Effects of Piper crocatum leaf extract-based ointments on bacteria associated with diabetic ulcers: an in vitro study

Yudha Rizky Nuari, Mila Abusri, Wahyu Yuntari, Oca Maharani Tryadi, Fiarriescha Marra Ardhiana

Abstract


Diabetic patients with poor blood glucose control are highly susceptible to developing secondary infections, which can lead to the development of prolonged diabetic ulcers. Therefore, a suitable medication that may effectively prevent the occurrence of secondary infections is crucial to shorten the closure of diabetic ulcers. Red betel leaf (Piper crocatum Ruiz & Pav) reportedly possesses antimicrobial activity due to the presence of flavonoids. This study aimed to evaluate the effect of ethanolic extract of red betel leaf (EERBL) ointments against the most prevalent bacteria associated with diabetic foot ulcers (DFU): Staphylococcus aureus and Pseudomonas aeruginosa. The EERBL was prepared by macerating powdered red betel leaf with 96% ethanol and was screened for the presence of flavonoids and the determination of total flavonoid content (TFC) by thin layer chromatography and UV-Vis spectrophotometry, respectively. This study examined three hydrophilic-based ointments containing 10%, 20%, and 30% EERBL, respectively, followed by characterization for pH, spreadability, stickiness, and viscosity. The EERBL ointments' effect on the bacteria was evaluated using the well-diffusion method by observing inhibition zone formation after 24-hour incubation. The results showed that varying the EERBL concentrations in the formulations led to different spreadability, stickiness and viscosity (p<0.05). Furthermore, all EERBL ointments demonstrated the formation of an inhibition zone on cultured media, indicating the presence of antimicrobial activity. The ointment with 30% EERBL had the largest diameter of the inhibition zone against both bacteria (p<0.05). The findings suggest enhanced antimicrobial activity was observed with an increase in the concentration of EERBL within the ointments.

Keywords


Red betel leaf (Piper crocatum); flavonoids; ointment formulation; antimicrobial activity; diabetic ulcers

References


Anonim. (2020). Farmakope Indonesia Edisi VI. Kemenkes RI.

Balouiri, M., Sadiki, M., & Ibnsouda, S. K. (2016). Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis, 6(2), 71–79. https://doi.org/10.1016/j.jpha.2015.11.005

Beulens, J. W. J., Yauw, J. S., Elders, P. J. M., Feenstra, T., Herings, R., Slieker, R. C., Moons, K. G. M., Nijpels, G., & van der Heijden, A. A. (2021). Prognostic models for predicting the risk of foot ulcer or amputation in people with type 2 diabetes: a systematic review and external validation study. Diabetologia, 64(7), 1550–1562.

Blando, F., Russo, R., Negro, C., De Bellis, L., & Frassinetti, S. (2019). Antimicrobial and Antibiofilm Activity against Staphylococcus aureus of Opuntia ficus-indica (L.) Mill. Cladode Polyphenolic Extracts. Antioxidants (Basel, Switzerland), 8(5). https://doi.org/10.3390/antiox8050117

Chammas, N. K., Hill, R. L. R., & Edmonds, M. E. (2016). Increased Mortality in Diabetic Foot Ulcer Patients: The Significance of Ulcer Type. Journal of Diabetes Research, 2016, 2879809. https://doi.org/10.1155/2016/2879809

Cho, N. H., Shaw, J. E., Karuranga, S., Huang, Y., da Rocha Fernandes, J. D., Ohlrogge, A. W., & Malanda, B. (2018). IDF Diabetes Atlas: Global estimates of diabetes prevalence for 2017 and projections for 2045. Diabetes Research and Clinical Practice, 138, 271–281. https://doi.org/10.1016/j.diabres.2018.02.023

Conti-Silva, A. C., Ichiba, A. K. T., Silveira, A. L. da, Albano, K. M., & Nicoletti, V. R. (2018). Viscosity of liquid and semisolid materials: Establishing correlations between instrumental analyses and sensory characteristics. Journal of Texture Studies, 49(6), 569–577. https://doi.org/10.1111/jtxs.12358

Dallo, M., Patel, K., & Hebert, A. A. (2023). Topical Antibiotic Treatment in Dermatology. Antibiotics (Basel, Switzerland), 12(2). https://doi.org/10.3390/antibiotics12020188

Datta, P., Chander, J., Gupta, V., Mohi, G. K., & Attri, A. K. (2019). Evaluation of various risk factors associated with multidrug-resistant organisms isolated from diabetic foot ulcer patients. Journal of Laboratory Physicians, 11(1), 58–62. https://doi.org/10.4103/JLP.JLP_106_18

Erwin DZ, C. P. (2024). Mupirocin. [Updated 2024 Jan 11]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2024 Jan. In StatPearls. https://www.ncbi.nlm.nih.gov/books/NBK599499/

Farmaki, P., Damaskos, C., Garmpis, N., Garmpi, A., Savvanis, S., & Diamantis, E. (2020). Complications of the Type 2 Diabetes Mellitus. Current Cardiology Reviews, 16(4), 249–251. https://doi.org/10.2174/1573403X1604201229115531

