Sains Malaysiana 52(12)(2023): 3485-3495
http://doi.org/10.17576/jsm-2023-5212-11
Cytotoxic and Antibacterial Properties of Piper betle Combined with Selected Root Canal Sealers
(Sifat Sitotoksik dan Antibakteria Piper betle Digabungkan dengan Pengedap Saluran Akar Terpilih)
CHEN WEI
QI, HUWAINA ABD GHANI* & SUHARNI MOHAMAD
School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia
Received:
7 September 2023/Accepted: 8 December 2023
Abstrct
Exploration in endodontic material had been made to maximise the elimination of microorganisms in root canal
system to improve the treatment success rate. Inappropriate use of antibiotics
can cause antibiotic resistance hence alternative is needed. Piper betle (PB),
a medicinal herb to be used for its effective antibacterial properties against Enterococcus faecalis. Therefore,
this study aims to investigate the
cytotoxicity and antibacterial
effects of ethanolic extract of PB in combination with AH Plus (AH) and BioRoot RCS (BR) and sealers alone. Four tested materials were used: AH Plus
(AH); PB and AH plus (PBAH); BioRoot RCS (BR); and PB and BioRoot RCS (PBBR). At 24, 48, and 72 h, MTT assay on HPdLF was performed to measure the cytotoxicity.
Antibacterial effect was evaluated with modified direct contact test (MDCT)
against E. faecalis at fresh, day1-set and day-7-set state of tested materials. Data were analysed with SPSS version 25 with Tukey’s post-test at the
level of significance of p = 0.05.
PBAH showed significant lower cytotoxicity than AH (p = 0.028) at 48 h, with a higher percentage of cell viability compared AH
(26.35%). PB reduce antibacterial effect of BR at all times at significant
level (p = 0.000, 0.014, 0.032).
Conclusion, PBAH has significant reduced cytotoxicity as compared to AH on HPdLF at 48 h. PBBR was cytotoxic to HPdLF at 24 and 48 h. Both AH and PBAH exhibited antibacterial property against E. faecalis, followed by BR and PBBR. In conclusion,
PBAH is a potential enhanced root canal sealer and may be further studied to
fully elicit its profound properties.
Keywords: AH Plus; Bioroot RCS; natural product
Abstrak
Penerokaan dalam bahan endodontik telah dilakukan untuk memaksimumkan penyingkiran mikroorganisma dalam sistem saluran akar bagi meningkatkan kadar kejayaan rawatan. Penggunaan antibiotik secara tidak wajar boleh menyebabkan rintangan ketahanan terhadap antibiotik. Oleh itu, penggantian diperlukan. Piper betle (PB) ialah herba perubatan yang digunakan kerana keberkesanan sifat antibakterianya terhadap Enterococcus faecalis. Oleh itu, penyelidikan ini bertujuan untuk mengkaji kesan sitotoksik dan antibakteria ekstrak etanol PB dalam gabungan dengan AH Plus (AH) dan BioRoot RCS (BR) serta pengedap AH dan BR sahaja. Empat bahan yang diuji adalah; AH Plus (AH); PB dan AH plus (PBAH); BioRoot RCS
(BR); serta PB dan BioRoot RCS (PBBR). Asai MTT ke atas HPdLF dijalankan untuk mengukur kesitoksikan pada 24, 48 dan 72 jam. Kesan antibakteria dinilai dengan ujian sentuhan langsung diubah suai (MDCT) terhadap E. faecalis pada keadaan bahan yang diuji yang segar, set pada hari pertama dan set pada hari ketujuh. Data dianalisis menggunakan SPSS versi 25 dengan ujian pasca Tukey pada taraf keertian P = 0.05. PBAH menunjukkan kesitotoksik yang lebih rendah secara signifikan berbanding AH (P = 0.028) pada 48
jam dengan peratusan kelangsungan sel yang lebih tinggi berbanding dengan AH (26.35%). PB mengurangkan kesan antibakteria BR pada semua masa pada tahap kepentingan yang signifikan (P = 0.000, 0.014, 0.032). Kesimpulannya, PBAH mempunyai pengurangan sitotoksisiti secara signifikan berbanding dengan AH terhadap HPdLF selepas 48 jam. PBBR menunjukkan kesan sitotoksik kepada HPdLF pada 24 dan 48 jam. Kedua-dua AH atau PBAH menunjukkan sifat antibakteria terhadap E. faecalis, diikuti oleh BP dan PBBR. Kesimpulannya, PBAH adalah pengedap kanal akar yang berpotensi ditingkatkan dan boleh dikaji lebih lanjut untuk mengenal pasti sepenuhnya sifatnya yang mendalam.
