In vitro Wound Healing Potential of Petroleum Ether Extracts from Litsea garciae Fruit Pulp and Seeds
Keywords:
collagen type 1, EGF, Litsea garciae, petroleum ether extract, wound healingAbstract
This study explores Litsea garciae’s (L. garciae) untapped wound healing potential, despite its traditional medicinal use. The current study aimed to elucidate the essential role of lipids in the wound healing process by evaluating the effects of L. garciae petroleum ether (PE) lipid extracts on human dermal fibroblast (HDF) cell line. The cytotoxicity of PE lipid extracts on the HDF was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. An in vitro study on the migration of HDF was performed using the wound scratch assay while the chemotactic motility of HDF was conducted by the transwell assay. The effects of L. garciae extracts on the expression levels of collagen type I and epidermal growth factor (EGF) were assessed using the enzyme-linked immunosorbent assays (ELISA). The MTT assay showed that PE lipid extracts (for pulp, 7.5 – 250 µg/mL and for seed, 7.5 to 500 µg/mL) were non-cytotoxic to HDF cells. Both extracts promoted cell migration, with 31.25 µg/mL pulp extract exhibited the highest wound closure rate (91.27 ± 8.35%) at 24 h, and 7.5 µg/mL seed extract significantly enhanced closure (90.62 ± 3.12%). The transwell assay indicated increased chemotactic motility when the HDF cells were treated with lipid extracts. Collagen type I synthesis improved in both pulp and seed lipid extracts. The pulp extracts enhanced EGF secretion at various concentrations, while the seed extracts showed no improvement. The study highlights L. garciae extracts’ potential in wound healing by facilitating cell migration, collagen synthesis, and EGF secretion.
References
Addis, R., Cruciani, S., Santaniello, S., Bellu, E., Sarais, G., Ventura, C., Maioli, M. & Pintore, G. (2020). Fibroblast proliferation and migration in wound healing by phytochemicals: evidence for a novel synergic outcome. International Journal of Medical Sciences, 17 (8): 1030–1042. DOI: 10.7150/ijms.43986.
Ado, M.A., Abas, F., Mohammed, A.S. & Ghazali, H. M. (2013). Anti-and pro-lipase activity of selected medicinal, herbal and aquatic plants, and structure elucidation of an anti-lipase compound. Molecules, 18 (12): 14651-14669. DOI: 10.3390/molecules181214651.
Amit, Z. & Ling, Z. (2021). A mini review on the nutritional compositions and pharmacological properties of Litsea garciae. Malaysian Applied Biology, 50 (1): 29–39. DOI: 10.55230/mabjournal.v50i1.10.
Aoki, M., Aoki, H., Mukhopadhyay, P., Tsuge, T., Yamamoto, H., Matsumoto, N.M., Toyohara, E., Okubo, Y., Ogawa, R. & Takabe, K. (2019). Sphingosine-1-phosphate facilitates skin wound healing by increasing angiogenesis and inflammatory cell recruitment with less scar formation. International Journal of Molecular Sciences, 20 (14): 1–16. DOI: 10.3390/ijms20143381.
Broughton, G., Janis, J.E. & Attinger, C.E. (2006). Wound healing: An overview. Plastic & Reconstructive Surgery, 117 (7S): 1–32. DOI: 10. 1097/01.prs.0000222562.60260.f9.
Bodnar, R.J. (2013). Epidermal growth factor and epidermal growth factor receptor: the yin and yang in the treatment of cutaneous wounds and cancer. Advances in Wound Care, 2 (1): 24–29. DOI: 10.1089/wound.2011.0326.
Bolla, S.R., Al-Subaie, A.M., Al-Jindan, R.Y., Balakrishna, J.P., Ravi, P.K., Veeraraghavan, V. P., Pillai, A.A., Gollapalli, S.S.R., Joseph, J.P. & Surapaneni, K.M. (2019). In vitro wound healing potency of methanolic leaf extract of Aristolochia saccata is possibly mediated by its stimulatory effect on collagen-1 expression. Heliyon, 5 (5):1–7. DOI: 10.1016/j.heliyon.2019.e01648.
Buranasukhon, W., Athikomkulchai, S., Tadtong, S. & Chittasupho, C. (2017). Wound healing activity of Pluchea indica leaf extract in oral mucosal cell line and oral spray formulation containing nanoparticles of the extract. Pharmaceutical Biology, 55 (1): 1767–1774. DOI: 10.1080/13880209.2017.1326511.
Caplan, A.I. (2005). Review: Mesenchymal stem cells: Cell–based reconstructive therapy in orthopedics. Tissue Engineering, 11 (7–8): 1198–1211. DOI: 10.1089/ten.2005.11.1198.
Cardoso, C.R.B., Souza, M.A., Ferro, E.a.V., Favoreto, S. & Pena, J.D.O. (2004). Influence of topical administration of n‐3 and n‐6 essential and n‐9 nonessential fatty acids on the healing of cutaneous wounds. Wound Repair and Regeneration, 12 (2): 235–243. DOI: 10.1111/j.1067-1927.2004.012216.x.
Cen, R., Wang, L., He, Y., Yue, C., Tan, Y., Li, L. & Lei, X. (2022). Dermal fibroblast migration and proliferation upon wounding or lipopolysaccharide exposure is mediated by stathmin. Frontiers in Pharmacology, 12: 1–14. DOI: 10.3389/fphar.2021.781282.
Choi, S., Yoon, M. & Choi, K. (2022). Approaches for regenerative healing of cutaneous wound with an emphasis on strategies activating the Wnt/β-Catenin Pathway. Advances in Wound Care, 11 (2): 70–86. DOI: 10.1089/wound.2020.1284.
Comino-Sanz, I.M., López-Franco, M.D., Castro, B. & Pancorbo-Hidalgo, P.L. (2021). The role of antioxidants on wound Healing: A review of the current evidence. Journal of Clinical Medicine, 10 (16): 1-22. DOI: 10.3390/jcm10163558.
de Albuquerque, P.B.S., Rodrigues, N.E.R., Silva, P.M.D.S., De Oliveira, W.F., Correia, M.T.D.S. & Coelho, L.C.B.B. (2023). The use of proteins, lipids, and carbohydrates in the management of wounds. Molecules, 28 (4): 1–23. DOI: 10.3390/molecules28041580.
de Oliveira, A.P., De Souza Franco, E., Barreto, R.R., Cordeiro, D.P., De Melo, R.G., De Aquino, C.M. F., Silva, A.A.R.E., De Medeiros, P.L., Da Silva, T.G., Da Silva Góes, A.J. & De Sousa Maia, M.B. (2013). Effect of semisolid formulation of Persea americana mill (avocado) oil on wound healing in rats. Evidence-based Complementary and Alternative Medicine, 2013: 1–8. DOI: 10.1155/2013/472382.
Frank, S., Kämpfer, H., Wetzler, C. & Pfeilschifter, J. (2002). Nitric oxide drives skin repair: Novel functions of an established mediator. Kidney International, 61 (3): 882–888. DOI: 10.1046/j.1523-1755.2002.00237.x.
Guidoni, M., De Christo Scherer, M., Figueira, M., Schmitt, E., De Almeida, L., Scherer, R., Bogusz, S. & Fronza, M. (2019). Fatty acid composition of vegetable oil blend and in vitro effects of pharmacotherapeutical skin care applications. Brazilian Journal of Medical and Biological Research, 52 (2): 1–8. DOI: 10.1590/1414-431x20188209.
Guo, S. & DiPietro, L. (2010). Factors affecting wound healing. Journal of Dental Research, 89 (3): 219–229. DOI: 10.1177/0022034509359125.
Häkkinen, L., Larjava, H. & Koivisto, L. (2011). Granulation tissue formation and remodeling. Endodontic Topics, 24 (1): 94–129. DOI: 10.1111/etp.12008.
Hassan, S.H.A., Fry, J.R. & Bakar, M.F.A. (2013). Antioxidant and phytochemical study on pengolaban (Litsea garciae), an edible underutilized fruit endemic to Borneo. Food Science and Biotechnology, 22 (5): 1–7. DOI: 10.1007/s10068-013-0202-x.
Ili, P. & Sari, F. (2023). Egg yolk oil accelerates wound healing in streptozotocin induced diabetic rats. Biotechnic & Histochemistry, 98 (2): 94–111. DOI: 10.1080/10520295.2022.2115554.
James, O. & Friday, E.T. (2010). Phytochemical composition, bioactivity and wound healing potential of Euphorbia heterophylla (Euphorbiaceae) leaf extract. International Journal on Pharmaceutical and Biomedical Research, 1 (1): 54-63.
