XTRA LINE

EFFICACY
STUDIES
SIMILDIET LABORATORIOS
 IN VITRO ANALYSIS OF THE
EFFECTS OF XTRA FACE
ANTIAGING ON HYALURONIC
ACID RELEASE IN HUMAN
KERATINOCYTES

Dr. Carlos Aznar (MD)

Simildiet
Laboratorios
 Product tested
The tested product was received in Bionos on
22/02/2017 at room temperature, and labelled as
indicated:

P.0628: Xtra Face Antiaging

The compound was stored at room temperature in

Bionos Biotech (dependent entity and endorsed by The
Polytechnic University of Valencia), to avoid alteration
of the product.
Composition
• 13 Vitamins

• 24 Aminoacids

• 4 Coenzymes

• 4 Minerals

• 6 Nucleic acids

• 2 Antioxidants

• Sodium hyaluronate

• DMAE

• 8 Biomimetic peptides
 Goal

• To determine the effects of Xtra Face Antiaging

upon hyaluronic acid release in vitro after
treatment during 72 hours on human
keratinocytes (HaCaT).
 Methodology

HaCaT cells were treated during 72 hours with Xtra Face Antiaging at different
concentrations. After this incubation period, the effects on hyaluronic acid release
were evaluated. Protein content was measured in the supernatants of all samples
(treatment vs untreated control) using ELISA. One biological replicates (4 technical
replicates per condition) were performed.
 Result
s

In this study, we assessed the capacity of the product Xtra Face Antiaging to
increase the content of hyaluronic acid on human keratinocytes in vitro,
after 72 hours of treatment.
 Result
s

Graph 1. Graph representing hialuronic acid levels at 2 different concentrations of Xtra

Face Antiaging (10ug/ml and 1mg/ml).
 Result
s
• MTT essay was performed in parallel to confirm the increase of Hyaluronic acid
levels in cells was not due to an increase of cells in culture, in the amount of
hyaluronic acid are not due to changes in cell amount.

• Results showed that treatment with Xtra Face Antiaging at 10μg/ml and
1mg/ml, were not statistically significant in number of cells, compared with non
treated control.
 Result
s

Graph 2. Graph representing average cell viability by Absorbance at 550nm, after treatment at 2
different concentrations of Xtra Face Antiaging (10ug/ml and 1mg/ml) compared with non treated
control.
 Conclusion
Results showed that treatment during 72 hours with Xtra Face Antiaging at
10μg/ml, significantly increased hyaluronic acid levels by 95,6 ± 15,5% on human
keratinocytes; meanwhile the treatment at 1mg/ml increased hyaluronic acid by
50,4 ± 32,5%.

In conclusion, the treatment with Xtra Face Antiaging during 72 hours on human
keratinocytes in vitro significantly increased hyaluronic acid levels, compared to
the control without treatment.
ANTI-WRINKLES EFFICACY
ASSESSMENT OF THE
TREATMENT WITH XTRA
FACELIFT DURING 4 WEEKS
ON HUMAN VOLUNTEERS

Dr. Carlos Aznar (MD)

Simildiet
Laboratorios
 Product tested
Anti-wrinkles efficacy assessment of the treatment
with XTRA Facelift during 4 weeks on human
volunteers.

P.0631: Xtra Facelift

The compound was stored at room temperature in

Bionos Biotech (dependent entity and endorsed by
The Polytechnic University of Valencia), to avoid
alteration of the product.
Composition
• Sodium hyaluronate

• Hydrolysed collagen

• 3 Biomimetic peptides:

• Acetyl hexapeptide-8

• Acetyl hexapeptide-30

• Pentapeptide-18
 Material and methods

A capture technique of 3D hyperspectral image was used. It combines the

hyperspectral image technology and 3D digitalization in visible and infrared ranges.
 Procedur
Data register and treatment application

• Experimental area was defined as: forehead (glabellar lines) and periorbital lines, since
they are optimal areas to detect and quantify wrinkles, main indication of this product.

• Hyperspectral images were taken from volunteers at the start (day 0) and at the end
(day 28) to identify the presence and localization of wrinkles.
 Procedur
The analysis performed is a curvature analysis. Wrinkles correspond to the areas with
minimum curvature. The total area, the length and the depth of these areas is obtained
from the 3D hyperspectral images.