Kata kunci:
AH plus; Bioroot RCS; produk semula jadi
REFERENCES
Al-Haddad, A. & Che Ab Aziz, Z.A 2016. Bioceramic-Based Root Canal Sealers: A Review. International Journal of Biomaterials 2016: 1-10. https://doi.org/10.1155/2016/9753210.
Alsubait, S., Albader, S., Alajlan, N., Alkhunaini, N., Niazy, A. & Almahdy, A. 2019.
Comparison of the antibacterial activity of calcium silicate- and epoxy
resin-based endodontic sealers against Enterococcus faecalis biofilms: A confocal laser-scanning microscopy analysis. Odontology 107(4): 513-520.https://doi.org/10.1007/s10266-019-00425-7
Alsubait, S.A., Al Ajlan, R., Mitwalli, H., Aburaisi, N., Mahmood, A., Muthurangan, M., Almadhri, R., Alfayez, M. & Anil, S. 2018. Cytotoxicity of different concentrations of three root canal sealers on human mesenchymal stem cells. Biomolecules 8(3):
68. https://doi.org/10.3390/biom8030068
Ameena Ali, Xiao, Y. Lim & Puteri F. Wahida. The fundamental study of antimicrobial
activity of Piper betle extract in commercial
toothpastes. Journal of Herbal Medicine 14: 29-34. https://doi.org/10.1016/j.hermed.2018.08.001
Azahar, N.I. Nadzirah Mohd Mokhtar & Mohd Azmir Arifin. 2020. Piper betle: A review on its bioactive compounds,
pharmacological properties, and extraction process. IOP Conference Series:
Materials Science and Engineering 999(1): 012044.
Azwanida, N.N. 2015. A review on the extraction methods use in
medicinal plants, principle, strength and
limitation. Medicinal & Aromatic Plants 4: 196. https://doi.org/10.4172/2167-0412.1000196
Bhayya, D.P, More, S.B,. Prabhat, K., Saurabh, T., Anushka, W., Aditi, M. & Anushika, S. 2021. Comparative Evaluation of the Antimicrobial Activity of Piper betel Leaf, Phyllanthus Emblica, Lawsonia Inermis Extracts as Irrigating Solutions: An In-Vitro Study. International Journal of Innovative Science and Research Technology 6(7): 1-5.
Buggapati, L. 2016. Herbs in dentistry. International Journal of Pharmaceutical
Science Invention 5(6): 7-12.
Camilleri, J. 2011. Characterization and hydration kinetics of tricalcium silicate cement for use as a dental biomaterial. Dental Materials 27(8): 836-844. https://doi.org/10.1016/j.dental.2011.04.010
Candeiro, G.T.M., Cacio, M‐N., D'Almeida‐Couto, R.S., Azambuja‐Júnior, N., Marques, M.M., Cai, S. & Gavini, G. 2016.
Cytotoxicity, genotoxicity and antibacterial
effectiveness of a bioceramic endodontic sealer. International Endodontic Journal 49(9):
858-864. https://doi.org/10.1111/iej.12523
Collado‐González, M., García‐Bernal,
D., Oñate‐Sánchez, R.E., Ortolani‐Seltenerich,
P.S., Lozano, A., Forner, L., Llena, C. & Rodríguez‐Lozano, F.J. 2017. Biocompatibility of three new calcium
silicate‐based endodontic sealers on human periodontal ligament stem
cells. International Endodontic Journal 50(9):
875-884.
Colombo, M., Poggio, C., Dagna, A., Meravini, M-V., Riva,
P., Trovati, F. & Pietrocola,
G. 2018. Biological and physico-chemical properties of new root canal sealers. Journal of Clinical and Experimental Dentistry 10(2): 120-126. https://doi.org/10.4317/jced.54548
Del Socorro, M.M.L., Bendoy, C.P. & Dacayana, C.M.L. 2014.
