The Wound Healing and Antibacterial Activity of Five Ethnomedical Calophyllum inophyllum Oils: An Alternative Therapeutic Strategy to Treat Infected Wounds

Type Article
Date 2015-09
Language English
Author(s) Leguillier Teddy1, Lecso-Bornet Marylin, Lemus Christelle3, Rousseau-Ralliard Delphine4, Lebouvier Nicolas5, Hnawia Edouard5, Nour Mohammed5, Aalbersberg William6, Ghazi Kamelia7, Raharivelomanana Phila8, Rat Patrice1
Affiliation(s) 1 : Univ Paris 05, UMR CNRS COMETE 8638, Lab Chim Toxicol Analyt & Cellulaire, Sorbonne Paris Cite,Fac Pharm, Paris, France.
2 : Univ Paris 05, Sorbonne Paris Cite, Fac Pharm, Lab Ecosyst Intestinal Probiot Antibio EA 4065, Paris, France.
3 : Univ Paris 05, UMR CNRS COMETE 8638, Lab Pharmacognosie, Sorbonne Paris Cite,Fac Pharm, Paris, France.
4 : INRA, Biol Dev & Reprod, UMR 1198, Jouy En Josas, France.
5 : Univ Nouvelle Caledonie, Lab Insuiaire Vivant & Environm, EA 4243, Noumea, New Caledonia.
6 : Univ S Pacific, Inst Appl Sci, Suva, Fiji.
7 : Univ Technol Compiegne, CNRS UMR 7338, Ctr Rech BioMecan & Bioingenierie, Compiegne, France.
8 : Univ Polynesie Francaise, UMR EIO 241, Equipe Etude Integree Metabolites Secondaires, FAAA, Tahiti, France.
Source Plos One (1932-6203) (Public Library Science), 2015-09 , Vol. 10 , N. 9 , P. e0138602. (20p.)
DOI 10.1371/journal.pone.0138602
WOS© Times Cited 16
Abstract

Background Calophyllum inophyllum L. (Calophyllaceae) is an evergreen tree ethno-medically used along the seashores and islands of the Indian and Pacific Oceans, especially in Polynesia. Oil extracted from the seeds is traditionally used topically to treat a wide range of skin injuries from burn, scar and infected wounds to skin diseases such as dermatosis, urticaria and eczema. However, very few scientific studies reported and quantified the therapeutic properties of Calophyllum inophyllum oil (CIO). In this work, five CIO from Indonesia (CIO1), Tahiti (CIO2, 3), Fiji islands (CIO4) and New Caledonia (CIO5) were studied and their cytotoxic, wound healing, and antibacterial properties were presented in order to provide a scientific support to their traditional use and verify their safety. Methods The safety of the five CIO was ascertained using the Alamar blue assay on human keratinocyte cells. CIO wound healing properties were determined using the scratch test assay on human keratinocyte cells. CIO-stimulated antibacterial innate immune response was evaluated using ELISA by measuring beta defensin-2 release in human derivative macrophage cells. CIO antibacterial activity was tested using oilogramme against twenty aerobic Gram-bacteria species, twenty aerobic Gram+ bacteria species, including a multi-drug resistant Staphylococcus aureus strain and two anaerobic Gram+ bacteria species e.g. Propionibacterium acnes and Propionibacterium granulosum. To detect polarity profile of the components responsible of the antibacterial activity, we performed bioautography against a Staphylococcus aureus strain. Results Based on Alamar Blue assay, we showed that CIO can be safely used on keratinocyte cells between 2.7% and 11.2% depending on CIO origin. Concerning the healing activity, all the CIO tested accelerated in vitro wound closure, the healing factor being 1.3 to 2.1 higher compared to control when keratinocytes were incubated after scratch with CIO at 0.1%. Furthermore, our results showed that CIO exhibit two distinct antibacterial effects: one against Gram + bacteria by direct inhibition of mitotic growth and another potent effect against Gram-bacteria due to increased release of beta-defensin 2 peptide by macrophages. Interestingly, the needed concentrations of CIO to inhibit bacteria growth and to promote wound healing are lower than concentrations exhibiting cytotoxic effects on keratinocyte cells. Finally, we performed bioautography assay against Staphylococcus aureus to determine polarity profile of the components responsible for CIO antibacterial activity. Our results showed for the five tested CIO that components responsible of the bacterial growth inhibition are the more polar one on the TLC chromatographic profile and are contained in the resinous fraction of the oil. Conclusions This study was conducted to evaluate cytotoxicity, wound healing and antibacterial properties of five CIO traditionally used to treat infected wounds. Using cell and bacteria cultures, we confirmed the pharmacological effects of CIO as wound healing and antimicrobial agent. Moreover, we showed that concentration of CIO needed to exhibit therapeutic effects are lower than concentrations exhibiting cytotoxic effects in vitro. For the first time, this study provides support for traditional uses of CIO. These wound healing and antibiotic properties make CIO a valuable candidate to treat infected wounds especially in tropical areas.

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S1 Fig. Aerobic Gram-negative and Gram-positive bacteria repartition on the plates. 1 26 KB Open access
S1 Table. References, geographic origins and characteristics of CIO. 1 114 KB Open access
S2 Table. Fatty acid composition of CIO. 1 303 KB Open access
S3 Table. Gram-negative bacterial strains tested in preliminary assay. 1 335 KB Open access
S4 Table. Aerobic Gram-negative and Gram-positive bacteria tested against CIO. 2 1 MB Open access
S5 Table. Anaerobic Gram-positive bacteria tested against CIO. 1 535 KB Open access
S6 Table. Bacterial strains resistant to antibiotics. 1 261 KB Open access
Publisher's official version 20 32 MB Open access
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Leguillier Teddy, Lecso-Bornet Marylin, Lemus Christelle, Rousseau-Ralliard Delphine, Lebouvier Nicolas, Hnawia Edouard, Nour Mohammed, Aalbersberg William, Ghazi Kamelia, Raharivelomanana Phila, Rat Patrice (2015). The Wound Healing and Antibacterial Activity of Five Ethnomedical Calophyllum inophyllum Oils: An Alternative Therapeutic Strategy to Treat Infected Wounds. Plos One, 10(9), e0138602. (20p.). Publisher's official version : https://doi.org/10.1371/journal.pone.0138602 , Open Access version : https://archimer.ifremer.fr/doc/00667/77934/