Potential Wound Healing Activity of Citrus micrantha Rut. (Biasong) Ethanolic Peel Extract on Excised Cutaneous Wounds in Male Albino Mice


Authors : Potri Kalmarjan M. Alango; Moamar Uthman B. Datumanong; Kaimah A. Galma; Nihara T. Macapintal; Patricia Mae B. Jarabe; Junnin Gay L. Garay

Volume/Issue : Volume 10 - 2025, Issue 7 - July

Google Scholar : https://tinyurl.com/mrsm3byu

Scribd : https://tinyurl.com/nhkcub55

DOI : https://doi.org/10.38124/ijisrt/25jul1301

PlumX Metrics

Semantic Scholar

ResearchGate

Google Scholar

Note : A published paper may take 4-5 working days from the publication date to appear in PlumX Metrics, Semantic Scholar, and ResearchGate.

Note : Google Scholar may take 30 to 40 days to display the article.

Abstract : Many Citrus plants, such as C. reticulata, C. aurantium, C. aurantifolia, C. sinensis, C. maxima, C. medica, C. clementina, C. paradisi, and C. limon, are recognized for their wound healing properties [36] [8] [6] [2] [3]. However, there is a lack of research specifically focused on Citrus micrantha (Biasong)—a wild plant native to the Philippines, known for its culinary and medicinal applications [19]. Therefore, this study aims to evaluate the wound healing activity of the plant’s peels using ethanol as the extraction solvent, focusing on excised cutaneous wounds in male Swiss albino mice. To accomplish this, toxicity testing based on OECD 402: Acute Dermal Toxicity guidelines was conducted before preparing the ethanolic peel extract at varying concentrations (1%, 5%, and 10%) and incorporating it into ointment formulations deemed safe for the test subjects. These formulations were topically applied to the dorsal area of the mice, with wound contraction measurements serving as the parameter to evaluate healing progress over a 14-day observation period. A significant difference in wound healing activity between treatment groups was most evident by Day 14, with the 10% extract showing the highest contraction (99.194%) and smallest final wound area (0.2278 mm2 ), followed by the 5% extract (98.772%, 0.347 mm2 ), 1% extract (98.678%, 0.37 mm2 ), negative control (97.22%, 0.7854 mm2 ), and positive control (94.616%, 1.522 mm2 ). The study concluded that the application of C. micrantha ethanolic peel extract ointment promoted wound healing and demonstrated a significant improvement compared to the positive control (Povidone-Iodine ointment).

Keywords : Ethanolic Peel Extract, Excised Cutaneous Wounds, Wound Healing Activity.

References :