Cytotoxic effects of betel vine, Piper betle Linn. leaf extracts using Artemia salina leach (brine shrimp lethality assay). Journal of Multidisciplinary Studies 3(1): 100-111. https://doi.org/10.7828/jmds.v3i1.629
Eldeniz, A.U., Shehata, M., Högg, C. & Reichl, F.X. 2016.
DNA double‐strand breaks caused by new and contemporary endodontic sealers. International Endodontic Journal 49(12): 1141-1151. https://doi.org/10.1111/iej.12577
Eldeniz, A.U.,
Mustafa, K., Ørstavik, D. & Dahl, J.E. 2007.
Cytotoxicity of new resin‐, calcium hydroxide‐and
silicone‐based root canal sealers on fibroblasts derived from human
gingiva and L929 cell lines. International Endodontic Journal 40(5):
329-337. https://doi.org/10.1111/j.1365-2591.2007.01211.x
Harshitha, R.T., Ramya, V. & Bagavad Gita. 2020. Efficacy of Piper betel-Based mouthwashes on dental plaque and gingival inflammation: A
systematic review. European Journal of
Molecular & Clinical Medicine 7(3): 1978-1992.
Huang, Y., Li, X., Preeti, M., Wu, Y., Liu, L., Huihua, G. & Liu, J. 2019. The in vitro antimicrobial activities of four endodontic sealers. BMC Oral Health 19(118): 1-7. https://doi.org/10.1186/s12903-019-0817-2
Jung, S., Libricht, V., Sielker, S., Hanisch, M.R., Schäfer, E. & Dammaschke, T.
2019. Evaluation of the biocompatibility of root canal sealers on human
periodontal ligament cells ex vivo. Odontology 107(1): 54-63.
Jung, S., Sielker, S., Hanisch, M.R., Libricht, V., Schäfer, E. and Dammaschke, T.
2018. Cytotoxic effects of four different root canal sealers on human
osteoblasts. PLoS ONE 13(3): 1-14. https://doi.org/10.1371/journal.pone.0194467
Karak, S., Acharya, J., Begum, S., Mazumdar,
I., Kundu, R. & De, B. 2018. Essential oil of Piper betle L. leaves: Chemical composition, anti-
acetylcholinesterase, anti-β-glucuronidase and
cytotoxic properties. Journal of Applied
Research on Medicinal and Aromatic Plants 10: 85-92. https://doi.org/10.1016/j.jarmap.2018.06.006
Karimjee, C.K., Koka, S., Rallis,
D.M. & Gound, T.G. 2006. Cellular toxicity of
mineral trioxide aggregate mixed with an alternative delivery vehicle. Oral Surgery, Oral Medicine, Oral Pathology,
Oral Radiology, and Endodontology102(4):
e115-e20. https://doi.org/10.1016/j.tripleo.2005.12.020
Kayaoglu, G., Erten, H., Alaçam, T. & Ørstavik, D. 2005. Short‐term antibacterial activity of root canal sealers towards Enterococcus faecalis. International
Endodontic Journal 38(7): 483-488. https://doi.org/10.1111/j.1365-2591.2005.00981.x
Khalifa, L., Shlezinger, M., Beyth, S., Houri-Haddad, Y., Coppenhagen-Glazer, S., Beyth, N. & Hazan, R. 2016. Phage Therapy against Enterococcus faecalis in dental root canals. Journal of Oral Microbiology 8(1): 32157. https://doi.org/10.3402/jom.v8.32157.
Komabayashi, T., Colmenar, D., Cvach, N., Bhat,
A., Primus, C. & Imai, Y. 2020.
Comprehensive review of current endodontic sealers. Dental Materials
Journal 39(5): 703-720. https://doi.org/10.4012/dmj.2019-288
Lee, J.K., Kwak, S.W., Ha,
J-H., Lee, W.C. & Kim, H-C. 2017. Physicochemical properties of epoxy resin-based and bioceramic-based root canal
sealers. Bioinorganic Chemistry and
Applications 2017: 2582849. https://doi.org/10.1155/2017/2582849
Leonardo,
M.R., Bezerra da Silva, L.A., Filho,
M.T. & Santana da Silva, R. 1999. Release of formaldehyde by 4 endodontic sealers. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology 88(2):
221-225. https://doi.org/10.1016/s1079-2104(99)70119-8
Mak, S.T., Leong, X.F, Tew, I.M., Kumolosasi, E. & Wong, L. 2012. In vitro Evaluation of the Antibacterial Activity of Endoseal MTA, iRoot SP, and AH Plus against Planktonic Bacteria. Materials 15(6): 2012.
Nair, R., Kalariya, T. & Chanda, S. 2008. Antibacterial Activity of Some Plant Extracts Used in Folk Medicine. Journal of Herbal Pharmacotherapy 7(3-4): 191-201.
Ng Pek Leng, Nor Fadilah Rajab, Sue Mian Then, Yasmin Anum Mohd Yusof, Wan Zurinah Wan Ngah, Kar Yong Pin & Mee Lee Looi. 2014. Piper betle leaf extract enhances the cytotoxicity effect of 5-fluorouracil in inhibiting
the growth of Ht29 and Hct116 colon cancer cells. Journal of Zhejiang University 15(8): 692-700. https://doi.org/10.1631/jzus.B1300303
Nirupama, D.N., Nainan, M.T., Ramaswamy, R., Muralidharan,
S., Usha, H.H.L., Sharma, R. & Gupta, S. 2014. In
vitro evaluation of the antimicrobial efficacy of four endodontic
biomaterials against Enterococcus faecalis,
Candida albicans, and Staphylococcus aureus. International Journal of Biomaterials 2014:
383756. https://doi.org/10.1155/2014/383756
Pizzo, G., Giammanco, G.M., Cumbo, E., Nicolosi, G. & Gallina, G. 2006. In vitro antibacterial activity of
endodontic sealers. Journal of Dentistry 34(1):
35-40. https://doi.org/10.1016/j.jdent.2005.03.001
Poggio, C., Riva, P., Chiesa, M., Colombo, M. & Pietrocola, G. 2017. Comparative cytotoxicity evaluation of eight root canal
sealers. Journal of Clinical and Experimental
Dentistry 9(4): e574-e578. https://doi.org/10.4317/jced.53724
Rafi, A. 2021. In-vitro study on antibacterial,
pH and cytotoxic evaluation of calcium hydroxide, propolis and Piper betle as intracanal medicament. MSc. Thesis. Universiti Sains Malaysia
(Unpublished). http://eprints.usm.my/50408/
Rao, S.P., Byrappa, K., Keerthiraj, N., Chatterjee, J. & Mustak,
M.S. 2018. Phyto-fabrication of Zno nanoparticles using Piper betel aqueous extract and evaluation of its
applicability in dentistry. Pharmaceutical
Nanotechnology 6(3): 201-208. https://doi.org/10.2174/2211738506666180611101852
Sakagami, H., Kishino, K., Amano,
O., Kanda, Y., Kunii, S., Yokote, Y., Oizumi, H. & Oizumi, T. 2009.
Cell death induced by nutritional starvation in mouse macrophage-like Raw264. 7 cells. Anticancer Research 29(1): 343-347.
Sari, R. & Isadiartuti,
D. 2006. Studi efektivitas sediaan gel antiseptik tangan ekstrak daun sirih (Piper betle Linn.). Majalah Farmasi Indonesia 17(4): 163-169.
Schweikl, H., Schmalz, G. & Federlin, M. 1998. Mutagenicity of the root canal sealer AHPlus in the Ames Test. Clinical Oral Investigations 2(3): 125-29. https://doi.org/10.1007/s007840050057.
Sharad, C.B. 2011. Evaluation of apical sealing ability and
antibacterial efficacy against E. faecalis of
two resin based sealers mixed with propolis: An in
vitro study. Master of Dental Surgery, Tamilnadu Government Dental College and Hospital, Chennai (Unpublished).
Sharma, D., Grover, R., Pinnameneni, P.S., Dey, S. & Raju,
P.R. 2014. Evaluation of efficacy of combinations of five endodontic sealers
with five antibiotics against Enterococcus faecalis - An in-vitro study. Journal of
International Oral Health 6(2): 90-95.
