Iranian Journal of Veterinary Surgery

Iranian Journal of Veterinary Surgery

Effects of Ceratonia siliqua L. Extract-Loaded Nanoliposomes on a Rat Model of Excisional Wound Healing

Articles in Press

Document Type : Original Article

Authors
Mehran Mohammadpour 1
Rahim Mohammadi 1
Ghader Jalilzadeh-Amin 2
Seyede Soraya Mahmoudi 3
Mohammad Shahraki 4
1 Department of Surgery and Diagnostic Imaging, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
2 Department of Internal Medicine and Clinical Pathology, Faculty of Veterinary Medicine, Urmia University, Urmia University , Urmia , Iran.
3 Department of Pathobiology, Faculty of Veterinary Medicine, Division of Pathology, Urmia University, Urmia, Iran.
4 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran.
10.30500/ivsa.2025.547784.1464
Abstract
Wound healing consists of inflammatory, proliferation, and remodeling phases. The objective of the current study was to investigate the effect of Ceratonia siliqua L. extract-loaded liposomes on wound healing. Ceratonia siliqua L. commonly called carob, is an evergreen tree that belongs to the Leguminosae family widely cultivated in Mediterranean countries. Twenty-five healthy adult male Wistar rats were randomized into five groups of five animals each: SHAM group with only wound creation. EUCRN group: As a control group with wound creation as well as 5 g eucerin administered topically to the wound bed. NLPSM group: As a group with wound creation, as well as 5 ml nanoliposome (20 mg). EUCRN/NLPSM group: As a group with wound creation as well as 5 g eucerin containing 5 ml nanoliposome (20 mg). TRTMNT group: As a group with wound creation as well as 5 ml eucerin (5 g) containing nanoliposome (20 mg) loaded with hydroalcoholic extract of Ceratonia siliqua L. The excisional wound model (8 mm in diameter) was used for biochemical (total antioxidant capacity, total oxidant status, Malondialdehyde levels, and glutathione peroxidase activity), histopathologic, and planimetric assessments. The wound area was significantly reduced in the TREATMENT group compared to other groups (p < 0.05). Biochemical and quantitative histopathological analyses revealed a significant difference between TREATMENT and other groups (p < 0.05). Ceratonia siliqua L. extract-loaded nanoliposomes showed the potential to improve wound healing significantly. This appeared to work by angiogenesis stimulation, fibroblast proliferation, inflammation reduction, and granulation tissue formation during the initial stages of the healing process. This accelerated healing led to earlier wound area reduction of the damaged area due to the reorganization of granulation tissue and collagen fibers. Topical administration of Ceratonia siliqua L. extract-loaded nanoliposomes could be recommended for wound healing due to reducing wound healing acceleration.
Keywords
Ceratonia siliqua L.
Extract
Nanoliposomes
Wound healing
Rat

