Bael Leaf Oil Increases AQP3 Expression and Exerts Wound-Healing Effect in Human Immortalized Keratinocytes


Budsarin Kesornnoi Orawin Prangsaengtong Duangratana Chuvisitkul Worapan Sitthithaworn


        Bael (Aegle marmelos L. Corrêa, Family Rutaceae) leaf oil has been reported to promote wound healing in keratinocyte cell models. The oil key components, according to GC-MS analysis, are trans-caryophyllene and limonene. In this study, we investigated the mechanism by which bael leaf oil and its key components aid wound healing by focusing on the water channel aquaporin-3 (AQP3), which is crucial for maintaining skin hydration and identifying the active component responsible for the activity. To this objective, keratinocyte (HaCaT) cells were grown in media containing 2% FBS and subjected to a wound healing assay. The expression of AQP3 mRNA in the cells was observed using RT-real-time PCR. The results demonstrated that trans-caryophyllene and bael leaf oil significantly stimulated keratinocytes to close the wound (p<0.05 vs. vehicle group) and enhanced the level of AQP3 mRNA expression. It suggests that trans-caryophyllene, at least among the components of bael leaf oil, enhances the wound healing action instigated by the bael leaf oil, and that this effect is associated with the water channel aquaporin-3.

Keywords: bael leaf oil, trans-caryophyllene, HaCaT, wound healing, aquaporin-3


Ahmad, M., Khan, T. H., Ansari, M. N., & Ahmad, S.F. (2014). Enhanced wound healing by topical administration of d-limonene in alloxan induced diabetic mice through reduction of pro-inflammatory markers and chemokine expression. BMC Genomics, 15(2), 29.
Boury-Jamot, M., Sougrat, R., Tailhardat, M., Le Varlet, B., Bonté, F., Dumas, M., & Verbavatz, J.M. (2006). Expression and function of aquaporins in human skin: Is aquaporin-3 just a glycerol transporter? Biochim Biophysic Acta, 1758(8), 1034-1042.
Boury-Jamot, M., Daraspe, J., Bonté, F., Perrier, E., Schnebert, S., Dumas, M., & Verbavatz, J.M. (2009). Skin aquaporins: function in hydration, wound healing, and skin epidermis homeostasis. Handbook of Experimental Pharmacology, 190, 205-17.
d'Alessio, P. A., Mirshahi, M., Bisson, J. F., & Bene, M. C. (2014). Skin repair properties of d-limonene and perillyl alcohol in murine models. Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry, 13(1), 29-35.
Francomano, F., Caruso, A., Barbarossa, A., Fazio, A., La Torre, C., Ceramella, J., … Sinicropi, M. S. (2019). β-Caryophyllene: a sesquiterpene with countless biological properties. Applied Sciences, 9(24), 5420.
Grada, A., Otero-Vinas, M., Prieto-Castrillo, F., Obagi, Z., & Falanga, V. (2017). Research techniques made simple: analysis of collective cell migration using the wound healing assay. Journal of Investigative Dermatology, 137(2), e11-e16.
Gushiken, L. F. S., Beserra, F. P., Hussni, M. F., Gonzaga, M. T., Ribeiro, V. P., de Souza, P. F., … Pellizzon, C. H. (2022). Beta-caryophyllene as an antioxidant, anti-inflammatory and re-epithelialization activities in a rat skin wound excision model. Oxidative Medicine and Cellular Longevity, 3, 9004014.
Kadam, S., Vandana, M., & Kaushik, K. S., (2020). Reduced serum methods for contact-based coculture of human dermal fibroblasts and epidermal keratinocytes. BioTechniques, 69, 00–00.
Kesornnoi, B., Prangsaengtong, O., & Sitthithaworn, W. (2022). Volatile oil from Aegle marmelos leaf facilitates wound healing in a human epidermal keratinocyte cell model. Proceeding on International Conference and Exhibition on Pharmaceutical Science and Technology held in Thammasat University, Patumthanee, Thailand 23-24 June 2022. Thailand: Faculty of Pharmacy, Thammasat University.
Koyama, S., Purk, A., Kaur, M., Soini, H. A., Novotny, M. V., Davis, K., … Mescher, A. (2019) Beta-caryophyllene enhances wound healing through multiple routes. PLoS ONE, 14(12), e0216104.
Jonkman, J. E., Cathcart, J. A., Xu, F., Bartolini, M. E., Amon, J. E., Stevens, K. M., & Colarusso, P. (2014). An introduction to the wound healing assay using live-cell microscopy. Cell Adhesion & Migration, 8(5), 440-451.
Liang, C. C., Park, A., & Guan, J. L. (2007). In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro, Nature Protocol, 2, 329–333.
Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2ΔΔCt method. Methods, 25, 402-408.
Martinotti, S., Pellavio, G., Laforenza, U., & Ranzato, E. (2019). Propolis induces AQP3 expression: a possible way of action in wound healing. Molecules, 24(8), 1544.
Stamm, A., Reimers, K., Strauß, S., Vogt, P., Scheper, T., & Pepelanova, I. (2016). In vitro wound healing assays-state of the art. BioNanoMaterials, 17(1), 79–87.

Research Articles


How to Cite
KESORNNOI, Budsarin et al. Bael Leaf Oil Increases AQP3 Expression and Exerts Wound-Healing Effect in Human Immortalized Keratinocytes. Naresuan University Journal: Science and Technology (NUJST), [S.l.], v. 31, n. 3, p. 67-75, aug. 2023. ISSN 2539-553X. Available at: <>. Date accessed: 29 nov. 2023. doi: