Influence of adding zeolite loaded with different charges to semen extender on sperm quality in rabbits after cryopreservation.

The aim of the present study was to investigate the effect of supplementing rabbit semen extender with zeolite loaded with different charges (Z+ or Z-, Z±) on sperm cryopreservation. Semen was collected from six healthy, fertile New Zealand rabbit bucks using an artificial vagina. The collected ejaculates were pooled and diluted with a tris-yolk fructose (TYF) extender supplemented with Z± (+16, +12, +8, -16, -12, and -8) at a concentration of 1% for a final sperm concentration of 25 × 106 sperm cells/mL. The diluted semen samples were then cryopreserved in 0.25 mL straws and stored in liquid nitrogen for 1 month. To evaluate sperm quality, we examined sperm progressive motility, vitality, morphological abnormalities, and plasma membrane integrity. In addition, apoptotic rates were determined using flow cytometry and by examining sperm ultrastructure under a transmission electron microscope (TEM). Moreover, total antioxidant capacity and markers of lipid peroxidation were measured in the extender after thawing. Addition of Z± had a positive effect on progressive motility, vitality, and membrane integrity after an equilibration period and post-thawing as compared with the controls (P < 0.05). Z± supplementation, particularly with a strong negative charge, also decreased the percentages of apoptotic and necrotic sperm cells compared to controls (P < 0.05), as shown both by flow cytometry and TEM. This was not associated with any marked effects on the oxidative biomarkers in the extender. In conclusion, addition of Z± to semen extender improved post-thawing sperm quality by improving sperm characteristics, decreasing apoptosis, and minimizing sperm damage during cryopreservation.

Effects of silver nanoparticles-polysaccharide on bleomycin-induced pulmonary fibrosis in rats.

OBJECTIVES: The first goal of this study was to synthesize the silver nanoparticles Alcaligenes xylosoxidans exopolysaccharide (Ag-AXEPS). The second objective was to analyse the role of Ag-AXEPS nanoparticles (NPS) in treating bleomycin (BLM)-induced lung fibrosis. METHODS: Intratracheal bleomycin (2.5 U/kg) was administered to prompt pulmonary fibrosis in rats, and pulmonary fibrosis was treated with Ag-AXEPS nanoparticles (100 ppm/twice a week for four weeks). KEY FINDINGS: Ag-AXEPS nanoparticles significantly decreased the diversity of pulmonary inflammatory agents in rats with BLM-induced fibrosis. Reduced levels of respiratory tumor necrosis factor-alpha, monocyte chemotactic protein-1, matrix metalloproteinases (MMP-2 and MMP-9) were observed on treatment with synthesized Ag-AXEPS. Similarly, the treatment decreased IL-12, mRNA levels of BAX and plasma fibrosis markers like N-terminal procollagen III propeptide and transforming growth factor-ß1. On the other hand, the treatment increased mRNA BCL2 and total antioxidant capacity. It also lowered the level of fibrosis, as was shown by a quantified pathologic study of hematoxylin-eosin-stained lung parts. The treatment, however, ensured that lung collagen was restored, as assessed by Masson's trichrome stain, and that overall survival was increased and enhanced. CONCLUSIONS: Our work showed that nanoparticles could be obtained at 37°C and may be a possible pulmonary fibrosis therapeutic agent.

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.