In vitro cytotoxic effect of annona squamosa pulp ex-tract as a mouthwash for children on human normal cell line

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Sumaia Hussein Ali
Zainab Juma Jafar


Background: Fruits and their by-products are the primary sources of bioactive chemicals in plants. Because of its phytochemical richness, Annona squamosa fruits have gained the alertness of people willing in health-promoting diets. The purpose of this in vitro study was to evaluate the cytocompatibility effect of ethanolic crude extract of Annona squamosa pulp against a human normal cell line as a mouthwash for children. Material and method: The ethanolic extract of Annona squamosa pulp was extracted using the ultrasonic method and then lyophilized to make it powder. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) test was performed to investigate the cytotoxic activity of the pulp extract on a human normal cell line derived from human dermal fibroblast, neonatal (HDFn). Plates were then incubated with 5% CO2 at 37°C For the following concentrations (400, 200, 100, 50, 25, 12.5, 6.25 g/ml). Each concentration, as well as the positive control chlorhexidine, and the negative control cells without tested material, were tested in triplicate. Results: No significant difference was found between the cytotoxicity of the ethanol crude extract of Annona squamosa and a chlorhexidine (P = >0.05) against human dermal fibroblast of neonate cells, with IC50 (50% growth inhibition of cells) values of 235.4 μg/mL while chlorhexidine had an IC50 of 97.8 μg/mL. Conclusion: Annona squamosa extract is more safe and has less cytotoxicity than chlorhexidine. So, to overcome the problems of chlorhexidine, herbal mouthwash formulations could be utilized as an al-ternative mouthwash.

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Ali SH, Jafar ZJ. In vitro cytotoxic effect of annona squamosa pulp ex-tract as a mouthwash for children on human normal cell line. J Bagh Coll Dent [Internet]. 2022 Mar. 15 [cited 2022 May 29];34(1):60-6. Available from:
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Deshpande, A., Deshpande, N., Raol, R, et al. Effect of green tea, ginger plus green tea, and chlorhexidine mouthwash on plaque-induced gingivitis: A randomized clinical trial. J Indian Soc Periodontol. 2021; 25(4):307.

Vinod, KS., Sunil, KS., Sethi, P., et al. A novel herbal formulation versus chlorhexidine mouthwash in efficacy against oral microflora. J Int Soc Prev Community Dent. 2018; 8(2):184.

Sharma, H., Kumar, A. Ethnobotanical studies on medicinal plants of Rajasthan (India): A review. J Med Plants Res. 2011;5(7):1107-1112.

Wang, DS., Rizwani, GH., Guo, H., et al. Annona squamosa Linn: cytotoxic activity found in leaf extract against human tumor cell lines. Pak J Pharm Sci. 2014; 27(5):1559-1563.

Gurib-Fakim, A. Medicinal plants: traditions of yesterday and drugs of tomorrow. Mol Aspects Med. 2006; 27(1):1-93.

Gajalakshmi, S., Vijayalakshmi, S., Devi, RV. Phytochemical and pharmacological properties of Annona muricata: a review. Int J Pharm Pharm Sci. 2012; 4(2):3-6.

Wunderlin, R., Hansen, B. Synonyms of Annona squamosa. Atlas of Florida Vascular Plants. 2008:24-27.

Das, S., Bhattacharya, A., Ghosh, B., et al. Analytical and Phytochemical Exploration of the Seeds of Annona squamosa. J Anal Pharm Res 2016; 3(4):00065-00070.

Vyas, K., Manda, H., Sharma, RK., et al. An update review on Annona Squamosa. Int J Pharm Therapeut. 2012; 3(2):107-118.

Pandey, N., Barve, D. Phytochemical and pharmacological review on Annona squamosa Linn. Int J Res Pharm Biomed Sci. 2011; 2(4):1404-1412.

Chimbevo, LM. Efficacy and Safety, Nutritional and Antioxidant Activity of Kenyan Annona muricata (L.) and Annona squamosa (L.) Fruits Extracts in BALB/c Mice Model of L. major Leshmaniasis (Doctoral dissertation, JKUAT-COHES), 2020.

Gupta, RK., Kesari, AN., Watal, G., et al. Nutritional and hypoglycemic effect of fruit pulp of Annona squamosa in normal healthy and alloxan-induced diabetic rabbits. Ann Nutr Metab. 2005; 49(6):407-413.

Tong, SY., Davis, JS., Eichenberger, E., et al. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev. 2015; 28(3):603-661

Ahmad, H., Ali, N., Ahmad, B., et al. Screening of solanum surrattense for antibacterial, antifungal, phytotoxic and hae-magglutination. J Tradit Chin Med. 2012; 32(4):616-620.

Adan, A., Kiraz, Y., Baran, Y. Cell proliferation and cytotoxicity assays. Curr Pharm Biotechnol. 2016; 17(14):1213-1221.

Decker, T., Lohmann-Matthes, ML. A quick and simple method for the quantitation of lactate dehydrogenase release in measurements of cellular cytotoxicity and tumor necrosis factor (TNF) activity. J Immunol Methods. 1988; 115(1):61-69.

Fotakis, G., Timbrell, JA. In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett. 2006; 160(2):171-177.

Rogero, SO., Lugão, AB., Ikeda, TI., et al. Teste in vitro de citotoxicidade: estudo comparativo entre duas metodologias. Mater Res.. 2003; 6(3): 317-320.

Trintinaglia, L., Bianchi, E., Silva, LB., et al. Cytotoxicity assays as tools to assess water quality in the Sinos River basin. Braz J Biol. 2015; 75(2):75-80.

Trindade, ML., Radünz, M., Ramos, AH., et al. Chemical characterization, antimicrobial and antioxidant activity of sug-ar-apple (Annona squamosa L.) pulp extract. Rev Chil Nutr. 2020; 47(2):281-285.

Freshney, RI. 'Culture of Animal Cells, A Manual Of Basic Technique And Specialized Applications, sixth ed., Wiley- Blackwell, 2010.

Mosmann, T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assay. J Immunol Methods. 1983; 65: 55–63.

Ebisawa, K., Kato, R., Okada, M., et al. Gingival and dermal fibroblasts: their similarities and differences revealed from gene expression. J Biosci Bioeng. 2011; 111(3): 255-258.

Berridge, MV., Herst, PM., Tan, AS. Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Bi-otechnol Annu Rev. 2005; 11: 127-152.

FDA: Pesticide Analytical Manual Volume I (PAM) 3rd Edition find in

Karbach, J., Ebenezer, S., Warnke, PH., et al. Antimicrobial effect of Australian antibacterial essential oils as alternative to common antiseptic solutions against clinically relevant oral pathogens. Clin Lab.2015;61(1-2):61-68.