Silva, E.J., Accorsi-Mendonça, T., Pedrosa, A.C., Granjeiro, J.M. & Zaia, A.A. 2016.
Long-term cytotoxicity, pH and dissolution rate of AH Plus and MTA Fillapex. Brazilian
Dental Journal 27: 419-423. https://doi.org/10.1590/0103-6440201600735
Singh, G., Gupta, I., Elshamy, F.M.M., Boreak, N. & Homeida, H.E. 2016. In vitro comparison of
antibacterial properties of bioceramic-based sealer,
resin- based sealer and zinc oxide eugenol based sealer and two mineral
trioxide aggregates. European Journal of
Dentistry 10(03): 366-369. https://doi.org/10.4103/1305-7456.184145
Subbiya, A., Kumar, E., Anuradha,
B. & Mitthra, S. 2020. Properties and clinical
application of resin based sealers: A review. European Journal of Molecular & Clinical Medicine 7(5): 1287-1292.
Taraslia, V., Anastasiadou, E., Lignou, C., Keratiotis, G., Agrafioti, A. & Kontakiotis,
E.G. 2018. Assessment of cell viability in
four novel endodontic sealers. European
Journal of Dentistry 12(02): 287-291. https://doi.org/10.4103/ejd.ejd_9_18
Tomer, A.K., Lungdin, L.C., Mushtaq, H., Tyagi, A., Ahlawat, S., Jose, T. & Khandelwal,
A. 2021. Emerging trends in endodontic irrigation. International Journal of Applied Dental Sciences 7(1): 468-471. https://doi.org/10.7759/cureus.24833
Valentão, P., Rui, F.G., Cristóvão, B., Paula, G.d.P., Paula, B.A. & Federico, F. 2010. Improving the Knowledge on Piper betle: Targeted Metabolite Analysis and Effect on Acetylcholinesterase. Journal of Separation Science 33(20): 3168-76. https://doi.org/10.1002/jssc.201000429.
Valle Jr., D.L., Cabrera, E.C., Puzon,
J.J.M. & Rivera, W.L. 2016. Antimicrobial activities of methanol, ethanol
and supercritical CO2 extracts of Philippine Piper betle L. on clinical isolates of Gram positive and Gram negative bacteria with
transferable multiple drug resistance. PLoS ONE 11(1):
e0146349. https://doi.org/10.1371/journal.pone.0146349
Wainstein, M., Morgental, R.D., Waltrick, S.B.G., Oliveira, S.D., Vier-Pelisser, F.V., Figueiredo, J.A.P., Steier, L., Tavares, C.O. & Scarparo, R.K. 2016. In vitro antibacterial activity of a silicone-based endodontic sealer and two conventional sealers. Brazilian Oral Research 30(1): S1806-83242016000100216. https://doi.org/10.1590/1807-3107BOR-2016.vol30.0018
Wuersching, S.N., Diegritz, C., Hickel, R., Huth, K.C. & Kollmuss, M. 2022. A comprehensive in vitro comparison
of the biological and physicochemical properties of bioactive root canal
sealers. Clinical Oral Investigations 26(10): 6209-6222. https://doi.org/10.1007/s00784-022-04570-2
Zeenath, A. & Chinappa, A.
2014. Go green-keep the root canal clean. International
Journal of Dental Sciences and Research 2(6B): 21-25. https://doi.org/10.12691/ijdsr-2-6B-7
Zhang, H., Shen, Y., Ruse, N.D. & Haapasalo, M. 2009. Antibacterial activity of endodontic
sealers by modified direct contact test against Enterococcus faecalis. Journal
of Endodontics 35(7): 1051-1055. https://doi.org/10.1016/j.joen.2009.04.022
Zhou, H-M., Du, T-F., Shen, Y., Wang, Z-J., Zheng,
Yu-F. & Haapasalo, M. 2015. In vitro cytotoxicity of calcium silicate–Containing endodontic sealers. Journal of
Endodontics 41(1): 56-61. https://doi.org/10.1016/j.joen.2014.09.012
*Corresponding author; email: huwaina@usm.my
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