Journal of Ethnopharmacology 171 (2015) 330–334

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Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jep

Sacha Inchi Oil (Plukenetia volubilis L.), effect on adherence

of Staphylococus aureus to human skin explant and keratinocytes
in vitro
German Gonzalez-Aspajo a,b, Haouaria Belkhelfa c, Laïla Haddioui-Hbabi c,
Geneviève Bourdy a,b, Eric Deharo a,b,n
a
Institut de Recherche pour le Développement (IRD), UMR 152 Pharma-DEV, F-31062 Toulouse cedex 09, France
b
Université de Toulouse 3, UMR 152 Pharma-DEV, Faculté des Sciences Pharmaceutiques, F-31062 Toulouse cedex 09, France
c
Fonderephar, Université Toulouse 3, Faculté des Sciences Pharmaceutiques, F-31062 Toulouse cedex 09, France

art ic l e i nf o a b s t r a c t

Article history: Ethnopharmacological relevance: Plukenetia volubilis L. (Euphorbiaceae) is a domesticated vine distributed
Received 17 April 2015 from the high-altitude Andean rain forest to the lowlands of the Peruvian Amazon. Oil from the cold-
Received in revised form pressed seeds, sold under the commercial name of Sacha Inchi Oil (SIO) is actually much in favour
4 June 2015
because it contains a high percentage of omega 3 and omega 6, and is hence used as a dietary
Accepted 5 June 2015
Available online 16 June 2015
supplement. SIO is also used traditionally for skin care, in order to maintain skin softness, and for the
treatment of wounds, insect bites and skin infections, in a tropical context where the skin is frequently
Keywords: damaged.
Sacha Inchi Oil Aims of the study: This study was designed in order to verify whether the traditional use of SIO for skin
Staphylococcus aureus
care would have any impact on Staphylococcus aureus growth and skin adherence, as S. aureus is involved
Human skin explant
in many skin pathologies (impetigo, folliculitis, furuncles and subcutaneous abscesses) being one if the
Keratinocytes cells
Plukenetia volubilis L. main pathogens that can be found on the skin. Therefore, our objective was to assess SIO bactericidal
activity and interference with adherence to human skin explants and the keratinocyte cell line.
Cytotoxicity on that cells was also determined. The activity of SIO was compared to coconut oil (CocO),
which is widely used for skin care but has different unsaturated fatty acids contents.
Materials and methods: Laboratory testing with certified oil, determined antibacterial activity against
radio labelled S. aureus. Cytotoxic effects were measured with XTT on keratinocyte cells and with neutral
red on human skin explants; phenol was used as cytotoxic control. Adherence assays were carried out by
mixing H3-labelled S. aureus bacteria with keratinocyte cells and human skin explants, incubated with
oils 2 h before (to determine the inhibition of adherence, assimilated to a preventive effect) or 2 h after
the contact of the biological material with S. aureus (to assess the detachment of the bacteria, assimilated
to a curative effect). Residual radioactivity measured after washings made it possible to determine the
adherence intensity. Bactericidal effect was determined by colony counting on trypticase soy agar.
Results: Laboratory assays showed that SIO and CocO, tested undiluted, were not cytotoxic on
keratinocytes nor human explants and were not bactericidal neither. SIO was more active as antiadherent
(preventive) than CocO on keratinocytes. There was no significant difference between detachment effects
(curative) of both oils on keratinocytes but SIO was almost 5 times more active on the detachment of
S. aureus from human skin explants.
Conclusion: From that study it can be concluded that the use of SIO on dermal cells is safe and efficient in
the inhibition of S. aureus adherence. Our results tend to support the traditional use of undiluted SIO in
skin care.
& 2015 Elsevier Ireland Ltd. All rights reserved.

