratio of (SiO(2) + MgO)/(CaO + P(2)O(5) + Na(2)O) in the glass chemical composition indicating significant bioactivity. Critical size bone defects created in the femurs of albino adult female rats, and grafted with the glass particles for 12 weeks post implantation, were completely healed by filling with mineralized bone matrix without infection showing a strong potential for new bone formation in vivo. Osteoblasts and osteocytes were observed close to the surface of the granular implants with active areas of bone deposition, resorption and remodelling. The bioglass with lowest (SiO(2) + MgO)/(CaO + P(2)O(5) + Na(2)O) ratio has shown the highest bioactivity while the bioglass with the highest (SiO(2) + MgO)/(CaO + P(2)O(5) + Na(2)O) has shown the lowest bioactivity. The newly formed bone in vivo has shown a similar structure to that of the original bone as indicated by the histology and microstructural results. In addition, Ca/P molar ratio of the newly formed bone was found to be (~1.67), which is similar to that of the original bone.