Abstract :
The study implemented a descriptive classic experimental design within a laboratory, utilizing the Chorioallantoic Membrane (CAM) assay to investigate the teratogenic potential of Blue Ternate (Clitoria ternatea) leaf extract using the different extract concentrations, and the phytochemical analysis screening unveiled the presence of flavonoids, saponins, tannins, alkaloids, and steroids in the leaves. The primary objectives encompassed the assessment of teratogenicity at various extract concentrations, comparing results with controls, and exploring the plant’s impact on embryonic development. While no significant differences emerged in primary and secondary blood vessels among treatments, a conspicuous variation in tertiary blood vessels indicated potential teratogenic effects at specific concentrations. Nevertheless, the study concluded that Clitoria ternatea exhibits promising therapeutic attributes. Recommendations include the implementation of public awareness programs elucidating the health benefits of Blue Ternate and further investigations into refining, storage practices, and potential teratogenicity in other plant parts. These insights, of considerable value to medical practitioners and future researchers, underscore the importance of caution and consultation, especially for pregnant individuals. In summary, the research significantly contributes to understanding Blue Ternate’s safety profile, presenting avenues for future exploration. The emphasis on careful consideration in medicinal applications, particularly during pregnancy, reinforces the need for responsible and informed decision-making in healthcare practices.
Keywords :
Blue ternate, Chorioallantoic Membrane, Experimental, Philippines., Phytochemical analysis, SurigaoReferences :
- Addis, G. T., Workneh, B. D., & Kahissay, M. H. (2021). Herbal medicines use and associated factors among pregnant women in Debre Tabor town, north West Ethiopia: a mixed method approach. BMC Complementary Medicine and Therapies, 21(1). https://doi.org/10.1186/s12906-021-03439-3
- Ain, Q., Khan, H., Mubarak, M. S., & Pervaiz, A. (2016). Plant alkaloids as antiplatelet agent: Drugs of the future in the light of recent developments. Frontiers in Pharmacology, 7. https://doi.org/10.3389/fphar.2016.00292
- Álvarez-Vergara, M. I., Rosales-Nieves, A. E., March-Díaz, R., Rodríguez-Periñán, G., Lara-Ureña, N., Luis, C. O. S., Sanchez-Garcia, M. A., Martin-Bornez, M., Gómez-Gálvez, P., Vicente‐Munuera, P., Fernández-Gómez, B., Marchena, M., Bullones-Bolaños, A., Dávila, J. C., González-Martínez, R., Trillo-Contreras, J. L., Sánchez-Hidalgo, A. C., Toro, R., Scholl, F. G., Herrera, E., Trepel, M., Korbelin, J., Escudero, L., Villadiego, J., Echevarria, M., De Castro, F., Gutierrez, A., Robano, A., Vitorica, J., & Pascual, A. (2021). Non-productive angiogenesis disassembles Aß plaque-associated blood vessels. Nature Communications, 12(1). https://doi.org/10.1038/s41467-021-23337-z
- American College of Obstretricians and Gynocologists. Early Pregnancy Loss. (2015). https://www.acog.org/clinical/clinical-guidance/practice-bulletin/articles/2018/11/early-pregnancy-loss
- Arensman, R. M., & Stolar, C. J. (1979). Vitamin A effect on tumor angiogenesis. Journal of Pediatric Surgery, 14(6), 809–813. https://doi.org/10.1016/s0022-3468(79)80271-7
- Ariel, B., Bland, M., & Sutherland, A. (2022). Experimental Designs. SAGE Publications. https://www.perlego.com/book/3277493/experimental-designs-pdf (Original work published 2022)
- Arnon, J., Meirow, D., Lewis-Roness, H., & Ornoy, A. (2001). Genetic and teratogenic effects of cancer treatments on gametes and embryos. Human Reproduction Update, 7(4), 394–403. https://doi.org/10.1093/humupd/7.4.394
- Bacanli, M., Dilsiz, S. A., Başaran, N., & Başaran, A. A. (2019). Effects of phytochemicals against diabetes. Advances in Food and Nutrition Research, 209–238. https://doi.org/10.1016/bs.afnr.2019.02.006