1. Introduction

Plukenetia volubilis L. (Euphorbiaceae) is a perennial oleaginous

n
Corresponding author at: Université de Toulouse 3, UMR 152 Pharma-DEV, woody vine belonging to the Euphorbiaceae family. It grows in
Faculté des Sciences Pharmaceutiques, F-31062 Toulouse cedex 09, France. Amazonian rainforests at an altitude ranging from 200 to 1500 m.
Tel.: þ 33 5 62 25 68 65; fax: þ 33 5 62 25 98 02.
E-mail address: eric.deharo@ird.fr (E.7 Deharo).
It has a star-shaped fruit, which contains dark oval seeds (Duke

http://dx.doi.org/10.1016/j.jep.2015.06.009
0378-8741/& 2015 Elsevier Ireland Ltd. All rights reserved.
 Gonzalez-Aspajo et al. / Journal of Ethnopharmacology 171 (2015) 330–334 331

and Vasquez, 1994). Its presence was reported in pre-Hispanic Among microorganisms, S. aureus is the predominant causative
times in Peru, and there is also evidence of cultivation by the Incas, agent of skin disorders. It can be the cause of minor problems,
as some seeds have been found in tombs together with ceramic such as impetigo, or serious ones i.e. scalded skin syndrome, and
representations (Brack, 1999, 2005; Ugent and Ochoa, 2006). can also induce life-threatening conditions such as septicaemia
Seeds are edible, when lightly toasted, or when boiled for a (Krut et al., 2003). Moreover, S. aureus contributes to the persis-
while in water, and are part of many local dishes, sometimes tence and the exacerbation of skin infections due to its ability to be
mixed with the leaves from the same plant (Hamaker et al., 1992). internalized by human keratinocytes (Kintarak et al., 2004).
Seed oil (“Sacha Inchi” oil or “Inca Inchi” oil) is traditionally Therefore, we determined in vitro the capacity of SIO to impair
used as an everyday skin care oil applied regularly in order to adherence of S. aureus to keratinocytes (preventive effect) and also
preserve skin softness and healthiness. It has been also observed to remove S. aureus from keratinocytes and human skin explants
that in some places, the whole seed is ground to a floury paste to (curative effect). Cytotoxicity (against keratinocytes and human
which some pure oil is added, and applied on the skin for skin explants) and bactericidy were also assessed.
rejuvenation purposes (Brack, 1999, 2005; Maxwell, 1990). One
of the authors of our study (Gonzalez-Aspajo, personal commu-
nication) also noted, through a survey performed in the Peruvian 2. Material and methods
Amazonia, that people living in the Plukenetia area of cultivation
(Mariscal Ramón Castilla, Loreto, Maynas, in the Loreto depart- 2.1. Chemical
ment, and Lamas, San Martin and Bellavista in the San Martin
department) used SIO when suffering from a skin lesion, corro- The extra virgin SIO was provided by José Anaya from AgroIn-
borating previous local reports (Correa and Bernal, 1989). dustrias Amazónicas Inc. The vegetal origin was certified by the
Actually, Plukenetia volubilis L. is considered a promising crop in commercial Registration no. RUC 20531294042/RSC1304308N/
Amazonia and is cultivated in many places in Peru and Bolivia, as UIARAA. Batch number: AI-SM-068. SIO is extracted by cold
the oil is said to be one of the richest plant sources of omega fatty pressing of almonds, subsequently decanted and passed through
acids (Chirinos et al., 2013; Guillén et al., 2003). Therefore SIO is filters to remove impurities. The fatty acids composition of the
now sold in Europe for its high dietetic properties, and is also tested oil is documented in Table 1, and was determined in the
formulated in packaged day-care skin cream with other ingredi- MetaToul-Lipidomique center Toulouse (France). Coconut oil and
ents (S.I.P.O., 2012). Phenol were from Sigma-Aldrich (C1758, P5566).
Because people living in places where the Sacha Inchi vine
grows naturally use it as an everyday skincare treatment and also 2.2. Keratinocyte cell-line
when suffering from a skin lesion, we have undergone research
aiming to assess the activity of SIO against Staphylococcus aureus. Human keratinocyte cell line was maintained in RPMI 1640
culture medium supplemented with 10% inactivated foetal bovine
serum 1% non-essential amino acids, 1% glutamine, and 1%
Table 1
Lipid profile of Sacha Inchi Oil.
penicillin and streptomycin at 37 1C in a 5% CO2 humidified
atmosphere. After 48 h, the mono-cellular layer obtained was
Common name Abbreviation Sacha Inchi Oil nmol/ml removed after incubating the cells with 5 ml trypsin-EDTA for
(%) 5 min at 37 1C. Trypsin was then inactivated by the addition of
10 mL of RPMI 1640 medium with inactivated foetal bovine serum.
Saturated fatty acid (SAFA)
Myristic acid 14:0 0 0 The cells were harvested by centrifugation (350g/5 min/20 1C). Cell
Palmitic acid 16:0 5 159,096 suspension was checked with the trypan blue test and then
Stearic acid 18:0 2 71,888 adjusted to 2  105 viable cells/ml. The cell suspensions were
arachidic acid 20:0 0 0 placed in adherence microtiter plates. All chemicals were from
behenic acid 22:0 0 0
Lignoceric acid 24:0 0 0
Dutscher, Brumath, France.
Mono-Unsaturated fatty acid
(MUFA) 2.3. Human skin explants
Oleic acid 18:1w9 9 298,341
vaccenic acid 18:1w7 0 0
Acid 11-eicosenoic 20:1w9 0 0
All subjects gave their consent for the use of their skin explants.
Poly-unsaturated fatty acid 12-mm-diameter human skin explant discs EXP001120AK008
(PUFA) obtained from Biopredic International, France, were placed in 24-
Linoleic Acid 18:2w6 35 1,139,775 well plates with 300 μl of MIL207 culture medium (Biopredic). The
Acid alpha-linolenic 18:3w3 48 1,541,927
explants were then incubated for 24 h at 32 1C prior to being
SAFA 7 230,984 processed. Skin explants are closer to what happens in real life
MUFA 9 298,341 than isolated keratinocytes as they contain all cell types resident in
PUFA 84 2,681,702
Total 100 3,211,027
the skin (dermis and epidermis) exerting their physiological
effects on the tissue and adjacent cells (Yasuoka et al., 2008).

