International Journal of Clinical Pediatric Dentistry

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VOLUME 17 , ISSUE 2 ( February, 2024 ) > List of Articles


Evaluation of Cytotoxicity of Allium sativum (Garlic Extract) against Human Dental Pulp Fibroblasts

Raghu Devaraju, Divya Reddy, Santhosh T Paul, Umme Azher, Keerthy Umashankar, Likhith Srinivas

Keywords : Allium sativum, Cytotoxicity, Garlic extract, Human dental pulp fibroblasts

Citation Information : Devaraju R, Reddy D, Paul ST, Azher U, Umashankar K, Srinivas L. Evaluation of Cytotoxicity of Allium sativum (Garlic Extract) against Human Dental Pulp Fibroblasts. Int J Clin Pediatr Dent 2024; 17 (2):143-148.

DOI: 10.5005/jp-journals-10005-2781

License: CC BY-NC 4.0

Published Online: 22-04-2024

Copyright Statement:  Copyright © 2024; The Author(s).


Background: Vital pulp therapy procedures in primary dentition focuses on preservation and maintenance of pulp tissue that has been compromised due to caries, trauma, etc. Several pulp dressing materials have been used in primary teeth and some natural materials from the field of traditional medicine have also been introduced as medicaments in vital pulp therapy. The understanding of biologic and cytotoxic properties of newer materials is important for safe clinical usage. The biologic compatibility of these newer materials is imperative to limit or avoid tissue irritation or degeneration. Aim: To evaluate the cytotoxic effects of Allium sativum on cultured human primary dental pulp fibroblasts. Materials and methods: Primary pulp fibroblasts were cultured from the pulp tissue obtained from extracted deciduous primary canines and central incisor teeth. The freshly prepared concentrations of 1000, 500, 250, 125, and 62.5 µg/mL A. sativum extract were added to the 96-well plate in triplicates to which culture medium containing fourth passage cell suspension was added previously. Cells without treatment served as control, while cells treated with 5% dimethyl sulfoxide (DMSO) served as toxic control. After the addition of experimental and control agents, cells were incubated for 24 and 48 hours at 37°C in 5% CO2 atmosphere. After the incubation period, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to determine the number of viable cells. Absorbance was read with a microplate reader at 570 nm wavelength and the relative viability of dental pulp fibroblasts at various concentrations was expressed as color intensity of the experimental wells relative to that of control. The percentage of cell viability was also calculated accordingly. Results: The MTT assay results revealed that A. sativum extract, in all the concentrations tested at both the time intervals maintained a cell viability of greater than 90%. At 24 hours, the mean absorbance value of untreated control wells was recorded as 0.84400 ± 0.00916 with 100% cell viability. Among all the concentrations of garlic extract tested, highest mean absorbance value of 0.83933 ± 0.00550 with 99.44% cell viability was recorded for 62.5 µg/mL concentration. At 48 hours, the mean absorbance value of untreated control wells was recorded as 1.22767 ± 0.01106 with 100% cell viability, and the highest mean absorbance value of 1.22567 ± 0.01006 with 99.83% cell viability was recorded for 62.5 µg/mL concentration. The cell viability did not seem to be affected by the concentration of A. sativum extract at 24 hours. However, at 48 hours, the sensitivity of the cells was observed to be dependent on the concentration of A. sativum with a decrease in the viability of cells noted with the increase in concentration. Conclusion: A. sativum extract is noncytotoxic in nature and preserves the vitality of cultured human primary dental pulp fibroblasts making it a suitable material for use in vital pulp therapy procedures of primary teeth.

