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Wound-healing Properties of the Oils of Vitis vinifera and Vaccinium macrocarpon


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Vitis vinifera (grape) and Vaccinium macrocarpon (cranberry) are well known medicinal plants; most of the pharmacologically active phytochemicals have been isolated from the skin, fruit juice, fermented extract and alcohol fractions of the plants above. Here, the pharmacological properties of the phytochemical constituents present in oils of cranberry and grape were investigated. The oil of grape and cranberry has been evaluated for their wound healing activity by using an excision wound model in rats. The animals were divided into four groups of six each (n = 6). The experimental group 1 and 2 animals were treated topically with the grape and cranberry oil (100 mg/kg body weight), respectively. The controls were treated with petroleum jelly. The standard group of animals were treated with mupirocin ointment (100 mg/kg body weight). The healing was assessed by the rate of wound contraction and hydroxyproline content. On day 13, animals treated with cranberry oil exhibited a (88.1%) reduction in the wound area compared with grape-oil treated (84.6%), controls (74.1%) and standard group animals (78.4%) (p < 0.001). The hydroxyproline content of the granulation tissue was significantly higher in the animals treated with cranberry and the grape-oil (p < 0.000). Comparative investigation of the curative properties of the oils of V. vinifera and V. macrocarpon revealed a significant result which suggests their wound-healing potential.
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Woundhealing Properties of the Oils of
Vitis vinifera and Vaccinium macrocarpon
B. Shivananda Nayak,
*D. Dan Ramdath,
Julien R. Marshall,
Godwin Isitor,
Sophia Xue
and John Shi
Department of Preclinical Sciences, Faculty of Medical Sciences, The University of the West Indies, St Augustine, Trinidad
Guelph Food Research Centre, Agriculture and AgriFood Canada, 93 Stone Road West, Guelph, Ontario N1G 5C9, Canada
Vitis vinifera (grape) and Vaccinium macrocarpon (cranberry) are well known medicinal plants; most of the
pharmacologically active phytochemicals have been isolated from the skin, fruit juice, fermented extract and
alcohol fractions of the plants above. Here, the pharmacological properties of the phytochemical constituents
present in oils of cranberry and grape were investigated. The oil of grape and cranberry has been evaluated for
their wound healing activity by using an excision wound model in rats. The animals were divided into four
groups of six each (n= 6). The experimental group 1 and 2 animals were treated topically with the grape and
cranberry oil (100 mg/kg body weight), respectively. The controls were treated with petroleum jelly. The
standard group of animals were treated with mupirocin ointment (100 mg/kg body weight). The healing was
assessed by the rate of wound contraction and hydroxyproline content. On day 13, animals treated with
cranberry oil exhibited a (88.1%) reduction in the wound area compared with grapeoil treated (84.6%),
controls (74.1%) and standard group animals (78.4%) (p< 0.001). The hydroxyproline content of the granulation
tissue was signicantly higher in the animals treated with cranberry and the grapeoil (p< 0.000). Comparative
investigation of the curative properties of the oils of V. vinifera and V. macrocarpon revealed a signicant result
which suggests their woundhealing potential. Copyright © 2011 John Wiley & Sons, Ltd.
Keywords: hydroxyproline; excision wound; wound area.
Ethnomedicinal research has searched arduously for
remedies that can be derived from the abundant
resources of nature. Recent discoveries into the wound
healing potential of Vitis vinifera,Morinda citrifolia and
Vinca rosea has prompted further work; which seeks to
increase the knowledge of the medicinal properties of
the wide variety of plants. Grape and cranberry are two
well known plants within ethnomedicine. Research has
highlighted the potential curative properties of antho-
cyanins, tannins and resveratrol found in grape extract.
Research has also discovered signicant concentrations
of polyphenols such as catechins, cyanidin3glucoside
and peonindin3glucoside in cranberry fruit. Intense
research has been directed to the discovery of the
phytochemical constituents of the skin and fruit of those
The common grape V. v i n i f e r a is a historically
important plant. The fruit is a berry, known as a grape.
In the wild species, it is 6 mm diameter and ripens dark
purple to blackish with a pale wax bloom. In cultivated
plants, it is usually much larger, up to 3 cm long and can
be green, red, or purple. Red wines from several red
grape varieties have been reported to have health
benets based on the signicant amount of polyphenolic
derivatives present (PastranaBonilla et al., 2003).
Resveratol, a nonavanoid biomolecule is found in
signicant quantities in red wine and has been reported
to possess antioxidant, antiinammatory and anticancer
properties (Elliott and Jirousek, 2008; Asensi et al.,
2002).Additionally, the topical application of a high
resveratol from grape seed extract was shown to
accelerate wound healing in mice, which was attributed
to modulation of the redoxsensitive processes that
drive dermal tissue repair (Khanna et al., 2002).
Cranberries are a group of evergreen creeping shrubs
or vines up to 2 m long and 520 cm in high. It is edible
with an acidic taste that can overwhelm its sweetness.
Cranberries and cranberry juice are abundant food
sources of the anthocyanidin, avonoids, cyanidin,
peonidin and quercetin (Duthie et al., 2006). Cranberry
juice is used in the prevention and treatment of urinary
system infections (Hood et al., 2004; Jenkins et al.,
2005), as well as in the treatment of periodontitis
(Ratcliffe and ShacharHill, 2005; Rios et al., 2002) and
other disorders.
It has been proposed that alkaloids and terpenoids
may provide astringent, antifungal or antimicrobial
properties that may be of benet to the progression of
the wound healing cascade (Scortichini and Pia Rossi,
1991; Miot et al., 2004; Roy and Saraf, 2006). Antho-
cyanins, leucoanthocyanins and other polyphenols have
been demonstrated to have signicant antioxidant
properties which also can be of benet in the progression
of wound healing (Shetty etal.,2007; Nayak et al., 2009;
Parry et al., 2006; Shi et al., 2003). It is reasonable to
expect that the phytochemical constituents in oil
fractions of grape and cranberry would have signicant
wound healing properties. However, there appear to be
* Correspondence to: B. Shivananda Nayak, Department of Preclinical
Sciences, Faculty of Medical Sciences, The University of the West Indies,
St Augustine. Trinidad and Tobago.