Table 2
Activities of oils on keratinocytes, human skin explant and S. aureus.

Oils Viability % Antiadhesive effect (preventive) % Detachment effect (curative) %

S. aureus Keratinocytes Explant Keratinocytes Keratinocytes Explant

SIO 88.17 5.7 94.2 7 2.6 96.9 73.0 39.2 73.4 33.971.8 33.6 70.4
CocO 88.3 7 7.5 80.8 7 3.0 ND 25.5 72.2 29.1 71.9 7.1 75

S. aureus: Staphylococcus aureus.

Phenol killed 100% of keratinocytes and S. aureus at 0.5% (data not shown).
ND, not determined, experiment repeated 3 times.
332 Gonzalez-Aspajo et al. / Journal of Ethnopharmacology 171 (2015) 330–334
2.4. Bacteria
S. aureus CIP 53154 cultures (2  108 cells/mL) were inoculated
in 10 mL of both RPMI 1640 and MIL207 culture media with 30 ml
of tritium (H3) adenine (Perkin Elmer), and incubated under
aerobic conditions at 37 1C until reaching the stationary phase.
To eliminate non-incorporated radioactivity, the bacteria were
washed three times using PBS buffer (centrifugation 2500g/
10 min/5 1C).
2.5. Cytotoxicity assays
Determination of cytotoxic effects on keratinocyte and skin
explants were performed as described below with XTT and neutral
red assays, respectively.
2.6. XTT assay
100 μl of trypsinized cell suspension (1  106 cells/mL) were
plated into flat bottomed 96-well microtiter plates and were
incubated for 2 h at 37 1C with 250 μl of undiluted SIO, CocO
and phenol (0.5% in culture medium). Medium was used as
control. Then, 50 μl of XTT reagent was added to each well and
incubated at 37 1C for 2 h.
2.7. Neutral red assay
250 μl of undiluted SIO, CocO or phenol (0.5% in culture
medium) were applied on the surface of the skin explants for
2 h at 32 1C. Then, skin explants were mixed with 0.4% red neutral
solution for 3 h at 32 1C, and washed 3 times with PBS. Neutral red
was extracted using 1% acetic acid-50% ethanol, and the optical
densities were measured at 550 nm.
After incubation, absorbance was measured at 450 nm. The % of
viability was calculated as follows:
ODcontrol–ODtreatment
% viability ¼  100
ODcontrol
All experiments were conducted in triplicate.
2.8. Bactericidal assay
Bacterial suspensions at 108 CFU/ml in MIL207 were mixed v/v
with undiluted SIO, CocO or phenol at 0.5% (in culture medium)
and returned to incubation at 37 1C for 48 h. Then the mixture was
deposited in Petri dishes on trypticase soy agar. After an additional
24 h at 37 1C, colonies were counted and the inhibition percentage
was calculated related to the control dish receiving the medium
without oil.
2.9. Oil challenge
H3-Labelled S. aureus bacteria were washed once with PBS and
re-suspended in 5 ml RPMI 1640 medium. After removing the
RPMI, 500 ml of the bacterial suspension was deposited on the
keratinocytes cell layer or human skin explants in an atmosphere
containing 5% CO2 for 2 h at 37 or 32 1C, respectively.
2.10. Preventive effect
keratinocytes cells received 250 ml of undiluted SIO, CocO or
phenol (0.5% in culture medium) in RPMI 1640 medium and
returned to incubation at 32 1C for 2 h. The explants and cells
were washed three time using PBS. Then 2  108 of bacterial
suspension was added in each well (500 μl/well) and incubated
at 32 1C for 2 h more. Then, the explants and cells were washed
three times using PBS to eliminate non-adherent bacteria.
2.11. Curative effect
500 μl/well of 2  108 bacteria suspension was added to the
human skin explants or keratinocyte cells. After 2 h of contact,
non-adherent bacteria were eliminated by PBS washing (3 times).
Then explants or cells were treated with 250 ml of undiluted SIO,
CocO or phenol (0.5% in culture medium) and incubated for 2 h.
Finally cells were washed 3 times with PBS.
At the end of the preventive or curative treatment, the human