Figure 1. Images showing the 3D areas selected and assessed (left), curve analysis of
selected area (centre) and detected wrinkle area analysis (right).
 Use

• Volunteers self-applied the product in the designed area twice per day (morning and
night) by giving a linear massage following the path of the wrinkle after cleansing the
skin.

• A small reusable pipette was given to each of the volunteers to make easier the
application of the product and determine an approximate application of 1 ml/cm2.
 Pane
Inclusion criteria Exclusion criteria

• Presence of wrinkles (particularly at • The number of volunteers according to

forehead and crow’s feet). the client’s need was 20.

• Relative frequency in the use of skin care • 22 volunteers were included in the study.
cosmetic product.
• 2 volunteers did not complete the study,
• Age between 35 and 65 years. since they could not come to the second

visit.
 Study
• Do not apply other similar treatments from the tested one in the experimental area.

• Do not apply exfoliating treatments or clinical peelings in the experimental area from 15 days before the
start of the experiment.

• To respect use conditions of the tested treatment.

• Do not apply self-tanning products in the experimental area.

• Do not take drugs of dietetic supplements containing carotene.

• Conservation of the higiene and /or makeup habits.

• Do not receive any treatment based on Vit-A or its derivatives during study period (in case of necessity,
exclusion from the study).

• Do not use maekup in the experimental area (Face) during control days in the research centre.
• To avoid intense sun exposure (directly to the sun or in tanning studio), during study period.
 Parameter

Wrinkles Wrinkles Wrinkles

area length depth
 Efficacy

The results obtained before and after the treatment were analysed to assess the total area,
the depth and the length in every wrinkle the system recognised.

WRINKLES AREA

• Results showed that treatment with XTRA FACELIFT during 4 weeks significantly reduced
the total area of frontal and lateral wrinkles by 19.8 ± 12.3% and 20.5 ± 21.3%,
respectively.
 Efficacy

Figure 2. Representation of total wrinkle area.

 Efficacy

Figure 3. Representation of lateral wrinkle area.

 Efficacy

WRINKLES LENGTH

• Results showed that treatment with XTRA FACELIFT during 4 weeks significantly reduced
the length of frontal and lateral wrinkles by 15.4 ± 12.6 and 23.6 ± 21.4% respectively
comparing each volunteer with its own control at day 0 as show in following figures.
 Efficacy

Figure 4. Representation of frontal winkle length.

 Efficacy

Figure 5. Representation of lateral wrinkle length.

 Efficacy

WRINKLES DEPTH

Results showed that treatment with XTRA Facelift during 4 weeks significantly reduced the
depth of frontal and lateral wrinkles by 4.7 ± 1.3% and 4.1 ± 2.1%, respectively,
compared with each volunteer its control day 9, as shown in the following figures.
 Efficacy

Figure 6. Representation of frontal wrinkle depth.

 Efficacy

Figure 7. Representation of lateral wrinkle depth.

 Self assessment questionnaire
For a significant percentage of volunteers, it is considered that the treatment:

• Has an easy application.

• Has a pleasant texture.

• Has a nice colour.

• Is not oily for the skin .

 Conclusion

We assessed the anti-wrinkles efficacy of the product Xtra Facelift through

Hyperspectral 3D Technology and subjective evaluation by self assessment
questionnaire on 20 human volunteers, during 4 weeks of treatment with 2
applications per day (morning and night).
 Conclusion
• 4 weeks application of Xtra Facelift:

• significantly reduced the total area of frontal and lateral wrinkles by 19.8 ± 12.3% and 20.5 ± 21.3%.

• reduced the length of frontal and lateral wrinkles by 15.4 ± 12.6% and 23.6 ± 21.4% respectively.

• reduced the depth of frontal and lateral wrinkles by 4.7 ± 1.3% and 4.1 ± 2.1% respectively
compared with its control at day 0.

• In conclusión, in vivo topical treatment with the product XTRA FACELIFT during 28 days (twice application
per day) in 20 human volunteers significantly reduced the total area, the length and the depth of frontal
wrinkles.

