IN VITRO ANTIMICROBIAL ACTIVITY OF PUNICA GRANATUM L. EXTRACT AND ANTI-INFLAMMATORY ACTIVITY OF NANO GEL IN VIVO INWISTAR RATS GINGIVA

ARMIA SYAHPUTRA; IRMA ERVINA; AIDA FADHILLA DARWIS; HAFID SYAHPUTRA; EMI SARAH NOVITA MARPAUNG; INDAH JAYANTI SITORUS; REZA ADITYA

doi:10.22159/ijap.2025v17i3.53619

Authors

ARMIA SYAHPUTRA Department of Periodontology, Faculty of Dentistry, Universitas Sumatera Utara, Medan-20115 Indonesia https://orcid.org/0000-0002-6474-897X
IRMA ERVINA Department of Periodontology, Faculty of Dentistry, Universitas Sumatera Utara, Medan-20115 Indonesia https://orcid.org/0000-0002-8705-4022
AIDA FADHILLA DARWIS Departement of oral medicine, faculty of dentistry, Universitas Sumatera Utara, Medan, north sumatera Indonesia
HAFID SYAHPUTRA Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Sumatera Utara, Medan-20115 Indonesia https://orcid.org/0000-0001-6628-7727
EMI SARAH NOVITA MARPAUNG Department of Periodontology, Faculty of Dentistry, Universitas Sumatera Utara, Medan-20115 Indonesia https://orcid.org/0009-0001-5240-9399
INDAH JAYANTI SITORUS Department of Periodontology, Faculty of Dentistry, Universitas Sumatera Utara, Medan-20115 Indonesia
REZA ADITYA Department of Periodontology, Faculty of Dentistry, Universitas Sumatera Utara, Medan-20115 Indonesia

DOI:

https://doi.org/10.22159/ijap.2025v17i3.53619

Keywords:

Anti-inflammatory, Antimicrobial activity, Gingivitis, Nano gel, Punica granatum L

Abstract

Objective: This study evaluates the antimicrobial activity of pomegranate (Punica granatum L.) peel extract in vitro and the anti-inflammatory effects of its nano gel formulation in vivo on gingival inflammation in male wistar rats.

Methods: The antimicrobial activity was tested against Aggregatibacteractinomycetemcomitans, Porphyromonasgingivalis, and Fusobacterium nucleatum using ethanol, ethyl acetate, and n-hexane extracts at concentrations of 0.8–25%. The anti-inflammatory effect was assessed using pomegranate peel extract nano gel (6.25%, 12.5%, and 25%) in a ligature-induced gingivitis model in Wistar rats, with Gengigel as a positive control. Evaluation parameters included inhibition zone diameters for antimicrobial activity, while histological analysis of lymphocyte, neutrophil, and macrophage counts was conducted on days 3 and 7 to assess anti-inflammatory effects.

Results: The 25% ethanol extract exhibited the highest antimicrobial inhibition zones (16.95 mm for A. actinomycetemcomitans, 16.83 mm for P. gingivalis, and 16.35 mm for F. nucleatum, p<0.05). The 25% nano gel formulation significantly reduced inflammatory cell counts compared to the control (p<0.05), showing reductions in lymphocytes (4.33±1.53), neutrophils (1.00±0.00), and macrophages (1.67±0.57) by day 7.

Conclusion: Pomegranate peel extract demonstrated potent antimicrobial properties, especially in ethanol-based formulations. The nano gel formulation exhibited significant anti-inflammatory effects, with the 25% concentration proving the most effective in reducing gingival inflammation. These findings highlight its potential as a natural therapeutic agent for periodontal disease management.

References

Dioguardi M, Ballini A, Sovereto D, Spirito F, Cazzolla AP, Aiuto R. Application of the extracts of Punica granatum in oral cancer: scoping review. Dent J (Basel). 2022;10(12):234. doi: 10.3390/dj10120234, PMID 36547050.

Moga MA, Dimienescu OG, Balan A, Dima L, Toma SI, Bigiu NF. Pharmacological and therapeutic properties of Punica granatum phytochemicals: possible roles in breast cancer. Molecules. 2021;26(4):1054. doi: 10.3390/molecules26041054, PMID 33671442.

Lampakis D, Skenderidis P, Leontopoulos S. Technologies and extraction methods of polyphenolic compounds derived from pomegranate (punica granatum) peels a mini-review. Processes. 2021;9(2):236. doi: 10.3390/pr9020236.

Masfria M, Sumaiyah S, Syahputra H, Jenifer J. Anti-inflammatory and antiaging activity of hydrogel with active ingredient Phyllantusemblica L. fruit nanosimplicia. J Med Plants. 2023;22(28):49-64.

