Skip to main content Skip to main navigation menu Skip to site footer

The effect of ethanol extract of green meniran leaves (Phyllanthus niruri Linn) in a mixture of calcium hydroxide and 2% chlorhexidine digluconate on matrix metalloproteinase-9, transforming growth factor-1 levels, osteoclast, and osteoblast cells in Wist

  • pdf  |
  • Published: 2022-12-02


Background: Infections of the dental pulp and periapical tissues involve bacteria which can induce and enhance the immune response of the dental pulp but are not effective in eliminating root canal bacteria. A study was conducted to determine the effect of giving ethanol extract of green meniran leaves (Phyllanthus niruri Linn) in a mixture of calcium hydroxide and 2% chlorhexidine digluconate on the chronic inflammatory response in periapical tissue by measuring MMP-9, TGF-β1 levels, osteoclast, and osteoblast cells count.

Methods: An experimental design with a post-test-only control group design involving 24 right molars of male Wistar rats divided into 4 groups. Each group consisted of 6 rats (K-) bacteria, (K+) bacteria, calcium hydroxide and 2% chlorhexidine digluconate, (P1) bacteria, calcium hydroxide and 10% meniran extract, (P2) bacteria, calcium hydroxide, 2% chlorhexidine digluconate and 10% meniran extract}. Examination of MMP-9, TGF-β1 levels, and the number of osteoclasts and osteoblasts were evaluated until day 14.

Results: The administration of ethanol extract of green meniran leaves (Phyllanthus niruri Linn) in a mixture of calcium hydroxide as an intracanal medicament (P1) which was evaluated for 14 days overall showed a significant difference between groups (p<0.005). This condition can be seen in the group (P1), which has a lower MMP-9 level of 2465.88 pg/mL compared to the group (P2), (K+), and (K-). The levels of TGF-β1 in the group (P1) were 181.57pg/mL, lower than the group (P2), (K+), and (K-). The number of osteoclast cells in the group (P1) was 1.67, which was lower than the group (P2), (K+), and (K-) was 5.00. The number of osteoblast cells in the (P1) group was 52.67, higher than the 32.17 group (P2), (K+), and (K-).

Conclusion: Green meniran extract (Phyllanthus niruri Linn) and its role as an intracanal medicament in teeth with chronic apical periodontitis.