• This means that the product has a clear effect on each volunteer, without taking into account randomness
between volunteer’s variation.
ANALYSIS OF THE EFFECTS OF
XTRA HAIR RESTORER ON CELL
VIABILITY UPON HUMAN
FOLLICLE DERMAL PAPILLA
CELLS

Dr. Carlos Aznar (MD)

 Simildiet
Laboratorios
 Product tested
The tested product was received in Bionos on
22/02/2017 at room temperature and it was
labelled as indicated:

P.0629: XTRA Hair Restorer

The compound was stored at room temperature

in Bionos Biotech, and dilutions were freshly
prepared each time.
Composition
• 12 Vitamins

• 24 Aminoacids

• 4 Coenzymes

• 4 Minerals

• 4 Nucleic acids

• 6 Antioxidants

• Sodium hyaluronate

• Troxerutin

• Ginkgo biloba extract

• Chamomilla recutita extract

• 5 Biomimetic peptides
Objective

• To determine the effects of Xtra Hair

Restorer on cell viability and proliferation
upon Human Follicle Dermal Papilla Cells.
 Methodolog

• Human Follicle Dermal Papilla Cells (HFDPC) were cultured in the presence of
different dilutions of the Xtra Hair Restorer during 24 hours. After treatment,
MTT assay was performed to assess cell viability, in order to determine
proliferation potential.
 Conclusion
Results showed that treatment with Xtra Hair Restorer at 1% concentration
during 24 hours, increase cell viability by 6.1 ± 3.9%, upon Human Follicle
Dermal Papilla Cells, although results are not statistically significant (p value =
0.1413).
 Figur

Graph 1. Proliferation.
 IN VITRO ANALYSIS OF THE
DEPIGMENTING EFFECTS OF XTRA
LIGHTENING PLUS THROUGH
MELANIN QUANTIFICATION IN
HUMAN MELANOCYTES

Dr. Carlos Aznar (MD)

Simildiet Laboratorios
 Tested products
Bionos Biotech (entity depending and supported by the
Polytechnic University of Valencia) received the product
to be analysed on 16/10/2019 at room temperature and
it was labelled as indicated:

P.1487: XTRA Lightening Plus

The compound was stored at room temperature in

Bionos Biotech until the beginning of the experiment
and dilutions were always freshly prepared.
Composition
• Tranexamic acid

• Glycyrrhiza glabra extract

• Ascorbic acid

• CromabrightTM

• Glutathione

• Decapeptide-15

• Oligopeptide-34

• Oligopeptide-50

• Oligopeptide-51
 Objective

• To asses the in vitro depigmenting effects of XTRA

Lightening Plus through melanin quantification
after treatment during 72 hours in Normal Human
Epidermal Melanocytes (NHEM).
 Methodolog
NHEM cells were cultured in different dilutions of XTRA Lightening Plus for 72 hours. After
treatment, MTT assay was performed to assess the effects on cell viability.

For melanin quantification, cells were cultured for 72 hours with XTRA Lightening Plus diluted at
0.1% and 1%.

During incubation period, cells were irradiated with UVA 8 times, 10-12 minutes for 3 days (total
dose = 6.95 J/cm2). After the last radiation, samples were processed for melanin quantification.

Melanin extraction: NaOH 1M at 60ºC for 1 hour. Samples were measured by spectrophotometry at
405 nm.
 Conclusion

Results show that in vitro treatment with XTRA Lightening Plus diluted at 0.1% and
1% for 72 hours in NHEM show:

• Significant reduction of melanin synthesis by 47.3 ± 1.7% and 51.5 ± 2.1%,

respectively.