Diyatri I, Juliastuti WS, Ridwan RD, Ananda GC, Waskita FA, Juliana NV. Antibacterial effect of a gingival patch containing nanoemulsion of red dragon fruit peel extract on porphyromonas gingivalis aggregatibacter actinomycetemcomitans and fusobacterium nucleatum assessed in vitro. J Oral Biol Craniofac Res. 2023;13(3):386-91. doi: 10.1016/j.jobcr.2023.03.011.

Bienvenue D, Bong D, Brian N, Staphane J, Hortense G, Charles M. In vitro evaluation of the efficacy of an aqueous extract of Allium sativum as an antibacterial agent on three major periodontal pathogen. J Oral Dent Health Res. 2021;3(1):2121.

Bamashmous S, Kotsakis GA, Kerns KA, Leroux BG, Zenobia C, Chen D. Human variation in gingival inflammation. Proc Natl Acad Sci USA. 2021;118(27):e2012578118. doi: 10.1073/pnas.2012578118, PMID 34193520.

Garcia Garcia ML, Tovilla Zarate CA, Villar Soto M, Juarez Rojop IE, Gonzalez Castro TB, Genis Mendoza AD. Fluoxetine modulates the pro-inflammatory process of IL-6, IL-1β and TNF-α levels in individuals with depression: a systematic review and meta-analysis. Psychiatry Res. 2022;307:114317. doi: 10.1016/j.psychres.2021.114317, PMID 34864233.

Teixeira QE, Ferreira D DE C, DA Silva AM, Goncalves LS, Pires FR, Carrouel F. Aging as a risk factor on the immunoexpression of pro-inflammatory IL-1β, IL-6 and tnf-α cytokines in chronic apical periodontitis lesions. Biology. 2021;11(1):14. doi: 10.3390/biology11010014, PMID 35053012.

Fernandes T, D Souza AD, Sawarkar SP. Ocimum sanctum L: promising agent for oral health care management. Nat Oral Care Dent Ther. 2020:ch16. doi: 10.1002/9781119618973.

Rosaini H, Halim A, Sandi IE, Makmur I, Asra R, Sidoretno WM. Formulation and antioxidant activity of nano gel ethanol extract of kepok banana peel (musa X paradisiaca L.). Int Res J Pharm. 2021;12(1):21-5. doi: 10.7897/2230-8407.1201112.

Alotaibi G, Alharthi S, Basu B, Ash D, Dutta S, Singh S. Nano gels: recent advancement in fabrication methods for mitigation of skin cancer. Gels. 2023;9(4):331. doi: 10.3390/gels9040331, PMID 37102943.

Masfria S, Hafid Syahputra S. Antibacterial and anti-inflammatory activities of nanosimpliciaphyllantusemblica L. fruit in suspension formulation. Int J Appl Pharm. 2025;17(2). doi: 10.22159/ijap.2025v17i2.52608.

Masfria, Sumaiyah, Syahputra H, Fani V. Antiinflammation and antifungal effects of silver nanoparticles greenly synthesized using phyllanthus emblica L. extract as a reducing agent against dermatophytosis. International Journal of Applied Pharmaceutics. 2024;16(4):29. doi: 10.22159/ijap.2024.v16s4.06.

Evaluation of anti-inflammatory activity of solanum nigrum infused selenium nanoparticles an in vitro study. Int J Pharm Res. 2020;12:(sp1). doi: 10.31838/ijpr/2020.sp1.426.

Kumaran H, Gajendran PL, Arjunkumar R, Rajesh S. Anti inflammatory activity of lycopene and its based chitosan nanoformulation an in vitro study. Plant Cell Biotechnol Mol Biol. 2020;21:(51-2):91-100.

Masfria M, Sumaiyah S, Syahputra H, Witarman M. Formulation and evaluation of antibacterial and anti-inflammatory capsules containing phyllanthus emblica L. fruit nanoparticles. Sci Technol Indones. 2023;8(4):607-15. doi: 10.26554/sti.2023.8.4.607-615.

Yulistina Y. Potential nanogel extract of advocate fruit (Persea americana Mill.) as alternative in prevention of inflammation in white rats white rats post-extraction wounds. Journal Research of Social Science. Econ Manag. 2022;2(2). doi: 10.59141/jrssem.v2i02.293.

Amalia M, Nisa Brkemit GM, Kahfi M, Syahputra A, Ervina I, Nasution RO. In vitro evaluation of andaliman (Zanthoxylum acanthopodium DC) extract on the growth of periodontal pathogenic bacteria. Malaysian Journal of Microbiology. 2023;19(5). doi: 10.21161/MJM.220110.

Dimitriu T, Bolfa P, Daradics Z, Suciu S, Armencea G, Catoi C. Ligature-induced periodontitis causes atherosclerosis in rat descending aorta: an experimental study. Med Pharm Rep. 2021;94(1):106-11. doi: 10.15386/mpr-2044, PMID 33629057.