Phytother. Res. (2011)
Published online in Wiley Online Library
( DOI: 10.1002/ptr.3363
Copyright © 2011 John Wiley & Sons, Ltd.
Received 30 March 2010
Revised 22 October 2010
Accepted 26 October 2010
very few reports on the wound healing potential and
antimicrobial activities of Vitis vinifera and Vaccinium
macrocarpon in literature. Therefore, this study was the
rst attempt to report the efcacy of Vitis vinifera and
Vaccinium macrocarpon for their possible role in the
treatment and management of wounds.
Grape seed oil production. The grape seeds oil was
obtained by compressing grape seeds (supplied by
Josephs Natural Products Inc.) using a screw extrusion
press (Komet S 87 G, IBG Monforts, Mönchengladbach,
Germany). Grape seed pressing was performed without
heating (cold pressed).
Cranberry oil production. The cranberry oil was
obtained by compressing cranberry (supplied by Josephs
Natural Products Inc.) using a screw extrusion press
(Komet S 87 G, IBG Monforts, Monchengladbach,
Germany). Pressing was performed without heating (cold
Lipid extraction, separation, methylation and GC
analysis. Grape seed oil (10 mL) was extracted with
140 mL chloroform: methanol (1:1, v/v) using an Ultra
Turrax homogenizer (T25, Ika Works Inc., USA). After
homogenization, it was stirred for 5 min to obtain a
uniform consistency in 63 mL water that was added to
the solution to avoid any miscellaneous phase (Bligh
and Dyer, 1959). The chloroform phase was collected
and partitioned again with some chloroform. The
combined extracts were taken to dryness by a vacuum
rotary evaporator (<30 °C) until constant weight and
then stored at 20 °C until analysis.
The total crude lipids obtained were dissolved in
15 mL chloroform. Fatty acid methyl esters (FAME)
analysis was done to determine the total fatty acids. In
this FAME analysis, 0.25 mL (1012 mg) of each lipid
extract was methylated with 1.5 mL 5% anhydrous
HCl/methanol (w/v) in a 15 mL culture tube equipped
with a Teonlined screw cap at 80 °C for 1 h. After
cooling to room temperature, a few drops of water and
2 mL hexane was added. The hexane layer was
collected and reduced in volume before the FAME
mixture was applied to TLC plates. The loaded TLC
plates were developed in the solvent mixture of hexane/
ethyl ester/acetic acid (85:15:1, v/v/v), sprayed with
2,7′‐dichlorouoroscein/methanol (0.1% w/v), and
viewed under UV light (254 nm). The corresponding
FAME band was removed and eluted with chloroform.
The ltrate was blown dry under nitrogen and the
FAME mixture was dissolved in 1.5 mL hexane and
analysed by GC.
For separating the lipid classes, 0.4 mL (at least
20 mg) of each crude extract was dried under a stream
of nitrogen, then added to chloroform and applied
onto TLC plates before methylation of the FA. TLC
plates were developed and viewed under UV light as
above. Major lipid class bands were scraped from the
plates. The rst two bands near the bottom were
eluted with methanol: chloroform (1:1, v/v), and the
upper three bands were eluted with chloroform. After
removal of the chloroform with nitrogen, the lipid
classes were methylated for 1 h, and then water
(5 mL) and 2 mL hexane were added to collect the
FAME in hexane as above. Calibration and identica-
tion of the different lipid classes were made by running
standards in parallel with the samples. The extractable
lipids were separated into ve classes: polar lipids
(phospholipids and glycolipids, PL), diacylglycerols
(DAG), free fatty acids (FFA), triacylglycerols (TAG)
and steryl esters (SE).
A GC (model 5890; HewlettPackard, Palo Alto, CA)
equipped with a ame ionization detector and an auto
sampler (model 7673, HewlettPackard, Palo Alto, CA,
USA), a 100 m CPSil 88 fused capillary column (Varian
Inc., Mississauga, ON, Canada) and ChemStation
software system (version A.09, HewlettPackard,
Palo Alto, CA, USA) were used for analysing FAME.
The injector and detector temperatures were 250 °C.
The temperature programme for the column was: hold
at 45 °C for 4 min, increase by 13 °C/min to 175 °C,
hold again at 175 °C for 27 min, increase at 4 °C/min to
215 °C, and then nally hold at 35 °C. A FAME
standard (mixture 463) was used to identify the FAME
and the FA amount was expressed as the percent of
total fatty acids.
The above procedure was repeated for cranberry oil
for lipid extraction, separation, methylation and GC
Determination of total phenols of cranberry oil and
green grape seed oil. Extraction of the phenols was
carried out according to a method presented by Parry et al.
(2005) with some modications. For the extraction of oils,
15 mL of methanol (methanol: water, 90:10 ) was added to
5.0 g oil, followed by vortex for 3 min and centrifugation
for 10 min at 3500 rpm. The supernatant was collected for
phenolic content measurements.
The quantitative determination of total phenols (TP)
was analysed by using the FolinCiocalteau colorimetric
method (Swain and Hillis, 1959). Five hundred micro-
litres of sample extract was placed in a test tube along
with 1.0 mL of FolinCiocalteau reagent and 2 mL of
distilled water. After leaving for 5 min, 0.5 mL of
sodium carbonate (10%) was added, mixed and allowed
to stand for 2 h in a dark place for reaction. Absorption
at 765 nm was measured in a UVVis spectrophotometer
against a blank sample. The TP content was expressed
as gallic acid equivalents in milligrams per gram of oil,
using a standard curve generated with 5 mg, 10 mg,
25 mg, 50 mg, 75 mg and 100 mg gallic acid per 100 mL
extraction solvent.
