ArticlePDF Available

Cosmeceutical effect of ethyl acetate fraction of Kombucha tea by intradermal administration in the skin of aged mice



Background/purpose: Natural ingredients have been always an interesting approach to prolong youthful appearance of skin. One of the natural compounds is Kombucha tea (KT), which has been mainly used as an energy drink in Asian countries for a long time. Previous reports indicated that it has pharmaceutical and favorable wound repairing effects. The beneficial properties of KT are thought to be mainly due to the presence of fermentation products such as flavonoids and other polyphenols with inhibition of hydrolytic and oxidative enzymes and anti-inflammatory effects. These properties prompted us to study the anti-aging potential of KT and investigate its effective fraction in aged mice, METHODS: Kombucha tea was fractionated into chloroform, butanol, and ethyl acetate, and flavonoid content was determined. Young and old mice were used as control. KT ethyl acetate fraction (KEAf), which had the highest flavonoid content, was intradermally administered to old mice. Results: Administration of KEAf significantly increased the collagen content, NAD(+) /NADH level, and concomitantly improved skin connective tissue abnormalities in the aged skin. No sensitivity or irritation was observed. Conclusion: This finding suggested that KEAf can be a suitable candidate as a cosmetic product to improve aging-related skin abnormalities and regeneration of aged skin.
Cosmeceutical effect of ethyl acetate fraction of Kombucha
tea by intradermal administration in the skin of aged mice
Nafiseh Pakravan PhD
Elaheh Mahmoudi PhD
Seyed-Ali Hashemi PhD
Jamal Kamali BSc
Reza Hajiaghayi PhD
Mitra Rahimzadeh PhD
Vajiheh Mahmoodi MSc
Division of Immunology, Medical School,
Alborz University of Medical Sciences,
Karaj, Iran
Department of Parasitology and Mycology,
Medical School, Alborz University of
Medical Sciences, Karaj, Iran
Department of Pathology, Medical School,
Alborz University of Medical Sciences,
Karaj, Iran
Department of Operating Room Nursing,
Paramedical School, Alborz University of
Medical Sciences, Karaj, Iran
Herbal Medicine Research Center, Institute
of Medicinal Plant, ACECR, Karaj, Iran
Health, Safety, and Environment Research
Center, Alborz University of Medical
Sciences, Karaj, Iran
Department of Food Science and
Technology, Faculty of Agriculture, Islamic
Azad University, Shahrekord, Iran
Elahe Mahmoudi, Department of
Parasitology and Mycology, Medical School,
Alborz University of Medical Sciences, Karaj
Funding information
Deputy of Research, Alborz University of
Medical Sciences
Background/purpose: Natural ingredients have been always an interesting approach
to prolong youthful appearance of skin. One of the natural compounds is Kombucha
tea (KT), which has been mainly used as an energy drink in Asian countries for a
long time. Previous reports indicated that it has pharmaceutical and favorable
wound repairing effects. The beneficial properties of KT are thought to be mainly
due to the presence of fermentation products such as flavonoids and other polyphe-
nols with inhibition of hydrolytic and oxidative enzymes and anti-inflammatory
effects. These properties prompted us to study the anti-aging potential of KT and
investigate its effective fraction in aged mice,
Methods: Kombucha tea was fractionated into chloroform, butanol, and ethyl acet-
ate, and flavonoid content was determined. Young and old mice were used as con-
trol. KT ethyl acetate fraction (KEAf), which had the highest flavonoid content, was
intradermally administered to old mice.
Results: Administration of KEAf significantly increased the collagen content, NAD
NADH level, and concomitantly improved skin connective tissue abnormalities in
the aged skin. No sensitivity or irritation was observed.
Conclusion: This finding suggested that KEAf can be a suitable candidate as a cos-
metic product to improve aging-related skin abnormalities and regeneration of aged
aging, collagen, ethyl acetate fraction, Kombucha tea, NAD
Skin aging is a biological process during which changes in the struc-
tural integrity and physiological function of skin are induced. Visible
sign of aging includes development of dyschromia, roughness, fine
wrinkles followed by persistent deeper folds.
These changes occur
following microscopic structural changes including epidermal thin-
ning, dermal atrophy, reduction in connective tissue, decreased num-
bers of keratinocytes, reduced levels of elastin and collagen
synthesis, increased level of collagenase/matrix metalloproteinase
(MMP), increased oxidants,
reduction in the number and size of
vascular vessels,
loss of subcutaneous fat and elasticity, and
increased melanogen.
Thinning and loss of collagen fibers, which
form 90% of wet weight of skin connective tissue, are other promi-
nent features of aging skin.
Anti-aging products are used to protect skin against aging pro-
cess. These agents are capable of protecting the skin matrix by inhi-
bition of enzymatic degradation or promotion of collagen synthesis
Accepted: 9 October 2017
DOI: 10.1111/jocd.12453
J Cosmet Dermatol. 2017;19. ©2017 Wiley Periodicals, Inc.
in the skin.
Several efforts have been made to replace synthetic
anti-aging agents with natural alternatives. Of particular interest is
the use of traditional foods and medicines containing active ingredi-
One of these natural compounds is Kombucha tea (KT).
This so-called long-life mushroom tea has been known as popular
natural remedy for a long time and mainly used as an energy drink
in Asian countries. It is prepared from the fermentation of sugared
black tea (BT) with a symbiotic culture of acetic bacteria and Kom-
bucha fungi.
Previous reports indicated that Kombucha has valu-
able cosmetic activities when applied topically. Kombucha
specifically fights against glycation and the various gaps of skin
metabolism leading to improvement in the aging signs and cutaneous
The beneficial properties of KT are thought to be
due to the presence of vitamins, amino acids, and a variety of
micronutrients produced during fermentation. KT is especially a good
source of flavonoid and vitamin B3.
Flavonoids are important
anti-aging components that their amount in KT is much more than
They are a group of polyphenolic compounds with free radical
scavenging, inhibition of hydrolytic and oxidative enzymes, and anti-
inflammatory activities and capable of physically blocking UV pene-
tration, influencing DNA repair, and induction of cytoprotective
pathways in skin.
Vitamin B3 is a precursor of nicotinamide ade-
nine dinucleotide (NAD), which declines during aging leading to
reduction in collagen expression within cells.
Apart from cosmeceutical effects of KT, it has also been claimed to
have pharmaceutical effects and favorable wound repairing effects
when applied topically.
However, there are reports attributing
side effects to this beverage
and specifically, to date no report has
determined the effective fraction of KT. To reduce possible side
effects, determination of the effective anti-aging fraction of KT can be
beneficial. To do so, in this study KT was fractionated using organic
solvents and the effective anti-aging fraction was determined.
BT and KT preparation
Black tea (Golestan, Tehran, Iran) was added to boiling water (1.2% w/
v), mixed, and left to brew for 5 minutes. The tea was then filtered
through a sterile sieve, and sucrose (10%) was dissolved in the tea. To
prepare KT, 200 mL of the cooled BT was inoculated with 3% w/v tea
fungus plus 10% v/v previously fermented KT liquid and left to fer-
ment by incubating the Kombucha culture at 28°C for 14 days. The
resultant fermented tea was centrifuged at 600 gfor 20 minutes.