Johnny, L., Yusuf, U.K. & Nulit, R. (2010). The effect of herbal plant extracts on the growth and sporulation of Colletotrichum gloeosporioides. Journal of Applied Biosciences, 34: 2218–2224.
Kim, D., Kim, S.Y., Mun, S.K., Rhee, S. & Kim, B.J. (2015). Epidermal growth factor improves the migration and contractility of aged fibroblasts cultured on 3D collagen matrices. International Journal of Molecular Medicine, 35 (4): 1017–1025. DOI: 10.3892/ijmm.2015.2088.
Kim, W., Park, B., Sung, J., Yang, J., Park, S., Kwak, S. & Park, J. (2007). Wound healing effect of adipose-derived stem cells: A critical role of secretory factors on human dermal fibroblasts. Journal of Dermatological Science, 48 (1): 15–24. DOI: 10.1016/j.jdermsci.2007.05.018.
Kutoi, C.J., Yen, K.H. & Seruji, N.M.U. (2012, April). Pharmacology evaluation of Litsea garciae (Lauraceae). In 2012 IEEE Business, Engineering & Industrial Applications Colloquium (BEIAC) IEEE. pp. 31–33. DOI: 10.1109/BEIAC.2012.6226075.
Lazarus, G.S., Cooper, D.M., Knighton, D.R., Margolis, D.J., Percoraro, R.E., Rodeheaver, G. & Robson, M.C. (1994). Definitions and guidelines for assessment of wounds and evaluation of healing. Wound Repair and Regeneration, 2 (3): 165–170. DOI: 10.1046/j.1524-475x.1994.20305.x.
Li, L., Wang, M., Yuan, T., Xu, X., Dad, H. A., Yu, C., Hou, J. & Peng, L. (2019). The crude ethanol extract of Periplaneta americana L. stimulates wound healing in vitro & in vivo. Chinese Medicine, 14 (1): 1–9. DOI: 10.1186/s13020-019-0259-4.
Ling, Z., Gabriele, R.a.F. & Zunika, A. (2022). Fatty acids composition and antimicrobial activities of Litsea garciae pulp and seed extracts. Medicinal Plants - International Journal of Phytomedicines and Related Industries, 14 (2): 301–311. DOI: 10.5958/0975-6892.2022.00035.1.
Mapoung, S., Umsumarng, S., Semmarath, W., Arjsri, P., Thippraphan, P., Yodkeeree, S. & Limtrakul, P. (2021). Skin wound-healing potential of polysaccharides from medicinal mushroom Auricularia auricula-judae (Bull.). Journal of Fungi, 7 (4): 1–16. DOI: 10.3390/jof7040247.
Mirfat, A.H.S., Amin, I., Kartinee, K.N., Muhajir, H. & Shukri, M.a.M. (2018). Underutilised fruits: a review of phytochemistry and biological properties. Journal of Food Bioactives, 1: 2–30. DOI: 10.31665/jfb.2018.1124.
Nguyen, T.L.A. & Bhattacharya, D. (2022). Antimicrobial activity of quercetin: An approach to its mechanistic principle. Molecules, 27 (8): 1–13. DOI: 10.3390/molecules27082494.
Park, S.M., Won, K.J., Hwang, D.I., Kim, D.Y., Kim, H.B., Li, Y. & Lee, H.M. (2020). Potential beneficial effects of Digitaria ciliaris flower absolute on the wound healing-linked activities of fibroblasts and keratinocytes. Planta Medica, 86 (05): 348–355. DOI: 10.1055/a-1101-9326.
Pils, V., Terlecki-Zaniewicz, L., Schosserer, M., Grillari, J. & Lämmermann, I. (2021). The role of lipid-based signalling in wound healing and senescence. Mechanisms of Ageing and Development, 198:1–15. DOI: 10.1016/j.mad.2021.111527.
Poli, F.M. & Assim, Z. (2019). Fatty acid profiles in the kernel oils of Artocarpus odoratissimus and Litsea garciae. Journal of Engineering and Applied Sciences, 14 (3): 6135–6138.
Postlethwaite, A.E., Keski-Oja, J., Moses, H.L. & Kang, A. H. (1987). Stimulation of the chemotactic migration of human fibroblasts by transforming growth factor beta. The Journal of Experimental Medicine, 165 (1): 251–256. DOI: 10.1084/jem.165.1.251.
