Investigation of Antimicrobial Properties of Mangifera indica L. Kernel Extract on Multispecies Enterococcus faecalis-Candida albicans Biofilm in an Ex Vivo Tooth Model.

Main Article Content

Warat Leelapornpisid
Worrapan Poomanee

Abstract

Objectives: This study aimedto develop a multi-species biofilm model that was representative of a root canal environment within dentinal tubules and to assess the antimicrobial efficacy of kernel extractderived from Mangifera indica L.grown in Northern Thailand (Kaew-Moragot cultivar) against multispecies Entercoccus faecalis-Candida albicansbiofilms in tooth model.


Methods: Agar diffusion and broth microdilution methods were performed to investigate the antibacterial activities of M. indica kernel extract against planktonic E. faecalisand C. albicans. Time-kill assay was conducted to evaluate time-point of the extract which showed bactericidal and fungicidal effects. Antimicrobial activity of M. indica kernel extract against E. faecalisand C. albicansbiofilms was investigated through the use of 25 mg/mL extract solutionand compared to traditional root canal medicaments. In order to form biofilms in root canal, the microbial suspension of E. faecalisand C. albicansin 100:1 was placed in the dental root canal, with the addition of media every other day for 21 days The tooth samples were divided into 3groups (n=40) according to the type of medicaments which were 0.009 mg/mL normal saline solution (as a negative control), 400 mg/mL calcium hydroxide, and 25 mg/ml M. indica kernel extract. The root canal medicaments were consequently applied onto the biofilms for 7 days.After 7 days, cylindrical steel burs with a diameter of 1.6 mm and 1.8 mm were used to collect samples from inner dentin and deeper dentin. The remaining tooth samples were collected as residual roots. The samples were cultured for 24 h, and 200 μL of suspension were place into 96-well microtiter plates for OD450measurement. The OD data were analyzed with two-way ANOVAand Tukey’s post hoc tests (p<0.05).


Results: M. indica kernel extract demonstrated the antimicrobial activity against multispecies E. faecalis-C. albicans in root canal.The extract was the most effective in eliminated the biofilms by 30.88% (p<0.001)in inner dentine layer. It was comparable tocalcium hydroxide. In residual root, the eliminating effects of calcium hydroxide and M. indica kernel extract were comparable with percent inhibitions of 30.45% and 26.99%, respectively.


Conclusions: M. indica kernel extract had potential to eliminate multispecies E. faecalis-C. albicansbiofilms up to the residual root. The antimicrobial activity of the extract was comparable to calcium hydroxide, which is a gold standard medicament used in root canal treatment.


 

Article Details

How to Cite
Leelapornpisid, W., & Poomanee , W. . (2021). Investigation of Antimicrobial Properties of Mangifera indica L. Kernel Extract on Multispecies Enterococcus faecalis-Candida albicans Biofilm in an Ex Vivo Tooth Model. Chiang Mai Dental Journal, 42(2), 65–74. Retrieved from https://he01.tci-thaijo.org/index.php/cmdj/article/view/247551
Section
Original article

References

Nair P. On the causes of persistent apical periodontitis: a review. Int Endod J 2006; 39(4): 249-281.

Torabinejad M, Corr R, Handysides R, Shabahang S. Outcomes of nonsurgical retreatment and endodontic surgery: a systematic review. J Endod 2009; 35(7): 930-937.

Siqueira Jr JF. Endodontic infections: concepts, paradigms, and perspectives. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2002; 94(3): 281-293.

Ricucci D, Siqueira Jr JF. Biofilms and apical periodontitis: study of prevalence and association with clinical and histopathologic findings. J Endod 2010; 36(8): 1277-1288.

Ricucci D, Siqueira Jr JF, Bate AL, Ford TRP. Histologic investigation of root canal–treated teeth with apical periodontitis: a retrospective study from twenty-four patients. J Endod 2009; 35(4): 493-502.

Gopikrishna V, Kandaswamy D, Jeyavel RK. Comparative evaluation of the antimicrobial efficacy of five endodontic root canal sealers against Enterococcus faecalis and Candida albicans. J Conserv Dent 2006; 9(1): 2.

Romano JD, Kolter R. Pseudomonas-Saccharomyces interactions: influence of fungal metabolism on bacterial physiology and survival. J Bacteriol 2005; 187(3): 940-948.

