فهرست نشریات علمی دسترسی آزاد ایران

In vitro maturation (IVM) efficiency in mice is not yet optimal (62.4 3.5 percent metaphase II rate) because of the presence of oxidative stress during culture. This is original experimental research which produced melatonin-impregnated chitosan nanoparticles (Mel-CS NPs), through the utilization of locally available Iraqi materials, and tested their effectiveness in promoting oocyte developmental competence in super ovulated BALB/c mice. Super ovulation of female BALB/c mice (n = 40, 810 weeks, 25 30g): (1) pregnant mare serum gonadotropin (PMSG, 5 IU, i.p.), which stimulates ovulation, was administered 48 hours before (2) human chorionic gonadotropin (hCG, 5 IU, i.p.), which induces fertilization. Cumulus-oocyte complexes (COCs, n = 400) were obtained 14 hours after the administration of hCG and randomly assigned to four experimental groups (n = 100 COCs/group): (1) Control (MEM- 0.4% BSA), (2) Free melatonin (10 -9 M), (3) Empty chitosan nanoparticles (50 0g/mL), and (4) Mel-CS NPs (equivalent melatonin concentration). Chitosan obtained through extracting the shell of Shrimp (Penaeus semisulcatus) was use in the ionic gelation process of nanoparticles synthesis at Shatt Al-Arab estuary. The oocytes were then matured, 16 hours at 37.0 C with 6% CO 2. Primary outcomes: metaphase II (MII) maturation rate (Hoechst 33342 staining), intracellular reactive oxygen species (ROS) concentration (DCFH-DA fluorescence) and mitochondrial membrane potential (JC-1 assay), and blastocyst formation following in vitro fertilization. Mel-CS NPs had the following morphological characteristics: spherical, hydrodynamic diameter of 83.7 -2.9 nm, polydispersity index of 0.16 -2.02, and zeta potential of +33.4 -1.6 mV. The efficiency of encapsulation was 88.2% with a standard deviation of 1.9. Mel-CS NPs enhanced maturation rate of MII by far 86.3% +/- 3.8% in comparison to control (62.4% +/- 3.5), free melatonin (71.8% +/- 3.9) and empty nanoparticles (64.1% +/- 3.6). There was a decrease in the levels of the ROS 44.7% (178.6 + 11.9 vs. 98.8 + 7.6 arbitrary units, p < 0.001), 51.2% increase of mitochondrial membrane potential (JC-1 ratio: 1.79 + 0.13 vs. 2.71 + 0.16, p < 0.001), 28.7% + 2.9% versus 45.3% The lowest concentration that had no cytotoxicity was 100 μg/mL and above (granulosa cell viability was above 96%). The overall price per IVM cycle was 1, 650 IQD (1.25) and 14, 520 IQD (11.00) on imported substitutes (88.6 cost-savings). Mel-CS NPs synthesized locally were found to be highly effective in promoting oocyte nuclear and cytoplasmic maturation in the BALB/c mice by having a strong antioxidant property and mitochondrial protection. This is a low-cost, reproducible protocol that uses locally available material that will serve as a basis of the application of nanotechnology in Iraqi reproductive biotechnology as well as a scaleable example of resource-constrained environments.