  1. Ademosun AO. 2022. Citrus peels odyssey: From the waste bin to the lab bench to the dining table. Applied Food Research. 2(1):100083. doi:10.1016/j.afres.2022.100083.
  2. Ahmad AA, Khalifa IIA, Abudayeh ZH. 2018. The role of pomelo peel extract for experimentally induced wound in diabetic rats. Pharmacognosy Journal. 10(5):885–891. doi:10.5530/pj.2018.5.149.
  3. Ayuningtyas NF, Hendarti HT, Soebadi B, Surboyo MDC, Hadi P, Ganesha R, Ernawati DS, Marsetyo RI. 2023. Expression of VEGF and CD-31 in traumatic ulcer of diabetic Wistar rats after application of Citrus limon peel essential oil. Journal of Oral Biology and Craniofacial Research. 13(3):380–385. doi:10.1016/j.jobcr.2023.03.009.
  4. Baygar T and Sarac N. 2018. Antimicrobial activity of clementine peel essential oil with its cytotoxic and in vitro wound healing potential on NIH-3T3 fibroblast cells. Mugla Journal of Science and Technology. 4(2):143–147. doi:10.22531/muglajsci.435610.
  5. Budiarto K, Andrini A, Budiyati E, Mariana BD, Martasari C, Mas’udah S, Yulia ND, Ikarini I, Yulianti F. 2024. Bioactive phytochemical contents on fruit peel of several Citrus species. BIO Web of Conferences. 91:01016. doi:10.1051/bioconf/20249101016. Available from: https://www.bio-conferences.org/articles/bioconf/abs/2024/10/bioconf_icgrc2024_01016/bioconf_icgrc2024_01016.html.
  6. Cherukuri S, Paramanayagam A, Prabakaran R, Mayakannan M, Vuppalapati L. 2023 Jan 27. Evaluation of cutaneous wound healing activity of Citrus aurantium fruit peel extract-based ointment in albino rats. Research Journal of Pharmacy and Technology. 16(1):250–254. doi:10.52711/0974-360x.2023.00046.
  7. Dahmani S, Chabir R, Errachidi F, Berrada W, Lansari H, Benidir M, Ghadraoui LE, Bour A. 2020. Evaluation of in vivo wound healing activity of Moroccan Citrus reticulata peel extract. Clinical Phytoscience. 6(1). doi:10.1186/s40816-020-00222-8.
  8. Fauzia M, Dewanti AP. 2022. The effect of lime (Citrus Aurantifola Swingle) peel extract in periodontal dressings on the number of fibroblasts in the gingival wound healing process. Dental Journal (Majalah Kedokteran Gigi). 55(2):81–87. doi:10.20473/j.djmkg.v55.i2.p81-87.
  9. Fernández-Guarino M, Hernández-Bule ML, Bacci S. 2023. Cellular and molecular processes in wound healing. Biomedicines. 11(9):2526. doi:10.3390/biomedicines11092526.
  10. Global Biodiversity Information Facility. 2024. Citrus micrantha Wester. Occurrence dataset. Royal Botanic Gardens, Kew. doi:10.15468/ly60bx. Available from: https://www.gbif.org/occurrence/912556804.
  11. Goldenheim PD 1993. An appraisal of povidone-iodine and wound healing. PubMed. https://pubmed.ncbi.nlm.nih.gov/8290466/.
  12. Ibrahim N, Wong S, Mohamed I, Mohamed N, Chin K-Y, Ima-Nirwana S, Shuid A. 2018. Wound healing properties of selected natural products. International Journal of Environmental Research and Public Health. 15(11):2360. doi:10.3390/ijerph15112360.
  13. Irawaty W, Ayucitra A. 2018. Assessment on antioxidant and in vitro antidiabetes activities of different fractions of Citrus hystrix peel. International Food Research Journal. 25(6):2467–2477. Available from: https://repository.ukwms.ac.id/id/eprint/18357/
  14. Ishfaq M, Akhtar B, Muhammad F, Sharif A, Akhtar MF, Hamid I, Sohail K, Muhammad H. 2020. Antioxidant and wound healing potential of essential oil from Citrus reticulata peel and its chemical characterization. Current Pharmaceutical Biotechnology. 22(8):1114–1121. doi:10.2174/1389201021999200918102123. Available from: http://research.riphah.edu.pk/id/eprint/1563/1/02%20current%20pharma%20biotechnology%20Antioxidant%20and%20Wound%20Healing%20Potential%20of%20Essential%20Oil%20from%20Citrus.pdf
  15. Kallel I, Tarhouni N, Elaguel A, Mekrazi S, Khabir A, Hadrich B, Bayoudh A. 2023. The phytochemical and pharmacological properties of Citrus sinensis ‘Maltese Half-blood’ essential oil peels extracted and optimized by response-surface methodology. Current Pharmaceutical Biotechnology. 24(15):1938–1951. doi:10.2174/1389201024666230405120520.
  16. Kancherla N, Dhakshinamoothi A, Chitra K, Komaram RB. 2019. Preliminary analysis of phytoconstituents and evaluation of anthelminthic property of Cayratia auriculata (In Vitro). MAEDICA – a Journal of Clinical Medicine. 14(4). doi:10.26574/maedica.2019.14.4.350.
  17. Karunya S, Al-Eqabi RSS, Jagathy K, Roseline F, Saranraj P. 2023. Efficiency of phytochemical compound from the peel extract of Citrus sinunsis and Citrus medica against clinical isolates from wound infection. Life Science Archives (LSA). 9(4):2721–2731. doi:10.22192/lsa.2024.9.4.1. Available from: https://www.academia.edu/120880826/efficiency_of_phytochemical_compound_from_the_peel_extract_of_citrus_sinunsis_and_citrus_medica_against_clinical_isolates_from_wound_infection.
  18. Liu E, Gao H, Zhao Y, Pang Y, Yao Y, Yang Z, Zhang X, Wang Y, Yang S, Ma X, et al. 2022. The potential application of natural products in cutaneous wound healing: A review of preclinical evidence. Frontiers in Pharmacology. 13. doi:10.3389/fphar.2022.900439.
  19. Madayag RE, Gentallan RJrP, Quiñones KJO, Bartolome MCB, Cruz JR a. V, Borromeo TH, Endonela LE, Timog EBS. 2022. The complete chloroplast genome of ‘biasong’ (Citrus micrantha Wester), a native species from the Southern Philippines. Mitochondrial DNA Part B. 7(11):1992–1996. doi:10.1080/23802359.2022.2144515.