Phenolic compound analyses by HPLC. An HP 1100
HPLC system equipped with an alphaBond C
column (4.6×250 mm, particle size 5 μm), coupled with
an Agilent 1100 series ChemStation software was used
for quantifying the individual phenolic compounds. The
mobile phases consisted of 2.0% acetic acid in distilled
water (A) and acetonitrile (B). The column was eluted
at 1.0 mL/min under a linear gradient from 5% mobile
phase B to 75% over 20 min, to 100% over 5 min,
isocratic 5 min at 100%, and then to 25% over 5 min
and to 5% over another 5 min. Sample injection
volumes were 20 μL. The compounds were detected
at 280 nm with an HP 1100 series ultraviolet (UV) diode
array detector. Standards of catechin and epicatechin
were injected for identication.
Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. (2011)
Animals. The study was approved by the Ethics
Committee for animal experimentation (AHC06/07/1)
by The Faculty of Medical Sciences, The University of
the West Indies, St Augustine. Healthy inbred male
Sprague Dawley rats weighing 180200 g were used for
the study. They were housed individually and main-
tained on normal food and water ad libitum. Animals
were periodically weighed before and after the experi-
ment. The rats were anaesthetized prior to and during
iniction of the experimental wounds. The surgical
interventions were carried out under sterile conditions
using ketamine anaesthesia (120 mg/kg body weight).
The animals were closely observed for any infection and
if they showed signs of infection were separated,
excluded from the study and replaced.
Woundhealing activity. An excision wound model was
used to evaluate the woundhealing activity of oils of
Vitis vinifera and Vaccinium macrocarpon.
Excision wound model. The animals were anaesthetized
prior to and during creation of the wounds. The rats
were inicted with excision wounds as described by
other researchers (Morton and Malone, 1972). The
dorsal fur of the animals was shaved with an electric
clipper and the area of the wound to be created was
outlined on the back of the animals with methylene blue
using a circular stainless steel stencil. A full thickness of
the excision wound of circular area 250 mm
and 2 mm
depth was created along the markings using toothed
forceps, a surgical blade and pointed scissors. The
wound closure rate was assessed by tracing the wound
on alternate days (day 1, 3, 5, 7, 9 etc.) using
transparency paper and a permanent marker. The
wound areas recorded were measured using graph
paper. The point at which the eschar fell off without
any residual raw wound was considered epithelization.
Estimation of hydroxyproline. Dry granulation tissue
from both the control and treated group was used for
the estimation of hydroxyproline. The hydroxyproline
present in the neutralized acid hydrolysate was
subsequently oxidized by sodium peroxide in the
presence of copper sulphate followed by complexing
with paradimethylaminobenzaldehyde to develop a
pink colour and that was measured at 540 nm by a
Histological study. The granulation tissue was obtained
on day 13 from the test and control group animals for
the histological study. Van Giesons and haematoxylin
and eosin stains were used to show the broblasts and
collagen deposition.
Antimicrobial activity. Pseudomonas aeruginosa
(ATCC 27853), Klebsiella pneumonia (ATCC 700603),
Enterococcus fecalis (ATCC 29212), Escherichia coli
(ATCC 25922), Staphylococcus aureus (ATCC25923)
and methicillin resistant Staphylococcus aureus
(MRSA) (ATCC 43300) were the organisms tested.
The bacterial strains were obtained from fresh colonies
grown on MacConkey and blood agar plates. The
sensitivity testing was done using Muller Hinton agar
plates. A known volume of bacterial suspension was
transferred to each microplate well. Ten microlitres of
(5 mg/mL) of V. v i n i f e r a and V. macrocarpon oils was
added to the microplate wells and incubated at 3537 °C
for 1820 h. The results were analysed visually.
Statistical analysis. The means of wound area measure-
ments between groups at different time intervals were
compared using oneway ANOVA, descriptive test,
followed by Tukeys posthoc test. Oneway ANOVA
was used to examine the mean difference in epitheliza-
tion period between the groups. Data were analysed
using the SPSS (Version 12.0, Chicago, USA) and a
value of p< 0.05 was used for all analyses.
Phytochemical analysis
Phytochemical analyses of grape and cranberry oils, by
quantitative methods have shown the presence of
signicant amounts of polyphenolic derivatives,
leucoanthocyanins and fatty acids (Table 1). Analysis
of the oils revealed a signicant amount of phenolic
derivatives, expressed as gallicacid equivalents and it
was 3.167 mg/g ± 0.124 and 3.330 mg/g ± 0.123 for
cranberry and grape oil, respectively (Table 2). A
quantitative evaluation of grape oil was done and the
total polyphenolic and leucoanthocyanin equivalents
were determined for grape seed. The seed contained
34.1 ± 5.3 mg of total polyphenolic equivalents. The
leucoanthocyanin equivalents were determined and it
was noted that the seed of the V. v i n i f e r a contained
368.6 mg/g ± 71.4 of cathechin and 276.3 mg/g ± 56.3 of
itsepimerepicathechin. These results suggest that
Table 1. Fatty acid composition of grape seed and cranberry oil
Fatty acid (file) Grape (Niagara) Grape W (WI) Cranberry
Total 95.15 97.79 93.73
Saturated 10.99 11.64 6.84
Monounsaturated 15.22 16.56 18.77
Polyunsaturated 68.94 69.59 68.12
Total fat (as TG) 95.63 98.29 94.21
Omega30.35 0.40 31.26
Omega668.59 69.19 36.86
Omega914.86 16.23 18.70
Palmitic acid 7.10 7.36 5.63
Stearic acid 3.67 3.96 1.21
Samples from Joseph Estate Wines, 1811 Niagara Stone Road,RR#, Niagaraonthelake, Ontario L0S 1 J0 Canada.
Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. (2011)
leucoanthocyanin and other polyphenols may exist in
the oil extracts of grape and cranberry, since these oils
were obtained from coldpressing of the seeds of these
Microbiological study
Microbial analysis to support the efcacy of grape and
cranberry oil revealed that concentrated oils of both
fruits exhibited some antibacterial effects. Concentrated
grape oil was proven to be effective against two strains
of Escherichia coli. Similarly concentrated cranberry oil
proved to be effective against methicillin resistant
Staphyloccocus aureus and two strains of Escherichia
coli and Klebsiella pneumonia (Table 3 and 4).