skin explant and keratinocytes received 500 ml of a lysis solution
[SDS 0.1% (w/v) in NaOH 0.5 mol/L] and were kept at 37 1C for 18 h.
Radioactivity was measured with a beta-liquid scintillation system
(Perkin Elmer, USA) in the 500 ml mixture of cells plus lysis
solution.
The inhibition percentage of the adherence was calculated as
follows:
ðCPM in controlÞ  ðCPM with treatmentÞ  100=ðCPM in controlÞ
CPM : counts per minute
2.12. Statistical analysis
Statistically significant difference Significance between control
and oil treated groups were determined using one-way analysis of
variance (ANOVA) through XLSTAT Software (Tukey range).
po 0.05 was considered significant.
3. Results
Chemical analysis (Table 1) shows that the major fatty acids
identified in SIO are polyunsaturated linoleic acid (35%) and
linolenic acid (48%). These findings are similar to the reported
fatty acid data on Sacha Inchi oil from China, with linoleic acid
(39%) and linolenic acid (43%) (Liu et al., 2014).
3.1. Impact on viability of S. aureus and cells (Table 2)
SIO and CocO did not show any antibacterial activity compared
with the control group, even at the maximal dose (while phenol
killed 100% of bacteria).
To discard the hypothesis that the effect of SIO or CocO on
adherence could be due to a possible cytotoxicity on keratinocytes
cells or human skin explants, assays on S. aureus, keratinocytes
and explants were performed under the same incubation condi-
tions as in the adherence test. After SIO challenge, more than 90%
of both keratinocytes and human skin explants cells were viable.
CocO was also non-toxic but was only tested on keratinocytes.
Almost 90% of bacteria were viable after oils exposure. On the
contrary phenol killed almost 100% of treated keratinocytes and
bacteria.
3.2. Pre- and post-adherence inhibition test (Table 2)
3.2.1. Pre-adherence inhibition test (preventive)
Preventive effect was only tested on keratinocytes.
Undiluted SIO was statistically more effective (almost 40%) than
CocO (25%) in inhibiting S. aureus attachment to keratinocyte.
3.2.2. Post-adherence inhibition test (curative)
3.2.2.1. On keratinocytes. Both oils were able to remove S. aureus
from keratinocytes with approximately 33% efficiency at 100%
 Gonzalez-Aspajo et al. / Journal of Ethnopharmacology 171 (2015) 330–334
concentration. No statistical significance between the activity of
both oils was observed.
3.2.2.2. On human skin explant. CocO was ineffective on the
detachment process with explants while SIO had the same
capacity to remove S. aureus from keratinocytes as from recently
extracted human skin explants (around 33% in both models).
4. Discussion
The skin is an intricate environment where commensal bacteria
protect the host from pathogenic bacteria (S. aureus, Streptococcus
pyogenes, enterobacteria), which form an occasional resident flora
responsible for a wide variety of bacterial pyodermas (Murakawa,
2004). These pathogens induce infection through the expression of
virulence gene products that promote, amongst other, bacterial
adherence to avoid clearance from their host (Chiller et al., 2001).
For bacteria it has been demonstrated that the sequence of the
adherence process starts with weak non-specific interactions with
the host cell surface, mediated by overall physicochemical proper-
ties of the bacterial and host surfaces, such as electric charge and
hydrophobicity (An et al., 2000). Adherence, allowing bacteria to
quickly and efficiently attach to host cells is thus a key process in
the interaction between a pathogen and its host, and also plays a
crucial role in biofilms formation. Biofilms, in turn, have peculiar
physico-chemical properties able to strongly impact antibiotic
sensitivity (Rennemeier et al., 2007).