- Ben-Haroush, A., Sirota, I., Salman, L., Son, W., Tulandi, T., Holzer, H., & Oron, G. (2018). The influence of body mass index on pregnancy outcome following single-embryo transfer. Journal of Assisted Reproduction and Genetics, 35(7), 1295–1300. https://doi.org/10.1007/s10815-018-1186-5
- Bilalissi, A., Meteyake, H., Kouame, Y., Oke, O., Lin, H., Onagbesan, O., Decuypere, E., & Tona, K. (2022). Effects of pre-incubation storage duration and nonventilation incubation procedure on embryonic physiology and post-hatch chick performance. Poultry Science, 101(5), 101810. https://doi.org/10.1016/j.psj.2022.101810
- Cancer Research UK. (2021). Drugs that block cancer blood vessel growth (Anti Angiogenics). https://www.cancerresearchuk.org/about-cancer/treatment/targeted-cancer-drugs/types/anti-angiogenics
- Cozma, G.v., Apostu,A., Macasoi, I., Dehelean,C.A., Cretu, O.M., Dihu,S., Gaita,D., Mane, A.(2020). In vitro and In Ovo Evaluation of the Potential Hepatoprotective Effect of Metformin. Medicina 2022, 58(6), 705; https://doi.org/10.3390/medicina58060705
- Doostkam, A., Iravani, K., & Bassiri-Jahromi, S. (2020). Punica granatum L. (Pomegranate): A Potential Anti-microbial Agent. Anti-infective Agents, 18(1), 2–14. https://doi.org/10.2174/2211352517666190215113232
- Ederio, N., Inocian, E., Calaca, N., & Espiritu, J.G. (2023). Ethical research practices in educational institutions: A literature review. International Journal of Current Science Research and Review, 06(5), 2709-2724. DOI: https://doi.org/10.47191/ijcsrr/V6-i5-02
- Ezzudin, R. M., & Rabeta. (2018). A potential of Telang tree (Clitoria ternatea) in human health. Food Research, 2(5), 415–420. https://doi.org/10.26656/fr.2017.2(5).073
- Fraga, L. R., Vianna, F. S. L., Del Campo, M., Sanseverino, M. T. V., & Schüler-Faccini, L. (2022). Editorial: Teratogenesis: Experimental models, Mechanisms and Clinical Findings in Humans. Frontiers in Genetics, 13. https://doi.org/10.3389/fgene.2022.901400
- Gandhi, G. R., Vasconcelos, A. B. S., Wu, D., Li, H., Antony, P. J., Li, H., Geng, F., Gurgel, R. Q., Narain, N., & Gan, R. (2020). Citrus flavonoids as promising phytochemicals Targeting diabetes and related complications: A Systematic review of in vitro and in vivo studies. Nutrients, 12(10), 2907. https://doi.org/10.3390/nu12102907
- Gomes, J. L. E., Olstad, E. W., Kowalski, T. W., Gervin, K., Vianna, F. S. L., Schüler‐Faccini, L., & Nordeng, H. (2021). Genetic Susceptibility to Drug Teratogenicity: A Systematic Literature review. Frontiers in Genetics, 12. https://doi.org/10.3389/fgene.2021.645555
- Gouda, A., Tolba, S. A., & Mahrose, K. (2021). Influences of vitamin A, L-carnitine, and folic acid in ovo feeding on embryo and hatchling characteristics and general health status in ducks. Animal Biotechnology, 33(1), 150–158. https://doi.org/10.1080/10495398.2020.1864389
- Han, D., Cho, S., Kwak, J., & Yoon, I. (2019). Medicinal plants and phytochemicals for diabetes mellitus: pharmacokinetic characteristics and herb-drug interactions. Journal of Pharmaceutical Investigation, 49(6), 603–612. https://doi.org/10.1007/s40005-019-00440-4
- Hoseinkhani, Z., Norooznezhad F, Rastegari-Pouyani M, Mansouri K. (2020) Medicinal Plants Extracts with Antiangiogenic Activity: Where Is the Link? Adv Pharm Bull, 10(3), 370-378. https://doi:10.34172/apb.2020.045.
- Inai, T., Mancuso, M. R., Hashizume, H., Baffert, F., Haskell, A., Baluk, P., Hu-Lowe, D. D., Shalinsky, D. R., Thurston, G., Yancopoulos, G. D., & McDonald, D. M. (2004). Inhibition of Vascular Endothelial Growth Factor (VEGF) Signaling in Cancer Causes Loss of Endothelial Fenestrations, Regression of Tumor Vessels, and Appearance of Basement Membrane Ghosts. American Journal of Pathology, 165(1), 35–52. https://doi.org/10.1016/s0002-9440(10)63273-7
- Indian, P. (2015) Dermal Leishmaniasi Experimental. Dermatology, 24(8), 215-239. https://doi:10.1111/EXD.12635.