Fractionation of KT
According to the method defined by Jayabalan,
3 fractions of KT
including KCf (Kombucha tea chloroform fraction), KBf (Kombucha tea
butanol fraction), and KEAf (Kombucha tea ethyl acetate fraction) (1/
2 v/v) were concentrated using a vacuum rotary evaporator (R-200
model of Buchi, Sigma-Aldrich, Taufkirchen, Germany). This process
led to preparation of a viscid mass, which was dissolved in distilled
water, filtered through 0.22-lMillipore membrane filter, lyophilized,
and stored at 20°C.
Content of total flavonoids in BT and KT
The aluminum chloride colorimetric method was used to calculate
the content of flavonoids.
Briefly, 1 mL (100 mg/mL) solution of
each of BT, KT, KEAf, KCf, and KBf was mixed with 3 mL methanol,
0.2 mL of 10% aluminum chloride, 0.2 mL potassium acetate (1 mol/L),
5.6 mL of distilled water and incubated for 30 minutes at room tem-
perature. Quercetin (Sigma-Aldrich, Taufkirchen, Germany) was pre-
pared in dilutions of methanol (250-1000 lg/mL) and used to
prepare standard curve using spectrophotometry at a wave length of
415 nm. Total content of flavonoid compound (lg of QE/mg sample)
was calculated by the formula: T=(C9V)/M,C=concentration of
quercetin (mg/mL), V=volume of solution (mL), M=weight of
methanolic extract (gr). KEAf had the highest flavonoid content and
was further used to test for its anti-aging effect.
Experimental groups and study design
A total of 54 female NMRI mice were purchased from Pasteur Institute
Experimental Animal Center (Tehran, Iran). All animals were kept under
standard conditions of temperature (23 2°C) and humidity
(50% 10%) with an alternating 12-h light/dark cycles at the conven-
tional animal house of Alborz University of Medical Sciences. Given
free access to food and water, mice were housed for 1 week before
experiments and maintained under standard environmental conditions.
All experiments were done according to Animal Care and Use Protocol
of Ethics Committee of Alborz University of Medical Sciences.
According to the age, 5 experimental groups (n =9) were consid-
ered including: Group 1 as control young group (aged 2 months),
Group 2 as old group (aged 15 months) treated with KEAf at a dose of
5 mg/mL, Group 3 as old group (aged 15 months) treated with KEAf
at a dose of 10 mg/mL, Group 4 as control old group (aged 15 months)
treated with saline, and Group 5 as control old group (aged 15 months)
without any treatment. Treatments were carried out for 14 days and
performed via the intradermal route after the induction of anesthesia
and shaving a 2 92 cm region behind the animals neck.
Evaluation of skin irritation
Each animal was examined visually for signs of skin reactions, such
as erythema and edema, approximately 24 and 72 hours after intra-
dermal injection.
Also, pathological damage such as inflammatory
cell infiltration and fibroplasias was evaluated in hematoxylin and
eosin (H&E)-stained sections.
Histological analysis
To perform histological evaluation of the skin, the animals were sac-
rificed under ether anesthesia 72 hours after the last injection. Skin
tissue samples, including epidermis, dermis, were taken using a scal-
pel No. 15. The skin samples from the control and test groups were
fixed in 10% buffered formalin and were processed for paraffin sec-
tioning. Sections of about 3 lm thickness were taken and stained
with H&E. To further assess the amount of total collagen, skin sam-
ples were stained with Massons trichrome. The stained sections
were examined with an Olympus cX41 microscope, (Olympus, Tokyo,
Japan), and photographed using an Olympus D330 digital camera.
The photographs were analyzed via Scion Image Software to analyze
markers of skin tissue alteration such as fiber length and width, fiber
fragmentation, number of fibroblasts, and the space between fiber
bundles of dermal connective tissue.
Evaluation of total collagen content
The content of hydroxyproline was measured according to the
method previously described by Kivirikko et al,
with some modifi-
cations. The skin tissue samples (0.3 g) were homogenized and
hydrolyzed in 6 mol/L HCl, at 105°C for 16 hours. Free hydroxypro-
line was oxidized using chloramine T. Then, Ehrlich`s reagent was
added and incubated at 60°C leading to formation of a chro-
mophore. Hydroxyproline product was further purified through
extraction of interfering chromophores with toluene and then acid
phase. The absorbance of the supernatant containing hydroxyproline
was measured by spectrophotometry at the wavelength of 543 nm.
Hydroxyproline content was calculated from the standard curve,
which was linear at 10-160 lg/mL.
Immunohistochemical evaluation of collagen
type I and III
Tissue sections of the skin samples were stained using antibody speci-
fic for collagen type I (1:100; Col 1A SC-59772, Santa Cruz, CA, USA)
and anti-collagen III antibody (Sigma-Aldrich, Taufkirchen, Germany)
as DAKO kit (Denmark). Anti-rabbit IgG antibody was used as sec-
ondary antibody, and the images were captured using a Zeiss LSM 5,
(Carl Zeiss, Tokyo, Japan) fluorescent microscope. Paraffin sections
obtained from each group were dewaxed and hydrated. Sections of
4lm were prepared from each sample, washed with PBS, incubated
in 3% H
for 20 minutes, washed once more with PBS, and then
incubated in a protein block solution for 10 minutes. Then, primary
antibody (diluted 1:100 in 0.01 mol/L PBS) was added to sections and
incubated for 1 hour, washed 3 times, and incubated with envision
solution (complex of secondary antibody diluted 1:200 in PBS, avidin,
and horseradish peroxidase). After 30-min incubation time, the
enzyme activity was visualized using 3, 30-diaminobenzidine (DAB). A
brown staining was regarded as a positive reaction. Data are pre-
sented as the mean of 3 randomly selected fields of microscopic view.
and NADH content
The content of dinucleotides NAD
and NADH was determined in
skin tissue samples using NAD
/NADH assay kit (Abcam, London,
UK), according to the manufacturers instructions. Briefly, about
20 mg of tissue sample was washed with cold PBS and homogenized
with NAD/NADH extraction buffer. The supernatant was split into 2
aliquots. For NADH detection, the aliquot was incubated at 60°C for
30 minutes as NAD needs to be decomposed before the reaction.
To determine NADt (NAD and NADH), the other aliquot was kept
on ice to preserve NAD and NADH. The samples were transferred
into 96-well plates, and then a mixture of NAD cycling enzyme/buf-
fer was added to each sample. Finally, NADH developer was added
to each reaction. NADH standard was prepared according to the
manufacturers instructions. Total NAD (NADt) and NADH were esti-
mated directly, whereas the value of NAD
was estimated by sub-
tracting NADH from NADt and expressed as pmol/lg of skin tissue.
Statistical analysis
Statistical analysis was performed using GraphPad Prism 7.01. Data
are presented as meansSD. ANOVA was used to indicate any
significant difference among the groups. Value of Pwas considered
statistically significant when it was less than .05.