Biochemical and wound area measurements
Animal specimens treated with cranberry oil showed
faster rates ofwound area contraction of 88.1% by day 13,
whereas the grape oil treated animals had 84.6% of
wound area contraction (Figs. 1 and 2). Biochemical
analysis of hydroxyproline content of granulation tissue
suggested that both grape and cranberry oils are effective
potential wound healing agents based on the recorded
hydroxyproline content. The results of the hydroxyproline
content of cranberry oil (21.22 mg/g±0.02) was slightly
higher compared with grape oil (20.10 mg/g ± 0.02)
which suggests that the cranberry has better wound
healing potential than grape oil (Fig. 3). These results
suggest that grape and cranberry oil have strong wound
healing potential.
Histological study
The histological study of the granulation tissue obtained
on day 13 from the experimental animals (grape and
cranberry treated) showed increased well organized
bands of collagen (Figs. 4 and 5) compared with the
controls, which showed scanty collagen bres (Fig. 6).
Mupirocin treated animals showed well organized
collagen bres (Fig. 7). Numerous broblasts in H & E
staining of the tissues obtained from the experimental
and standard suggesting a signicant amount of collagen
and bronectin deposition.
Antibiotic therapy represents one mechanism involved
in the progression of the inammatory pathway, in vivo
which is carried out by resident leucocytes. Exogenous
antibiotics are often given to augment the activity of
leucocytes. Initial antibiotics were derived from a
natural source, namely bacteria. Most antibiotics are
classied as bacteriostatic or bactericidal based on their
relative action on the bacterial organism. A bacterio-
static chemical usually arrests protein or carbohydrate
synthesis in the bacterium causing the organism to
remain in static growth, but does not kill the cell.
Whereas bactericides may inhibit the production of
essential material required for organism survival, e.g.
inhibition of peptidoglycan layer formation in Gram
positive bacteria, or via induction of DNA instability
(Walsh, 2003; Spratt, 1977; Kasten and Reski, 1997;
Widmer, 2008).
Possible role of phytochemical constituents on
wound healing
Both oils demonstrated signicant antimicrobial activity
against Escherichia coli, which suggest the intrinsic
effect of polyphenols modulating the integrity of the
peptidoglycan layer of Grampositive bacteria. It has
been also suggested that they act synergistically with
Table 2. Phenolic content of cranberry and grape oil
Sample Gallic acid eq. (mg/g) % content/(g) of oil
Cranberry oil 3.167± 0.124 0.32
Green grape oil 3.330± 0.123 0.33
Total phenol content was analysed as gallic acid equivalent mg/g
of oil, values were taken as the average of triplicates.
Table 3. Comparative antimicrobial analysis of 1 in 20 dilutions of oils of cranberry and grape
Sample QCAQCB123456789
Table 4. Comparative antimicrobial analysis of concentrated oils of cranberry and grape
Sample QCAQCB123456789
1, Pseudomonas aeruginosa ATCC 27853; 2, Staphylococcus aureus ATCC 25923; 3, Staphylococcus aureus ATCC 43300 (MRSA); 4,
Escherichia coli ATCC 25922; 5, Escherichia coli ATCC 35218; 6, Klebsiella pneumonia ATCC 700608; 7, Enterococcus faecalis ATCC
29212; 8, Candida albicans (bench strain yeast); 9, Clostridium perfringens (bench strain anaerobe bacteria); QCA, Muller Hinton agar (no
extract/bacterial challenge) Control; QCB, direct culture of extract (no bacterial challenge) Control; G, growth of bacterial colonies; NG, no
growth of bacterial colonies.
Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. (2011)
betalactams to act as betalactamase at a capacity
similar to clavulanic acid (Spratt, 1977; Gianfranco
et al., 2008). Phenolic compounds in products play
important roles for antioxidant, antiinammatory and
antimicrobial effects (Shi et al., 2003). At this stage, it
can be stated that the antioxidant activity of both oils
might have contributed more value to the antimicrobial
activity against Escherichia coli.
The demonstrated antimicrobial activity of cranberry
oil against Staphyloccocus aureus and Klebsiella
pneumonia may be due to the presence of a phyto-
chemical constituent in the oil. Polyphenols have been
investigated previously for their curative properties and it
has been indicated that cathechin, a monomeric avonoid
component of leucoanthocyanin; conjugates of cathechin
such as cathechin gallate, and its epimer epicathechin
may have inhibitory effects against (MRSA) methicillin
resistant Staphyloccocus aureus and Streptoccocus mutans
(Ikigai et al., 1993; ElGammal and Mansour, 1986;
Muroi and Kudo, 1993).
Recent research has suggested that in addition to
antimicrobial activity, polyphenols may possess protease
inhibitor properties. Matrix proteases are required for
the degradation of the extracellular matrix proteins,
brin and bronectin, in preparation for the deposition
of collagen by broblasts. This protease activity gradually
Figure 1. Woundhealing activity of grape and cranberry oil,in comparison with standard and control (n=6). Each column represents mean ± SE.
This figure is available in colour online at
Figure 2. (C) Excision wound on day 1 treated with petroleum jelly, (C13) excision wound on day 13 treated with petroleum jelly (74.2%
wound closure), (T) excision wound on day 1 treated with V. macrocarpon, (Cr13) excision wound on day 13 treated with V. macrocarpon
(88.1% wound closure), (T) excision wound on day 1 treated with V. vinifera, (Gr13) excision wound on day 13 treated with V. vinifera
(84.6% wound closure), (S) excision wound on day 1 treated with mupirocin ointment, (S13) excision wound on day 13 treated with
mupirocin (78.4% wound closure). This figure is available in colour online at
Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. (2011)
decreases as the more stable collagen type (III) bre
is deposited. If the protease activity is not curtailed in a
systematic pattern extracellular matrix repair, granulation
tissue development and wound area closure is affected
(Sartor et al., 2002).