Ha, J. H., Jin, H., & Park, J.-U. (2021). Association between socioeconomic position and diabetic foot ulcer outcomes: a population-based cohort study in South Korea. BMC Public Health, 21(1), 1395. https://doi.org/10.1186/s12889-021-11406-3

Hartini, Y. S., & Nugroho, L. H. (2020). Antibacterial effect of red betel (Piper crocatum) extract in combination with vancomycin against staphylococcus aureus. Biodiversitas, 21(7), 3271–3274. https://doi.org/10.13057/biodiv/d210750

Heliawati, L., Lestari, S., Hasanah, U., Ajiati, D., & Kurnia, D. (2022). Phytochemical Profile of Antibacterial Agents from Red Betel Leaf (Piper crocatum Ruiz and Pav) against Bacteria in Dental Caries. Molecules (Basel, Switzerland), 27(9). https://doi.org/10.3390/molecules27092861

Herbig, M. E., Evers, D.-H., Gorissen, S., & Köllmer, M. (2023). Rational Design of Topical Semi-Solid Dosage Forms-How Far Are We? Pharmaceutics, 15(7). https://doi.org/10.3390/pharmaceutics15071822

Hidayat, B., Ramadani, R. V., Rudijanto, A., Soewondo, P., Suastika, K., & Siu Ng, J. Y. (2022). Direct Medical Cost of Type 2 Diabetes Mellitus and Its Associated Complications in Indonesia. Value in Health Regional Issues, 28, 82–89. https://doi.org/10.1016/j.vhri.2021.04.006

Ibrahim, N. ’Izzah, Wong, S. K., Mohamed, I. N., Mohamed, N., Chin, K.-Y., Ima-Nirwana, S., & Shuid, A. N. (2018). Wound Healing Properties of Selected Natural Products. International Journal of Environmental Research and Public Health, 15(11). https://doi.org/10.3390/ijerph15112360

Ishikawa, J., & Horii, T. (2005). Effects of Mupirocin at Subinhibitory Concentrations on Biofilm Formation in Pseudomonas aeruginosa. Chemotherapy, 51(6), 361–362. https://doi.org/10.1159/000088962

Juliantina Rachmawaty, F., Mahardika Akhmad, M., Hikmah Pranacipta, S., Nabila, Z., & Muhammad, A. (2018). Optimasi Ekstrak Etanol Daun Sirih Merah (Piper Crocatum) sebagai Antibakteri terhadap Bakteri Staphylococcus Aureus. Mutiara Medika: Jurnal Kedokteran Dan Kesehatan, 18(1), 13–19. https://doi.org/10.18196/mm.180109

Jupiter, D. C., Thorud, J. C., Buckley, C. J., & Shibuya, N. (2016). The impact of foot ulceration and amputation on mortality in diabetic patients. I: From ulceration to death, a systematic review. International Wound Journal, 13(5), 892–903. https://doi.org/10.1111/iwj.12404

Kaul, K., Tarr, J. M., Ahmad, S. I., Kohner, E. M., & Chibber, R. (2013). Introduction to Diabetes Mellitus BT - Diabetes: An Old Disease, a New Insight (S. I. Ahmad (ed.); pp. 1–11). Springer New York. https://doi.org/10.1007/978-1-4614-5441-0_1

Kemenkes RI. (2017). Farmakope Herbal Indonesia Edisi 2. 561.

Li, Y., Ju, S., Li, X., Li, W., Zhou, S., Wang, G., Cai, Y., & Dong, Z. (2022). Characterization of the microenvironment of diabetic foot ulcers and potential drug identification based on scRNA-seq. Frontiers in Endocrinology, 13, 997880. https://doi.org/10.3389/fendo.2022.997880

Matheson, E. M., Bragg, S. W., & Blackwelder, R. S. (2021). Diabetes-Related Foot Infections: Diagnosis and Treatment. American Family Physician, 104(4), 386–394.

Maulina, L., & Sugihartini, N. (2015). FORMULASI GEL EKSTRAK ETANOL KULIT BUAH MANGGIS (Garcinia mangostana L.) DENGAN VARIASI GELLING AGENT SEBAGAI SEDIAAN LUKA BAKAR. Pharmaciana, 5(1), 43–52. https://doi.org/10.12928/pharmaciana.v5i1.2285

Navirius, F. J., Pamudji, G., & Herowati, R. (2023). Effect of Red Betel (Piper crocatum) Leaf Ethanol Extract On Increased Creatinine and Ureum Levels In White Rat Wistar Strain Induced Streptozotosin-Nikotinamid. Media Farmasi: Jurnal Ilmu Farmasi, 20(2), 57. https://doi.org/10.12928/mf.v20i2.25457

Nerdy, N., Barus, B. R., El-Matury, H. J., Ginting, S., Zebua, N. F., & Bakri, T. K. (2022). Comparison of flavonoid content and antioxidant activity in calyces of two roselle varieties (Hibiscus sabdariffa L.). IOP Conference Series: Earth and Environmental Science, 956(1). https://doi.org/10.1088/1755-1315/956/1/012001

