Comparative Evaluation of Cytotoxicity of Neem Leaf Extract, 2% Chlorhexidine, Nigella sativa Extract and 3% Sodium Hypochlorite
Vignesh Guptha Raju, A Shafie Ahamed, Madhuram Krishnamurthy, KE Selvendran, Hamed Ahmed Ali Alshawkani, Omaima Ahmed Hassam Allayl, Manar Ali Ibrahim Alhomood, Mohammed Ali Ali Sumayli
To assess and compare the cytotoxicity of different root canal irrigants, namely 25% aqueous neem leaf extract (Azadirachta indica), 2% chlorhexidine (CHX), 25% aqueous Nigella sativa extract (black seed) and 3% sodium hypochlorite (NaOCl). Dulbecco modified Eagle's medium containing glutamine, fetal bovine serum, and antibiotics were used to culture human periodontal ligament cells. After trypsinization, 1 × 105 cell suspension was seeded in 24 well culture plates with 100 µL of culture medium. Then 10 µl of each irrigant was added to the wells, sealed, and incubated in an oven at 37°C in an atmosphere of 5% CO2 and 95% humidity. Cytotoxicity was assessed at 1, 3, 6, and 24 hours by evaluating the cell viability using the trypan blue assay. A microscope was used for counting viable cells and subjected to statistical analysis. 25% aqueous Neem leaf extract was the most cytotoxic irrigant tested, followed by 2% CHX, while 25% aqueous N. sativa extract was the least cytotoxic irrigant tested. 25% aqueous N. sativa extract and 3% NaOCl solutions were significantly less toxic than 25% aqueous Neem leaf extract solution and cytotoxicity of 2% chlorhexidine was significantly more than 25% aqueous N. sativa extract and 3% NaOCl at all periods. Considering this encouraging in vitro data, herbal extracts could be an alternative root canal irrigant with the most negligible toxicity compared to conventional root canal irrigants, with undesirable effects.
Chandra S. Grossman's Endodontic Practice. Wolters Kluwer (India) 2014. https://books.google.co.in/books?id=tC3vDwAAQBAJ
Gernhardt CR, Eppendorf K, Kozlowski A, et al. Toxicity of concentrated sodium hypochlorite used as an endodontic irrigant. Int Endod J 2004;37(4):272–280. DOI: 10.1111/j.0143-2885.2004.00804.x
Salzgeber RM, Brilliant JD. An in vivo evaluation of the penetration of an irrigating solution in root canals. J Endod 1977;3(10):394–398. DOI: 10.1016/S0099-2399(77)80172-6
Zehnder M. Root canal irrigants. J Endod 2006;32(5):389–398. DOI: 10.1016/j.joen.2005.09.014
Pujar M, Patil CKA. Comparison of antimicrobial efficacy of Triphala, (GTP) green tea polyphenols and 3% of sodium hypochlorite onbiofilms formed on tooth substrate: in vitro Enterococcus faecalis. J Int Oral Heal 2011;3(2). DOI: 10.1016/j.joen.2009.09.040
Bohora A, Hegde V, Kokate S. Comparison of the antibacterial efficiency of neem leaf extract and 2% sodium hypochlorite against, and mixed culture – an in vitro study E. faecalisC. albicans. Endodontology 2009;22.
Ahmad A, Husain A, Mujeeb M, et al. A review on therapeutic potential of: a miracle herb Nigella sativa. Asian Pac J Trop Biomed 2013;3(5):337–352. DOI: 10.1016/S2221-1691(13)60075-1
Khodair A I HFA, A-AH. Phytochemical investigation of thymus de cassatus. Qutar Univ Sci J 1993;13(12):211–213.
Chang YC, Huang FM, Tai KW, et al. The effect of sodium hypochlorite and chlorhexidine on cultured human periodontal ligament cells. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2001;92(4):446–450. DOI: 10.1067/moe.2001.116812
Hauman CHJ, Love RM. Biocompatibility of dental materials used in contemporary endodontic therapy: a review. Part 1. Intracanal drugs and substances. Int Endod J 2003;36(2):75–85. DOI: 10.1046/j.1365-2591.2003.00631.x
Lessa FCR, Aranha AMF, Nogueira I, et al. Toxicity of chlorhexidine on odontoblast-like cells. J Appl Oral Sci 2010;18(1):50–58. DOI: 10.1590/s1678-77572010000100010
Faria G, Celes MRN, De Rossi A, et al. Evaluation of chlorhexidine toxicity injected in the paw of mice and added to cultured l929 fibroblasts. J Endod 2007;33(6):715–722. DOI: 10.1016/j.joen.2006.12.023
Aubut V, Pommel L, Verhille B, et al. Biological properties of a neutralized 2.5% sodium hypochlorite solution. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;109(2):e120-e125. DOI: 10.1016/j.tripleo.2009.09.022
Hidalgo E, Bartolome R, Dominguez C. Cytotoxicity mechanisms of sodium hypochlorite in cultured human dermal fibroblasts and its bactericidal effectiveness. Chem Biol Interact 2002;139(3):265–282. DOI: 10.1016/s0009-2797(02)00003-0
el Daly ES. Protective effect of cysteine and vitamin E, Crocus sativus andextracts on cisplatin-induced toxicity in rats Nigella sativa. J Pharm Belg 1998;53(2):87–93; (Discussion 93–5). http://www.ncbi.nlm.nih.gov/pubmed/9609969
Khanna T, Zaidi FA, Dandiya P. CNS and analgesic studies on Nigella sativa. Fitoterapia 1993;64:407–410.
Ali B, Blunden G. Pharmacological and toxicological properties of Nigella sativa. Phytother Res. 2003;17:299–305. DOI:10.1002/ptr.1309