Raduan, S.Z., Ahmed, Q.U., Kasmuri, A.R., Rusmili, M.R.A., Mia, M.A.R., Sulaiman, W.M.A.W., Mahmood, M.H. & Shaikh, M.F. (2022). Antioxidant capabilities of Litsea garciae bark extracts and their relation to the phytochemical compositions. Malaysian Applied Biology, 51 (1): 99–118. DOI: 10.55230/mabjournal.v51i1.2038.
Rangaraj, A., Harding, K. & Leaper, D. (2011). Role of collagen in wound management. Wounds UK, 7 (2): 54–63.
Ranzato, E., Mazzucco, L., Patrone, M. & Burlando, B. (2008). Platelet lysate promotes in vitro wound scratch closure of human dermal fibroblasts: different roles of cell calcium, P38, ERK and PI3K/AKT. Journal of Cellular and Molecular Medicine, 13 (8b), 2030–2038. DOI: 10.1111/j.1582-4934.2008.00467.x.
Rojo, L. E., Villano, C. M., Joseph, G., Schmidt, B., Shulaev, V., Shuman, J. L., Lila, M. A. & Raskin, I. (2010). Original Contribution: wound‐healing properties of nut oil from Pouteria lucuma. Journal of Cosmetic Dermatology, 9 (3): 185–195. DOI: 10.1111/j.1473-2165.2010.00509.x.
Schäfer, M. & Werner, S. (2007). Transcriptional control of wound repair. Annual Review of Cell and Developmental Biology, 23 (1): 69–92. DOI: 10.1146/annurev.cellbio.23.090506.123609.
Seo, S.H., Lee, S.H., Cha, P.H., Kim, M.Y., Min, D.S. & Choi, K.Y. (2016). Polygonum aviculare L. and its active compounds, quercitrin hydrate, caffeic acid, and rutin, activate the Wnt/β‐catenin pathway and healing. Phytotherapy Research, 30 (5): 848–854. DOI: 10.1002/ptr.5593.
Silva, J.R., Burger, B., Kühl, C.M.C., Candreva, T., Anjos, M.B.P.D. & Rodrigues, H.G. (2018). Wound healing and omega-6 fatty acids: From inflammation to repair. Mediators of Inflammation, 2018: 1–17. DOI: 10.1155/2018/ 2503950.
Singer, A.J. & Clark, R. A. (1999). Cutaneous wound healing. New England Journal of Medicine, 341 (10): 738–746. DOI: 10.1056/nejm199909023411006.
Smith, A.N., Muffley, L.A., Bell, A.N., Numhom, S. & Hocking, A.M. (2011). Unsaturated fatty acids induce mesenchymal stem cells to increase secretion of angiogenic mediators. Journal of Cellular Physiology, 227 (9): 3225–3233. DOI: 10.1002/jcp.24013.
Souto, E.B., Yoshida, C.M.P., Leonardi, G.R., Cano, A., Sanchez-Lopez, E., Zielinska, A., Viseras, C., Severino, P., Da Silva, C.F. & De M Barbosa, R. (2021). Lipid-Polymeric films: composition, production and applications in wound healing and skin repair. Pharmaceutics, 13 (8): 1–23. DOI: 10.3390/pharmaceutics13081199.
Stock, J.B. & Baker, M.D. (2009). Chemotaxis. In Encyclopedia of Microbiology. Third Edition. Elsevier. pp. 71–78.
Tracy, L.E., Minasian, R.A. & Caterson, E. (2016). Extracellular matrix and dermal fibroblast function in the healing wound. Advances in Wound Care, 5 (3): 119–136. DOI: 10.1089/wound.2014.0561.
Tschumperlin, D.J. (2013). Fibroblasts and the ground they walk on. Physiology, 28 (6): 380–390. DOI: 10.1152/physiol.00024.2013.
Vaidyanathan, L. (2021). Growth factors in wound healing – a review. Biomedical & Pharmacology Journal, 14 (3): 1469–1480. DOI: 10.13005/bpj/2249.
Wulandari, I., Kusuma, I.W. & Kuspradini, H. (2018). Antioxidant and antibacterial activity of Litsea garciae. IOP Conference Series Earth and Environmental Science, 144 (1): 1–7. DOI: 10.1088/1755-1315/144/1/012024.
Zhang, C., Barrios, M.P., Alani, R.M., Cabodi, M. & Wong, J.Y. (2016). A microfluidic transwell to study chemotaxis. Experimental Cell Research, 342 (2): 159–165. DOI: 10.1016/j.yexcr.2016.03.010.
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