  20. Mahato N, Sharma K, Sinha M, Cho MH. 2018. Citrus waste derived nutra-/pharmaceuticals for health benefits: Current trends and future perspectives. Journal of Functional Foods. 40:307–316. doi:10.1016/j.jff.2017.11.015. Available from: https://www.researchgate.net/publication/322012259_Citrus_waste_derived_nutra-pharmaceuticals_for_health_benefits_Current_trends_and_future_perspectives
  21. Masyita A, Sari RM, Astuti AD, Yasir B, Rumata NR, Emran TB, Nainu F, Simal-Gandara J. 2022. Terpenes and terpenoids as main bioactive compounds of essential oils, their roles in human health and potential application as natural food preservatives. Food Chemistry X. 13:100217. doi:10.1016/j.fochx.2022.100217. Available from: https://pubmed.ncbi.nlm.nih.gov/35498985/.
  22. Namutebi B, Bukke SPN, Jackim N, Moses K, Nicholas B, Goruntla N, Neel G. 2024. Evaluation of the antibacterial effect of a combination of the ethanolic extracts of Citrus limon peels and Eucalyptus globulus leaves against Streptococcus pneumoniae. Journal of Advances in Microbiology Research. https://www.researchgate.net/publication/384598482_Evaluation_of_the_antibacterial_effect_of_a_combination_of_the_ethanolic_extracts_of_Citrus_limon_peelsand_Eucalyptus_globulus_leaves_against_Streptococcus_pneumoniae
  23. Ollitrault P, Curk F, Krueger R. 2020. Citrus taxonomy. In: Elsevier eBooks. p. 57–81. doi:10.1016/b978-0-12-812163-4.00004-8.
  24. Organization of Economic Co-operation and Development. 2017. Test No. 402: Acute Dermal Toxicity. OECD Guidelines for the Testing of Chemicals, Section 4. OECD Publishing, Paris, doi:10.1787/9789264070585-en. Available from: https://www.oecd-ilibrary.org/environment/test-no-402-acute-dermal-toxicity_9789264070585-en?fbclid=IwZXh0bgNhZW0CMTEAAR1zIsfYrX7bBKPx9ckytV-Xj-LMh7pc9e73AH41hGsVCDvpiNeUMnWaU4c_aem_B4cBUlpjnWuLgb7c-IKvAw.
  25. Ratanachamnong P, Chunchaowarit Y, Namchaiw P, Niwaspragrit C, Rattanacheeworn P, Jaisin Y. 2023. HPLC analysis and in vitro antioxidant mediated through cell migration effect of C. hystrix water extract on human keratinocytes and fibroblasts. Heliyon. 9(2):e13068. doi:10.1016/j.heliyon.2023.e13068. Available from: https://www.researchgate.net/publication/367363537_HPLC_analysis_and_in_vitro_antioxidant_mediated_through_cell_migration_effect_of_Chystrix_water_extract_on_human_keratinocytes_and_fibroblasts
  26. Sachdeva C, Satyamoorthy K, Murali TS. 2022. Microbial interplay in skin and chronic wounds. Current Clinical Microbiology Reports. 9(3):21–31. doi:10.1007/s40588-022-00180-4.
  27. Shedoeva A, Leavesley D, Upton Z, Fan C. 2019. Wound healing and the use of medicinal plants. Evidence-based Complementary and Alternative Medicine. 2019:1–30. doi:10.1155/2019/2684108.
  28. Smit G. 2018. Male versus female mice in behavioral studies. Noldus. https://www.noldus.com/blog/male-mice-behavioral-studies.
  29. Sukmawati N, Kamri AM, Eni NSA. 2024. Effect of pomelo orange (Citrus maxima) peel ethanol extract on burn healing in white rats (Rattus novergicus). Pharmacy Reports. 3(3):54. doi:10.51511/pr.54. Available from: https://www.researchgate.net/publication/383568752_Effect_of_pamelo_orange_Citrus_maxima_peel_ethanol_extract_on_burn_healing_in_white_rats_Rattus_novergicus?_tp=eyJjb250ZXh0Ijp7ImZpcnN0UGFnZSI6ImhvbWUiLCJwYWdlIjoic2VhcmNoIiwicG9zaXRpb24iOiJwYWdlSGVhZGVyIn19
  30. Surboyo MDC, Mahdani FY, Ernawati DS, Hadi P, Hendarti HT, Parmadiati AE, Radithia D, Mardiyana SD, Syarifah IM. 2021. Number of macrophages and transforming growth factor β expression in Citrus limon L. Tlekung peel oil-treated traumatic ulcers in diabetic rats. Tropical Journal of Pharmaceutical Research. 18(7):1427–1433.
  31. doi:10.4314/tjpr.v18i7.9. Available from: https://www.researchgate.net/publication/334770201_Number_of_macrophages_and_transforming_growth_factor_b_expression_in_Citrus_limon_L_Tlekung_peel_oil-treated_traumatic_ulcers_in_diabetic_rats
  32. Tomic-Canic M, Burgess JL, O’Neill KE, Strbo N, Pastar I. 2020. Skin microbiota and its interplay with wound healing. American Journal of Clinical Dermatology. 21(S1):36–43. doi:10.1007/s40257-020-00536-w.
  33. UNC Eshelman School of Pharmacy. 2024. Ointments: preparation & evaluation of drug release. Pharmlabs. https://pharmlabs.unc.edu/labexercises/compounding/ointments/?fbclid=IwZXh0bgNhZW0CMTEAAR2xcqPlC0pIEzou1uc_SH9-_TtRl9e-lh3zfPtYdvoPaxk-UN7SifPERSc_aem_Y9r5JWZIlu2WuWuZWyLYIA.
  34. Wuhan Diao Pharmaceutical Co Ltd. 2023. CN102885852B - Povidone iodine ointment and preparation method thereof. Google Patents. https://patents.google.com/patent/CN102885852B/en.
  35. Yang Y, Huang J, Zeng A, Long X, Yu N, Wang X. 2023. The role of the skin microbiome in wound healing. Burns & Trauma. 12. doi:10.1093/burnst/tkad059.
  36. Yiblet TG, Tsegaw A, Ahmed N, Dagnew SB, Tadesse TY, Kifle ZD. 2022. Evaluation of wound healing activity of 80% methanol root crude extract and solvent fractions of Stephania abyssinica (Dill. & A. Rich.) Walp. (Menispermaceae) in mice. Journal of Experimental Pharmacology. Volume 14:255–273. doi:10.2147/jep.s364282. Available from:  https://www.dovepress.com/evaluation-of-wound-healing-activity-of-80-methanol-root-crude-extract-peer-reviewed-fulltext-article-JEP