  1. Zero DT, Zandona AF, Vail MM, Spolnik KJ. Dental caries and pulpal disease. Dent Clin North Am. 2011;55(1):29-46. doi:10.1016/j.cden.2010.08.010.
  2. Sutasmi Y and Natsir Y. Identification of bacteria in dental root canal diagnosed with chronic apicalis periodontitis. Journal of Dentomaxillofacial Science. 2014;13(3). p182-185.
  3. Indonesia Ministry of Health. 2011. Dental and Oral Health Profile in Indonesia 2010.
  4. Gomes BP, Pinheiro ET, Sousa EL, et al. Enterococcus faecalis in dental root canals detected by culture and by polymerase chain reaction analysis. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;102(2):247-253. doi:10.1016/j.tripleo.2005.11.031.
  5. Graves DT, Oates T, Garlet GP. Review of osteoimmunology and the host response in endodontic and periodontal lesions. J Oral Microbiol. 2011;3:10.3402/jom.v3i0.5304. Published 2011 Jan 17. doi:10.3402/jom.v3i0.5304.
  6. Herrera D, Retamal-Valdes B, Alonso B, Feres M. Acute periodontal lesions (periodontal abscesses and necrotizing periodontal diseases) and endo-periodontal lesions. J Periodontol. 2018;89 Suppl 1:S85-S102. doi:10.1002/JPER.16-0642.
  7. Graunaite I, Lodiene G, Maciulskiene V. Pathogenesis of apical periodontitis: a literature review. J Oral Maxillofac Res. 2012;2(4):e1. doi:10.5037/jomr.2011.2401.
  8. Kayaoglu G, Ørstavik D. Virulence factors of Enterococcus faecalis: relationship to endodontic disease. Crit Rev Oral Biol Med. 2004;15(5):308-320. Published 2004 Sep 1. doi:10.1177/154411130401500506.
  9. Wan C, Yuan G, Yang J, et al. MMP9 deficiency increased the size of experimentally induced apical periodontitis. J Endod. 2014;40(5):658-664. doi:10.1016/j.joen.2014.01.003.
  10. Ahmed GM, El-Baz AA, Hashem AA, Shalaan AK. Expression levels of matrix metalloproteinase-9 and gram-negative bacteria in symptomatic and asymptomatic periapical lesions. J Endod. 2013;39(4):444-448. doi:10.1016/j.joen.2012.11.009.
  11. Corotti MV, Zambuzzi WF, Paiva KB, et al. Immunolocalization of matrix metalloproteinases-2 and -9 during apical periodontitis development. Arch Oral Biol. 2009;54(8):764-771. doi:10.1016/j.archoralbio.2009.04.013.
  12. Hernádi K, Gyöngyösi E, Mészáros B, et al. Elevated tumor necrosis factor-alpha expression in periapical lesions infected by Epstein-Barr virus. J Endod. 2013;39(4):456-460. doi:10.1016/j.joen.2012.12.028.
  13. Indahyani DE, Santoso A, Utoro T, and Soesatyo MH. Fish oil regulates bone sialoprotein and osteopontin in alveolar bone resorption. Joint Scientific Meeting in Dentistry. 2010. p1-24.
  14. Carneiro E, Menezes R, Garlet GP, et al. Expression analysis of matrix metalloproteinase-9 in epithelialized and nonepithelialized apical periodontitis lesions. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009;107(1):127-132. doi:10.1016/j.tripleo.2008.07.030
  15. Vaday GG, Schor H, Rahat MA, Lahat N, Lider O. Transforming growth factor-beta suppresses tumor necrosis factor alpha-induced matrix metalloproteinase-9 expression in monocytes. J Leukoc Biol. 2001;69(4):613-621.
  16. Yuanita T, Hotmaria H, Effendy R, Suardita K. TGF-B and MMP-1 expression in chronic apical periodontitis due to Enterococcus faecalis induction on Wistar rats. Conservative Dentistry Journal. 2017;7(2). p86-94. DOI:10.20473/cdj.v7i2.2017.86-94.
  17. Álvares PR, Arruda JAA, Silva LPD, Nascimento GJFD, Silveira MFD, Sobral APV. Immunohistochemical expression of TGF-β1 and MMP-9 in periapical lesions. Braz Oral Res. 2017;31:e51. doi:10.1590/1807-3107BOR-2017.vol31.0051.
  18. Kim J, Lee J. Plasma MMP-9, TIMP-1, and TGF-β1 Responses to Exercise-Induced Muscle Injury. Int J Environ Res Public Health. 2020;17(2):566. doi:10.3390/ijerph17020566.
  19. Checchi V, Maravic T, Bellini P, et al. The Role of Matrix Metalloproteinases in Periodontal Disease. Int J Environ Res Public Health. 2020;17(14):4923. doi:10.3390/ijerph17144923.
  20. Paula-Silva FW, da Silva LA, Kapila YL. Matrix metalloproteinase expression in teeth with apical periodontitis is differentially modulated by the modality of root canal treatment. J Endod. 2010;36(2):231-237. doi:10.1016/j.joen.2009.10.030.
  21. 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. doi:10.1007/s00784-018-2719-0.
  22. Sathorn C, Parashos P, Messer H. Antibacterial efficacy of calcium hydroxide intracanal dressing: a systematic review and meta-analysis. Int Endod J. 2007;40(1):2-10. doi:10.1111/j.1365-2591.2006.01197.x.