Flavonoid composition of BT, KT, and KT
Total flavonoid content of each of BT, KT, KEAf, KCf, and KBf is
shown in Figure 1. The flavonoid content increased in BT after fer-
mentation by Kombucha culture. The rate of flavonoid concentration
in KT and its derivatives was about 21%, 38.7%, 21%, and 25.4% in
KT, KEAf, KBf, and KCf, respectively. As Figure 1 shows, concentra-
tion of flavonoid in KEAf is about 22.3% more than KT. As KEAf had
the highest flavonoid content, it was further tested for anti-aging
µg QE/mg Sample
FIGURE 1 Total flavonoid content of each sample (lg
QE/100mg); the flavonoid concentrations significantly increased in
Kombucha tea rather than unfermented black tea (BT). *P<.05 vs
BT, **P<.007 vs BT
Skin irritation assessment
Visual evaluation of skin reaction at 24 and 72 hours after ID injec-
tion did not show inflammatory reactions, such as edema and ery-
thema. The score of irritation in KEAf and PBS groups was 0%, and
the mean evaluation score was 0 (Table 1). Also, histological obser-
vation of the skin tissue sections did not show pathological damage
or accumulation of inflammatory cells in the treated animals (Fig-
ure 2) suggesting that KEAf is not cytotoxic and does not sensitize
the skin.
Histological observation
Histological view of H&E- and trichrome-stained skin tissue of mice
aged 2 and 15 months showed that age increment extends skin
tissue abnormalities represented as reduced fiber bundles width and
length, increased space between dermal connective fiber bundles
(P<.05, Figure 3A), and decreased numbers of fibroblasts (P<.05,
Figure 3B). Also, decreased number of the collagen bundles in the
upper layer of dermis, epidermal atrophy, and decreased interdigita-
tion between the epidermis and dermis were seen in old skin tissue
vs young samples (Figure 2). In the mice aged 2 months, the fibers
formed a fibrous network and were less regularly arranged compare
with in the older animals. Degrees of skin connective tissue abnor-
malities were significantly improved in samples treated with KEAf.
Figures 2 and 3 together show that treatment with KEAf on average
decreased skin connective tissue abnormalities approximately 48.2%
in aged skin sample (15 month at age) vs vehicle-treated old skin
(P<.007). There was no significant difference between samples
treated with 5 or 10 mg/mL KEAf.
TABLE 1 Evaluation of skin irritation in Kombucha tea ethyl acetate fraction (KEAf)- and PBS-treated animals; total score/number of mice
(n =9) =Ms
Groups Vehicle L (PBS) KEAf
Hours 24 72 24 72
Skin reaction Erythema Edema Erythema Edema Erythema Edema Erythema Edema
Mean score (Ms) 0 0.1 0 0 0 0 0 0
FIGURE 2 Formalin-fixed skin sections were stained with H&E and trichrome. (A) control 2-months old, (B) focus on (A), (C) control 15-
months old, (D) focus on (C), (D) 15-months old mice treated with 5mg/ml KEAf, (E) focus on (D), (F) 15-months old mice treated with 5mg/ml
KEAf, (E) focus on (F), (a) Trichrom staining of 2-months old, and (b) trichrom staining of 15-months old control mice skin. Mice aged 15
months showed significant increase in abnormalities in connective tissue structure: thin, disorganized fiber bundles present throughout the
dermis, and reduced numbers of fibroblasts. Also, patches of ground substance deposition increased in areas resembling degraded fiber
bundles. Skin connective tissue alterations due to age increment are partially improved by KEAf treatment. Data is presented as the mean of
three randomly selected fields of microscopic view. (Scale bar 50 lm).
KEAf stimulated type I and III collagen
synthesis in the skin
Based on hydroxyproline estimation, the total collagen content was
different in the skin tissue of the 2 different age groups. The aged
skin tissue samples showed a mean reduction of 35.5% in hydrox-
yproline content comparing to the young skin tissue samples
(P<.05, Table 2). Treatment with KEAf led to a 31.8% increase at
the hydroxyproline content of the skin connective tissue samples
of the old animals compared with those treated with PBS
(P<.007, Table 2). Immunohistochemical staining of the skin from
old and young animals with monoclonal antibody against collagen
type I revealed that type I collagen content decreased in the aged
skin compared to the skin tissue from young mice (Figure 4A).
Analysis of the images by Image Scan software showed that there
is about 29.3% lower collagen type I in the skin tissue samples
from old animals than the young animals (P<.05, Figure 4B). How-
ever, immunohistochemical staining of collagen type III showed no
prominent change in the skin connective tissue in mice aged
15 months (Figure 5). Based on hydroxyproline content and
immunohistochemical analysis of collagen type I, KEAf treatment
restored abnormalities (Figure 2), and increased total collagen in
the skin of aged mice (Table 2). Intradermal treatment of skin of
aged mice with KEAf revealed a significantly higher content of col-
lagen type I and III compared with that of the vehicle group (Fig-
ures 4 and 5).
/NADH content
In the final set of experiments, level of NAD
/NADH production
upon administration of KEAf was investigated. The findings showed
notable reduction in NAD
/NADH content in the aged skin vs skin
of the young animals. Parallel to this, the NAD
/NADH level in the
skin of old animals treated with KEAf was significantly higher than
the skin tissue samples of their age-matched treated with PBS. There
was no difference in NAD
/NADH level between 5 and 10 mg/mL
KEAf (P<.01, Figure 6).
The health benefits of KT have made some researchers consider it
as a probiotic product.
During the fermentation process, effective
components are formed and cause the beneficial properties of this
drink. Antioxidant and free radical scavenging properties of KT,
which are mainly attributed to flavonoid and polyphenols, are
formed during fermentation.
The result of the present study
indicated an increase in the flavonoid content of KT and especially
KEAf in BT by about 21% and 38.7%, respectively, after fermenta-
tion of BT by Kombucha culture. The increase in the flavonoid con-
tent of KT and especially KEAf might be due to the secretion of
some enzymes by the microbial biofilm that degrades the polyphe-
nols to flavonoids.
Anti-inflammatory effects of flavonoids
consistent with the lack of skin sensitization upon KEAf administra-
tion. Also, a large body of literature attributed anti-aging activities of
flavonoids to their potent antioxidant properties.
Histological comparison of the dermal sections between mice
aged 15 months and 2 months revealed that increasing of age is
associated by thinner fiber bundles, more open space within and
between bundles in the dermis, fragmentation of collagen fibers, epi-
dermal atrophy, and decreased numbers of fibroblasts. Previous
FIGURE 3 (A) Average of skin connective tissue abnormalities was measured with an image analysis (image J: ¤space
between dermal connective fiber bundles was scored using a scale of 1-10. (B) Number of fibroblast, per unit area: about 0.5 mm2 per skin
tissue sample. Correlation analysis between the numbers of fibroblasts in the dermis and age revealed a significant effect of age increment on
the decrease of fibroblasts in the dermis. The amount of abnormalities decreased significantly with KEAf treatment. Values are means of 9
measurements. *P < 0.05 vs. 2 month-old mice, **P < 0.007 vs. vehicle-treated skin.
TABLE 2 Total collagen content evaluated by hydroxyproline
Mice skin tissue Old mice skin tissue
Young Old Vehicle KEAf
Hydroxyproline (lg/0.1 g sample)
294.19 189.5* 173.14 253.93**
*P˂.05 vs. young skin connective tissue.