Subjects

  1. Xu Z, Han S, Gu Z, Wu J. Advances and impact of antioxidant hydrogel in chronic wound healing. Advanced Healthcare Materials. 2020; 9: 1901502. doi: 10.1002/adhm.201901502
  2. Farag MA, El-Kersh DM. Volatiles profiling in Ceratonia siliqua (Carob bean) from Egypt and in response to roasting as analyzed via solid-phase microextraction coupled to chemometrics. Journal of Advanced Research. 2017; 8: 379–385. doi: 10.1016/j.jare.2017.04.002
  3. Gioxari A, Amerikanou C, Nestoridi I, Gourgari E, Pratsinis H, Kalogeropoulos N, Andrikopoulos NK, Kaliora AC. Carob: A sustainable opportunity for metabolic health. Foods. 2022; 11: 2154. doi: 10.3390/foods11142154
  4. Stavrou IJ, Christou A, Kapnissi-Christodoulou CP. Polyphenols in carobs: a review on their composition, antioxidant capacity and cytotoxic effects, and health impact. Food Chemistry. 2018; 269: 355–374. doi: 10.1016/j.foodchem.2018.07.027
  5. Darwish WS, Khadr AES, Kamel MAEN, Abd Eldaim MA, El Sayed IET, Abdel-Bary HM, Ullah S, Ghareeb DA. Phytochemical characterization and evaluation of biological activities of Egyptian carob pods (Ceratonia siliqua L.) aqueous extract: in vitro study. Plants. 2021; 10: 2626. doi: 10.3390/plants10122626
  6. Farag MA, El-Kersh DM, Ehrlich A, Choucry MA, El-Seedi H, Frolov A, Wessjohann LA. Variation in Ceratonia siliqua pod metabolome in context of its different geographical origin, ripening stage and roasting process. Food Chemistry. 2019; 283: 675–687. doi: 10.1016/j.foodchem.2018.12.118
  7. Çavuşoğlu K, Kurt D, Yalçın E. A versatile model for investigating the protective effects of Ceratonia siliqua pod extract against 1,4-dioxane toxicity. Environmental Science and Pollution Research. 2020; 27: 27885–27892. doi: 10.1007/s11356-020-09194-2
  8. Peng ZT, Xia YJ, Yashiro T, Gao X, Dong TTX, Tsim KWK, Wang HY. Novel phenylpropanoids and isoflavone glycoside are isolated and identified from the carob pods (Ceratonia siliqua L.). Natural Product Research. 2022; 36: 1–7. doi: 10.1080/14786419.2022.2061482
  9. Ben Ayache S, Behija Saafi E, Emhemmed F, Flamini G, Achour L, Muller CD. Biological activities of aqueous extracts from carob plant (Ceratonia siliqua L.) by antioxidant, analgesic and proapoptotic properties evaluation. Molecules. 2020; 25: 3120. doi: 10.3390/molecules25143120
  10. Saratsi K, Hoste H, Voutzourakis N, Tzanidakis N, Stefanakis A, Thamsborg SM, Mueller-Harvey I, Hadjigeorgiou I, Sotiraki S. Feeding of carob (Ceratonia siliqua) to sheep infected with gastrointestinal nematodes reduces faecal egg counts and worm fecundity. Veterinary Parasitology. 2020; 284: 109200. doi: 10.1016/j.vetpar.2020.109200
  11. Alzoubi KH, Alibbini S, Khabour OF, El-Elimat T, Al-zubi M, Alali FQ. Carob (Ceratonia siliqua L.) prevents short-term memory deficit induced by chronic stress in rats. Journal of Molecular Neuroscience. 2018; 66: 314–321. doi: 10.1007/s12031-018-1168-5
  12. Rico D, Martín-Diana AB, Martínez-Villaluenga C, Aguirre L, Silván JM, Dueñas M, De Luis DA, Lasa A. In vitro approach for evaluation of carob by-products as source bioactive ingredients with potential to attenuate metabolic syndrome (MetS). Heliyon. 2019; 5:e01175. doi: 10.1016/j.heliyon.2019.e01175
  13. Valero-Muñoz M, Ballesteros S, Ruiz-Roso B, Pérez-Olleros L, Martín-Fernández B, Lahera V, de las Heras N. Supplementation with an insoluble fiber obtained from carob pod (Ceratonia siliqua L.) rich in polyphenols prevents dyslipidemia in rabbits through SIRT1/PGC-1_ pathway. European Journal of Nutrition. 2019; 58: 357–366. doi: 10.1007/s00394-017-1596-2
  14. Rtibi K, Selmi S, Grami D, Saidani K, Sebai H, Amri M, Eto B, Marzouki L. Ceratonia siliqua L. (immature carob bean) inhibits intestinal glucose absorption, improves glucose tolerance and protects against alloxan-induced diabetes in rat. Journal of the Science of Food and Agriculture. 2017; 97: 2664–2670. doi: 10.1002/jsfa.8091
  15. Qasem MA, Noordin MI, Arya A, Alsalahi A, Jayash SN. Evaluation of the glycemic effect of Ceratonia siliqua pods (Carob) on a streptozotocin-nicotinamide induced diabetic rat model. PeerJ. 2018; 6: e4788. doi: 10.7717/peerj.4788
  16. Mansouri FE, Silva JCGE, Cacciola F, Asraoui F, Tayeq H, Ben Amar YM, Lovillo MP, Chouaibi N, Brigui J. Evaluation of different extraction methods on the phenolic profile and the antioxidant potential of Ceratonia siliqua L. pods extracts. Molecules. 2022; 27: 6163. doi: 10.3390/molecules27196163
  17. Li Z, Liu M, Wang H, Du S. Increased cutaneous wound healing effect of biodegradable liposomes containing madecassoside: preparation optimization, in vitro dermal permeation, and in vivo bioevaluation. International Journal of Nanomedicine. 2016; 11: 2995. doi: 10.2147/IJN.S105401
  18. Nasab ME, Takzaree N, Saffari PM, Partoazar A. In vitro antioxidant activity and in vivo wound-healing effect of lecithin liposomes: a comparative study. Journal of Comparative Effectiveness Research. 2019; 8(8): 633–643. doi: 10.2217/cer-2018-0128
  19. Haddadi R, Tamri P, Jooni FJ. In vitro wound healing activity of Scrophularia striata hydroalcoholic extract. South African Journal of Botany. 2019; 121: 505–509. doi: 10.1016/j.sajb.2018.10.038
  20. Noudoost B, Noori N, Amo Abedini G, Gandomi H, Akhondzadeh Basti A, Jebeli Javan A, Ghadami F. Encapsulation of green tea extract in nanoliposomes and evaluation of its antibacterial, antioxidant and prebiotic properties. Journal of Medicinal Plants. 2015; 3(55): 66–78.
  21. Golmohammadzadeh S, Jaafari M, Khalili N, Greenoak G. Determination of SPF and moisturizing effects of liposomal and conventional formulations of octyl methoxycinnamate as a sunscreen. Iranian Journal of Basic Medical Sciences. 2007; 10(2): 99–110.
  22. Nolen RS. Revision process begins for AVMA euthanasia guidelines. Journal of the American Veterinary Medical Association. 2009; 235(3): 246–247.
  23. Das S, Singh S, Dowding JM, Oommen S, Kumar A, Sayle TX, Saraf S, Patra CR, Vlahakis NE, Sayle DC, Self WT. The induction of angiogenesis by cerium oxide nanoparticles through the modulation of oxygen in intracellular environments. Biomaterials. 2012; 33(31): 7746–7755. doi: 10.1016/j.biomaterials.2012.07.019
  24. Lawrence RA, Burk RF. Glutathione peroxidase activity in selenium-deficient rat liver. Biochemical and Biophysical Research Communications. 1976; 71(4): 952–958. doi: 10.1016/0006-291X(76)90747-6
  25. Qiu Z, Kwon AH, Kamiyama Y. Effects of plasma fibronectin on the healing of full-thickness skin wounds in streptozotocin-induced diabetic rats. Journal of Surgical Research. 2007; 138(1): 64–70. doi: 10.1016/j.jss.2006.07.034
  26. Lachkar N, Al-Sobarry M, El Hajaji H, Lamkinsi T, Lachkar M, Cherrah Y, Alaoui K. Anti-inflammatory and antioxidant effect of Ceratonia siliqua L. methanol barks extract. Journal of Chemical and Pharmaceutical Research. 2016; 8(3): 202–210.
  27. Garcia FP, Marin E, Canigueral S, Adzet T. Anti-inflammatory activity of plant extracts. Life Sciences. 1996; 59(24): 2033–2040. doi: 10.1016/S0024-3205(96)00507-8
  28. Bruneton J. Pharmacognosie: Phytochimie, Plantes médicinales. 4th ed. Paris: Editions Tec & Doc; 2009.
  29. Polya GM. Biochemical targets of plant bioactive compounds. In: A Pharmacological Reference Guide to Sites of Action and Biological Effects. Florida: CRC Press; 2003.
  30. Giri TK. Breaking the barrier of cancer through liposome loaded with phytochemicals. Current Drug Delivery. 2019; 16(1): 3–17. doi: 10.2174/1567201815666180925140835
  31. Takahashi M, Kitamoto D, Asikin Y, Takara K, Wada K. Liposomes encapsulating Aloe vera leaf gel extract significantly enhance proliferation and collagen synthesis in human skin cell lines. Journal of Oleo Science. 2009; 58(12): 643–650. doi: 10.5650/jos.58.643
  32. Li P, Wu G. Roles of dietary glycine, proline, and hydroxyproline in collagen synthesis and animal growth. Amino Acids. 2018; 50: 29–38. doi: 10.1007/s00726-017-2500-6
  33. Aziz Z, Hassan BAR. The effects of honey compared to silver sulfadiazine for the treatment of burns: a systematic review of randomized controlled trials. Burns. 2017; 43(1): 50–57. doi: 10.1016/j.burns.2016.07.004
  34. Carmeliet P. Angiogenesis in health and disease. Nature Medicine. 2003; 9(6): 653–660. doi: 10.1038/nm0603-653
  35. Caetano GF, Fronza M, Leite MN, Gomes A, Frade MAC. Comparison of collagen content in skin wounds evaluated by biochemical assay and by computer-aided histomorphometric analysis. Pharmaceutical Biology. 2016; 54(11): 2555–2559. doi: 10.3109/13880209.2016.1170866
  36. Dwivedi D, Dwivedi M, Malviya S, Singh V. Evaluation of wound healing, anti-microbial and antioxidant potential of Pongamia pinnata in Wistar rats. Journal of Traditional and Complementary Medicine. 2017; 7(1): 79–85. doi: 10.1016/j.jtcme.2015.12.002
  37. Süntar IP, Akkol EK, Yılmazer D, Baykal T, Kırmızıbekmez H, Alper M, Yeşilada E. Investigations on the in vivo wound healing potential of Hypericum perforatum L. Journal of Ethnopharmacology. 2010; 127(2): 468–477. doi: 10.1016/j.jep.2009.10.011
  38. Diegelmann RF, Evans MC. Wound healing: an overview of acute, fibrotic and delayed healing. Frontiers in Bioscience. 2004; 9(1): 283–289. doi: 10.2741/1184
  39. Marchete R, Oliveira S, Bagne L, Silva JI, Valverde AP, Aro AA, Figueira MM, Fronza M, Bressam TM, Goes VF, Gaspari de Gaspi FO. Anti-inflammatory and antioxidant properties of Alternanthera brasiliana improve cutaneous wound healing in rats. Inflammopharmacology. 2021; 29(5): 1443–1458. doi: 10.1007/s10787-021-00858-2
Iranian Journal of Veterinary Surgery

Articles in Press, Corrected Proof
Available Online from 28 December 2025

  • Receive Date 17 September 2025
  • Revise Date 15 December 2025
  • Accept Date 28 December 2025
  • First Publish Date 28 December 2025