Assessment of Antiangiogenic and Cytotoxic Effects of Moringa oleifera Silver Nanoparticles Using Cell Lines

Rolla Al-Shalabi^1,2, Nozlena Abdul Samad^1*, Ibrahim Al-Deeb², Julia Joseph¹ and Bassam M. Abualsoud³

¹Department of Toxicology, Sains@BERTAM, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Kepala Batas, Penang, Malaysia.

²Pharmacological and Diagnostic Research Center, Faculty of Pharmacy, Al-Ahliyya Amman University, Al-Salt.

³Department of Pharmaceutics and Pharmaceutical Technology, College of Pharmacy, Al-Ahliyya Amman University, Amman, Jordan.

Corresponding Author E-mail: nozlena@usm.my

Angiogenesis is the physiological process through which new blood vessels are developed from pre-existing vessels. It includes steps like migration, differentiation, and growth of endothelial cells. Furthermore, it plays a critical role in cancer formation and metastasis. Recently, Moringa oleifera (MO) has gained interest due to its properties in various fields, particularly in nanoparticle (NP) technology. This study aims to demonstrate the cytotoxic and antiangiogenic effects of MO silver nanoparticles (MO-AgNPs). Briefly, the cytotoxic effects of MO-AgNPs on Ea. hy 926 and HT 29 cell lines were observed in the Colony Formation Assay. The antiproliferative effects of MO-AgNPs were then further confirmed using MTT assay, in addition to morphological characterization, migration and closure percent calculation. To evaluate the antiangiogenic effect of MO, the chorioallantoic membrane (CAM) assay was done. Fertilized chicken eggs were divided into three groups: MO-AgNPs, MO ethanol extract and negative control. The results showed antiangiogenic effects were observed at 6-12 µg/mL concentration of MO-AgNPs. Whereas the plate efficiency was 45.7 % ± 1%. The surviving fraction at 12, 6, 3, and 1.5 μg/mL was 27.3± 0.5%, 30.1 ± 0.5 %, 41.8 ± 1.5%, and 69.8 ± 2.5 %, respectively. MO-AgNPs showed higher cytotoxic activity than MO ethanol extract on the Ea. hy926 cell line, where the IC50 of MO-AgNPs were 74, 35, and 12 µg/mL for 24, 48 and 72 hours, respectively. On the other hand, the IC50 of MO ethanol extract was above 200, 135 and 11 µg/mL for 24, 48 and 72 hours, respectively. Additionally, significant morphological changes were observed in Ea. hy cells. Whereas the closure percent average of Ea. hy cells at 1.5, 3, 6, and 12 µg/mL were 91.603, 88.507, 84.599, and 63.144%, respectively. Based on the results showed in this study, MO extract shows potential to be an effective anti-angiogenic agent and is a promising therapeutic option in cancer treatment.

Antiangiogenic effects; Cytotoxic effects; Cell line; Moringa oleifera; Silver Nanoparticles

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