**P˂.007 vs. vehicle.
study showed that decreased numbers of keratinocytes in aged skin
are largely responsible for the epidermal thinning.
Also, thinning of
dermis in aged skin is due to reduction in the amount and
organization of connective tissue comprising febrile collagen bundles
and elastic fibers along with a complex array of proteoglycan and
other extra cellular matrix molecules.
The rate of such abnormalities
FIGURE 4 (A) Immunostaining of the skin connective tissue from old and young animals with monoclonal antibody collagen type I revealed
that type I collagen content decreased in aged skin, compared with younger mice skin. Intradermal treatment of skin of aged mice with KEAf
revealed a significantly increased content of type I collagen (secondary antibody conjugate to FITC, magnification 2009). (B) Content of
collagen type I was quantified for each image using the ImageJ analysis ( 4 nonconsecutive tissue sections (*P<.05 vs. 2-
mo-old mice, **P<.007 vs. vehicle)
FIGURE 5 Immunohistological evaluation of type III collagen in old mice skin aged 15 mo revealed prominent staining vs. 2 mo old
(collagen fibers stained brown). This dermal staining was significantly increased in the skin of aged mice when treated with KEAf (n =9, Scale
bars =100 lm)
was significantly improved in aged mice skin treated with KEAf com-
pared to the control groups. It suggests that these alterations in the
dermal connective tissue are largely responsible for the thin, fragile,
and finely wrinkled quality of naturally aged skin.
Immunohistochemical evaluation of collagen type I in the skin
of old subjects revealed a decrease in collagen type I by 29.3%
compared with the young mice skin. Many studies have also
demonstrated that collagen type I content decreases with age.
However, there are differences in the collagen type I content of
naturally aged skin,
sun protected skin,
and photo-aged skin.
As for type III collagen, there was no significant difference
between skin of the old and young mice. This is consistent with
previous studies indicating that type III collagen content does not
change in covered skin with increasing age.
other studies have shown that type III collagen decreases with age
increment, although it is noteworthy that these researchers studied
skin of a sun exposed site.
In addition, it should be considered
that type III collagen constitutes a small portion of the total colla-
gen in the skin,
and type I collagen is the main constituent of
skin connective tissue by approximately 80% of skin wet weight.
Presumably, reduction in collagen content and declining trend in
collagen fiber number and width with increasing age are conse-
quences of destruction and impairment in the replacement of col-
lagen type I. Treatment of aged mice skin with KEAf led to a
measurable restoration of the dermal connective tissue abnormali-
ties as a consequence of the natural aging process. KEAf could
also significantly increase total collagen content of the skin and
increase in the type I and III collagen content. This could be
mainly attributed to flavonoid and polyphenols formed during fer-
mentation causing antioxidant and free radical scavenging proper-
ties of KEAf.
In addition, Jayaban et al
reported that ethyl
acetate fraction of KT contains dimethyl malonate and vitexin.
These compounds show strong radical scavenging activity with
anti-aging properties.
Alternatively, the present study demonstrated that ethyl acetate
fraction of KT can stimulate energy production. Accordingly, the
treatment of aged mice skin with KEAf significantly increased
/NADH content whose level normally decreased in old mice.
In parallel with these findings, a previous report demonstrated a cor-
relation between NADPH levels and type I collagen expression in
adult human skin fibroblast cell in culture. The type I collagen
expression was decreased in parallel with decrease in NAD
level and increased with treatment with a complex of niacinamide.
Gomes et al. directly measured that during aging, decline in NAD
content inducing a pseudohypoxic state led to distribution of
nuclear-mitochondrial communication and finally decline in mito-
chondrial function with age. They demonstrated that this phe-
nomenon can be apparently reversible with adding NAD
to cell.
When there is a sufficient level of NAD
/NADH on fibrob-
lasts, production of collagen is high.
Based on the findings of other
studies and the finding presented in this study, it is hypothesized
that the reduction in collagen content in aged animals occurs as a
consequence of reduced NAD
/NADH level. Hence, it is hypothe-
sized presumably that the beneficial effects of KEAf are due to fla-
vonoids and energy-producing compounds such as great source of
Vit B3, which can stimulate collagen production and repair abnormal-
ities of connective tissue in naturally aged skin. Finally, the results of
the current study suggest that KEAf treatment is useful to improve
thickness and flexibility of aged skins. Accordingly, KEAf can be
applied as a potential candidate for natural cosmeceuticals/functional
cosmetics either used in this study or as a topically supplement in an
attempt to prolong youthful skin appearance. However, more
research is required to elucidate the mechanisms underlying this
phenomenon and conduct an investigation on human skin.
Altogether, our results suggest that KEAf treatment is useful to
improve thickness and flexibility of aged skin. The beneficial effects
of KEAf are presumed to be due to flavonoids and energy-producing
compound capable of repairing connective tissue in naturally aged
skin. This study suggests KEAf as a potential candidate for natural
cosmeceuticals/functional cosmetics and long-term use in aged pop-
ulations. However, more research is required to elucidate the mecha-
nisms beneath this phenomenon.
This work was financially supported by the Deputy of Research,
Alborz University of Medical Sciences under reference No. 2427656.
The authors have no conflict of interest.
Nafiseh Pakravan
/NADH ratio is measured by spectrophotometric
assay. All KEAf-treated samples had significantly greater NAD
ratio compared with controls (N = 9, *P< .05 vs. 2-mo-old mice,
**P< 0.01 vs. vehicle)
1. West MD, Pereira-Smith OM, Smith JR. Replicative senescence of
human skin fibroblasts correlates with a loss of regulation and
overexpression of collagenase activity. Exp Cell Res. 1989;184:138-
2. Lavker R. Cutaneous aging: chronologic versus photoaging. Photo-
dam. 1995;1:123-135.
3. Glogau RG. Physiologic and structural changes associated with aging
skin. Dermatol Clin. 1997;15:555-559.
4. Varani J, Warner RL, Gharaee-Kermani M, et al. Vitamin A
antagonizes decreased cell growth and elevated collagen-degrading
matrix metalloproteinases and stimulates collagen accumulation
in naturally aged human skin. J Invest Dermatol. 2000;114:480-
5. Chung JH, Yano K, Lee MK, et al. Differential effects of photoaging
vs intrinsic aging on the vascularization of human skin. Arch Derma-
tol. 2002;138:1437-1442.
6. Yaar M. Clinical and histological features of intrinsic versus extrinsic
skin aging. In: Gilchrest BA, Krutmann J, eds. Skin Aging. Berlin:
Springer; 2006.
7. Cheng W, Yan-hua R, Fang-gang N, Guo-an Z. The content and ratio
of type I and III collagen in skin differ with age and injury. Afr J Bio-
tech. 2011;10:2524-2529.
8. Varani J, Dame MK, Rittie L, et al. Decreased collagen production in
chronologically aged skin: roles of age-dependent alteration in
fibroblast function and defective mechanical stimulation. Am J Pathol.
9. Lee H-S, Kim M-R, Park Y, et al. Fermenting red ginseng enhances
its safety and efficacy as a novel skin care anti-aging ingredient:
in vitro and animal study. J Med Food. 2012;15:1015-1023.