Seneviratne C, Jin L, Samaranayake L. Biofilm lifestyle of Candida: a mini review. Oral Dis 2008; 14(7): 582-590.

Kennedy J, Hussey D. The antimicrobial effects of root canal irrigation and medication. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2007; 103(4): 560-569.

Haapasalo M, Shen Y. Current therapeutic options for endodontic biofilms. Endod Topics 2010; 22(1): 79-98.

Vera J, Siqueira Jr JF, Ricucci D, et al. One-versus two-visit endodontic treatment of teeth with apical periodontitis: a histobacteriologic study. J Endod 2012; 38(8): 1040-1052.

Safavi KE, Spngberg LS, Langeland K. Root canal dentinal tubule disinfection. J Endod 1990; 16(5): 207-210.

Barbosa-Ribeiro M, Arruda-Vasconcelos R, de-Jesus-Soares A, et al. Effectiveness of calcium hydroxide-based intracanal medication on infectious/inflammatory contents in teeth with post-treatment apical periodontitis. Clin Oral Investig 2019; 23(6): 2759-2766.

Shah K, Patel M, Patel R, Parmar P. Mangifera indica (mango). Pharmacogn Rev 2010; 4(7): 42.

Jahurul M, Zaidul I, Ghafoor K, et al. Mango (Mangifera indica L.) by-products and their valuable components: A review. Food Chem 2015; 183: 173-180.

Lauricella M, Emanuele S, Calvaruso G, Giuliano M, D’Anneo A. Multifaceted health benefits of Mangifera indica L.(Mango): the inestimable value of orchards recently planted in Sicilian rural areas. Nutrients 2017; 9(5): 525.

Parvez GM. Pharmacological activities of mango (Mangifera Indica): A review. J Pharmacognosy Phytother 2016; 5(3): 1.

Singh YN. Traditional medicine in Fiji: some herbal folk cures used by Fiji Indians. J Ethnopharmacol 1986; 15(1): 57-88.

Sairam K, Hemalatha S, Kumar A, et al. Evaluation of anti-diarrhoeal activity in seed extracts of Mangifera indica. J Ethnopharmacol 2003; 84(1): 11-15.

Rajan S, Thirunalasundari T, Jeeva S. Anti—enteric bacterial activity and phytochemical analysis of the seed kernel extract of Mangifera indica Linnaeus against Shigella dysenteriae (Shiga, corrig.) Castellani and Chalmers. Asian Pac J Trop Dis 2011; 4(4): 294-300.

Khammuang S, Sarnthima R. Antioxidant and antibacterial activities of selected varieties of Thai mango seed extract. Pak J Pharm Sci 2011; 24(1): 37-42.

Poomanee W, Chaiyana W, Mueller M, Viernstein H, Khunkitti W, Leelapornpisid P. In-vitro investigation of anti-acne properties of Mangifera indica L. kernel extract and its mechanism of action against Propionibacterium acnes. Anaerobe 2018; 52: 64-74.

Clinical, Institute LS. Reference method for broth dilution antifungal susceptibility testing of yeasts. Clinical and Laboratory Standards Institute Wayne, PA; 2008.

Clinical, Institute LS. Performance standards for antimicrobial susceptibility testing. Clinical and Laboratory Standards Institute Wayne, PA; 2017.

Keepers TR, Gomez M, Celeri C, Nichols WW, Krause KM. Bactericidal activity, absence of serum effect, and time-kill kinetics of ceftazidime-avibactam against β-lactamase-producing Enterobacteriaceae and Pseudomonas aeruginosa. Antimicrob Agents Chemother 2014; 58(9): 5297-5305.

Zhang C, Du J, Peng Z. Correlation between Enterococcus faecalis and persistent intraradicular infection compared with primary intraradicular infection: a systematic review. J Endod 2015; 41(8): 1207-1213.

Ashraf H, Samiee M, Eslami G, Hosseini MRG. Presence of Candida albicans in root canal system of teeth requiring endodontic retreatment with and without periapical lesions. Iran Endod J 2007; 2(1): 24.

Reller LB, Weinstein M, Jorgensen JH, Ferraro MJ. Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Clin Infect Dis 2009; 49(11): 1749-1755.