There is much information on the antioxidant
properties of polyphenols. The antioxidant activity
may be signicant in accounting for the antioxidant
activity proposed by the varied literature that currently
exists. Research into the effect of free radicals on
wound healing has suggested that the presence of these
radicals inhibits the activity of broblasts and macro-
phages. Inhibition of these cells may prevent the
progression of the inammatory pathway. One method
of controlling the presence of free radicals is to solicit
the activity of radical scavenger molecules. Fatty acids
such as oleic acid, linoleic acid were proven to have a
signicant antioxidant prole when compared with
αtocopherol, and may be able adequately to scavenge
free radicals. Based on the quantitative results of grape
oil and cranberry, it is noted these oils contain
signicant amounts of omega6 and omega9 fatty acids.
Therefore the woundhealing activity could be attrib-
uted to the antioxidant activity of the fatty acids and
anthocyanins measured in the oil fractions (Gupta et al.,
2002; AlNaqeeb et al., 2009).
Proinammatory molecules are responsible for the
progression of the inammatory phase in would
healing. Arachidonic acid is considered to be a
signicant proinammatory molecule and it has been
suggested that its precursor, linoleic acid, may be
antiinammatory as it has been observed in recent
studies to weakly inhibit cyclooxygenase (Henry et al.,
Figure 3. Hydroxyproline content of the granulation tissue in
comparison with standard (mupirocin), control (petroleum jelly),
grape and cranberry oil (n=6). Each column represents mean ± SE
(p<0.000) versus control and standard (oneway ANOVA,
descriptive test). This figure is available in colour online at http://
Figure 4. (HE) H&E stain of granulation tissue of animal specimen treated with grape oil shows a matrix of collagen fibres, and fibroblast in the
vicinity. a, macrophages; b, fibroblast; c, collagen. (VG) Van Gieson stain of granulation tissue of animal specimen treated with grape oil shows
a dense matrix of wavy collagen fibres. a, collagen fibres. This figure is available in colour online at
Figure 5. (HE) H&E stain of granulation tissue of animal specimen treated with cranberry oil shows a matrix of collagen fibres, and fibroblast
in the vicinity. a, macrophages; b, fibroblast; c, collagen. (VG) Van Gieson stain of granulation tissue of animal specimen treated with
cranberry oil, the stain shows a dense matrix of wavy collagen fibres. a, collagen fibres. This figure is available in colour online at http://
Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. (2011)
The data demonstrate that grape and cranberry oil
facilitate signicant wound healing. Its wound healing
promoting activity could be due to a combination of
antimicrobial, antiinammatory and antioxidant activities,
by the respective constituents or it could be due to the
individual activity of active leucoanthocyanins, fatty acids
and other polyphenols. Further investigation is required to
isolate the active ingredients that promote wound healing,
before it can be considered for clinical application.
We extend our sincere thanks to Dr William Swanston and Mrs
Sabana Mayers for their excellent microbiological work.
Conflict of Interest
The authors state there was no conict of interest.
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Figure 6. (HE) H&E stain of granulation tissue of animal specimen treated with petroleum jelly shows sparse collagen deposition, numerous
infiltrates of macrophages and limited mature fibroblast colonies. a, immature fibroblast; b, collagen fibres; c, macrophages. (VG) Van
Giesons stain of granulation tissue of animal specimen treated with petroleum jelly shows wavy strands of sparse collagen deposition.
a, collagen fibres. This figure is available in colour online at
Figure 7. (HE) H&E stain of granulation tissue of animal specimen treated with mupirocin ointment shows a matrix of collagen fibres, and
fibroblast in the vicinity. a, macrophages; b, fibroblast; c, collagen. (VG) Van Gieson stain of granulation tissue of animal specimen treated
with mupirocin ointment shows a matrix of dense bundles of collagen fibres. c, collagen fibres. This figure is available in colour online at
Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. (2011)
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... This itself reduces inflammation, and gradually, the wound heals. The medicines also possess fast wound healing properties, especially Draksha, Jati, and Triphala [21][22][23]. ...
Background: Patients undergoing radiotherapy (RT) or concurrent chemo-radiation (CCRT) for head and neck squamous cell carcinoma (HNSCC) often suffer from side effects such as mucositis, xerostomia, pharyngitis, laryngitis, and pain, which are being managed symptomatically by alcohol-based mouthwashes. Objectives: To determine the effectiveness of Ayurvedic mouthwash “Draksha Guduchyadi Kashaya” in reducing the severity of oral side effects of chemo-radiation. Material and methods: This concurrent parallel randomized controlled study was conducted at Sir Sunderlal Hospital, BHU, on 70 HNSCC patients scheduled to undergo RT/CCRT. Patients who met the inclusion-exclusion criteria were enrolled, and 35 were randomly assigned to either the intervention group (Ayurveda) or the control group using a simple random technique (lottery method). Blinding was not implemented in this study. Patients in the intervention group (Ayurveda) were instructed to perform kavala with 50 ml of “Draksha Guduchyadi Kashaya” for 2 min, ten times daily, while the control group performed 2-min gargling with soda-salt mouthwash ten times daily. Results: Out of the 70 patients enrolled, data from 60 patients were analyzed, revealing statistically significant differences in the onset of mucositis (p = 0.049), pharyngitis (p = 0.034), laryngitis (p = 0.009) and intensity of variables such as mucositis (p = 0.000), xerostomia (p = 0.046), pharyngitis (p = 0.002), laryngitis (p = 0.035), and pain (p = 0.000). These findings indicate that Ayurvedic mouthwash may be beneficial in managing the oral side effects of chemo-radiation in HNSCC. Conclusion: This AYUSH financially supported trial (Reg No: CTRI/2020/04/024672) demonstrates promise as a safe and cost-effective alternative for managing oral complications of RT/CCRT, offering complementary treatment for comprehensive cancer care.