10. Mukherjee PK, Maity N, Nema NK, Sarkar BK. Bioactive com-
pounds from natural resources against skin aging. Phytomed.
11. Perricone NV. Treatment of skin damage using polyenylphos-
phatidylcholine. Google Patents; 2001.
12. Yadav T, Mishra S, Das S, Aggarwal S, Rani V. Anticedants and natu-
ral prevention of environmental toxicants induced accelerated aging
of skin. Environ Toxicol Pharmacol. 2015;39:384-391.
13. Gould L, Abadir P, Brem H, et al. Chronic wound repair and healing
in older adults: current status and future research. Wound Rep Regen.
14. Katz SE. Wild Fermentation: The Flavor, Nutrition, and Craft of Live-Cul-
ture Foods. White River Junction, VT: Chelsea Green Publishing; 2016.
15. Khan N, Mukhtar H. Tea and health: studies in humans. Curr Pharm.
16. Lintner K. Cosmetic or dermopharmaceutical compositions contain-
ing kombucha. Google Patents; 2008.
17. Baumann LS. Less-known botanical cosmeceuticals. Dermatol Ther.
18. Jayabalan R, Malbaˇsa RV, Lonˇcar LS, Vitas JS, Sathishkumar M. A
review on Kombucha teamicrobiology, composition, fermentation,
beneficial effects, toxicity, and tea fungus. Comp Rev Food Sci Food
Safety. 2014;13:538-550.
19. Jayabalan R, Chen PN, Hsieh YS, et al. Effect of solvent fractions of
kombucha tea on viability and invasiveness of cancer cellscharac-
terization of dimethyl 2-(2-hydroxy-2-methoxypropylidine) malonate
and vitexin. IJBT. 2011;10:75-82.
20. Jayabalan R, Marimuthu S, Swaminathan K. Changes in content of
organic acids and tea polyphenols during kombucha tea fermenta-
tion. Food Chem. 2007;102:392-398.
21. Sajilata M, Bajaj PR, Singhal R. Tea polyphenols as nutraceuticals.
Comp Rev Food Sci Food Safetys. 2008;7:229-254.
22. Schagen SK, Zampeli VA, Makrantonaki E, Zouboulis CC. Discovering
the link between nutrition and skin aging. Dermato-Endocrinol.
23. Saeed N, Khan MR, Shabbir M. Antioxidant activity, total phenolic
and total flavonoid contents of whole plant extracts Torilis lepto-
phylla L. BMC Complement Altern Med. 2012;12:221.
24. Gonz
alez S, Fern
andez-Lorente M, Gilaberte-Calzada Y. The latest
on skin photoprotection. Clin Dermatol. 2008;26:614-626.
25. Gao K, Henning SM, Niu Y, et al. The citrus flavonoid naringenin
stimulates DNA repair in prostate cancer cells. J Nut Biochem.
26. Srinivasan R, Smolinske S, Greenbaum D. Probable gastrointestinal
toxicity of kombucha tea: is this beverage healthy or harmful? J Gen
Intern Med. 1997;12:643-644.
27. Meda A, Lamien CE, Romito M, Millogo J, Nacoulma OG. Determi-
nation of the total phenolic, flavonoid and proline contents in Burk-
ina Fasan honey, as well as their radical scavenging activity. Food
Chem. 2005;91:571-577.
28. Draize JH, Woodard G, Calvery HO. Methods for the study of irrita-
tion and toxicity of substances applied topically to the skin and
mucous membranes. J Pharmacol Exp Ther. 1944;82:377-390.
29. Kivirikko K, Liesmaa M. A colorimetric method for determination of
hydroxyproline in tissue hydrolysates. Scand J Clin Lab Invest.
30. H
ainen M, Nieminen R, Vuorela P, Heinonen M, Moilanen E.
Anti-inflammatory effects of flavonoids: genistein, kaempferol, quer-
cetin, and daidzein inhibit STAT-1 and NF-jB activations, whereas
flavone, isorhamnetin, naringenin, and pelargonidin inhibit only NF-
jB activation along with their inhibitory effect on iNOS expression
and NO production in activated macrophages. Mediat Inflamm.
2007;2007:10 pp.
31. Lamming DW, Ye L, Sabatini DM, Baur JA. Rapalogs and mTOR inhi-
bitors as anti-aging therapeutics. J Clin Lab Invest. 2013;123:980-
32. Ami
c D, Davidovi
slo D, Trinajsti
c N. Structure-radical
scavenging activity relationships of flavonoids. Croat Chem Acta.
33. Masaki H. Role of antioxidants in the skin: anti-aging effects. J Der-
matol Sci. 2010;58:85-90.
34. Bernstein EF, Chen YQ, Kopp JB, et al. Long-term sun exposure
alters the collagen of the papillary dermis: comparison of sun-pro-
tected and photoaged skin by Northern analysis, immunohistochemi-
cal staining, and confocal laser scanning microscopy. J Am Acad
Dermatol. 1996;34:209-218.
35. Sakai S, Yamanari M, Miyazawa A, et al. In vivo three-dimensional
birefringence analysis shows collagen differences between young
and old photo-aged human skin. J Invest Dermatol. 2008;128:1641-
36. Lovell C, Smolenski K, Duance V, Light N, Young S, Dyson M. Type I
and III collagen content and fibre distribution in normal human skin
during ageing. Br J Dermatol. 1987;117:419-428.
37. Light ND. Estimation of types I and III collagens in whole tissue by
quantitation of CNBr peptides on SDS-polyacrylamide gels. BBA-Pro-
tein Struct Mol Enzym. 1982;702:30-36.
38. Wulf HC, Sandby-Møller J, Kobayasi T, Gniadecki R. Skin aging and
natural photoprotection. Micron. 2004;35:185-191.
39. Antonopoulou I, Varriale S, Topakas E, Rova U, Christakopoulos P,
Faraco V. Enzymatic synthesis of bioactive compounds with high
potential for cosmeceutical application. App Microbiol Biotechnol.
40. V
ına I, Semjonovs P, Linde R, Deninßa I. Current evidence on physio-
logical activity and expected health effects of kombucha fermented
beverage. J Med Food. 2014;17:179-188.
41. Osborne R, Carver R, Mullins L, Finlay D. Practical application of cel-
lular bioenergetics to the care of aged skin. Br J Dermatol.
42. Gomes AP, Price NL, Ling AJ, et al. Declining NAD+induces a pseu-
dohypoxic state disrupting nuclear-mitochondrial communication
during aging. Cell. 2013;155:1624-1638.
43. Wagner S, Hussain M, Beckert S, et al. Lactate down-regulates cellu-
lar poly (ADP-ribose) formation in cultured human skin fibroblasts.
Eur J Clin Invest. 2007;37:134-139.
How to cite this article: Pakravan N, Mahmoudi E, Hashemi S-A,
et al. Cosmeceutical effect of ethyl acetate fraction of Kombucha
tea by intradermal administration in the skin of aged mice. J
Cosmet Dermatol. 2017;00:19.