Therefore, in the fight against infectious disease it is of great
importance to search for substances able to prevent adherence
between hosts and pathogens. In this context, because SIO was
reported to be used topically on the skin, it seemed us pertinent to
study its capacity to impair S. aureus adherence on isolated
keratinocytes and skin explants, and compare it to CocO. The
latter is also widely used for skin care but has a different
composition, rich in highly saturated fatty acids (4 80%) with
almost no omega 3 and 6 fatty acids, while in SIO, unsaturated
fatty acids predominate (80%).
We showed that both oils were able to prevent attachment of S.
aureus to keratinocytes, but this effect was statistically higher for
SIO than for CocO. Also, SIO was almost 5 times more efficient in
supporting detachment from human skin explants, than CocO in
this model. Surprisingly there was no difference between SIO and
CocO in promoting detachment from keratinocytes (around 30% of
the bacteria population was removed with both oils). To the best of
our knowledge, there is no reference in the relevant literature to
the minimum percentage detachment considered to be efficient;
this study is therefore the first to examine this. In vivo assays are
obviously required to confirm the benefit of applying SIO.
Although it has been shown that lipids from the skin possess
antibacterial activity against S. aureus (Burtenshaw, 1942; Drake et
al., 2008), and that skin lipases participate in the production of
antibacterial short chain lipids (Verallo-Rowell et al., 2008), the
detachment effect we have observed cannot be related to any
direct cytotoxic effect of both oils as they did not display any
deleterious effect on S. aureus itself.
Unsaturated fatty acids such as linoleic acid (present in high
concentration in SIO) are known to increase membrane fluidity in
S. aureus, interfering with microbial adherence to the skin (Arsic
et al., 2012), and also to induce significant enhancement of the net
negative surface charge density of thylakoids, a membrane-bound
compartment inside cyanobacteria, thus having an impact on cell
surface electric charge and hydrophobicity (Doltchinkova and
Nikolov, 1997). In the case of CocO it can also be suggested that
lauric acid, the major component of this oil, could be partially
responsible for the detachment observed effect (at least in the
333
keratinocyte model) as it is known to inhibit the transduction
signal linked with S. aureus virulence and protein surface (Clarke
et al., 2007; Ruzin and Novick, 2000).
It must also be observed, as reported by Leug (2008), that the skin
of patients with atopic eczema shows increased adherence for S.
aureus, which is thought to be related to the underlying skin atopic
inflammation. As long-chain polyunsaturated fatty acids are often
associated with anti-inflammatory properties (Mullen et al., 2010),
SIO deserve special consideration for topical application providing
supplementary benefit because of its high polyunsaturated fatty acids
contents. To the best of our knowledge there is no reference in the
relevant literature on the minimum percentage detachment termed
as efficient; but, especially in atopic eczema, where the density of S.
aureus can reach 107 organisms per cm2 in the acute phase (Leug,
2008), 30% detachment could be very beneficial for patients. In this
pathology the classical treatments are corticoids (to reduce inflam-
mation) and antibiotics. However, prolonged use of corticoids can lead
to the emergence of resistance to the immunosuppressive effects of
corticosteroids induced by S. aureus itself. Moreover, the cutaneous
side effects of corticosteroids are numerous, including atrophy of the
skin, telangiectasias and skin pigment changes among others