... It is extremely light; therefore, it is comfortably absorbed into the skin and tends to leave no oil residue. 3 Microemulsion is a transparent colloidal system composed of oil, surfactant, co-surfactant, and water. The oil-water interfacial tension is drastically decreased by the surfactant, which is observable at the interface between the two phases. ...
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Grape seed oil, obtained from grape seeds (Vitis Vinifera L.), is used in cosmetic products. It demonstrates antifungal, antioxidant, anti-inflammation, and antibacterial activities. Microemulsions are a type of drug delivery system that has thermodynamic stability. Grape seed oil-loaded microemulsions were prepared by mixing water, oil, surfactant, and co-surfactant in an appropriate ratio. The objectives of this study were to develop grape seed oil-loaded microemulsions and to investigate the effects of surfactants (polysorbate 80 and polysorbate 20) and co-surfactants (ethanol, isopropyl alcohol, Cetiol® HE, and glycerin) on the physicochemical properties of the microemulsions. The stability was studied for 30 days at room temperature and protected from light. In addition, the antioxidant activity of grape seed oil and microemulsion formulations was analyzed. From the pseudo-ternary phase diagram, polysorbate 80, and Cetiol® HE at a ratio of 1:2 showed the largest microemulsion area. The amount of the loaded-grape seed oil and surfactant mixtures were 5-15% and 75-85%, respectively. All formulations were transparent light-yellow emulsions showing good thermodynamic stability. Moreover, surface charges, conductivity, pH, and viscosity were desirable. After the 30 day-stability tests, all microemulsion formulations presented good thermodynamic stability with no apparent changes observed. The antioxidant activity increased as grape seed oil was added in greater quantities. Despite the fact that the size of the particle was greater than 300 nm and the PDI value was high, the findings indicated that a microemulsion system containing grape seed oil, polysorbate 80, and Cetiol ® HE could be utilized with good antioxidant activity.
... Despite the existence of studies describing the in vitro and in vivo wound healing potential of extracts of different V. vinifera organs (leaves, fruits, seeds, and grape skin) [24][25][26][27][28][29][30][31] and their in vitro anti-MRSA inhibition activity, 32,33 there is little information available about its liposomal preparations, which alter the crude extract activity and mechanism of action in wound healing potential and anti-MRSA inhibition activities. Additionally, our preliminary zone-of-inhibition-based antibacterial screening against MRSA showed that the plantleaf-derived extract was the most biologically active one. ...
Vitis vinifera Egyptian edible leaf extract loaded on a soybean lecithin, cholesterol, and Carbopol gel preparation (VVL-liposomal gel) was prepared to maximize the in vivo wound healing and anti-MRSA activities for the crude extract, using an excision wound model and focusing on TLR-2, MCP-1, CXCL-1, CXCL-2, IL-6 and IL-1β, and MRSA (wound infection model, and peritonitis infection model). VVL-liposomal gel was stable with significant drug entrapment efficiency reaching 88% ± 3, zeta potential value ranging from -50 to -63, and a size range of 50-200 μm nm in diameter. The in vivo evaluation proved the ability of VVL-liposomal gel to gradually release the drugs in a sustained manner with greater complete wound healing effect and tissue repair after 7 days of administration, with a significant decrease in bacterial count compared with the crude extract. Phytochemical investigation of the crude extract of the leaves yielded fourteen compounds: two new stilbenes (1, 2), along with twelve known ones (3-14). Furthermore, a computational study was conducted to identify the genes and possible pathways responsible for the anti-MRSA activity of the isolated compounds, and inverse docking was used to identify the most likely molecular targets that could mediate the extract's antibacterial activity. Gyr-B was discovered to be the best target for compounds 1 and 2. Hence, VVL-liposomal gel can be used as a novel anti-dermatophytic agent with potent wound healing and anti-MRSA capacity, paving the way for future clinical research.
... Animal models using grape seed oil have demonstrated wound healing activity [233,234], efficacy against ulcerative colitis [235], protection against carbon tetrachloride-induced liver inflammation [236]. In cell lines, pancreatic β-cell apoptosis induced by hyperglycemia was reduced [237]. ...
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Oxidative stress causes various diseases, such as type II diabetes and dyslipidemia, while antioxidants in foods may prevent a number of diseases and delay aging by exerting their effects in vivo. Phenolic compounds are phytochemicals such as flavonoids which consist of flavonols, flavones, flavanonols, flavanones, anthocyanidins, isoflavones, lignans, stilbenoids, curcuminoids, phenolic acids, and tannins. They have phenolic hydroxyl groups in their molecular structures. These compounds are present in most plants, are abundant in nature, and contribute to the bitterness and color of various foods. Dietary phenolic compounds, such as quercetin in onions and sesamin in sesame, exhibit antioxidant activity and help prevent cell aging and diseases. In addition, other kinds of compounds, such as tannins, have larger molecular weights, and many unexplained aspects still exist. The antioxidant activities of phenolic compounds may be beneficial for human health. On the other hand, metabolism by intestinal bacteria changes the structures of these compounds with antioxidant properties, and the resulting metabolites exert their effects in vivo. In recent years, it has become possible to analyze the composition of the intestinal microbiota. The augmentation of the intestinal microbiota by the intake of phenolic compounds has been implicated in disease prevention and symptom recovery. Furthermore, the “brain–gut axis”, which is a communication system between the gut microbiome and brain, is attracting increasing attention, and research has revealed that the gut microbiota and dietary phenolic compounds affect brain homeostasis. In this review, we discuss the usefulness of dietary phenolic compounds with antioxidant activities against some diseases, their biotransformation by the gut microbiota, the augmentation of the intestinal microflora, and their effects on the brain–gut axis.
