Investigating the Effects of Herbal Nanoparticle Endodontic Irrigants on Candida albicans and Enterococcus faecalis: An In Vitro Study

Hesham Hassan; Mohamed Zaazou; Doaa Sadony; Tarek Mohamed

doi:10.26538/tjnpr/v8i2.6

Authors

Hesham Hassan Restorative and Dental Materials Department, National Research Centre, Dokki, Giza, Egypt.
Mohamed Zaazou Restorative and Dental Materials Department, National Research Centre, Dokki, Giza, Egypt.
Doaa Sadony Restorative and Dental Materials Department, National Research Centre, Dokki, Giza, Egypt.
Tarek Mohamed Laser Institute for Research and Applications, Beni-Suef University, Beni-Suef, Egypt.

DOI:

https://doi.org/10.26538/tjnpr/v8i2.6

Keywords:

Root canal irrigants, antibacterial and antifungal effect, Nano-curcumin, Nano-neem

Abstract

Sodium hypochlorite irrigant (NaOCl) has many disadvantages although it possesses antibacterial activity and having the capability to disband the pulp tissue.. The aim of this study was to compare the effect of two nanoparticle herbal irrigants to that of Sodium hypochlorite (NaOCl) on Enterococcus faecalis and Candida albicans. Sixty single-rooted teeth were mechanically prepared. Thirty teeth were inoculated with Enterococcus faecalis, and the other 30 with Candida albicans. These teeth were divided into three groups: Group 1 contained 20 Enterococcus faecalis inoculated teeth, which were further divided into 10 teeth irrigated with nanocurcumin and 10 teeth irrigated with nanoneem. Group 2 contained 20 Candida albicans-inoculated teeth which were irrigated like Group 1. Group 3 contained ten Enterococcus faecalis-inoculated teeth and ten Candida albicans-inoculated teeth which were irrigated with 2.5 % NaOCl. Microbial swabs were collected from the teeth before and after irrigation and cultured. Pre-operative and post-operative colony forming units (CFU) were compared in each sub-group using the Mann-Whitney test and the Kruskal-Wallis compared the difference between the groups. Significant post-operative reduction of the CFUs was shown for the incubated bacteria and fungi in both the nanoneem and nanocurcumin groups. However, the NaOCl showed a significantly greater reduction of the CFU than the nanoparticle herbal irrigants. The use of nanoneem and nanocurcumin in conjunction with sodium hypochlorite as root canal irrigants is advantageous due to their antibacterial and antifungal effect.

References

Liu R, Kaiwar A, Shemesh H. Incidence of apical root cracks and apical dentinal detachments after canal preparation with hand and rotary files at different instrumentation lengths. J Endod 2013;39:129–132.

Paqu F, Laib A, Gautschi H, Zehnder M. Hard-tissue debris accumulation analysis by high-resolution computed tomography scans. J Endod 2009;35:1044–1047.

Love R, Jenkinson H. Invasion of dentinal tubules by oral bacteria. Critical Reviews in Oral Biology and Medicine 2002;13:171–183.

Arslan S, Ozbilge H, Kaya EG, Er O. In vitro antimicrobial activity of propolis, BioPure MTAD, sodium hypochlorite, and chlorhexidine on Enterococcus faecalis and Candida albicans. Saudi Med. J 2011;32:479-483.

Ramani N, Mathew S. Comparative evaluation of antimicrobial efficacy of chlorhexidine digluconate and propolis when used as intracanal medicament. JIOH 2012;4:17-23.

Trope M, Bergenholtz G. Microbiological basis for endodontic treatment: can a maximal outcome be achieved in one visit? Endodontic Topics 1. 2002; 40–53.

Suzuki S, Masuda Y, Morisaki H, et al. The study of chitosan-citrate solution as a root canal irrigant: a preliminary report. J Oral Hyg Health 2014;2:142.

Clegg MS, Vertucci FJ, Walker C, Belanger M, Britto LR. The effect of exposure to irrigant solutions on apical dentin biofilms in vitro. J Endod 2006;32:434–437.

Garg P, Tyagi SP, Sinha DJ et al. Comparison of antimicrobial efficacy of propolis, morinda citrifolia, Azadirachta indica, triphala, green tea polyphenols and 5.25% sodium hypochlorite against Enterococcus fecalis biofilm. Saudi Endod J 2014;4:122-127.