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  1. Primosch RE, Glomb TA, Jerrell RG. Primary tooth pulp therapy as taught in predoctoral pediatric dental programs in the United States. Pediatr Dent 1997;19(2):118–122.
  2. The Reference Manual of Pediatric Dentistry. American Academy of Pediatric Dentistry: Chicago, Illinois; 2021. pp. 399–407.
  3. Mandrol PS, Bhat K, Prabhakar AR. An in vitro evaluation of cytotoxicity of curcumin against human dental pulp fibroblasts. J Indian Soc Pedod Prev Dent 2016;34(3):269–272. DOI: 10.4103/0970-4388.186757
  4. Waterhouse PJ, Nunn JH, Whitworth JM. An investigation of the relative efficacy of Buckley's formocresol and calcium hydroxide in primary molar vital pulp therapy. Br Dent J 2000;188(1):32–36. DOI: 10.1038/sj.bdj.4800380
  5. Garcia-Godoy F. A 42 month clinical evaluation of glutaraldehyde pulpotomies in primary teeth. J Pedod 1986;10(2):148–155.
  6. Fuks AB, Eidelman E, Cleaton-Jones P, et al. Pulp response to ferric sulfate, diluted formocresol and IRM in pulpotomized primary baboon teeth. ASDC J Dent Child 1997;64(4):254–259.
  7. Kakarla P, Avula JS, Mellela GM, et al. Dental pulp response to collagen and pulpotec cement as pulpotomy agents in primary dentition: a histological study. J Conserv Dent 2013;16(5):434–438. DOI: 10.4103/0972-0707.117525
  8. Bossù M, Iaculli F, Di Giorgio G, et al. Different pulp dressing materials for the pulpotomy of primary teeth: a systematic review of the literature. J Clin Med 2020;9(3):838. DOI: 10.3390/jcm9030838
  9. Schwartz RS, Mauger M, Clement DJ, et al. Mineral trioxide aggregate: a new material for endodontics. J Am Dent Assoc 1999;130(7):967–975. DOI: 10.14219/jada.archive.1999.0337
  10. Stringhini Junior E, Dos Santos MGC, Oliveira LB, et al. MTA and biodentine for primary teeth pulpotomy: a systematic review and meta-analysis of clinical trials. Clin Oral Investig 2019;23(4):1967–1976. DOI: 10.1007/s00784-018-2616-6
  11. Tsao SM, Yin MC. In-vitro antimicrobial activity of four diallyl sulphides occurring naturally in garlic and Chinese leek oils. J Med Microbiol 2001;50(7):646–649. DOI: 10.1099/0022-1317-50-7-646
  12. Bakri IM, Douglas CW. Inhibitory effect of garlic extract on oral bacteria. Arch Oral Biol 2005;50(7):645–651. DOI: 10.1016/j.archoralbio.2004.12.002
  13. Narendhirakannan RT, Rajeswari K. In vitro antioxidant properties of three varieties of Allium sativum L. extracts. E-J Chem 2010;7(S1):S573–S579. DOI: 10.1155/2010/283627
  14. Oosthuizen C, Arbach M, Meyer D, et al. Diallyl polysulfides from Allium sativum as immunomodulators, hepatoprotectors, and antimycobacterial agents. J Med Food 2017;20(7):685–690. DOI: 10.1089/jmf.2016.0137
  15. Farahpour MR, Hearaki S, Faraji D, et al. Hydroethanolic Allium sativum extract accelerates excision wound healing: evidence for roles of mast-cell infiltration and intracytoplasmic carbohydrate ratio. Braz J Pharm Sci 2017;53:e15079.
  16. Elheeny AAH. Allium sativum extract as an irrigant in pulpectomy of primary molars: a 12-month short-term evaluation. Clin Exp Dent Res 2019;5(4):420–426. DOI: 10.1002/cre2.197
  17. Prabhakaran P, Mariswamy AB. A scanning electron microscope evaluation of efficacy of sodium hypochlorite and Allium sativum in smear layer removal in root canals with the use of modified evacuation system: an ex vivo study. J Conserv Dent 2018;21(4):401–407. DOI: 10.4103/JCD.JCD_373_16
  18. Koppolu M, Mathew VB, Thangala V, et al. Evaluation of effect of Allium sativum on smear layer removal in root canals - an ex vivo study. Ann Essences Dent 2012;4(3):17–22.
  19. Eswar K, Venkateshbabu N, Rajeswari K, et al. Dentinal tubule disinfection with 2% chlorhexidine, garlic extract, and calcium hydroxide against Enterococcus faecalis by using real-time polymerase chain reaction: in vitro study. J Conserv Dent 2013;16(3):194–198. DOI: 10.4103/0972-0707.111312
  20. Groppo FC, Ramacciato JC, Motta RH, et al. Antimicrobial activity of garlic against oral streptococci. Int J Dent Hyg 2007;5(2):109–115. DOI: 10.1111/j.1601-5037.2007.00230.x
  21. Mohammad SG, Raheel SA, Baroudi K. Clinical and radiographic evaluation of Allium sativum oil as a new medicament for vital pulp treatment of primary teeth. J Int Oral Health 2014;6(6):32–36.
  22. Kahvand M, Mehran M, Haghgoo R, et al. Clinical and radiographic evaluation of Allium sativum oil (garlic oil) in comparison with formocresol in primary molar pulpotomy. J Int Soc Prevent Communit Dent 2019;9(4):390–395. DOI: 10.4103/jispcd.JISPCD_145_19
  23. Mjor IA. Biologic assessment of restorative dental materials: interrelationship of biologic and technologic properties. Oper Dent 1978;3:9–13.
  24. Kamiloglu S, Sari G, Ozdal T, et al. Guidelines for cell viability assays. Food Frontiers 2020;1:332–349. DOI: 10.1002/fft2.44
  25. van Wyk CW, Olivier A, Maritz JS. Cultured pulp fibroblasts: are they suitable for in vitro cytotoxicity testing? J Oral Pathol Med 2001;30(3):168–177. DOI: 10.1034/j.1600-0714.2001.300307.x
  26. Benkeblia N. Antimicrobial activity of essential oil extracts of various onions (Allium cepa) and garlic (Allium sativum). Lebensm-Wiss u-Technol 2004;37(2):263–268. DOI: 10.1016/j.lwt.2003.09.001
  27. Siddik K, Mahmood AA, Salmah I. Acceleration of wound healing by aqueous extract of Allium sativum in combination with honey on cutaneous wound healing in rats. Int J Mol Med Adv Sci 2006;2(2):231–235.
  28. Ejaz S, Chekarova I, Cho JW, et al. Effect of aged garlic extract on wound healing: A new frontier in wound management. Drug Chem Toxicol 2009;32(3):191–203. DOI: 10.1080/01480540902862236
  29. Szychowski KA, Binduga UE, Rybczyńska-Tkaczyk K, et al. Cytotoxic effects of two extracts from garlic (Allium sativum L.) cultivars on the human squamous carcinoma cell line SCC-15. Saudi J Biol Sci 2018;25(8):1703–1712. DOI: 10.1016/j.sjbs.2016.10.005
  30. Wagner H, Wierer M, Fessler B. Effects of garlic constituents on arachidonate metabolism. Planta Med 1987;53(3):305–306. DOI: 10.1055/s-2006-962719
  31. Bhandari PR. Garlic (Allium sativum L.): a review of potential therapeutic applications. Int J Green Pharm 2012;6(2):118–129. DOI: 10.4103/0973-8258.102826
  32. Motamayel FA, Hassanpour S, Alikhani MY, et al. Antibacterial effect of eucalyptus (globulus Labill) and garlic (Allium sativum) extracts on oral cariogenic bacteria. J Microbiol Res Rev 2013;1(2):12–17.
  33. Houshmand B, Mahjour F, Dianat O. Antibacterial effect of different concentrations of garlic (Allium sativum) extract on dental plaque bacteria. Indian J Dent Res 2013;24(1):71–75. DOI: 10.4103/0970-9290.114957
  34. Kshirsagar MM, Dodamani AS, Karibasappa GN, et al. Antibacterial activity of garlic extract on cariogenic bacteria: an in vitro study. Ayu 2018;39(3):165–168. DOI: 10.4103/ayu.AYU_193_16
  35. Mohammad SG, Baroudi K. Assessment of the potential of Allium sativum oil as a new medicament for non-vital pulpotomy of primary teeth. J Int Soc Prevent Communit Dent 2015;5(4):314–320. DOI: 10.4103/2231-0762.161762
  36. Schmalz G. Concepts in biocompatibility testing of dental restorative materials. Clin Oral Investig 1997;1(4):154–162. DOI: 10.1007/s007840050027
  37. Tabatabaei FS, Moezizadeh M, Javand F. Effects of extracts of Salvadora persica on proliferation and viability of human dental pulp stem cells. J Conserv Dent 2015;18(4):315–320. DOI: 10.4103/0972-0707.159740
  38. Mohammad SG, Raheel SA, Baroudi K. Histological evaluation of Allium sativum oil as a new medicament for pulp treatment of permanent teeth. J Contemp Dent Pract 2015;16(2):85–90. DOI: 10.5005/jp-journals-10024-1641
  39. Bramanti I, Sudarso ISR, Wahyuningsih MSH, et al. Ethanolic garlic extract (Allium sativumL) increased viability and proliferation of human gingival fibroblast in vitro. Bangladesh J Med Sci 2018;17(4):556–561.
  40. Ozan F, Ozdem M, Ozan U, et al. Evaluation of cytotoxic effect of garlic on human gingival fibroblasts: a preliminary study. Gulhane Med J 2013;55(4):276–280. DOI: 10.5455/gulhane.28177
  41. Shrivastava S, Ganesh N. Tumor inhibition and cytotoxicity assay by aqueous extract of onion (Allium cepa) & garlic (Allium sativum): an in-vitro analysis. Int J Phytomedicine 2010;2:80–84. DOI: 10.5138/ijpm.2010.0975.0185.02013
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