... O número de melanócitos tende a diminuir com a idade e, então, a pele fica menos protegida contra os raios ultravioleta. Com essas mudanças, a pele torna-se mais sensível às lesões e a cura torna-se mais lenta (PAKRAVAN et al., 2018). ...
... A administração de fração kombucha de acetato de etila aumentou significativamente o conteúdo de colágeno, e melhorou concomitantemente as anormalidades do tecido conjuntivo da pele envelhecida. Não foi observada nenhuma sensibilidade ou irritação o que sugeriu que a fração kombucha de acetato de etila pode ser um candidato adequado como produto cosmético para melhorar as anormalidades da pele relacionadas ao envelhecimento e a regeneração da pele envelhecida (PAKRAVAN et al., 2018). ...
... In addition, vitamin B3 stimulates epidermal ceramide synthesis and helps to eliminate hyperpigmentation. Thus, it is an agent used in the treatment of skin blemishes (Pakravan et al., 2018). In a study investigating the inhibition of the enzymes, e.g. ...
Full-text available
Kombucha, originated in China 2000 years ago, is a sour and sweet-tasted drink, prepared traditionally through fermentation of black tea. During the fermentation of kombucha, consisting of mainly acidic compounds, microorganisms, and a tiny amount of alcohol, a biofilm called SCOBY forms. The bacteria in kombucha has been generally identified as Acetobacteraceae. Kombucha is a noteworthy source of B complex vitamins, polyphenols, and organic acids (mainly acetic acid). Nowadays, kombucha is tended to be prepared with some other plant species, which, therefore, lead to variations in its composition. Pre-clinical studies conducted on kombucha revealed that it has desired bioactivities such as antimicrobial, antioxidant, hepatoprotective, anti-hypercholestorelomic, anticancer, anti-inflammatory, etc. Only a few clinical studies have been also reported. In the current review, we aimed to overhaul pre-clinical bioactivities reported on kombucha as well as its brief compositional chemistry. The literature data indicate that kombucha has valuable biological effects on human health.
... Kombucha maintains the quality of minced beef, significantly slowing down the growth of microbes, extending the shelf life of minced beef to 3 days. This makes it a potential food storage material [34]. ...
Full-text available
Nowadays, people's awareness of the role of diet in maintaining well-being and good health is increasing. Consumers expect that products will not only provide them with essential nutrients, but will also be a source of biologically active substances that are beneficial to their health. One of the "health trends" that has emerged among consumers around the world is kombucha with high antioxidant potential, obtained through the activity of a consortium of acetic acid bacteria and osmophilic yeast, also called "tea fungus". Kombucha is characterized by its health properties. Consumption of kombucha has always been associated with various health benefits. There are many personal experiences and feedback from kombucha drinkers around the world about the kombucha's ability to protect against a huge number of metabolic and infectious diseases. The promising results in in vitro and in vivo studies prompted us to create a new type of kombucha concentrat, which can be used directly for consumption, can be added to other drinks (juises, teas, herbal decoctions), might be used for preparation of carbonated and non-carbonated drinks, for food preservation, for vinegar preparation, as an additive in cosmetics and antiinflammatory products. The aim of this article is to review research in search of experimental confirmation of the many health aspects of kombucha. The analysis of the literature data is performed in accordance with the latest concepts of health protection requirements. Attention is paid to the active compounds in kombucha, responsible for the specific effect and the mechanisms of their action. It has been shown that kombucha can be effective in health prevention and recovery due to four main properties: detoxification, antioxidant, energizing potentials and stimulation of suppressed immunity. In recent years, its consumption has increased due to its useful properties and ingredients, immunostimulatory, anti-inflammatory, anti-tumor, antioxidant and other properties. Recent experimental studies of kombucha consumption suggest that the drink is suitable for prophylaxis against broad-spectrum metabolic and infectious disorders. This makes kombucha attractive as a drink for health prevention. In this article we describe the benefits of consuming the ancient traditional Chinese drink kombucha and describe the production of our new product using modern technology. Our combucha extract can be used for direct consumption and as a raw material for the production of various products containing kombucha. KOMBUCHA EXTRACT of DALVITA is made by a special technology and is 100 times more concentrated than kombucha tea, which makes its action a hundred times stronger and reduces the required daily amounts a hundred times
... Healing effects of KT, on the gastrointestinal system, particularly its extraordinary effect on the healing of intestinal ulcers has been purported traditionally and reported scientifically (20)(21)(22). Also, we previously demonstrated that KT had rejuvenating effects on the skin of old mice (23). It prompted us to investigate if KT can also have therapeutic or relieving effects on the mice model of colitis. ...
Full-text available
Objectives: Zonula occludens proteins (ZO-1 and ZO-2) are important intracellular tight junction (TJ)-associated proteins that link the cell cytoskeleton to the trans-membrane TJ proteins. Destruction of TJ proteins is called the "leaky gut syndrome" and has been observed in some of the gastrointestinal diseases such as the inflammatory bowel disease (IBD). So, therapeutic approaches aim to restore the expression of TJ proteins and reduce intestinal permeability. Healing effect of Kombucha tea (KT), so-called long-life mushroom, on the gastrointestinal system, particularly its extraordinary healing effects on intestinal ulcers has been purported traditionally and rarely reported scientifically. This study aimed to investigate the therapeutic effect of filtered KT (fKT) in young and old mice model of colitis. Materials and methods: Leaky gut was induced in two groups of young and old age using dextran sodium sulfate in drinking water for seven days. Then, fKT was administered to the mice affected by colitis and compared with the age-matched normal and untreated animals with colitis. Results: Survival rate of the fKT-treated young and old animals with colitis increased and weight loss decreased. Accordingly, digestive disorders characterized by bleeding and diarrhea were improved in fKT-treated mice. Molecular and histological examination indicated that expression of ZO-1 and ZO-2 was significantly improved in fKT-treated mice. Conclusion: Our results suggest KT as a promising therapeutic candidate to reduce intestinal permeability. Young animals with colitis showed more severe clinical signs and less survival rate than old mice with colitis, but this group responded better to fKT treatment than the old mice.
... Recent research on antibacterial (Bhattacharya et al. , 2018Sknepnek et al. 2018), antifungal, (Mahmoudi et al. 2016) antiviral (Fu et al. 2015), and antioxidant properties (Sun et al. 2015;Gamboa-Gomez et al. 2016 of plant-based fermented beverage 'Kombucha' states that the antioxidant property is mediated by microbial metabolism of tea polyphenols . This report has been confirmed by performing both in vitro Vazquez-Cabral et al. 2017;Sknepnek et al. 2018) and in vivo (Bellassoued et al. 2015;Gamboa-Gomez et al. 2017;Salafzoon et al. 2017;Pakravan et al. 2017) tests. ...
... Recent research on antibacterial (Bhattacharya et al. , 2018Sknepnek et al. 2018), antifungal, (Mahmoudi et al. 2016) antiviral (Fu et al. 2015), and antioxidant properties (Sun et al. 2015;Gamboa-Gomez et al. 2016 of plant-based fermented beverage 'Kombucha' states that the antioxidant property is mediated by microbial metabolism of tea polyphenols . This report has been confirmed by performing both in vitro Vazquez-Cabral et al. 2017;Sknepnek et al. 2018) and in vivo (Bellassoued et al. 2015;Gamboa-Gomez et al. 2017;Salafzoon et al. 2017;Pakravan et al. 2017) tests. ...