Pascon FM, Kantovitz KR, Sacramento PA et al. Effect of sodium hypochlorite on dentine mechanical properties. A review. J Dent 2009;37:903–908.

Zehnder M. Root canal irrigants. J Endod 2006;32:389–398.

Susan AC, Bharathraj AR, Praveen M, Kumar NS, Karunakaran JV. Intraradicular smear removal efficacy oftriphala as a final rinse solution in curved canals: A scanning electron microscope study. J Pharm Bioallied Sci 2019;11:420‑428.

Pratishta J, Ranjan M. Role of herbs in root canal irrigation‑A review. IOSR J Pharm Biol Sci 2014;9:6‑10.

Afshan T, Parwez A, Prasanna PL, et al. Comparison of antimicrobial efficacy of herbal root canal irrigants (Azadirachta indica, Morinda citrifolia) against Enterococcus faecalis. World J Dent 2020;11:206–210.

Ambareen Z, Konde S, Raj SN, Kumar NC. Antimicrobial efficacy of herbal extracts. Int J Oral Health Dentistry 2015;1:108.

Neelakantan P, Jagannathan N, Nazar N. Ethnopharmacological approach in endodontic treatment: A focused review. Int J Drug Dev Res 2011;3:68‑77.

Ireson C, Orr S, Jones DJ. Characterization of metabolites of the chemopreventive agent curcumin in human and rat hepatocytes and in the rat in vivo, and evaluation of their ability to inhibit phorbol ester-induced prostaglandin E2 production. Cancer Res 2001;61:1058–1064.

Jamalzadeh L, Ghafoori H, Sariri R, Rabuti H, Nasirzade J, Hasani H, et al. Cytotoxic Effects of some common organic solvents on MCF-7, RAW-264.7 and human umbilical vein endothelial cells. Avicenna J Med Biochem 2016;4:e33453.

Lakshmi T, Krishnan V, Rajendran R, et al. Azadirachta indica: a herbal panacea in dentistry – an update. Pharmacogn Rev 2015;9:41–44.

Biswas K, Chattopadhyay I, Banerjee RK, Bandyopadhyay U. Biological activities and medicinal properties of neem (Azadirachta indica). Curr Sci 2002;82:1336‑1345.

John P, Gopalakrishnan, Dinesh K, Romel J. Herbal root canal irrigants: A review. J Odontol Res 2015;3:9‑14.

Sundaram D, Narayanan K, Vadakkepurayil K. A Comparative evaluation on antimicrobial effect of honey, neem leaf extract and sodium hypochlorite as intracanal irrigant: An ex‑vivo study. J Clin Diagn Res 2016;10:88‑91.

Jose J, Shoba K, Aman S, et al. Comparative evaluation of antimicrobial activity of green tea extract, garlic extract, neem leaf extract and sodium hypochlorite as root canal irrigants against E. faecalis and C. albicans an in vitro study. Int J Curr Microbiol Appl Sci 2015;4:384‑391.

Karunaratne DN. Nanotechnology in medicine. J Natn Sci Foundation Sri Lanka 2007;35:149-152.

Mima EG, Pavarina AC, Dovigo LN, et al. Susceptibility of Candida albicans to photodynamic therapy in a murine model of oral candidosis. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;109:392-401.

Sjostrom, B. and B. Bergenståhl. Preparation of submicron drug particles in lecithin-stabilized o/w emulsions I. Model studies of the precipitation of cholesterylacetate. Int. J. Pharm 1992;88:53–62.

Siekmann, B. and Westesen K. Investigations on solid lipid nanoparticles prepared by precipitation in o/w emulsions. Eur. J. Pharm. Biopharm 1996;43:104-109.

Asnawi, S., A. R. Abd. Aziz and A. K. Khamis. Formulation of geranium oil loaded solid lipid nanoparticles for mosquito repellent application. Jor J. Chem. Natural, Res. Engineering 2008;2:90-99.

Adel, M.M., Yoseif, N.S and Ibrahim, S.S: Laboratory and field studies of geranium oil and GO-SLNs against the cotton leaf worm spodoptera littoralis (Boisd.) (Lep. Noctuidae). International Review of Humanities and Scientific Research by International Scientific Indexing ISSN (Online): 2519-5336.

