Assessment of the Effect of Moringa Oleifera Leaves Extract on Angiogenesis using the Novel Yolk Sac Model: An In-ovo Experimental Study

Abstract

Introduction: In recent times, the focus of cancer research has shifted to naturally occurring compounds derived from plants that may have the potential for anticancer activity. Moringa oleifera is a softwood tree whose fruits, roots, and leaves have been advocated for medicinal and industrial uses. Angiogenesis plays an important role in many physiological and pathological conditions and is considered a hallmark of tumour development and progression. An efficient anticancer drug is expected to have a significant inhibitory effect on angiogenesis. However, there are a limited number of studies reported to explore the anti-angiogenic activities of Moringa Oleifera Leaf extract (MOL). Aim: To assess the effect of MOL on angiogenesis using the Yolk Sac Model (YSM). Materials and Methods: A preliminary in-ovo study was conducted in the in the specialised Regenerative laboratory of Dr. D.Y. Patil Dental College and Hospital, Pimpri, Pune, Maharashtra, India, between September 2022 and February 2023 after obtaining necessary scientific and ethical permissions. The calculated sample size for YSM analysis was seven in each study and control group (total of 14); however, the authors used a total of 28 YSMs (seven in each of the three study concentration groups and seven in the control group). In this method, freshly fertilised White Leghorn chicken (Gallus domesticus) eggs, procured from the hatchery, were incubated and treated with different concentrations (10 μg/mL, 100 μg/mL, and 500 μg/mL) of the treatment substance (MOL) along with a control group (Avastin). The anti-angiogenic effect of MOL extract was determined by calculating vessel density, total vessel network length, total branching points, total segments, mean segment length, and width in the three groups compared to Avastin after 48 hours of treatment using WimCam software for analysing the images. Descriptive statistical analysis and one-way Analysis of Variance (ANOVA) were then applied to compare the parameters in the four groups. Results: Statistical analysis by one-way ANOVA showed a significant (p<0.05) percentage reduction in the number of blood vessels in each treatment group after 48 hours of treatment. Among the different concentrations of MOL used, 500 μg/mL showed greater anti-angiogenic capacity. Conclusion: The results indicate that MOL could suppress abnormal angiogenesis in a dose-dependent manner and may be considered a potential anti-angiogenic agent in various diseases, including cancer.

Keywords

• anti-angiogenic
• anticancer
• in-ovo model