Puspita, P. J., Safithri, M., & Sugiharti, N. P. (2019). Antibacterial Activities of Sirih Merah (Piper crocatum) Leaf Extracts. Current Biochemistry, 5(3), 1–10. https://doi.org/10.29244/cb.5.3.1-10

Rubio, J. A., Jiménez, S., & Lázaro-Martínez, J. L. (2020). Mortality in Patients with Diabetic Foot Ulcers: Causes, Risk Factors, and Their Association with Evolution and Severity of Ulcer. Journal of Clinical Medicine, 9(9). https://doi.org/10.3390/jcm9093009

Sadeghpour Heravi, F., Zakrzewski, M., Vickery, K., G Armstrong, D., & Hu, H. (2019). Bacterial Diversity of Diabetic Foot Ulcers: Current Status and Future Prospectives. Journal of Clinical Medicine, 8(11). https://doi.org/10.3390/jcm8111935

Safithri, M., Andrianto, D., Gaisani Arda, A., Hawa Syaifie, P., Mardia Ningsih Kaswati, N., Mardliyati, E., Ramadhan, D., Miftah Jauhar, M., Wahyu Nugroho, D., Anggraini Septaningsih, D., Tria Melati, L., Hudayanti, M., Sarah, E., Alifibi Putera Irsal, R., & Taufiqu Rochman, N. (2023). The effect of red betel (Piper crocatum) water fraction as tyrosinase inhibitors: In vitro, molecular docking, and molecular dynamics studies. Journal of King Saud University - Science, 35(10), 102933. https://doi.org/10.1016/j.jksus.2023.102933

Setyawati, A., Sri, M., Wahyuningsih, H., Aris, D., Nugrahaningsih, A., Effendy, C., & Ibeneme, S. (2023). Piper crocatum Ruiz & Pav as a commonly used typically medicinal plant from Indonesia: What do we actually know about it? Scoping review. Indonesian Contemporary Nursing Journal, 7(2), 61–78.

Setyawati, A., Wahyuningsih, M. S. H., Nugrahaningsih, D. A. A., Effendy, C., Fneish, F., & Fortwengel, G. (2021). Piper crocatum Ruiz & Pav. ameliorates wound healing through p53, E-cadherin and SOD1 pathways on wounded hyperglycemia fibroblasts. Saudi Journal of Biological Sciences, 28(12), 7257–7268. https://doi.org/10.1016/j.sjbs.2021.08.039

Shamsudin, N. F., Ahmed, Q. U., Mahmood, S., Ali Shah, S. A., Khatib, A., Mukhtar, S., Alsharif, M. A., Parveen, H., & Zakaria, Z. A. (2022). Antibacterial Effects of Flavonoids and Their Structure-Activity Relationship Study: A Comparative Interpretation. Molecules (Basel, Switzerland), 27(4). https://doi.org/10.3390/molecules27041149

Shigeyama, M., Ohgaya, T., Kawashima, Y., Takeuchi, H., & Hino, T. (1999). Mixed base of hydrophilic ointment and purified lanolin to improve the drug release rate and absorption of water of minocycline hydrochloride ointment for treatment of bedsores. Chemical & Pharmaceutical Bulletin, 47(6), 744–748. https://doi.org/10.1248/cpb.47.744

Suri, M. A., Azizah, Z., & Asra, R. (2021). A Review: Traditional Use, Phytochemical and Pharmacological Review of Red Betel Leaves (Piper Crocatum Ruiz & Pav). Asian Journal of Pharmaceutical Research and Development, 9(1), 159–163. https://doi.org/10.22270/ajprd.v9i1.926

Tanoey, J., & Becher, H. (2021). Diabetes prevalence and risk factors of early-onset adult diabetes: results from the Indonesian family life survey. Global Health Action, 14(1), 2001144. https://doi.org/10.1080/16549716.2021.2001144

USP. (2007). Hydrophilic Ointment: In United States Pharmacopeia Convention Committee of Revision (Ed.), United States Pharmacopeia (USP29-NF4 ed.). In United States Pharmacopeial Convention Committee of Revision (Ed.).

Yan, X., Song, J.-F., Zhang, L., & Li, X. (2022). Analysis of risk factors for multidrug-resistant organisms in diabetic foot infection. BMC Endocrine Disorders, 22(1), 46. https://doi.org/10.1186/s12902-022-00957-0




DOI: http://dx.doi.org/10.12928/pharmaciana.v14i2.28653

Refbacks

  • There are currently no refbacks.


Copyright (c) 2024 Yudha Rizky Nuari, Mila Abusri, Wahyu Yuntari, Oca Maharani Tryadi, Fiarriescha Marra Ardhiana

Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

 
Pharmaciana
ISSN Print: 2088-4559 | ISSN Online: 2477-0256
Website: http://journal.uad.ac.id/index.php/PHARMACIANA
Office: Faculty of  Pharmacy, Universitas Ahmad Dahlan
Jl. Prof. Dr. Soepomo, S.H., Janturan, Warungboto, Umbulharjo, Yogyakarta, Indonesia
Kode pos 55164
Email: pharmaciana@pharm.uad.ac.id