- Jahromi, S. B., Doostkam, A., & Punica, L. (2019): A Potential Anti-Microbial Agent Anti-Infective Agents 17(1) https://doi:10.2174/2211352517666190215113232
- Kamath, M. S., Mascarenhas, M., Kirubakaran, R., & Bhattacharya, S. (2020). Number of embryos for transfer following in vitro fertilisation or intra-cytoplasmic sperm injection. The Cochrane Library, 8(8). https://doi.org/10.1002/14651858.cd003416.pub5
- Kang, J. H., Park, E. C., Yang, Y., Park, M., & Park, W. S. (2013). Crown-Rump Length Measured in the Early First Trimester as a Predictor of Low Birth Weight. Yonsei Medical Journal, 54(4), 1049. https://doi.org/10.3349/ymj.2013.54.4.1049
- Kennedy, D. C., Barbara, C., Antony M. W., & Karl J. A., (2022). “Microvascular Experimentation in the Chick Chorioallantoic Membrane as a Model for Screening Angiogenic Agents including from Gene-Modified Cells. International Journal of Molecular Sciences23(1), 452. https://doi.org/10.3390/ijms23010452
- Khater, M., Greco, F., Osborn HMI. (2020), Antiangiogenic Activity of Flavonoids: A Systematic Review and Meta-Analysis. Molecules, 25(20), 4712. https://doi:10.3390/molecules25204712.
- Lee, J.S., Shukla, S., Kim JA, Kim, M. (2015). Anti-angiogenic effect of Nelumbo nucifera leaf extracts in human umbilical vein endothelial cells with antioxidant potential. PLoS One, 10(2). https://doi:10.1371/journal.pone.0118552.
- Lee, S.H., Cai, M., & Yang, EJ. (2021) Anti-inflammatory Effects of a Novel Herbal Extract in the Muscle and Spinal Cord of an Amyotrophic Lateral Sclerosis Animal Model. Neurosci., Sec. Neuropharmacology, 15(11), 743-705. https://doi.org/10.3389/fnins.2021.743705
- Lewicki, S., Skopińska-Różewska, E., Bałan, B.J., Kalicki, B., Patera, J., Wilczak, J., Wasiutyński, A., & Zdanowski, R. (2017). Morphofunctional Renal Alterations in Progeny of Mice Fed Rhodiola kirilowii Extracts or Epigallocatechin During Pregnancy and Lactation. J. Food, 20, 86–92. https://www.liebertpub.com/doi/abs/10.1089/jmf.2016.0126
- Lokman, N. A., Elder, A. S., Ricciardelli, C., & Oehler, M. K. (2012). Chick Chorioallantoic Membrane (CAM) assay as an in vivo model to study the effect of newly identified molecules on ovarian cancer invasion and metastasis. International Journal of Molecular Sciences, 13(8), 9959–9970. https://doi.org/10.3390/ijms13089959
- Lu, K., Bhat, M., Basu, S., (2016) Plants and their active compounds: natural molecules to target angiogenesis. Angiogenesis, 19(3), 287-95. https://doi:10.1007/s10456-016-9512-y.
- Lumsangkul, C., Fan, Y., Chang, S., Ju, J., & Chiang, H. (2018). Characterizing early embryonic development of Brown Tsaiya Ducks (Anas platyrhynchos) in comparison with Taiwan Country Chicken (Gallus gallus domestics). PLOS ONE, 13(5), e0196973. https://doi.org/10.1371/journal.pone.0196973
- Maity, N., Nema, N.K., Sarkar,B.K., Mukherjee,P.K.(2012). Standard Clitoria ternatea leaf extractas hyaluronidase, elastase and matrix-metalloproteinase – 1 inhibitor. Indian J Pharmacol, 44(5), 584-7. https://doi:10.4103/00253-7653.100381.
