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. 2010 Sep;9(3):185-95.
doi: 10.1111/j.1473-2165.2010.00509.x.

Wound-healing Properties of Nut Oil From Pouteria Lucuma

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Free PMC article

Wound-healing Properties of Nut Oil From Pouteria Lucuma

Leonel E Rojo et al. J Cosmet Dermatol. .
Free PMC article

Abstract

Background: Cell migration, angiogenesis, inflammation, and extracellular matrix remodeling are key events in wound healing. Natural products, including fatty acids (FAs), can accelerate wound healing by modulating the aforementioned events.

Aims: This study aims to evaluate the effect of lucuma (Pouteria lucuma O Kezte) nut oil (LNO) on fibroblasts migration, angiogenesis, inflammation, bacterial and fungal growth, and wound healing. Methods GC-MS analysis of FAs methyl esters (FAMES) was used for chemical characterization of LNO. In vitro studies were carried out with LNO investigating the induction of cell migration, cytoskeleton remodeling of human fibroblasts, inhibition of LPS-induced nitric oxide production in macrophages, and antibacterial and antifungal effects. Two in vivo studies were carried out to study LNO's effect on angiogenesis and wound healing: (i) tail fin regeneration in transgenic zebrafish larvae expressing enhanced green fluorescent protein (EGFP) in vascular endothelial cells was used to study vessel sprouting and wound healing and (ii) the closure of wounds was evaluated in CD-1 mice after topical applications of LNO-containing formulations.

Results: Lucuma nut oil is a mixture of FAs, 99.7% of which were characterized. Major components of LNO (w/w) are linoleic acid (38.9%), oleic acid (27.9%), palmitic acid (18.6%), stearic acid (8.9%), and γ linolenic acid (2.9%). In vitro studies showed that LNO significantly promoted migration and vinculin expression in human fibroblasts. LNO decreased LPS-induced nitric oxide production and did not display significant antibacterial or antifungal effects. LNO induced tail fin regeneration in transgenic zebrafish larvae 48 h after tail fin amputation and significantly accelerated cutaneous wound closure in CD-1 mice.

Conclusions: Natural FAs from P. lucuma nut promote skin regeneration and, thus, may have applications in medicine and skin care.

Figures

Figure 1
Figure 1
Chromatographic profile of FAMEs from lucuma nut oil (LNO) analyzed by GC–MS. (a) Chromatograms of LNO 10 dilution and (b) food industry standard mixture of 37 FAMEs; both LNO and standards were injected at 100 µg/mL.
Figure 2
Figure 2
Effect of lucuma nut oil (LNO) on human fibroblasts migration. Human neonatal fibroblasts were labeled with the green fluorescent probe Calcein-AM. The effect of LNO on fibroblast migration was evaluated using the ORIS° cell migration assay as described in the experimental section. Digital images were taken 72 h after starting the experiment. From left to right: time zero (t = 0), Vehicle, LNO 40 µg/mL (LNO 40) and LNO 60 µg/mL (LNO 60).
Figure 3
Figure 3
Effect of lucuma nut oil (LNO) on scratched monolayer of human fibroblasts. Monolayers of fibroblasts were scratched and treated with LNO 60 µg/mL (n = 3) or with equivalent concentrations of pure fatty acids (FAs), oleic acid (OA, n = 4), linoleic acid (LA, n = 3) or 10% fetal bovine serum (FBS, n = 4) as positive control. The effect of LNO and FAs was evaluated as described in the experimental section. Values are the mean ± standard error (**P < 0.01 vs. FBS, one-way anova, Newman–Keuls multiple comparison test).
Figure 4
Figure 4
Effect of lucuma nut oil (LNO) on vinculin expression and actin fibers organization in fibroblasts. Human dermal fibroblasts were treated with LNO 60 µg/mL or vehicle and doubly stained with rhodamine phalloidin (red) and mAb anti-vinculin (green). LNO caused mild decrease in stress fibers (F-actin) and increased the immunoreactivity to vinculin.
Figure 5
Figure 5
Effect of lucuma nut oil (LNO) on LPS-induced nitric oxide production in RAW 264.7 cells. RAW cells were pretreated with either vehicle alone or LNO for 2 h. Subsequently, LPS (1 µg/mL) was added to each well and incubated for 8 h. LPS significantly increased NO production with respect to the nonstimulated cells (control). LNO decreased LPS-induced NO production in a concentration-dependent manner. Results are expressed as mean ± standard error (***P < 0.001 vs. control **P < 0.01 vs. control, one-way anova, Newman–Keuls multiple comparison test).
Figure 6
Figure 6
Effect of lucuma nut oil (LNO) on tail fin regeneration and angiogenesis in transgenic zebrafish larvae. (a) Representative fluorescent micrographs of GFP-positive endothelial cells during tail fin regeneration induced by 100 µg/mL of LNO (LNO 100). (b) Quantitation of tail endothelial cells regeneration 48 h after amputation (hpa) of tail fin. Results were expressed as mean ± standard error of nine replicates (*P = 0.003, P = 0.079 vs. control).
Figure 7
Figure 7
Dose-dependent effect of lucuma nut oil on tail fin endothelial repopulation of transgenic zebrafish larvae. The increase in fluorescent GFP-positive-endothelial cells 48 h after amputation (hpa) of tail fin was dose dependent. Results were expressed as mean ± standard error of nine replicates.
Figure 8
Figure 8
Time course of in vivo wound-healing effect of lucuma nut oil (LNO). The percent of wound closure was evaluated during 11 days of topical application of LNO 200, 500, and 1000 µg/wound. Values are presented as mean ± standard error of five replicates (*P < 0.05 vs. vehicle). One-way anova followed by Dunnett’s test was applied for comparisons between the treated and vehicle.
Figure 9
Figure 9
Progression of the dermal wound healing induced by lucuma nut oil (LNO). Representative images of the wound-healing progression in mice after daily application of LNO (500 and 1000 µg/wound), vehicle (CMC 50.5%, PBS pH7.4) and CGS (10 µg/mouse). Photographs of the wounds were taken every other day during 11 days postskin wounding (DPSW).

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