El-boraey, A. N., Abo-Almaged, H. H., El-Ashmawy, A. et al. Biological and mechanical properties of denture base material as a vehicle for novel hydroxyapatite nanoparticles Loaded with Drug. Advanced Pharmaceutical Bulletin 2021;11:86.

Sultan, M., Nagieb, Z. A., El-Masry, H. M., & Taha, G. M. Physically-crosslinked hydroxyethyl cellulose-g-poly (acrylic acid-co-acrylamide)-Fe3+/silver nanoparticles for water disinfection and enhanced adsorption of basic methylene blue dye. International Journal of Biological

Macromolecules 2022;196:180-193.

El-Anssary, A. A., Abdel Raoof, G. F., Saleh, D. O., & ElMasry, H. M.Bioactivities, physicochemical parameters and GC/MS profiling of the fixed oil of Cucumis melo L. seeds: A focus on anti-inflammatory, immunomodulatory, and antimicrobial activities. J Herbmed Pharmacol 2021;10:476-485.

Mah TFC, O Toole GA. Mechanisms of biofilm resistance to antimicrobial agents. Trends Microbiol 2001;8:34–39.

Kho P, Baumgartner JC. A comparison of the antimicrobial efficacy of NaOCl/EDTA against Enterococcus faecalis. J Endod 2006;32:652-655.

M.-K. Wu, P. R. Wesselink, and R. E. Walton. Apical terminus location of root canal treatment procedures. Oral Surg Oral Med Oral Pathol Oral Radiol 2000;89:99–103.

Panchal V, Gurunathan D, Muralidharan NP. Comparison of antibacterial efficacy of cinnamon extract, neem extract as irrigant and sodium hypochlorite against enterococcus fecalis: An in vitro study. Indian J Dent Res 2020;31:124-128.

Balasenthil S, Arivazhagan S, Ramachandran CR, Ramachandran V, Nagini S. Chemopreventive potential of neem (Azadirachta indica) on 7, 12-dimethylbenz (a) anthracene (DMBA) induced hamster buccal pouch carcinogenesis. J Ethnopharmacol 1999;67:189–195.

Arun P, Pare P. A review: Antimicrobial activity of Azadirachta indica (Neem). Int J of Pharm Life Sci 2018;9:5755-5756.

Dubey S, Chaodary M, Gupta P. Comparative study of the antimicrobial efficiency of neem leaf extract, sodium hypochlorite and biopure MTAD - An in vitro study. IJDA 2012; 4;740-43.

Mustafa M. Antibacterial efficacy of neem (Azadirachta indica) extract against Enterococcus faecalis: an in vitro study. J Contemp Dent Pract 2016;17:791‑794.

Kamath U, Sheth H, Ramesh S, Singla K .Comparison of the antibacterial efficacy of tea tree oil with 3% sodium hypochlorite and 2% chlorhexidine against E. faecalis: An in vitro J Contemp Dent 2013;3;117-120.

Thosar N, Basak S, Bahadure R N, Rajurkar M. Antimicrobial efficacy of five essential oils against oral pathogens: An in vitro study. Eur J Dent 2013;7:71–77.

Vinothkumar TS, Rubin MI, Balaji L, Kandaswamy D. In vitro evaluation of five different herbal extracts as an antimicrobial endodontic irrigant using real time quantitative polymerase chain reaction. J Conserv Dent 2013;16:167-170.

Kumar A., A. S. Sarthaj, and S. M. Antony. Comparative evaluation of antibacterial and layer removal efficacy of two different herbal irrigants: an in vitro study. Int Healthc Res J. 2018;1:350–354.

Chandrappa PM, Dupper A, Tripathi P. et al. Antimicrobial activity of herbal medicines (tulsi extract, neem extract) and chlorhexidine against Enterococcus faecalis in endodontics: an in vitro study. J Int Soc Prev Community Dent 2015;5:89-92.

Rafi Shaik M, Sharaf MAF, Li X, Yousuf S, Pan SS. In vitro antimicrobial activity and comparison of the herbal extracts and sodium hypochlorite against primary plaque colonizers. FEMS Microbiol Lett 2021;3:368-371.

