Anti-Angiogenic Screening of Moringa Oleifera Leaves Extract Using Chorioallantonic Membrane Assay


  • Nisha Shri Chengama Raju Segi University
  • Wong Wing Kei



Anti-angiogenic screening, Moringa oleifera leaves, Chick Chorioallantoic Membrane (CAM) assay.


Background: Angiogenesis is defined as the formation of new blood vessels. However, angiogenesis in cancer will lead to tumour growth and metastasis. Therefore, anti-angiogenesis is one of the ways to slow down growth and spreading of tumour. Moringa oleifera is also known as a “Miracle tree” which has high nutritive value and various therapeutics effect in different parts of the plant. This study aims to determine the anti-angiogenic property of Moringa oleifera leaves extract by using chick chorioallantoic membrane (CAM) assay. Materials and Methods: The extracts were prepared by decoction method using methanol and water. The qualitative phytochemical screening was carried out for both methanol and aqueous extracts. The fertilised chicken eggs were divided into six groups which include negative control group (phosphate-buffer saline with pH 7.4), positive control group (sunitinib), 50% and 100% methanol extract, 50% and 100% aqueous extract. The anti-angiogenic effect of Moringa oleifera leaves extract was determined by calculating the number and percentage decrease in blood vessels in post-24 and post-48 hours of treatment. Results: Statistical analysis by one-way ANOVA has shown significant (p<0.05) percentage reduction in the blood vessels between each treatment group after 48 hours of treatment. Among all the extracts, 100% aqueous extract of Moringa oleifera was found to have highest anti-angiogenic effect with the greater percentage decrease in blood vessels (81.33%) in post-48 hours of treatment. Furthermore, the anti-angiogenic effect of Moringa oleifera leaves was found to increased when the concentration of the Moringa oleifera extract was increased. Conclusion: Moringa oleifera leaves with various phytochemicals was found to possess anti-angiogenic potential.


Yoo SY, Kwon SM. Angiogenesis and its therapeutic opportunities. Mediators Inflamm. 2013 Jul;2013: doi: 10.1155/2013/127170.

Ucuzian AA, Gassman AA, East AT, Greisler HP. Molecular mediators of angiogenesis. J Burn Care Res. 2010 Jan;31(1):158–75.

Gupta K, Zhang J. Angiogenesis: A curse or cure?. Postgrad Med J. 2005 Apr;81(954):236–42.

Harris AL, Generali D. Inhibitors of Tumor Angiogenesis. Cancer Drug Design and Discovery, 2nd Edition. Academic press. 2013;275–317.

Makrilia N, Lappa T, Xyla V, Nikolaidis I, Syrigos K. The role of angiogenesis in solid tumours: an overview. Eur J Intern Med. 2009 Nov;20(7):663–71.

Cook KM, Figg WD. Angiogenesis Inhibitors: Current Strategies and Future Prospects. CA Cancer J Clin. 2010 Jul;60(4):222–43.

Hoseinkhani Z, Norooznezhad F, Rastegari-Pouyani M, Mansouri K. Medicinal plants extracts with antiangiogenic activity: Where is the link?. Adv Pharm Bull. 2020 Jul;10(3):370–8.

Sutar NG, Bonde CG, Patil VV, Narkhede SB, Patil AP, Kakade RT. Analgesic activity of seeds of Moringa oleifera Lam. Int J Green Pharm. 2008;2(2):doi.10.4103/0973-8258.41182.

Kumbhare M, Sivakumar T. Anti-inflammatory and analgesic activity of stem bark of Moringa oleifera. Pharmacology online. 2011;3:641–50.

Dorairaj S. Antimicrobial Activity of Moringa oleifera (Lam.) Root Extract. J Pharm Res. 2011;4(5):1426–7.

Edoga CO, Njoku OO, Amadi EN, Okeke JJ. Blood Sugar Lowering Effect of Moringa Oleifera Lam in Albino Rats. Int J Sci Technology. 2013 Jan;3(1):88-90.