(Coondoo et al., 2014). Therefore, a topical non-steroidal preparation
containing SIO could be of great interest, as a maintenance therapy in
conjunction with topical corticosteroids for acute flares. In vivo assays
are obviously required to confirm this assertion.
Moreover, as fatty acids are ordinary constituents of the tissue,
it could be anticipated that they would not cause irritation or
sensitization, which is often the case when using topical exogen-
ous compounds.
Finally, in an early study, Zurita and Hay (1987) suggested that
the effect of fatty acids on adherence, between dermatophyte
conidia and human keratinocytes, could be an important regula-
tory factor. It would therefore be interesting to test the activity of
SIO against mycosis, because skin mycoses are a worldwide health
concern particularly in tropical area (Lupi et al., 2005).
In conclusion, for the first time we have highlighted the
relevance of SIO in both preventive and curative treatments
against staphylococcal skin adherence in vitro. Our results tend
to support the traditional use of undiluted SIO in skin care.
Bioguided fractionation and identification of the most-effective
anti-adherent fatty acid could therefore lead to new safe and cost-
effective strategies for the treatment of, or prophylaxis against,
certain types of skin infections. Omega 3–6 could be responsible,
at least partly, for the detachment effect detected on human skin
explant, but further experiments are obviously necessary to
confirm this hypothesis and also to determinate the best omega
3–6 ratio leading to the best anti-adherence effect.
Conflict of interest
The authors have declared that there is no conflict of interest.
Acknowledgements
We would like to thank José Anaya from AgroIndustrias Amazó-
nicas Inc. (Peru) for supplying SIO. German Gonzalez received Grant
from the IRD (ARTS). We are also grateful to Nada Matas-Runquist and
Elizabeth Elliott for valuable editorial assistance.
References
An, Y.H., Dickinson, R.B., Doyle, R.J., 2000. Mechanisms of bacterial adhesion and
pathogenesis of implant and tissue infections. In: An, Y.H., Friedman, R.J. (Eds.),
Handbook of Bacterial Adhesion Principles, Methods, and Applications.
Humana Press, pp. 1–27.
334 Gonzalez-Aspajo et al. / Journal of Ethnopharmacology 171 (2015) 330–334
Arsic, B., Zhu, Y., Heinrichs, D.E., McGavin, M.J., 2012. Induction of the staphylo-
coccal proteolytic cascade by antimicrobial fatty acids in community acquired
methicillin resistant Staphylococcus aureus. PLoS One 7, e45952.
Brack, A., 1999. Plukenetia volubilis L. Diccionario Enciclopédico de Plantas Útiles del
Perú. PNUD. Cuzco – Perú. p. 550.
Brack, A., 2005. Perú: Legado milenario – Millenary Legacy. Lima. Publicado por
Universidad de San Martín de Porres, Escuela Profesional de Turismo y
Hotelería, p.168.
Burtenshaw, J.M.L., 1942. The mechanism of self-disinfection of the human skin and
its appendages. J. Hyg. 42, 184–210.
Chiller, K., Selkin, B.A., Murakawa, G.J., 2001. Skin microflora and bacterial
infections of the skin. J. Investig. Dermatol. Symp. Proc. 6, 170–174.
Chirinos, R., Zuloeta, G., Pedreschi, R., Mignolet, E., Larondelle, Y., Campos, D., 2013.
Sacha inchi (Plukenetia volubilis): a seed source of polyunsaturated fatty acids,
tocopherols, phytosterols, phenolic compounds and antioxidant capacity. Food
Chem. 141, 1732–1739.
Clarke, S.R., Mohamed, R., Bian, L., Routh, A.F., Kokai-Kun, J.F., Mond, J.J., Tarkowski,
A., Foster, S.J., 2007. The Staphylococcus aureus surface protein IsdA mediates
resistance to innate defenses of human skin. Cell Host Microbe 1, 199–212.
Coondoo, A., Phiske, M., Verma, S., Lahiri, K., 2014. Side-effects of topical steroids: a
long overdue revisit. Indian Dermatol. Online J. 5, 416–425.