37. Yu X, Liu Z, Yu Y, Qian C, Lin Y, Jin S, Wu L, Li S. 2024. Hesperetin promotes diabetic wound healing by inhibiting ferroptosis through the activation of SIRT3. Phytotherapy Research. 38(3):1478–1493. doi:10.1002/ptr.8121. Available from: https://www.researchgate.net/publication/377488465_Hesperetin_promotes_diabetic_wound_healing_by_inhibiting_ferroptosis_through_the_activation_of_SIRT

Many Citrus plants, such as C. reticulata, C. aurantium, C. aurantifolia, C. sinensis, C. maxima, C. medica, C. clementina, C. paradisi, and C. limon, are recognized for their wound healing properties [36] [8] [6] [2] [3]. However, there is a lack of research specifically focused on Citrus micrantha (Biasong)—a wild plant native to the Philippines, known for its culinary and medicinal applications [19]. Therefore, this study aims to evaluate the wound healing activity of the plant’s peels using ethanol as the extraction solvent, focusing on excised cutaneous wounds in male Swiss albino mice. To accomplish this, toxicity testing based on OECD 402: Acute Dermal Toxicity guidelines was conducted before preparing the ethanolic peel extract at varying concentrations (1%, 5%, and 10%) and incorporating it into ointment formulations deemed safe for the test subjects. These formulations were topically applied to the dorsal area of the mice, with wound contraction measurements serving as the parameter to evaluate healing progress over a 14-day observation period. A significant difference in wound healing activity between treatment groups was most evident by Day 14, with the 10% extract showing the highest contraction (99.194%) and smallest final wound area (0.2278 mm2 ), followed by the 5% extract (98.772%, 0.347 mm2 ), 1% extract (98.678%, 0.37 mm2 ), negative control (97.22%, 0.7854 mm2 ), and positive control (94.616%, 1.522 mm2 ). The study concluded that the application of C. micrantha ethanolic peel extract ointment promoted wound healing and demonstrated a significant improvement compared to the positive control (Povidone-Iodine ointment).

Keywords : Ethanolic Peel Extract, Excised Cutaneous Wounds, Wound Healing Activity.

CALL FOR PAPERS


Paper Submission Last Date
31 - December - 2025

Video Explanation for Published paper

Never miss an update from Papermashup

Get notified about the latest tutorials and downloads.

Subscribe by Email

Get alerts directly into your inbox after each post and stay updated.
Subscribe
OR

Subscribe by RSS

Add our RSS to your feedreader to get regular updates from us.
Subscribe