... Estudos relataram que essa capacidade se deve principalmente à capacidade do ácido glicurônico em se ligar a moléculas tóxicas e de aumentar sua excreção (26). Ainda, Pakravan e cols (2017) avaliaram o uso intradérmico do kombucha com finalidades estéticas como antiaging, com resultados promissores em modelo animal (33). ...
Full-text available
A busca por um estilo de vida mais saudável tem levado as pessoas a procurarem e escolherem produtos mais saudáveis. O kombucha é um probiótico, consumido no mundo todo, como uma bebida caseira refrescante, sendo também comercializada por algumas empresas. Esse trabalho avaliou o kombucha, relacionando composição, efeitos benéficos e aspectos toxicológicos causados por seu consumo. Foi realizada uma revisão crítica da literatura, baseada em artigos científicos, livros e documentos oficiais, nas bases de dados de artigos científicos: SciELO, Lilacs, Medline, e Science Direct, nos idiomas português, inglês e espanhol publicados entre 2014 e 2018, onde foi utilizado o descritor “kombucha”. A busca bibliográfica foi feita sem restrição de população e tema para não limitar o número de artigos. Os estudos apontaram prováveis benefícios relacionados à composição química do kombucha. Os benefícios relatados foram: atividade antioxidante, antibacteriana, antifúngica, efeito probiótico, potencial anticarcinogênico, efeito hipoglicemiante, além de outros efeitos terapêuticos e de prevenção de doenças crônicas não transmissíveis. Poucas pesquisas foram feitas com humanos para avaliar com mais precisão seus benefícios, toxicidade, e usos terapêuticos. Portanto, mais estudos devem ser realizados, com o intuito de identificar os metabólitos produzidos, entender sua relação com as atividades biológicas, assim como sua aplicação em humanos.
Tea polyphenol (TP) loaded alginate‐collagen microspheres were prepared by ionic gelation method by using Ca2+ as cation and alginate as anion. The morphology of the prepared microspheres was characterized by scanning electron microscopy. TP appeared in an amorphous state with an average size of 510.26 μm when encapsulated in the microspheres. The loading capacity (LC%) and the encapsulation efficiency (EE%) were about 5.11% (w/w) and 33.51%, respectively. These microencapsulated beads were evaluated as a pH‐sensitive system for the delivery of tea polyphenol. At pH 7.4, the amounts of TP released increased significantly as compared with those released at pH 1.2. The 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH ) radical‐scavenging activity and the stability of the microencapsulated TP were higher than that of free TP. Overall, the TP microspheres could improve antioxidant activity and the storage stability of TP at room temperature for 6 months.
Full-text available
The striving of the modern man towards overall well-being includes healthy food as an essential element. According to modern perceptions, food not only needs to satisfy the body's needs for energy and nutrients, but it also needs to have a preventive function. In recent years the Balkans have been in the focus of ethnobotanical research that reveals the traditional knowledge of the model of diet and use of the means of healing in the light of cultural diversity, placing it in a comparative matrix as part of the traditional knowledge on the Balkans and Europe. Diet on the Balkan Peninsula is close to modern requirements for a healthy diet. Some recent studies aim to isolate the yeast strains that could be used effectively as baker's yeast and compared them with the commercial baker's yeast available in the market and to identify these strains which carried a possibility of being used as industrial baker's yeast. An essential part of the food of people on the Balkans is fermented food and fermented beverages. Fermented food includes bread, leavened pasta, dairy products, pickles and fruit pickles. Bread is considered as leading food, and at the same time, it is much more than just ordinary food. Despite the long history of making leavened bread, where all its components and processes are technologically streamlined, today there is a real revival of interest in its traditional production. As a result of the ethnobotanical studies in the Balkan region, more than forty (40) plant taxa have been established which are used in traditional practices as a fermentation element. Over thirty (30) are used in making bread yeast. Twenty-five (25) taxa of them have a high index of citation in documentary sources and by informants in field studies. They are representatives of 24 families and 17 genera. The plants of the Poaceae (5), Leguminosae (3), Solanaceae (2) and Amaryllidaceae (2) predominate. Dicotyledon plant taxa are dominant (72%). All taxa are well-known to the local population as plant species used for food, spices or medicinal plants. They are related to people's lifestyle, used in everyday life, easily accessible and well-recognisable. The number of wild and cultivated plants used for yeast is almost the same. This is yet another confirmation that cultural characteristics are seen as the leading reason for choosing the use of the plant at the expense of local flora and biogeographical features. Flowers, fruits and seeds are the most commonly used parts. Fresh unripe apples and grapes are used for yeast, while ripe fruit is used for food. Chickpeas and hops (Cicer arietinum and Humulus lupulus), are of primary importance as a fermenting element for the study area, which is also used today.
Full-text available
Cosmeceuticals are cosmetic products containing biologically active ingredients purporting to offer a pharmaceutical therapeutic benefit. The active ingredients can be extracted and purified from natural sources (botanicals, herbal extracts, or animals) but can also be obtained biotechnologically by fermentation and cell cultures or by enzymatic synthesis and modification of natural compounds. A cosmeceutical ingredient should possess an attractive property such as anti-oxidant, anti-inflammatory, skin whitening, anti-aging, anti-wrinkling, or photoprotective activity, among others. During the past years, there has been an increased interest on the enzymatic synthesis of bioactive esters and glycosides based on (trans)esterification, (trans)glycosylation, or oxidation reactions. Natural bioactive compounds with exceptional theurapeutic properties and low toxicity may offer a new insight into the design and development of potent and beneficial cosmetics. This review gives an overview of the enzymatic modifications which are performed currently for the synthesis of products with attractive properties for the cosmeceutical industry.
Full-text available
The relationship between the structural characteristics of 29 flavonoids and their antiradical activity was studied. The obtained results suggest that the free radical scavenger potential of these polyphenolic compounds closely depends on the particular substitution pattern of free hydroxyl groups on the flavonoid skeleton. The possible mechanism of action of flavonoids lacking B ring OHs as free radical scavengers has been proposed.
Full-text available
In the present study, cytotoxic and anti-invasive properties of solvent fractions of kombucha tea were studied. Kombucha tea was fractionated with chloroform, ethyl acetate and butanol. Three solvent extracts and the final aqueous phase were concentrated and used for anti-cancer study at 25, 50, 75 and 100 μg/mL concentrations. Results revealed that ethyl acetate fraction at a concentration of 100 μg/mL caused cytotoxic effect on 786-O and U2OS cells; reduced the activities of matrix metalloproteinase-2 (MMP-2) and-9 in 786-O cells and MMP-2 activity in A549 cells; and significantly reduced the cell invasion and cell motility in A549, U2OS and 786-O cells. Thus, ethyl acetate fraction was further purified by chromatographic studies and presence of dimethyl 2-(2-hydroxy-2-methoxypropylidene) malonate and vitexin were confirmed through IR, NMR and mass spectroscopic studies, which might be responsible for the observed anticancer property of kombucha tea.