- Mangir, N., Dikici, S., Claeyssens, F., & MacNeil, S. (2019). Using ex ovo chick chorioallantoic membrane (CAM) assay to evaluate the biocompatibility and angiogenic response to biomaterials. ACS Biomaterials Science & Engineering, 5(7), 3190–3200. https://doi.org/10.1021/acsbiomaterials.9b00172
- Mani, J. S., Johnson, J. B., Steel, J. C., Broszczak, D. A., Neilsen, P. M., Walsh, K. B., & Naiker, M. (2020). Natural product-derived phytochemicals as potential agents against Coronaviruses: A Review. Virus Research, 284, 197989. https://doi.org/10.1016/j.virusres.2020.197989
- Manjula,P., Mohan,C.H., Sreekanth,P., Keerthi,B., Prathibha,P.D.(2013).Phytochemical Analysis of Clitoria Ternatea Linn., A Valuable Medicinal Plant J Indian bot.Soc. 92(3&4) 2013:173-178. https://www.researchgate.net/publication/277247891
- Minda, D., Ghiulai, R., Banciu, C., Magyari-Pavel, I. Z., Danciu, C., Racoviceanu, R., Şoica, C., Budu, O., Muntean, D., Diaconeasa, Z., Dehelean, C., & Avram, Ş. (2022). Phytochemical profile, antioxidant and wound healing potential of three artemisia species: in vitro and in ovo evaluation. Applied Sciences, 12(3), 1359. https://doi.org/10.3390/app12031359
- Mohammad, A., Armen, A., Soroush, E., Mona, M., Nader, S., (2017). Vitamins and regulation of angiogenesis: [A, B1, B2, B3, B6, B9, B12, C, D, E, K], Journal of Functional Foods, 38, 180-196. https://doi.org/10.1016/j.jff.2017.09.005.
- Moreno-Jiménez, I., Hulsart-Billstrom, G., Lanham, S. A., Janeczek, A. A., Kontouli, N., Kanczler, J. M., Evans, N. D., & Oreffo, R. O. C. (2016). The chorioallantoic membrane (CAM) assay for the study of human bone regeneration: A refinement animal model for tissue engineering. Scientific Reports, 6(1). https://doi.org/10.1038/srep32168
- Moses, M. A. (2013). Chorioallantoic membrane (CAM) assay. In Springer eBooks (pp. 400–401). https://doi.org/10.1007/978-1-4419-9863-7_1536
- Mukherjee,S., Mukhopadhyay,D., Braun,C., Barbhuiya,J.N., Das, N.K., Chatterjee,U., Stebut, E.V., Chatterjee, M.(2015). Decreased presence of Langerhans cells is a critical determinant for Indian Post Kala-azar dermal leishmaniasis. Experimental Dermatology, 24(6), 215-239. https://doi:10.1111/EXD.12635.
- Müller, T. D., Blüher, M., Tschöp, M. H., & DiMarchi, R. D. (2021). Anti-obesity drug discovery: advances and challenges. Nature Reviews Drug Discovery, 21(3), 201–223. https://doi.org/10.1038/s41573-021-00337-8
- Naik, P. D., Srinivas, B. K., Krishnappa, D. K., Jayaram, S. (2021). Apoptosis-inducing and antiangiogenic activity of partially purified protein from the pericarp of Zanthoxylum rhetsa in vitro and in vivo. http://www.phcog.com/text.asp?2021/17/5/96/318031
- Naito, H., Iba, T., Takakura, N. (2020). Mechanisms of new blood-vessel formation and proliferative heterogeneity of endothelial cells. Int Immunol 32 (5), 295-305. https://doi: 10.1093/intimm/dxaa008.
- Nazarali, A. J., & Puthucode, R. N. (2003). In vivo animal models of teratogenicity. In Humana Press eBooks (pp. 253–294). https://doi.org/10.1385/0-89603-511-5:253
- Niapour, A., Miran, M., Seyedasli, N., & Norouzi, F. (2022). Anti-angiogenic effects of aqueous extract from Agrostemma githago L. seed in human umbilical vein endothelial cells via regulating Notch/VEGF, MMP2/9, ANG2, and VEGFR2. Environmental Science and Pollution Research, 30(9), 22413–22429. https://doi.org/10.1007/s11356-022-23510-x
- Nithianantham, K., Shyamala, M., Chen, Y., Latha, L. Y., Jothy, S. L., & Sasidharan, S. (2011). Hepatoprotective Potential of Clitoria ternatea Leaf Extract Against Paracetamol Induced Damage in Mice. Molecules, 16(12), 10134–10145. https://doi.org/10.3390/molecules161210134
- Nowak-Śliwińska, P., Segura, T., & Iruela‐Arispe, M. L. (2014). The chicken chorioallantoic membrane model in biology, medicine and bioengineering. Angiogenesis, 17(4), 779–804. https://doi.org/10.1007/s10456-014-9440-7
- Ngo, T. H., Park, J., Jo, Y.D., Jin, C.H., Jung, C.H., Nam, B., Han, A.R., & Nam, J.W. (2022). Content of Two Major Steroidal Glycoalkaloids in Tomato (Solanum lycopersicum cv. Micro-Tom) Mutant Lines at Different Ripening Stages. Plants (Basel), 11(21);2895. https://doi: 10.3390/plants11212895.