... Rats with excision wounds were used to test the wound-healing properties of cranberry and grape oil Animals that were administered cranberry and grape oil had considerably more hydroxyproline in their granulation tissue [108] Analyze the anti-ulcerogenic and anti-inflammatory properties of Vitis vinifera seed extracts (BGSE) and oil (BGSO) in rat experimental colitis After oral treatment, the hydroalcoholic extract and black grape seed oil displayed protective and prophylactic actions on the acute model of experimental ulcerative colitis, and this effect was highly dosage dependent [109] This investigation looked at the impact of GSO on acute liver damage brought on by carbon tetrachloride in rats exposed to γ radiation (7Gy) Due to its powerful antioxidant, anti-apoptotic and anti-inflammatory properties, GSO has protective effects on CCl4 -induced acute liver injury in γ-irradiated rats [110] Used an excision wound model to investigate the in vivo wound healing ability of Vitis vinifera seed extract with emphasis on wound healing therapeutic targets In contrast to the Mebo ® -treated group, the wound healing data showed that V. vinifera seed extract increased wound closure rates, increased VEGF and TGF-β levels, and considerably decreased IL-1β and TNF-α levels [111] Cells culture By assessing insulin levels and cell apoptosis rates, this study intended to assess the impact of TGSO on elevated glucose-induced Rattus pancreatic β-cell death and identify its signal transduction pathway processes. ...
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Seeds' abundant biologically active compounds make them a suitable primary platform for the production of natural extracts, innovative foods, medicines, and cosmetics. High levels of industrial and agricultural residues and byproducts are generated during the processing of grapes, although some parts can also be repurposed. This paper examines the phytochemical composition, manufacturing processes, and health-improving attributes of many varieties of grape oil derived using various extraction methods. Since the results are influenced by a range of factors, they are expressed differently among studies, and the researchers employ a variety of measuring units, making it difficult to convey the results. The primary topics covered in most papers are grape seed oil's lipophilic fatty acids, tocopherols, and phytosterols. In addition, new methods for extracting grape seed oil should therefore be designed; these methods must be affordable, energy-efficient, and environmentally friendly in order to increase the oil's quality by extracting bioactive components and thereby increasing its biological activity in order to become part of the overall management of multiple diseases.
... Is obtained from the seeds of grapes, and it was a by-product of winemaking and had many uses ranging from cooking cosmetics in controlling several diseases and wound healing potential [55][56][57] . Vitis vinifera L., which is commonly called grape used as a food and a beverage 58 . ...
... Supplementary Materials: The next supporting information are available to be downloaded at:, Figure S1: GC-MS spectrum for V. vinifera seed oil; Figures S2-S21:1D and 2D NMR spectra of compounds 28-36. See [19,[50][51][52][53][54][55][56][57][58]. ...
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This study explored the in vivo wound healing potential of Vitis vinifera seed extract using an excision wound model with focus on wound healing molecular targets including TGFBR1, VEGF, TNF-α, and IL-1β. The wound healing results revealed that V. vinifera seed extract enhanced wound closure rates (p < 0.001), elevated TGF-β and VEGF levels, and significantly downregulated TNF-α and IL-1β levels in comparison to the Mebo®-treated group. The phenotypical results were supported by biochemical and histopathological findings. Phytochemical investigation yielded a total of 36 compounds including twenty-seven compounds (1–27) identified from seed oil using GC-MS analysis, along with nine isolated compounds. Among the isolated compounds, one new benzofuran dimer (28) along with eight known ones (29–36) were identified. The structure of new compound was elucidated utilizing 1D/2D NMR, with HRESIMS analyses. Moreover, molecular docking experiments were performed to elucidate the molecular targets (TNF-α, TGFBR1, and IL-1β) of the observed wound healing activity. Additionally, the in vitro antioxidant activity of V. vinifera seed extract along with two isolated compounds (ursolic acid 34, and β-sitosterol-3-O-glucopyranoside 36) were explored. Our study highlights the potential of V. vinifera seed extract in wound repair uncovering the most probable mechanisms of action using in silico analysis.
Ethnopharmacological relevance: Pinus halepensis Mill. seed (Pinaceae), commonly known as "Zgougou," is widely consumed in the Mediterranean region and has long been used to treat diseases including bronchitis, rheumatism, infection, and inflammation. Aim of the study: The present study was focused on the investigation of some pharmacological activities, such as analgesic, anti-inflammatory, and wound healing effects, of cold-pressed oil of Pinus halepensis Mill. seed (COPHS). Materials and methods: The fixed oil of Pinus halepensis seed was extracted by a cold pressing process. The analgesic activity of COPHS was examined by acetic acid and formalin-induced contortion and pain tests. The anti-inflammatory effects were evaluated in carrageenan and formalin induced paw and ear edema models. Wound healing potential was assessed on an experimental skin wound, and the expression levels of inflammation mediators were determined by RT-qPCR in skin wound healing. Results: The results showed analgesic efficacy through significant inhibition of abdominal cramps (59.15%) and pain (75.91%). In addition, this oil exerted an anti-inflammatory effect by inhibiting ear (62.25%) and paw (70.00%) edema. The COPHS stimulated wound contraction in experimental skin wound healing with a contraction rate of 89.23% with notable reduction of TNF- and NF-kB expression levels in the treated groups. Conclusions: This study provided for the first time the pharmacological profile, particularly the analgesic, anti-inflammatory, and healing effects, of fixed oil extracted by cold pressing from the seed of Pinus halepensis Mill.
Research Proposal
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The present proposal of research aims to develop eco-friendly skin care formulations using microemulsions of fixed oils and extracts rich in bioactive compounds recovered from GCO pressing. To accomplish this overarching aim, this research proposal seeks to test different mixtures using oils from green coffee beans (Coffea arabica L.), cacay (Caryodendron orinocense), sacha inchi (Plukenetia volubilis L.), guava (Psidium guajava) and passion fruit (Passiflora edulis). The potential of microemulsion of novel fixed oils and antioxidants extracts from GCO pressing will be study to achieve results on (1) to improve the release and permeation of skin active, (2) to improve stability of formulations, (3) as a natural alternative to synthetic antioxidant, (4) as a self-preserving system, and (5) to improve both any skin disorder of interest and skin microbiome.