Correa, J.E., Bernal, H.Y., 1989. In Convenio Andrés Bello. Programa de Recursos
Vegetales. Edition SECAB. Bogota, Colombia, 489 pp.
Doltchinkova, V., Nikolov, R., 1997. Unsaturated fatty acids induced changes in
surface charge density and light – scattering in pea thylakoids. Bulg. J. Plant
Physiol. 23, 3–11.
Drake, D.R., Brogden, K.A., Dawson, D.V., Wertz, P.W., 2008. Thematic review series:
skin lipids. Antimicrobial lipids at the skin surface. J. Lipid Res. 49, 4–11.
Duke, J.A., Vasquez, R., 1994. Amazonian ethnobotanical dictionary. Boca Raton, Ann
Arbor, London, Tokyo, CRC Press, 215 pp.
Guillén, M.D., Ruiz, A., Cabo, N., Chirinos, R., Pascual, G., 2003. Characterization of
sacha inchi (Plukenetia volubilis L.) oil by FTIR spectroscopy and 1H NMR.
Comparison with linseed oil. J. Am. Oil Chem. Soc. 80, 755–762.
Hamaker, B.R., Valles, C., Gilman, R., Hardmeier, R.M., Clark, D., García, H.H.,
Gonzales, A.E., Kohlstad, I., Castro, M., 1992. Amino acid and fatty acid profiles
of the Inca peanut (Plukenetia volubilis L.). Cereal Chem. 69, 461–463.
Kintarak, S., Whawell, S.A., Speight, P.M., Packer, S., Nair, S.P., 2004. Internalization
of Staphylococcus aureus by Human Keratinocytes. Infect. Immun. 72,
5668–5675.
Krut, O., Utermöhlen, O., Schlossherr, X., Krönke, M., 2003. Strain-specific associa-
tion of cytotoxic activity and virulence of clinical Staphylococcus aureus isolates.
Infect. Immun. 71, 2716–2723.
Leug, D.Y.M., 2008. The role of Staphylococcus aureus in atopic eczema. Acta Derm.
Venereol. 216, 21–27.
Liu, Q., Xu, Y.K., Zhang, P., Na, Z., Tang, T., Shi, Y.X., 2014. Chemical composition and
oxidative evolution of Sacha Inchi (Plukenetia volubilis L.) oil from Xishuang-
banna (China). Grasas y Aceites 65, e012.
Lupi, O., Tyring, S.K., McGinnis, M.R., 2005. Tropical dermatology: fungal tropical
diseases. J. Am. Acad. Dermatol. 53, 931–951.
Maxwell, N., 1990. Witch Doctor's Apprentice, Hunting for Medicinal Plants in the
Amazonia, 3rd edition Citadel Press, New York (391 pp).
Mullen, A., Loscher, C.E., Roche, H.M., 2010. Anti-inflammatory effects of EPA and
DHA are dependent upon time and dose-response elements associated with
LPS stimulation in THP-1-derived macrophages. J. Nutr. Biochem. 21, 444–450.
Murakawa, G.J., 2004. Common pathogens and differential diagnosis of skin and
soft tissue infections. Cutis 73, 7–10.
Rennemeier, C., Hammerschmidt, S., Niemann, S., Inamura, S., Zähringer, U., Kehrel, B.E.,
2007. Thrombospondin-1 promotes cellular adherence of gram-positive pathogens
via recognition of peptidoglycan. FASEB J. 21, 3118–3132.
Ruzin, A., Novick, R.P., 2000. Equivalence of lauric acid and glycerol monolaurate as
inhibitors of signal transduction in Staphylococcus aureus. J. Bacteriol. 182,
2668–2671.
S.I.P.O., 2012. Market Brief for Sacha Inchi. An introduction to the European market
for Peruvian exporters. Osec. Swiss Import Promotion Programme. 〈www.
ThisIsProFound.com〉.
Ugent, D., Ochoa, C.M., 2006. La Etnobotánica del Perú desde la Prehistoria al
Presente. Lima, Peru, Consejo Nacional de Ciencias, Technología e Innovación
Tecnológica-CONCYTEC.
Verallo-Rowell, V.M., Dillague, K.M., Syah-Tjundawan, B.S., 2008. Novel antibacter-
ial and emollient effects of coconut and virgin olive oils in adult atopic
dermatitis. Dermatitis 19, 308–315.
Yasuoka, H., Larregina, A.T., Yamaguchi, Y., Feghali-Bostwick, C.A., 2008. Human skin
culture as an ex vivo model for assessing the fibrotic effects of insulin-like
growth factor binding proteins. Open Rheumatol. J. 2, 17–22. http://dx.doi.org/
10.2174/1874312900802010017.
Zurita, J., Hay, R.J., 1987. Adherence of dermatophyte microconidia and arthroco-
nidia to human keratinocytes in vitro. J. Investig. Dermatol. 89, 529–534.