Pujiastuti P, Sakinah NN, DA AT AYM, Wahyukundari MA, Praharani D, Sari DS. The potential of toothpaste containing robusta coffee bean extract in reducing gingival inflammation and dental plaque formation. Dent J. 2023;56(2):109-14. doi: 10.20473/J.DJMKG.V56.I2.

Garcia VG, Miessi DM, Esgalha DA, Rocha T, Gomes NA, Nuernberg MAA, Cardoso J DE M, Ervolino E, Theodoro LH. The effects of Lactobacillus reuteri on the inflammation and periodontal tissue repair in rats: a pilot study. Saudi Dental Journal. 2022;34(6):516-26. doi: 10.1016/j.sdentj.2022.05.004.

Suryono S, Wulandari F, Andini H, Widjaja J, Nugraheni T. Methodology in wistar rats periodontitis induction: a modified ligation technique with injection of bacteria. Int J Oral Health Sci. 2020;10(1). doi: 10.4103/ijohs.ijohs_44_19.

Setiawatie EM, Gani MA, Rahayu RP, Ulfah N, Kurnia S, Augustina EF. Nigella sativa toothpaste promotes anti-inflammatory and anti-destructive effects in a rat model of periodontitis. Arch Oral Biol. 2022;137:105396. doi: 10.1016/j.archoralbio.2022.105396, PMID 35286946.

Sehari NH, Wafa S, Sehari NH, Siouda W, Sehari M, Merinas L. Phytochemical profile FTIR studies antioxidant and antifungal activity of extracts of pomegranate peel (Punica granatum L.). Ukr J Ecol. 2022;12(12):8-16.

Rahmat D, Budiati A. Formulation and activity of gel containing nanoparticles of javanese turmeric extract as antiacne (formulasi dan aktivitas gel yang mengandungnanopartikelekstraktemulawaksebagai antiacne). J Ilmu Kefarmasian Indones. 2020;18(1).

Azarashkan Z, Motamedzadegan A, Ghorbani Hasan Saraei A, Biparva P, Rahaiee S. Investigation of the physicochemical antioxidant rheological and sensory properties of ricotta cheese enriched with free and nano encapsulated broccoli sprout extract. Food Sci Nutr. 2022;10(11):4059-72. doi: 10.1002/fsn3.3001, PMID 36348770.

Amalia M, Yosuana PS, Mohammad IS Binti Nababan FSR Zulkarnain, Wulandari P, Nasution AH, Syahputra A. Inhibitory test of andaliman (Zanthoxylum achantopodium DC) extract mouthwash against dental plaque bacteria. Dental Journal. 2023;56(2):92-7. doi: 10.20473/J.DJMKG.V56.I2.P92-97.

Pattanashetti JI, Kalburgi NB, Patil KA, Sulakod K, Hannahson P. Antibacterial activity of abutilon indicum leaf extract against periodontal pathogens: an in vitro study. Int J Res Ayurveda Pharm. 2023;14(1):72-5. doi: 10.7897/2277-4343.140117.

Quintao Manhanini Souza E, Felipe Toro L, Franzao Ganzaroli V, DE Oliveira Alvarenga Freire J, Matsumoto MA, Casatti CA, Tavares Angelo Cintra L, Leone Buchaim R, Mardegan Issa JP, Gouveia Garcia V. Peri implantitis increases the risk of medication-related osteonecrosis of the jaws associated with osseointegrated implants in rats treated with zoledronate. Scientific Reports. 2024;14(1). doi: 10.1038/s41598-023-49647-4.

Isnurhakim A, Suhartono B, Putranto R. Comparison for Carica papaya and gengigel leaves extraction for gingivitis healing effectiveness in orthodontic application. MEDALI. 2021;3(1). doi: 10.30659/medali.v3i1.17055.

Sayed S, Alotaibi SS, El Shehawi AM, Hassan MM, Shukry M, Alkafafy M. The anti-inflammatory, anti-apoptotic and antioxidant effects of a pomegranate peel extract against acrylamide induced hepatotoxicity in rats. Life (Basel). 2022;12(2):224. doi: 10.3390/life12020224, PMID 35207511.

Diyatri I, Kusumaningsih T, Tantiana HAR. Analysis of the expression of macrophage among periodontitis rat model after treatment with graptophyllum pictum (L.) Griff. leaves extract gel. Malays J Med Health Sci. 2020;16(4):92-6.

Saidah M, Oktiani BW, Taufiqurrahman I. The effect of flavonoid propolis kelulut (trigona spp) extract on macrophage cell number in periodontitis (in vivo study in male wistar rate (rattus novergicus) gingiva). Dentino :Jurnal Kedokteran Gigi. 2020;5(1):28. doi: 10.20527/dentino.v5i1.8117.