Dumani A, Yoldas O, Yilmaz S et al. In vitro susceptibility of E. faecalis and C. albicans isolates from apical periodontitis to common antimicrobial agents, antibiotics and antifungal medicaments. J Clin Exp Dent 2012;4:1–7.

Saxena D, Saha SG, Saha MK, Dubey S, Khatri M. An in vitro evaluation of antimicrobial activity of five herbal extracts and comparison of their activity with 2.5% sodium hypochlorite against Enterococcus faecalis. Indian J Dent Res 2015;26:524-527.

Tatekalva P, Subbaiyan H, Kumar SR. Comparative evaluation of antimicrobial potential of herbal extracts on Streptococcus mutans and Enterococcus faecalis: An in vitro study. Braz. Dent. Sci 2021;24:1-7.

Babaji P, Jagtap K, Lau H, et al. Comparative evaluation of antimicrobial effect of herbal root canal irrigants (Morinda citrifolia, Azadirachta indica,Aloe vera) with sodium hypochlorite: Anin vitro study. J Int Soc Prev Community Dent 2016;6:196-199.

Dutta A, Kundabala M. Comparative anti‑microbial efficacy of Azadirachta indica irrigant with standard endodontic irrigants: A preliminary study. J Conserv Dent 2014;17:133‑137.

Sinha JD, Nandha K, Jaiswal N, Vasudeva A, Tyagi SP, Singh UP. Antibacterial effect of Azadirachta indica (Neem) or Curcuma longa (Turmeric) against Enterococcus faecaliscompared with that of 5% sodium hypochlorite or 2% chlorhexidine in vitro. Bull Tokyo Dent Coll 2017;58:103‑109.

Ghonmode WN, Balsaraf OD, Tambe VH, et al. Comparison of the antibacterial efficacy of neem leaf extracts, grape seed extracts and 3% sodium hypochlorite against E. faecalis - an in vitro study. J Int Oral Health 2013;5:61-66.

Hegde V, Kesaria DP. Comparative evaluation of antimicrobial activity of neem, propolis, turmeric, liquorice and sodium hypochlorite as root canal irrigants against Enterococcus faecalis and Candida albicans‑an in vitro study. Endodontology 2013;25:38‑45.

Damre PG. Comparative evaluation of antimicrobial activity of herbal vs chemical root canal irrigants against Enterococcus faecalis‑an in vitro study. Int J Adv Res 2015;3:1563‑1572.

Podar R, Kulkarni GP, Dadu SS, et al. In vivo antimicrobial efficacy of 6% Morinda citrifolia, Azadirachta indica, and 3% sodium hypochlorite as root canal irrigants. Eur J Dent 2015;9:529–534.

Bohora A, Hegde V, Kokate S. Comparison of the antibacterial efficiency of neem leaf extract and 2% sodium hypochlorite against Enterococcus faecalis, Candida albicans and mixed culture‑an in vitro study. Endodontology 2010;22:8‑12.

Tyagi SP, Sinha DJ, Garg P, et al. Comparison of antimicrobial efficacy of propolis, morinda citrifolia, Azadirachta indica (neem) and 5% sodium hypochlorite on Candida albicans biofilm formed on tooth substrate: An in‑vitro study. J Conserv Dent 2013;16:532‑535.

Bhargava KY, Aggarwal S, Kumar T, Bhargava S. Comparative evaluation of the efficacy of three anti‑oxidants vs NaOCl and EDTA: used for root canal irrigation in smear layer removal‑SEM study. Int J Pharm Pharm Sci 2015;7:366‑371.

Singh M, Singh S, Salgar AR, et al. An In vitro comparative evaluation of Antimicrobial Efficacy of propolis, morinda Citrifolia Juice, sodium hypochlorite and chlorhexidine on Enterococcus faecalis and Candida albicans. J Contemp Dent Pract 2019;20:40-45.

Raghavendra SS, Balsaraf KD. Antifungal efficacy of Azadirachta indica (neem)-An in vitro study. Brazilian Journal of Oral Sciences 2014;13:242-245.