• Significant inhibition of UVA-induced melanin synthesis by 91.5 ± 12.7% and

104.7 ± 15.3%, respectively.
Bar graph representing melanin levels after treatment during 72 hours with XTRA Lightening Plus at 0.1% and 1%
concentrations in Normal Human Epidermal Melanocytes submitted to UVA irradiation (6.95J/cm2), compared to Control
+ UVA. **** Represents statistical significance with p value < 0.0001.
Bar graph representing UVA-induced melanin levels after treatment during 72 hours with XTRA Lightening Plus at 0.1%
and 1% concentrations in Normal Human Epidermal Melanocytes submitted to UVA irradiation (6.95J/cm 2), compared to
Control + UVA. **** Represents statistical significance with p value < 0.0001.
 Conclusion

In vitro treatment with XTRA Lightening Plus for 72 hours in Normal Human
Epidermal Melanocytes shows depigmenting effects through significant inhibition
of UVA-induced melanin synthesis.
ANALYSIS OF THE EFFECTS
OF XTRA SKIN FIRMING
UPON COLLAGEN 1 AND
ELASTIN GENE EXPRESSION

Dr. Carlos Aznar (MD)

Simildiet
Laboratorios
 Products tested
Bionos Biotech (entity depending and
supported by the Polytechnic University of
Valencia) received the product to be analysed
on 03/31/2017 at room temperature and it
was labelled as indicated:

P.0630: XTRA Skin Firming

The component was stored at 4°C in Bionos

Biotech, to avoid alteration of the product.
Composition
• 12 Vitamins
• 24 Aminoacids
• 4 Coenzymes
• 4 Minerals
• 6 Nucleic acids
• 2 Antioxidants
• Sodium hyaluronate
• DMAE
• Hydrolysed collagen
• Methylsilanol mannuronate
• 4 Biomimetic peptides
Objective
• To determine the effects of the Xtra Skin
Firming product on collagen 1 and
Elastin gene expression, after topical
application on medaka (Oryzias latipes)
eleutheroembryos.
 Metodology
XTRA Skin Firming was applied topically on the medaka
eleutheroembryos during 5 minutes. After 2 hours (response time),
mRNA was purified and quantified and it was used to synthesize
complementary DNA (cDNA). This cDNA from treated and untreated
embryos (control) was used to determine the relative gene expression
of Col 1 and Eln through real-time quantitative PCR.
 Results
The results show that topical application with Xtra Skin Firming significantly
increased the expression of Col1 by 41.8% ± 13.5% as shown below.

Graph 1. Graph showing Col 1 increase.

 Conclusions

The results show that the topical application of Xtra Skin Firming fights
the effects of skin aging, significantly increasing the genetic
expression of Col 1 gene.
 ANÁLISIS IN VITRO DE LOS
EFECTOS DE XTRA SKIN
REPAIR EN LA CURACIÓN DE
HERIDAS EN
QUERATINOCITOS HUMANOS

Dr. Carlos Aznar (MD)

Simildiet
Laboratorios
 Productos testados
Bionos Biotech (entidad dependiente y avalada por la
Universidad Politécnica de Valencia) recibió el producto
testado el 22/02/2017 a temperatura ambiente y este se
etiquetó de la siguiente manera:

P.0627: XTRA Skin Repair

El componente se almacenó a 4ºC temperatura ambiente

en Bionos Biotech, para evitar la alteración del producto.
Composición
• Centella asiatica.

• Methylsilanol mannuronate.

• Troxerutin.

• Sodiun hyaluronate.

• Alaria esculenta.

• 4 Péptidos biomiméticos.
Objetivo

• Determinar los efectos del producto Xtra

Skin Repair en la curación de heridas en
24 horas en queratinocitos humanos
(HaCat) en cultivo.
 Metodología
Queratinocitos HaCat fueron insertados en dos pozos de cultivo. Se
plantaron 8.000 células durante la noche en DMEM enriquecido con suero
de feto de ternera. Se testaron distintas muestras a diferentes
concentraciones donde se añadieron a las HaCaT células suplementadas con
0,25 % de dicho suero.
 Resultados
En este estudio, evaluamos la capacidad del producto XTRA Skin Repair en la curación de
heridas en queratinocitos humanos in vitro, después de 24 horas de tratamiento.

Figura 1. Representación de la curación de heridas.

 Conclusiones
Los resultados revelan que el tratamiento durante 24 horas con XTRA Skin Repair
en una concentración del 0,01 % mejoró significativamente la curación de heridas
en un 21,6 ± 6,0 % sobre queratinocitos humanos.

En conclusión, el tratamiento con XTRA Skin Repair durante 24 horas en

queratinocitos humanos in vitro, mejoran significativamente la curación de
heridas, comparado con el control sin tratar.