  23. Ballal NV, Shavi GV, Kumar R, Kundabala M, and Bhat KS. In Vitro Sustained Release of Calcium Ions and pH Maintenance from Different Vehicles Containing Calcium Hydroxide. Journal of Endodontics. 2010;36(5). p862-866. DOI:10.1016/j.joen.2009.12.021.
  24. Guerreiro-Tanomaru JM, Chula DG, de Pontes Lima RK, Berbert FL, Tanomaru-Filho M. Release and diffusion of hydroxyl ion from calcium hydroxide-based medicaments. Dent Traumatol. 2012;28(4):320-323. doi:10.1111/j.1600-9657.2011.01112.x.
  25. Shuping GB, Orstavik D, Sigurdsson A, Trope M. Reduction of intracanal bacteria using nickel-titanium rotary instrumentation and various medications. J Endod. 2000;26(12):751-755. doi:10.1097/00004770-200012000-00022.
  26. da Silva RA, Leonardo MR, da Silva LA, Faccioli LH, de Medeiros AI. Effect of a calcium hydroxide-based paste associated to chlorhexidine on RAW 264.7 macrophage cell line culture. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2008;106(5):e44-e51. doi:10.1016/j.tripleo.2008.06.027.
  27. Signoretti FG, Gomes BP, Montagner F, Barrichello Tosello F, Jacinto RC. Influence of 2% chlorhexidine gel on calcium hydroxide ionic dissociation and its ability of reducing endotoxin. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2011;111(5):653-658. doi:10.1016/j.tripleo.2010.11.016.
  28. Paula-Silva FW, da Silva LA, Kapila YL. Matrix metalloproteinase expression in teeth with apical periodontitis is differentially modulated by the modality of root canal treatment. Journal of Endodontics. 2010 Feb;36(2):231-237. DOI:10.1016/j.joen.2009.10.030.
  29. Martinho FC, Teixeira FF, Cardoso FG, et al. Clinical Investigation of Matrix Metalloproteinases, Tissue Inhibitors of Matrix Metalloproteinases, and Matrix Metalloproteinase/Tissue Inhibitors of Matrix Metalloproteinase Complexes and Their Networks in Apical Periodontitis. J Endod. 2016;42(7):1082-1088. doi:10.1016/j.joen.2016.04.001.
  30. Dong M, Blobe GC. Role of transforming growth factor-beta in hematologic malignancies. Blood. 2006;107(12):4589-4596. doi:10.1182/blood-2005-10-4169.
  31. Dezerega A, Madrid S, Mundi V, et al. Pro-oxidant status and matrix metalloproteinases in apical lesions and gingival crevicular fluid as potential biomarkers for asymptomatic apical periodontitis and endodontic treatment response. J Inflamm (Lond). 2012;9(1):8. Published 2012 Mar 21. doi:10.1186/1476-9255-9-8.
  32. Lundmark A, Johannsen G, Eriksson K, et al. Mucin 4 and matrix metalloproteinase 7 as novel salivary biomarkers for periodontitis. J Clin Periodontol. 2017;44(3):247-254. doi:10.1111/jcpe.12670.
  33. Raheja J, Tewari S, Tewari S, Duhan J. Evaluation of efficacy of chlorhexidine intracanal medicament on the periodontal healing of concomitant endodontic-periodontal lesions without communication: an interventional study. J Periodontol. 2014;85(8):1019-1026. doi:10.1902/jop.2014.130430.
  34. Vianna ME, Gomes BP, Berber VB, Zaia AA, Ferraz CC, de Souza-Filho FJ. In vitro evaluation of the antimicrobial activity of chlorhexidine and sodium hypochlorite. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2004;97(1):79-84. doi:10.1016/s1079-2104(03)00360-3.
  35. Athanassiadis B, Abbott PV, Walsh LJ. The use of calcium hydroxide, antibiotics and biocides as antimicrobial medicaments in endodontics. Aust Dent J. 2007;52(1 Suppl):S64-S82. doi:10.1111/j.1834-7819.2007.tb00527.x.
  36. Yeung SY, Huang CS, Chan CP, Lin HN, and Lee PH. Antioxidant and Pro-Oxidant Properties of Chlorhexidine and its Interaction with Calcium Hydroxide Solutions. Int Endod J. 2007;40(11). p837-44. DOI:10.1111/j.1365-2591.2007.01271.x.
  37. Reynaud af Geijersstam A, Culak R, Molenaar L, et al. Comparative analysis of virulence determinants and mass spectral profiles of Finnish and Lithuanian endodontic Enterococcus faecalis isolates. Oral Microbiol Immunol. 2007;22(2):87-94. doi:10.1111/j.1399-302X.2007.00327.x.
  38. Nair PN. Pathogenesis of apical periodontitis and the causes of endodontic failures. Crit Rev Oral Biol Med. 2004;15(6):348-381. DOI:10.1177/154411130401500604.
  39. Manicone AM, McGuire JK. Matrix metalloproteinases as modulators of inflammation. Semin Cell Dev Biol. 2008;19(1):34-41. doi:10.1016/j.semcdb.2007.07.003.
  40. Kumar MS, Vamsi G, Sripriya R, Sehgal PK. Expression of matrix metalloproteinases (MMP-8 and -9) in chronic periodontitis patients with and without diabetes mellitus. J Periodontol. 2006;77(11):1803-1808. DOI:10.1902/jop.2006.050293.