- Podolak I, Galanty A, Sobolewska D. (2010). Saponins as cytotoxic agents: a review. Phytochem Review, (3);425-474. https://doi: 10.1007/s11101-010-9183-z.
- Somfai, T., Haraguchi, S., Dang‐Nguyen, T. Q., Kaneko, H., & Kikuchi, K. (2023). Vitrification of porcine immature oocytes and zygotes results in different levels of DNA damage which reflects developmental competence to the blastocyst stage. PLOS ONE, 18(3), e0282959. https://doi.org/10.1371/journal.pone.0282959
- Tamanoi, F. (2019). Recent excitements in the study of the CAM assay. In The Enzymes 46 (1), 1–9. https://doi.org/10.1016/bs.enz.2019.10.002
- Tangara, M., Chen, W., Xu, J., Huang, F., & Peng, J. (2010). Effects ofin ovofeeding of carbohydrates and arginine on hatchability, body weight, energy metabolism and perinatal growth in duck embryos and neonates. British Poultry Science, 51(5), 602–608. https://doi.org/10.1080/00071668.2010.520303
- Tanzina, S., Alfi K., Ahmed Q., Mohd H.,Nasir M., Supandi F., Taher M., Mohd Z., (2021). Preliminary Phytochemical Screening, In Vitro Antidiabetic, Antioxidant Activities, and Toxicity of Leaf Extracts of Psychotria malayana. Jack Journal: Plants, 10(12), 268. https://doi: 3390/plants10122688
- Tian, X. Y., Cheung, L. M., Leung, K. S., Chen, Q., Deng, B., Deng, P. X., & Xu, M. (2009). The effects of Scutellaria baicalensisextract on embryonic development in mice. Birth Defects Research, 86(2), 79–84. https://doi.org/10.1002/bdrb.20186
- Thilavech,T., Adisakwattana,S., Channuwong,P., Radarit,K., Jantarapat,K., Ngewlai,K., Sonprasan,N., Chusak,C.(2021). Clitoria ternatea Flower extract Attenuates Postprandial Lipemia and Increases Plasma Antioxidant Status Responses to a High-fat Meal Challenge in Overweight and Obsessed Participants. Biology (Basel), 10(10):975. https://doi:10.3390/biology/10100975.
- Vijaykumar G., Vivek N., Anant P., Priyanka M., Geetanjali R., (2022) Angiogenic potential of Bambusa vulgaris leaves: Results of an in-vitro study with chicken embryo chorioallantoic membrane (CAM) model. Journal of Ayurveda and Integrative Medicine, Volume 13, https://doi.org/10.1016/j.jaim.2022.100629.
- Yahya, A.L. (2017). Angiosuppressive and Teratogenic Influence of an Edible Medicinal herb, Insulin plant (Costus igneus) in Mallard Duck (Anas platyrhynchos) Embryos. United Research Association Advances in Engineering (URUAE) Journal 1(17), 17-25 https://doi.org/10.17758/uruae.ae0117303
- Yu-Qing T., Fei L., Yu-Kun D., Shuang D., Ying-Xin L., Yun-Shu Z., Jun L., Heng-Wen C. (2022). Pharmacological properties of total flavonoids in Scutellaria baicalensis for the treatment of cardiovascular diseases. Phytomedicine, Volume 107(10), 0944-7113 https://doi.org/10.1016/j.phymed.2022.154458. 5
- Zhang, A. L., Story, D. F., Lin, V., Vitetta, L., & Xue, C. C. (2008). A population survey on the use of 24 common medicinal herbs in Australia. Pharmacoepidemiology and Drug Safety, 17(10), 1006–1013. https://doi.org/10.1002/pds.1610