Cytotoxicity, Wound Healing Activity, and Antioxidant Potential of Peronema canescens Leaf Extract: An In vitro Study on B16 Melanoma Cells
Article Sidebar
Main Article Content
Abstract
Peronema canescens Jack, commonly known as sungkai, is a medicinal plant traditionally used due to its anti-inflammatory and wound-healing properties. Therefore, this study aims to evaluate cytotoxicity, wound healing activity, antioxidant potential, and total flavonoid content of P. canescens leaf extract using B16 melanoma cells in vitro. Cytotoxicity was assessed using the MTT assay, showing that concentrations below 25 µg/mL maintained cell viability above 80%, indicating the extract is non-cytotoxic at low doses. Wound healing activity was evaluated through the scratch assay, in which extract concentrations of 6.25, 12.5, and 25 µg/mL accelerated wound closure, with the 25 µg/mL treatment showing the most significant effect (42.37% closure at 24 h). ABTS assay showed moderate antioxidant activity, with an IC₅₀ value of 74.88 µg/mL, while vitamin C showed higher potency (IC₅₀ of 7.04 µg/mL). The total flavonoid content of the extract was determined to be 84.08 ± 0.33 mg QE/g. These results suggest that P. canescens leaf extract has potential as a natural therapeutic agent for promoting wound healing and protecting cells from oxidative stress.
Downloads
Article Details
Section
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
How to Cite
References
1. Diller RB, Tabor AJ. The Role of the Extracellular Matrix (ECM) in Wound Healing: A Review. Vol. 7, Biomimetics. MDPI; 2022; 4:1-15.
2. Papaccio F, D’arino A, Caputo S, Bellei B. Focus on the Contribution of Oxidative Stress in Skin Aging. Antioxidants. MDPI. 2022; 11(6): 1121.
3. Szliszka E, Czuba ZP, Domino M, Mazur B, Zydowicz G, Krol W. Ethanolic Extract of Propolis (EEP) Enhances the Apoptosis- Inducing Potential of TRAIL in Cancer Cells. Molecules. 2009; 4(2): 738–54.
4. Han G, Ceilley R. Chronic Wound Healing: A Review of Current Management and Treatments. Vol. 34, Adv Ther. Springer Healthcare; 2017. p. 599–610.
5. Rahardhian MRR, Sumiwi SA, Susilawati Y, Muchtaridi M. Immunomodulatory Potential of Kaempferol Isolated from Peronema canescens Jack. Leaves Through Inhibition of IL-6 Expression. Int J Mol Sci. 2025; 26(7): 3068
6. Rahardhian MRR, Susilawati Y, Sumiwi A, Muktiwardoyo M, Muchtaridi M, Sumiwi SA. A Review Of Sungkai (Peronema Canescens): Traditional Usage, Phytoconstituent, And Pharmacological Activities. Int J Appl Pharm. 2022;14 (Special issue 5):15–23.
7. Rahardhian MRR, Susilawati Y, Musfiroh I, Febriyanti RM, Muchtaridi M, Sumiwi SA. In Silico Study Of Bioactive Compounds From Sungkai (Peronema Canescens) As Immunomodulator. Int J Appl Pharm. 2022;14(4):1–7.
8. Fadilah NIM, Phang SJ, Kamaruzaman N, Salleh A, Zawani M, Sanyal A. Antioxidant Biomaterials in Cutaneous Wound Healing and Tissue Regeneration: A Critical Review. Vol. 12, Antioxidants. MDPI; 2023. 12(4):1-35.
9. Erharuyi O, Falodun A. Combinatorial chemistry and natural product: Recycling the past in the present. Vol. 2, Trop J Nat Prod Res. 2018. p. 1–2.
10. Hajialyani M, Tewari D, Sobarzo-Sánchez E, Nabavi SM, Farzaei MH, Abdollahi M. Natural product-based nanomedicines for wound healing purposes: Therapeutic targets and drug delivery systems. Vol. 13, Int J Nanomedicine. Dove Medical Press Ltd.; 2018. p. 5023–43.
11. Rahardhian MRR, Aryanti N, Susilawati Y, Sumiwi SA, Putri CN, Ramonah D. Exploring the α-Amylase Inhibitory Potential of Peronema canescens Jack: An In vitro and In Silico Study. Hayati. 2025;32(3):712–26.
12. Rahardhian MRR, Susilawati Y, Sumiwi SA, Ramonah D, Putri CN, Suharsanti R. In vitro and in Silico Analysis of Antilipase, Antioxidant, and Optimization of Granule Effervescent from Peronema canescens Jack. Trop J Nat Prod Res. 2025; 9(2): 545 – 553.
13. Jung GD, Yang JY, Song ES, Park JW. Stimulation of melanogenesis by glycyrrhizin in B16 melanoma cells. Exp Mol Med. 2001;33(3):131–5.
14. Martinotti S, Ranzato E. Scratch wound healing assay. In: Methods in Molecular Biology. Humana Press Inc.; 2020. p. 225–9.
15. Ilyasov IR, Beloborodov VL, Selivanova IA, Terekhov RP. ABTS/PP decolorization assay of antioxidant capacity reaction pathways. Int J Mol Sci. 2020. 21: 1-27
16. Imieje VO, Onochie CF, Nwaka B, Falodun A. In vitro Antioxidant and Antidiabetic Potentials of Extracts of the Stem Bark of Cylicodiscus gabunensis (Harms) Mimosaceae. Trop J Nat Prod Res. 2022; 6(11):1858–63.
17. Rahardhian MRR, Suharsanti R, Sugihartini N, Lukitaningsih E. In vitro assessment of total phenolic, total flavonoid and sunscreen activities of crude ethanolic extract of belimbing wuluh (Averrhoa bilimbi) fruits and leaves. J Glob Pharma Technol. 2019;11(4):308–13.
18. Syukur M, Prahasiwi MS, Nurkhasanah, Yuliani S, Purwaningsih Y, Indriyanti E. Profiling of Active Compounds of Extract Ethanol, n-Hexane, Ethyl Acetate and Fraction Ethanol of Star Anise (Illicium verum Hook. f.) and Determination of Total Flavonoids, Total Phenolics and Their Potential as Antioxidants. Sci Technol Indones. 2023; 8(2):219–26.
19. Suharsanti R, Sugihartini N, Lukitaningsih E, Rahardhian MRR. Effect of Different Solvent on Total Phenolic, Total Flavonoid, and Sun Protection Factor of Belimbing Wuluh (Averrhoa Bilimbi linn.) Fruits Fraction. J Glob Pharma Technol. 2019;11(1):154–62.
20. Rahardhian MR R, Ardiansyah F, Susilawati Y, Wilar G, Sumiwi SA, Levita J, Muchtaridi M. Immunomodulatory and Acute Toxicity Studies of Peronema canescens Jack Leaves: in vivo Hematological Analysis and in vitro IL-6 Gene Expression Inhibition. Indones Biomed J. 2025; 17(3): 295–306.
21. Bahuguna A, Khan I, Bajpai VK, Kang SC. MTT assay to evaluate the cytotoxic potential of a drug. Bangladesh J Pharmacol. 2017;12(2):115–8.
22. Eming SA, Martin P, Tomic-Canic M. Wound repair and regeneration: Mechanisms, signaling, and translation [Internet]. Available from: www.ScienceTranslationalMedicine.org
23. Huang Z, Li L, Cheng B, Li D. Small molecules targeting HDAC6 for cancer treatment: Current progress and novel strategies. Vol. 178, Biomed Pharmacother. Elsevier Masson; 2024: 1-41.
24. Kubo I, Nitoda T, Nihei KI. Effects of Quercetin on Mushroom Tyrosinase and B16-F10 Melanoma Cells. Molecules [Internet]. 2007;12:1045–56. Available from: www.mdpi.org/molecules
25. Harhaji L, Mijatović S, Maksimović-Ivanić D, Stojanović I, Momčilović M, Maksimović V, et al. Anti-tumor effect of Coriolus versicolor methanol extract against mouse B16 melanoma cells: In vitro and in vivo study. Food Chem Toxicol. 2008; 46(5):1825–33.
26. Rajasekar S, Park DJ, Park C, Park S, Park YH, Kim ST, et al. In vitro and in vivo anticancer effects of Lithospermum erythrorhizon extract on B16F10 murine melanoma. J Ethnopharmacol. 2012; 144(2):335–45.
27. Skandrani I, Pinon A, Simon A, Ghedira K, Chekir-Ghedira L. Chloroform extract from Moricandia arvensis inhibits growth of B16-F0 melanoma cells and promotes differentiation in vitro. Cell Prolif. 2010;43(5):471–9.
28. Wei FKBHLK. 5-Hydroxymethylfurfural Mitigates Lipopolysaccharide-Stimulated Inflammation via Suppression of MAPK, NF- κB and mTOR Activation in RAW 264.7 Cells. Molecules. 2019;24(275):1–16.
29. Fronza M, Heinzmann B, Hamburger M, Laufer S, Merfort I. Determination of the wound healing effect of Calendula extracts using the scratch assay with 3T3 fibroblasts. J Ethnopharmacol. 2009 126(3):463–7.
30. Khosravitabar F, Abrishamchi P, Bahrami AR, Matin MM, Ejtehadi H, Varasteh-Kojourian M. Enhanced Cutaneous Wound Healing by the Leaf Extract of Achillea eriophora D.C. Using the In vitro Scratch Assay. J Sci IR Iran. [Internet]. 2017;28(4):305–12. Available from: http://jsciences.ut.ac.ir
31. Okolie NP, Falodun A, Oluseyi D. Evaluation of the antioxidant activity of root extract of pepper fruit (Dennetia tripetala), and its potential for the inhibition of Lipid peroxidation. Afr J. Trad Compl and Altern Med. 2014; 11(3):221-227. Doi: 10.4314/ajtcam. v11i3.31.
32. Haddadi R, Tamri P, Javani Jooni F. In vitro wound healing activity of Scrophularia striata hydroalcoholic extract. S Afr J Bot. 2019;121: 505–9.
33. Saeed N, Khan MR, Shabbir M. Antioxidant activity, total phenolic and total flavonoid contents of whole plant extracts Torilis leptophylla L. BMC Complement Altern Med. 2012;12; 1-12
34. Suryanti V, Sariwati A, Sari F, Handayani DS, Risqi HD. Metabolite Bioactive Contents of Parkia timoriana (DC) Merr Seed Extracts in Different Solvent Polarities. Hayati. 2022;29(5):681–94.
35. Andriana Y, Xuan TD, Quy TN, Minh TN, Van TM, Viet TD. Antihyperuricemia, antioxidant, and antibacterial activities of tridax procumbens L. Foods. 2019;8(1): 1–12.
36. Oktiansyah R, Setiawan A, Widjajanti H. Combination Effect of Extracts and Pure Compounds of Endophytic Fungi Isolated from Sungkai (Peronema canescens) Leaves on Antioxidant Activity. Sci Technol Indones. 2024;9(1): 69-79
37. Nicolescu A, Bunea CI, Mocan A. Total flavonoid content revised: An overview of past, present, and future determinations in phytochemical analysis. Vol. 700, Anal Biochem. Academic Press Inc.; 2025.