Dedhia J, Mukharjee E, Luke AM, Mathew S, Pawar AM. Efficacy of andrographis paniculata compared to Azadirachta indica, Curcuma longa, and sodium hypochlorite when used as root canal irrigants against Candida albicans and Staphylococcus aureus: An in vitro antimicrobial study. J Conserv Dent 2018;21:642-645.

Sainudeen S, Nair VS, Zarbah M, et al. Can herbal extracts serve as antibacterial root canal irrigating solutions? antimicrobial efficacy of tylophora indica, curcumin longa, Phyllanthus amarus, and sodium hypochlorite on Enterococcus faecalis biofilms formed on tooth substrate: In Vitro Study. J Pharm Bioallied Sci 2020;12:423-429.

Salem WH, Moussa TA, Abou Abouraya NL. Cytotoxicity and antibacterial activity of bleached turmeric ahydroalcoholic extract versus sodium hypochlorite as an endodontic irrigant: in vitro study. Braz. Dent. Sci 2022;19:25.

Chaitanya BV, Somisetty KV, Diwan A, Pasha S, Shetty N, Reddy Y, Nadigar S. Comparison of antibacterial efficacy of turmeric Extract, morinda citrifolia and 3% sodium hypochlorite on Enterococcus faecalis: An In-vitro Study. J Clin Diagn Res 2016;10:55-57.

Suvarna R, Bhat SS, Hegde KS. Antibacterial activity of turmeric against Enterococcus faecalis‑An in vitro study. Int J Curr Microbiol App Sci 2014;3:498-504.

Hegde MN, Shetty S, Mahalaxmi Y, Patil AB. Evaluation of antimicrobial activity of aqueous and hydroalcoholic Curcuma longa extracts against endodontic pathogens. IOSR Journal of Pharmacy 2012;2:192-198.

Neelakantan P, Subbarao C, Subbarao CV. Analysis of antibacterial activity of curcumin against Enterococcus faecalis. Int J Curr Res Rev 2011;3:37‑42.

Kumar H. An in vitro evaluation of the antimicrobial efficacy of Curcuma longa, tachyspermum ammi, chlorhexidine gluconate, and calcium hydroxide on Enterococcus faecalis. J Conserv Dent 2013;16:144-147.

Lanicotti R, Gianotti A, Patrigani N, Belleti N, Guerzoni ME, Gardini F. Use of natural aroma compounds to improve shelf life of minimally processed fruits. Trends Food Sci Technol 2004;15:201‑208.

Eskandarinezhad M, Soroush Barhaghi MH, Allameh K, Sadrhaghighi A, Katebi K. The comparison of calcium hydroxide, curcumin, and Aloe vera antibacterial effects on 6-week-old Enterococcus faecalis biofilm as an intracanal medicament: An in vitro study. Dent Res J 2022;19:14.

Tyagi P, Singh M, Kumari H, Kumari A, Mukhopadhyay K. Bactericidal activity of curcumin I is associated with damaging of bacterial membrane. PLoS One. 2015;10:e0121313.

Chainani-Wu N. Safety and anti-inflammatory activity of curcumin: a component of turmeric. J Int Oral Health2003;9:161–168.

Menni AC, Ambati NR, Prasad MG. Clinical and radiographic evaluation of Curcumin Gel mixed Endoflas (CGE) powder as obturating material in primary molars. Braz. Dent. Sci 2020;23:1-8.

Gul N, Mujahid TY, Jehan N, Samia A. Studies on the antibacterial effect of different fractions of Curcuma longaagainst urinary tract infection isolates. Pak J Biol Sci 2004;7:2055–2060.

Rai D, Singh JK, Roy N, Panda D. Curcumin inhibits FtsZ assembly: an attractive mechanism for its antibacterial activity. Biochemical J 2008;410:147-155.

Swapnil SM, Sharma A, Shah N, Mandlik J, Ghogare AA. Antimicrobial efficacy of triphala and curcumin extract in comparison with calcium hydroxide against E. faecalis as an intracanal medicament a study. Paripex Indian J Res 2017;6:876-879.

ArianiN, AliM, NoorhamdaniN, Aulanni’amA, et al. Curcumin and Fluconazole to Resolve Fluconazole-resistant Candida albicans infection in HIV Patient. Trop J Nat Prod Res 2022; 6:1930-1935.