Cuenca-Estrella M, Gomez-Lopez A, Alastruey-Izquierdo A, et al. Comparison of the Vitek 2 antifungal susceptibility system with the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) broth microdilution reference methods and with the Sensititre YeastOne and Etest techniques for in vitro detection of antifungal resistance in yeast isolates. J Clin Microbiol 2010; 48(5): 1782-1786.

Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal 2016; 6(2): 71-79.

Pfaller M, Sheehan D, Rex J. Determination of fungicidal activities against yeasts and molds: lessons learned from bactericidal testing and the need for standardization. Clin Microbiol Rev 2004; 17(2): 268-280.

Sjögren U, Figdor D, Spångberg L, Sundqvist G. The antimicrobial effect of calcium hydroxide as a short‐term intracanal dressing. Int Endod J 1991; 24(3): 119-125.

Brändle N, Zehnder M, Weiger R, Waltimo T. Impact of growth conditions on susceptibility of five microbial species to alkaline stress. J Endod 2008; 34(5): 579-582.

Lang ML, Zhu L, Kreth J. Keeping the bad bacteria in check: interactions of the host immune system with oral cavity biofilms. Endod Topics 2010; 22(1): 17-32.

Basrani B, Tjäderhane L, Santos JM, et al. Efficacy of chlorhexidine-and calcium hydroxide–containing medicaments against Enterococcus faecalis in vitro. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2003; 96(5): 618-624.

George S, Kishen A, Song P. The role of environmental changes on monospecies biofilm formation on root canal wall by Enterococcus faecalis. J Endod 2005; 31(12): 867-872.

Zapata RO, Bramante CM, de Moraes IG, et al. Confocal laser scanning microscopy is appropriate to detect viability of Enterococcus faecalis in infected dentin. J Endod 2008; 34(10): 1198-1201.

Du T, Wang Z, Shen Y, Ma J, Cao Y, Haapasalo M. Combined antibacterial effect of sodium hypochlorite and root canal sealers against Enterococcus faecalis biofilms in dentin canals. J Endod 2015; 41(8): 1294-1298.

Sakko M, Tjäderhane L, Sorsa T, Hietala P, Rautemaa R. 2‐Hydroxyisocaproic acid is bactericidal in human dental root canals ex vivo. Int Endod J 2017; 50(5): 455-463.

Evans M, Davies JK, Sundqvist G, Figdor D. Mechanisms involved in the resistance of Enterococcus faecalis to calcium hydroxide. Int Endod J 2002; 35(3): 221-228.

Vaghela DJ, Kandaswamy D, Venkateshbabu N, Jamini N, Ganesh A. Disinfection of dentinal tubules with two different formulations of calcium hydroxide as compared to 2% chlorhexidine: As intracanal medicaments against Enterococcus faecalis and Candida albicans: An in vitro study. J Conserv Dent 2011; 14(2): 182.

Sakko M, Tjäderhane L, Sorsa T, Hietala P, Rautemaa R. Antimicrobial 2‐hydroxyisocaproic acid and chlorhexidine resist inactivation by dentine. Int Endod J 2016; 49(4): 352-360.

Haapasalo H, Sirén E, Waltimo T, Orstavik D, Haapasalo M. Inactivation of local root canal medicaments by dentine: an in vitro study. Int Endod J 2000; 33(2): 126-131.

Portenier I, Haapasalo H, Rye A, Waltimo T, Ørstavik D, Haapasalo M. Inactivation of root canal medicaments by dentine, hydroxylapatite and bovine serum albumin. Int Endod J 2001; 34(3): 184-188.

Zand V, Mokhtari H, Hasani A, Jabbari G. Comparison of the penetration depth of conventional and nano-particle calcium hydroxide into dentinal tubules. Iran Endod J 2017; 12(3): 366.

Siqueira Jr JF, Rôças IN, Alves FR, Santos KR. Selected endodontic pathogens in the apical third of infected root canals: a molecular investigation. J Endod 2004; 30(9): 638-643.

Pourhajibagher M, Ghorbanzadeh R, Bahador A. Culture-dependent approaches to explore the prevalence of root canal pathogens from endodontic infections. Braz Oral Res 2017; 31.