The aim of this study is to examine type I and III collagen content and distribution in skin within the contexts of patient age and injury, as well as to elucidate possible mechanisms of hypertrophic scar formation. Normal human skin and hypertrophic scar specimens were obtained from spontaneously aborted fetuses and burn patients of different ages (adolescent group, ≤18 years; adult group, >19 and ≤50 years; elderly group, >50 years). Total collagen content was indirectly determined by quantification of hydroxyproline. Levels of type I and III collagen as well as the ratio of type I/III were determined by immunohistochemistry and image analysis. Results obtained showed that the mean content of type I and III and type I/III ratio in normal skin differed significantly among age groups (P0.05), with the lowest levels of type I, III, and the highest ratio of type I/III observed in the elderly age group. Differences between normal uninjured skin and hypertrophic scar tissue were significant for all investigated parameters within any of the three age groups examined (adolescent, adult, elderly; P<0.05). Content of type I, III collagen and type I/III ratio also exhibited age-dependent differences during recovery in hypertrophic scar specimens. Thus, type III collagen synthesis decreases with age resulting in a skewed type I/III ratio and changes in skin tension, elasticity, and healing. Also, the content of type I, III collagen and type I/III ratio are significantly altered in hypertrophic scar tissue compared to uninjured age-matched controls, resulting in a different structural organization that is also influenced by patient age.
Skin is frequently exposed to a variety of environmental and chemical agents that accelerate ageing. External stress such as UV radiations (UVR) and environmental pollutants majorly deteriorate the skin morphology, by activating certain intrinsic factors such as Reactive Oxygen Species (ROS) which trigger the activation of Matrix Metalloproteinases (MMPs) and inflammatory responses hence damaging the extracellular matrix (ECM) components. To counter this, an exogenous supply of anti-oxidants, is required since the endogenous anti-oxidant system cannot alone suffice the need. Bio-prospecting of natural resources for anti-oxidants has hence been intensified. Immense research is being carried out to identify potential plants with potent anti-oxidant activity against skin ageing. This review summarizes the major factors responsible for premature skin ageing and the plants being targeted to lessen the impact of those. Copyright © 2014 Elsevier B.V. All rights reserved.
Older adults are more likely to have chronic wounds than younger people, and the effect of chronic wounds on quality of life is particularly profound in this population. Wound healing slows with age, but the basic biology underlying chronic wounds and the influence of age-associated changes on wound healing are poorly understood. Most studies have used in vitro approaches and various animal models, but observed changes translate poorly to human healing conditions. The effect of age and accompanying multimorbidity on the effectiveness of existing and emerging treatment approaches for chronic wounds is also unknown, and older adults tend to be excluded from randomized clinical trials. Poorly defined outcomes and variables; lack of standardization in data collection; and variations in the definition, measurement, and treatment of wounds also hamper clinical studies. The Association of Specialty Professors, in conjunction with the National Institute on Aging and the Wound Healing Society, held a workshop, summarized in this article, to explore the current state of knowledge and research challenges, engage investigators across disciplines, and identify research questions to guide future study of age-associated changes in chronic wound healing. © 2015 by the American Geriatrics Society and the Wound Healing Society.
Fermentation of sugared tea with a symbiotic culture of acetic acid bacteria and yeast (tea fungus) yields kombucha tea which is consumed worldwide for its refreshing and beneficial properties on human health. Important progress has been made in the past decade concerning research findings on kombucha tea and reports claiming that drinking kombucha can prevent various types of cancer and cardiovascular diseases, promote liver functions, and stimulate the immune system. Considering the widespread reports on kombucha, we recognized the need to review and update the research conducted in relation to kombucha tea, its products and tea fungus. Existing reports have suggested that the protective effects of kombucha tea are as good as those of black tea, however, more studies on kombucha tea and its composition are needed before final conclusions can be made.
In human skin fibroblasts in vitro, procollagen-1 and NAD+/NADH were reduced in three strains of adult fibroblasts compared with neonatal fibroblasts. The levels of both procollagen-1 and NAD+/NADH were increased in the adult fibroblasts by treatment for 24 (NAD energy) or 48 h (procollagen-1) with a complex containing niacinamide, Pal-KTTKS peptide and an olive oil fatty acid derivative (Olivem®), especially in combination with a natural extract from dill (Lys'lastine V®). In one of the adult fibroblast strains evaluated, these changes in procollagen-1 and NAD+/NADH in response to the complex of bioactives were in parallel with increased expression of mRNA biomarkers related primarily to dermal matrix and basement membrane structure, including COL1A1, COL3A1, COL5A1, COL14A1, ELN and LOXL2, in addition to SOD2, NAMPT and TGFBR3; MMP1 was decreased in expression. In general, these mRNA biomarker effects were maintained or boosted by the addition of Lys'lastine V, particularly at 1%, and were similar to the fold changes in mRNA expression in neonatal compared with adult fibroblasts. These results indicate that the complex of niacinamide, Pal-KTTKS and Olivem, especially with addition of Lys'lastine V, increases the NAD+/NADH bioenergy level of adult skin fibroblasts in parallel with increased expression of skin structure biomarkers in vitro to levels similar to those in younger fibroblasts. Thus, niacinamide, Pal-KTTKS, Olivem and Lys'lastine V are promising bioactive candidates for inclusion in cosmetic formulations.
Ever since eukaryotes subsumed the bacterial ancestor of mitochondria, the nuclear and mitochondrial genomes have had to closely coordinate their activities, as each encode different subunits of the oxidative phosphorylation (OXPHOS) system. Mitochondrial dysfunction is a hallmark of aging, but its causes are debated. We show that, during aging, there is a specific loss of mitochondrial, but not nuclear, encoded OXPHOS subunits. We trace the cause to an alternate PGC-1α/β-independent pathway of nuclear-mitochondrial communication that is induced by a decline in nuclear NAD(+) and the accumulation of HIF-1α under normoxic conditions, with parallels to Warburg reprogramming. Deleting SIRT1 accelerates this process, whereas raising NAD(+) levels in old mice restores mitochondrial function to that of a young mouse in a SIRT1-dependent manner. Thus, a pseudohypoxic state that disrupts PGC-1α/β-independent nuclear-mitochondrial communication contributes to the decline in mitochondrial function with age, a process that is apparently reversible.
Abstract Consumption of kombucha fermented tea (KT) has always been associated with different health benefits. Many personal experiences and testimonials of KT drinkers are available throughout the world on the ability of KT to protect against a vast number of metabolic and infectious diseases, but very little scientific evidence is available that validates the beneficial effects of KT. The aim of this review is to give an overview of the recent studies in search of experimental confirmation of the numerous KT health-promoting aspects cited previously. Analysis of the literature data is carried out in correspondence to the recent concepts of health protection's requirements. Attention is given to the active compounds in KT, responsible for the particular effect, and to the mechanisms of their actions. It is shown that KT can efficiently act in health prophylaxis and recovery due to four main properties: detoxification, antioxidation, energizing potencies, and promotion of depressed immunity. The recent experimental studies on the consumption of KT suggest that it is suitable for prevention against broad-spectrum metabolic and infective disorders. This makes KT attractive as a fermented functional beverage for health prophylaxis.