Sulaiman MR, Zakaria ZA, Bujarimin AS, Somchit MN, Israf DA, Moin S. Evaluation of Moringa oleifera aqueous extract for antinociceptive and anti-inflammatory activities in animal models. Pharm Biol. 2008 Jan;46(12):838–45.

Jaja-Chimedza A, Graf BL, Simmler C, Kim Y, Kuhn P, Pauli GF, et al. Biochemical characterization and anti-inflammatory properties of an isothiocyanate-enriched moringa (Moringa oleifera) seed extract. PLoS One. 2017 Aug;12(8):e0182658.

Purwal L, Pathak AK, Jain UK. In vivo anticancer activity of the leaves and fruits of Moringa oleifera on mouse melanoma. Pharmacology online. 2010;1:655–65.

Krishnamurthy PT, Vardarajalu A, Wadhwani A, Patel V. Identification and characterization of a potent anticancer fraction from the leaf extracts of Moringa oleifera L. Indian J Exp Biol. 2015 Feb;53(2):98-103.

Shaikh JR, Patil M. Qualitative tests for preliminary phytochemical screening: An overview. Int J Chem Stud. 2020;8(2):603–8.

Evans WC. Pharmacognosy 16th edition. W.B. Sanders Company Ltd., New York. 2009;42–44. 221–229, 246–249, 304–306, 331–332, 391–393 p.

BAM R59: Phosphate-Buffered Saline (PBS), pH 7.4 | FDA [Internet]. [cited 2021 Jan 19]. Available from:

Nowak-Sliwinska P, Segura T, Iruela-Arispe ML. The chicken chorioallantoic membrane model in biology, medicine and bioengineering. Angiogenesis. 2014 Oct;17(4):779–804.

Archer GS, Cartwright AL. EPS-001-Incubating-and-Hatching-Eggs1. 2017;1–13. Available from:

West DC, Thompson WD, Sells PG, Burbridge MF. Angiogenesis Assays Using Chick Chorioallantoic Membrane. Methods Mol Med. 2001;46:107–29.

Ribatti D. Chicken Chorioallantoic Membrane Angiogenesis Model. Methods Mol Biol. 2012;843:47–57.

Pachava VR, Krishnamurthy T, Dahabal SP, Wadhwani A, Chinthamaneni PK. Anti-Angiogenesis Potential of Ethyl Acetate Extract of Moringa oleifera Lam Leaves in Chick Chorioallantoic Membrane (CAM) Assay. J Nat Prod Plant Resour. 2017;7(4):18–22.

Thorat AA, Suryanarayanan R. Characterization of Phosphate Buffered Saline (PBS) in Frozen State and after Freeze-Drying. Pharm Res. 2019 Jul;36(7):98.

Mena AC, Pulido EG, Guillén-Ponce C. Understanding the molecular-based mechanism of action of the tyrosine kinase inhibitor: Sunitinib. Anticancer Drugs. 2010;21(SUPPL.1):3–11.

Christensen JG. A preclinical review of sunitinib, a multitargeted receptor tyrosine kinase inhibitor with anti-angiogenic and antitumour activities. Ann Oncol. 2007;18:3-10.

Dharani S, Pabba S, Madhusudanrao Y. Evaluation of biological activities using ethanolic crude extract of Moringa oleifera. Octa J Biosci. 2014 Dec;2(2):91–3.

Lin M, Zhang J, Chen X. Bioactive flavonoids in Moringa oleifera and their health-promoting properties. Journal of Functional Foods. 2018 Jun;47:469–79.

Zhang Y, Chen AY, Li M, Chen C, Yao Q. Ginkgo biloba Extract Kaempferol Inhibits Cell Proliferation and Induces Apoptosis in Pancreatic Cancer Cells. J Surg Res. 2008 Jul;148(1):17–23.

Shields M. Chemotherapeutics. In: Pharmacognosy: Fundamentals, Applications and Strategy. Academic Press. 2017 May;p. 295–313.