  41. Gomes BP, Ferraz CC, Vianna ME, et al. In vitro antimicrobial activity of calcium hydroxide pastes and their vehicles against selected microorganisms. Braz Dent J. 2002;13(3):155-161. doi:10.1590/s0103-64402002000300002.
  42. Pereira Faustino IS, Azevedo RS, Takahama A Jr. Metalloproteinases 2 and 9 Immunoexpression in Periapical Lesions from Primary Endodontic Infection: Possible Relationship with the Histopathological Diagnosis and the Presence of Pain. J Endod. 2016;42(4):547-551. doi:10.1016/j.joen.2015.12.020.
  43. Kehrl JH, Wakefield LM, Roberts AB, et al. Production of transforming growth factor beta by human T lymphocytes and its potential role in the regulation of T cell growth. J Exp Med. 1986;163(5):1037-1050. doi:10.1084/jem.163.5.1037.
  44. Stevens WW, Lee RJ, Schleimer RP, Cohen NA. Chronic rhinosinusitis pathogenesis. J Allergy Clin Immunol. 2015;136(6):1442-1453. doi:10.1016/j.jaci.2015.10.009.
  45. Bagalkotkar G, Sagineedu SR, Saad MS, Stanslas J. Phytochemicals from Phyllanthus niruri Linn. and their pharmacological properties: a review. J Pharm Pharmacol. 2006;58(12):1559-1570. doi:10.1211/jpp.58.12.0001.
  46. Cushnie TP, Lamb AJ. Recent advances in understanding the antibacterial properties of flavonoids. Int J Antimicrob Agents. 2011;38(2):99-107. doi:10.1016/j.ijantimicag.2011.02.014.
  47. Lee NY, Khoo WK, Adnan MA, Mahalingam TP, Fernandez AR, Jeevaratnam K. The pharmacological potential of Phyllanthus niruri. J Pharm Pharmacol. 2016;68(8):953-969. doi:10.1111/jphp.12565.
  48. Tjandrawinata RR, Susanto LW, and Nofiarny D. The use of Phyllanthus niruri L. as an immunomodulator for the treatment of infectious diseases in clinical settings. Asian Pacific Journal of Tropical Disease. 2017;7(3). p132-140. DOI:10.12980/apjtd.7.2017D6-287.
  49. Beg S, Swain S, Hasan H, Barkat MA, Hussain MS. Systematic review of herbals as potential anti-inflammatory agents: Recent advances, current clinical status and future perspectives. Pharmacogn Rev. 2011;5(10):120-137. doi:10.4103/0973-7847.91102.
  50. Beutler BA. TLRs and innate immunity. Blood. 2009;113(7):1399-1407. doi:10.1182/blood-2008-07-019307.
  51. Tang YQ, Jaganath IB, Manikam R, Sekaran SD. Phyllanthus spp. Exerts Anti-Angiogenic and Anti-Metastatic Effects Through Inhibition on Matrix Metalloproteinase Enzymes. Nutr Cancer. 2015;67(5):783-795. doi:10.1080/01635581.2015.1040518.
  52. Tibúrcio-Machado CS, Michelon C, Zanatta FB, Gomes MS, Marin JA, Bier CA. The global prevalence of apical periodontitis: a systematic review and meta-analysis. Int Endod J. 2021;54(5):712-735. doi:10.1111/iej.13467.
  53. Metzger Z, Huber R, Slavescu D, Dragomirescu D, Tobis I, Better H. Healing kinetics of periapical lesions enhanced by the apexum procedure: a clinical trial. J Endod. 2009;35(2):153-159. doi:10.1016/j.joen.2008.11.019.
  54. Arruda MEF, Neves MAS, Diogenes A, et al. Infection Control in Teeth with Apical Periodontitis Using a Triple Antibiotic Solution or Calcium Hydroxide with Chlorhexidine: A Randomized Clinical Trial. J Endod. 2018;44(10):1474-1479. doi:10.1016/j.joen.2018.07.001.
  55. Tervit C, Paquette L, Torneck CD, Basrani B, Friedman S. Proportion of healed teeth with apical periodontitis medicated with two percent chlorhexidine gluconate liquid: a case-series study. J Endod. 2009;35(9):1182-1185. doi:10.1016/j.joen.2009.05.010.

How to Cite

Widiadnyani, N. K. E., I Nyoman Mantik Astawa, I Wayan Putu Sutirta Yasa, I Dewa Made Sukrama, I Made Bakta, Ida Bagus Putra Manuaba, I Wayan Suardana, & I Made Sudarmaja. (2022). The effect of ethanol extract of green meniran leaves (Phyllanthus niruri Linn) in a mixture of calcium hydroxide and 2% chlorhexidine digluconate on matrix metalloproteinase-9, transforming growth factor-1 levels, osteoclast, and osteoblast cells in Wist. Bali Medical Journal, 11(3), 1870–1878. Retrieved from




Search Panel

Ni Kadek Eka Widiadnyani
Google Scholar
BMJ Journal

I Nyoman Mantik Astawa
Google Scholar
BMJ Journal

I Wayan Putu Sutirta Yasa
Google Scholar
BMJ Journal

I Dewa Made Sukrama
Google Scholar
BMJ Journal

I Made Bakta
Google Scholar
BMJ Journal

Ida Bagus Putra Manuaba
Google Scholar
BMJ Journal

I Wayan Suardana
Google Scholar
BMJ Journal

I Made Sudarmaja
Google Scholar
BMJ Journal