In the past few decades, the spray drying technique has gained wide acceptance as an alternative drying technique for transferring liquid forms into stable dry powder forms with application in the food, pharmaceuticals and cosmetic industry. Spray drying technique enables short operation time, drying of heat sensitive materials, desirable moisture content and desirable particle size of obtained powder. For the centuries, medicinal plants are considered as one of the most important sources of bioactive compounds. Different plant parts including leaves, flowers, stalks, fruits, seeds and roots but also whole plants are used for producing herbal pharmaceuticals, herbal cosmetic and dietary supplements. One of the most common used form are herbal extracts, produced with different extraction techniques (conventional and green novel techniques). Herbal extracts are highly concentrated in bioactive compounds such as phenolic compounds, tannins, alkaloids, pigments, vitamins, bioactive polysaccharides, peptides and volatile compounds with different biological properties. Liquid form of herbal extracts suffers from many disadvantages, such as short shelf life, higher cost of transport and storage. This chapter provides an overview on the principal applications related to encapsulation of bioactive compounds from herbal extracts with special attention on the choice of encapsulation agent on the quality of final dry extract. The application of spray drying represents a new promising drying technique for the production of dry extracts rich in bioactive compounds, which can be easily incorporated into new highly-valuable, bioactive compounds-rich products
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Nigella sativa (N. sativa) is popularly known as the black seed, a herb that has traditionally been used for centuries in many parts of the world. It has gained popularity due to its potential health benefits. However, more scientific data is needed to support the various health claims. This study was carried out to determine the fatty acid profile, α-tocopherol content and to evaluate the antioxidant activity of seed oil samples from three different regions in Yemen namely Marib, Sadah and Taiz. N. sativa seeds oil was extracted using three different solvents (n-hexane, petroleum ether and chloroform: methanol 2:1 v/v) and the fatty acids composition was analyzed using gas chromatography, while the α-tocopherol was determined using HPLC. Ferric thiocyanate (FTC) and thiobarbituric acid (TBA) methods were used to evaluate the antioxidant activity of the seeds oil. Results indicated that N. sativa seeds contain high amount of oil (30-48%) and the major unsaturated fatty acids were linoleic acid (57.96, 58.04 and 57.04%) followed by oleic acid (21.49, 20.87 and 20.60%), while the main saturated fatty acids were palmitic (11.56, 11.23 and 11.22%), followed by stearic and myristic acids in Marib, Taiz and Sadah samples respectively. Oil extracts exhibited strong antioxidant properties when compared to α-tocopherol with 78-82% inhibition in the FTC method and 70-80% in the TBA assays. The oil extracts were found to be rich in α-tocopherol content 290±1.5, 170±0.40 and 120±0.15 mg/100 g, in Marib, Sadah and Taiz samples, respectively. Present results suggest that N. sativa seeds contain high amount of antioxidants that are essential for health and preventing numerous diseases.
Photoautotrophic suspension cultures of moss protonema of Physcomitrella patens (Hedw.) B.S.G. and of tomato (Lycopersicon esculentum L.), respectively, were treated with three different β-lactam antibiotics. Ampicillin, cefotaxime and penicillin in the range from 10 μmol/L to 1 mmol/L had no visible effects on tomato cells but led to moss cells with giant chloroplasts. According to electron micrographs these cells are morphologically normal but contain 2–3 macrochloroplasts with well developed inner membranes. As only chloroplasts in dividing tip cells were affected, we conclude that β-lactam antibiotics specifically inhibit the plastid division process in the moss. Major proteins were not affected by antibiotic treatments as judged by SDS-PAGE and subsequent silver stain. The inhibitory effect on chloroplast division could not be compensated for by exogenous cytokinin (5 × 10−6 mol/L i6 Ade). However, upon removal of the antibiotics, macrochloroplasts started to divide. As division of bacteria and of cyanelles is sensitive to β-lactam antibiotics, we suggest that mosses conserved an ancient component of the chloroplast division mechanism that was subsequently modified or substituted during plastid evolution in land plants.
Novel 4-alkylidene-beta-lactams with a polyphenolic side chain were synthesized and evaluated as radical scavengers. We have undertaken a detailed study of the antioxidant activity in vitro with chemical and biological testing of the new beta-lactams and of the corresponding methyl polyhydroxy benzoates. Antioxidant activity of beta-lactams and methyl benzoates was measured with the Briggs-Rauscher oscillating reaction, the TEAC (Trolox( Equivalent Antioxidant Capacity) assay, and as ability to inhibit ROS (=Reactive Oxygen Species) production on myoblast H9c2 cells. The results were discussed with regard to mechanism and correlated with structural parameters.
Combinations of green tea flavor compounds with indole were tested for antibacterial activity against Streptococcus mutans, a bacterium responsible for causing dental caries. Synergism was found in the combination of sesquiterpene hydrocarbons (delta-cadinene and beta-caryophyllene) with indole; their bactericidal activities increased from 128-fold to 256-fold. The combination effect was confirmed by time-kill curve assay which showed that 6.25 mug/mL delta-cadinene combined with 400 mug/mL (subinhibitory concentration) indole was bactericidal against S. mutans. Furthermore, the combination of 25 mug/mL delta-cadinene and 400 mug/mL indole reduced the number of viable cells at any stage of growth.
Methods for the quantitative analysis of anthocyanins, leuco-anthocyanins, flavanols and total phenols in plant tissue extracts are critically examined and suitable modifications of existing methods are described.
The antimicrobial activity of 20 10% (v/v) solutions in ethanol of terpenes and terpenoids at several concentrations was tested against Erwinia amylovora NCPPB 595 in the liquid medium 523. The test organism responded differently to the chemicals. At 600, 900 and 1200 mg/l, none of the compounds reduced the growth of the bacterium. At 1500 mg/l, only some of the chemicals significantly inhibited growth x-Pinene. β-terpinene, dihydrocarveol, isopulegol and linalool reduced growth of suspensions of 1 × 103 cfu/ml, whereas β-pinene was more effective when challenged with larger numbers of cells (i.e. 1 × 105 cfu/ml and 1 × 107 cfu/ml). At 1500 mg/l, geraniol and citronellol exerted a bactericidal activity regardless of the concentrations of the test organism.