Ali et al (2021): Production and characterization of nanoparticles lipid carrier (NLCs) © Annals of Tropical Medicine & Public Health DOI: http://doi.org/10.36295/ASRO.2021.24468 580-594| Page Production and characterization of nanoparticles lipid carrier (NLCs) loaded with red clover isoflavones extract for menopause therapy Athraa H. Ali 1 ; Muna T. Al-Musawi 2 ; Zainab J. Shanan 3 1Department of Biotechnology, College of Science, University of Baghdad, Baghdad, Iraq 2 Department of Biology, College of Science for Women, University of Baghdad, Baghdad, Iraq 3 Department of Physics, College of Science for Women, University of Baghdad, Baghdad, Iraq Corresponding author: Athraa H. Ali athraahussain2002@gmail.com Abstract Plant-derived edible nanoparticles (PDNPs) are nano-sized membrane vesicles released by edible plants. They are non-toxic, have tissue-specific targeting properties, and can be mass-produced.Our aim is to assess the effect of phytoestrogens (PEs) for menopause therapy using nanoparticles lipid carrier (NLCs) loaded with red clover isoflavones extract (RCIE).Study was conducted using fifty adult female mice model for menopause using 4-Vinylcyclohexene dioxide(VCD) and handled as follows for 6 weeks. Twoexperiments were preformed, the first experiment included 20 adult female mice it lasted two weeks and divided into two groups (10 mice/ group)Group A: Control group has injected 0.1 ml D.W intraperitoneal (IP) daily. Group B: This group has injected IP daily 160 mg/kg B.W of VCD.The second experiment was included 30 adult female mice have injected IP daily 160 mg/kg of VCD for inducing menopause for two weeks, then shaved at the last third of the back and divided into three groups (C, D and E), 10 mice in each one, then treated with dermal sticker for six weeks: Group C: The group was treated with dermal sticker saturated with10ug/ kg B.W of estradiol benzoate (EB) diluted with virgin coconut oil (VCNO).Group D: Control group was treated with dermal sticker saturated with 0.1 ofVCNO. Group E: The group was treated with dermal sticker saturated with 0.1ml of RCIE-NLCs. A red clover extract isoflavones standardized by HPLC to contain (2.7, 6.6 and 1.04) mg/g of genistein, glyciteinand malonate respectively. Thecharacterization of RCIE-NLCs was done by transmission electron microscopy (TEM), Scanning probe microscopy (SPM), Zeta potential. TEM images showed that shape of particles wasmostly spherical and few cylindrical with average diameter (53.70) nm. SPM images showed the average grain size was 38.78 nm. Encapsulation Efficiency (EE) was 83.3%. Zetapotential was -41.1 Mv.The results of statistical analysis showed a significant decrease (P≤0.01) in the level of estrogen in group B (23.767) ng/ml, compared with group A( 33.154) ng/ml, no significant decrease(P≤0.01) compared with group D( 22.728) ng/ml,and significant increase (P≤0.01) in the levels of FSH and LH in group B (4.0688 and 3.0839) mIU/ml compared with groups A (2.9206 and 2.1469) mIU/ml respectively, and no significant change(P≤0.01) compared with group D (3.9974 and 3.1049) mIU/ml respectively, and significantincrease(P≤0.01) in the level of estrogen in groupsC, E(36.444 and 30.008) ng/ ml respectively compared with groupsB and D (23.767 and 22.728 ) ng/ ml respectively, and significant decrease (P≤0.01) in the levels of FSH and LH in group C (2.5556 and 0.0485)mIU/ml and groupE ( 3.2116 and 2.4825)mIU/ml respectively, compared with group B(4.0688 and 3.0839)mIU/ml and group D( 3.9974 and 3.1049)mIU/ml respectively.The results of this study revealed that administration RCIE-NLCs shows an effective in compensating for estrogen deficiency resulting from loss of primordial and primary follicles in the ovary via impact of VCD, compared to EB.This study strongly suggests that RCIE-NLCs have potential as alternative to HRT for menopause therapy Keywor ds : Red clover, Trifoliumpratense, Menopause, NLCs, isoflavones, Estrogen benzoate,