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In vitro study of RRS HA injectable mesotherapy/biorevitalization product on human skin fibroblasts and its clinical utilization

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Mesotherapy/biorevitalization with hyaluronic acid (HA) is a treatment approach currently used for skin rejuvenation. Various products with a wide range of polycomponent formulations are available on the market. Most of these formulations contain noncross-linked HA in combination with a biorevitalization cocktail, formed by various amounts of vitamins, minerals, amino acids, nucleotides, coenzymes, and antioxidants. Although ingredients are very similar among the different products, in vitro and clinical effects may vary substantially. There is a real need for better characterization of these products in terms of their action on human skin or in vitro skin models. In this study, we analyzed the effect of the RRS(®) (Repairs, Refills, Stimulates) HA injectable medical device on human skin fibroblasts in vitro. Skin fibroblast viability and its capacity to induce the production of key extracellular matrix were evaluated in the presence of different concentrations of RRS HA injectable. Viability was evaluated through colorimetric MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay, and key extracellular matrix genes, type I collagen and elastin, were quantified by quantitative polymerase chain reaction. Results demonstrated that RRS HA injectable could promote human skin fibroblast viability (+15%) and increase fibroblast gene expression of type I collagen and elastin by 9.7-fold and 14-fold in vitro, respectively. These results demonstrate that mesotherapy/biorevitalization products can, at least in vitro, effectively modulate human skin fibroblasts.
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ORIGINAL RESEARCH
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Open Access Full Text Article
http://dx.doi.org/10.2147/CCID.S95108
In vitro study of RRS HA injectable mesotherapy/
biorevitalization product on human skin
broblasts and its clinical utilization
Pierre-Antoine Deglesne*
Rodrigo Arroyo*
Evgeniya Ranneva
Philippe Deprez
Research and Development, SKIN
TECH PHARMA GROUP, Castelló
d’Empúries, Spain
*These authors contributed equally
to this work
Correspondence: Philippe Deprez
Research and Development, SKIN TECH
PHARMA GROUP, C/Pla de l’Estany 29,
17486 Castelló d’Empúries, Spain
Tel +34 6 6438 5219
Email drdeprez@clinicahera.es
Abstract: Mesotherapy/biorevitalization with hyaluronic acid (HA) is a treatment approach
currently used for skin rejuvenation. Various products with a wide range of polycomponent
formulations are available on the market. Most of these formulations contain noncross-linked
HA in combination with a biorevitalization cocktail, formed by various amounts of vitamins,
minerals, amino acids, nucleotides, coenzymes, and antioxidants. Although ingredients are very
similar among the different products, in vitro and clinical effects may vary substantially. There
is a real need for better characterization of these products in terms of their action on human
skin or in vitro skin models. In this study, we analyzed the effect of the RRS® (Repairs, Refills,
Stimulates) HA injectable medical device on human skin fibroblasts in vitro. Skin fibroblast
viability and its capacity to induce the production of key extracellular matrix were evaluated in
the presence of different concentrations of RRS HA injectable. Viability was evaluated through
colorimetric MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay, and
key extracellular matrix genes, type I collagen and elastin, were quantified by quantitative
polymerase chain reaction. Results demonstrated that RRS HA injectable could promote human
skin fibroblast viability (+15%) and increase fibroblast gene expression of type I collagen and
elastin by 9.7-fold and 14-fold in vitro, respectively. These results demonstrate that mesotherapy/
biorevitalization products can, at least in vitro, effectively modulate human skin fibroblasts.
Keywords: mesotherapy, medical device, RRS, collagen, elastin, extracellular matrix
Introduction
Mesotherapy is a minimally invasive technique that consists of local intradermal
therapy (LIT) with pharmaceuticals or other substances. These substances are given
in small quantities through multiple dermal punctures, where the injection site corre-
sponds to the area of the pathological condition.1 The term “meso” from mesotherapy
comes from the Greek word “mesos” referring to the mesoderm (or middle germ cell
layer) in the early embryo, which differentiates into tissues and structures, including
bone, cartilage, muscle, and connective tissue.2
Pharmacological products have been given intradermally for over a century, but it
was not until much later that this method became more widely accepted.1–3 A French
physician, Michel Pistor, reported encouraging results with small doses of drugs
administered intradermally to patients with a variety of clinical conditions.1,2 Apart
from the discovery of the method itself, one of the most relevant points is the fact that,
for instance, in 1988, the French Academy of Medicine confirmed1 that this method
is an integral part of classic medicine.
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Deglesne et al
Since the description of the technique, the use of LIT
increased slowly during the first 20 years (1971–1991) but
exponentially during the last 20 years.1 Clinical studies con-
cerning LIT procedures used to treat pain confirm the efficacy
and tolerability of anti-inflammatory/analgesic drugs admin-
istered using this technique.3 Nevertheless, the increased
number of LIT procedures nowadays appears to be related in
great part to skin rejuvenation purpose. Indeed, it is difficult
to evaluate the number of aesthetic physicians using LIT pro-
cedures, which may vary from 10,000 to 40,000 worldwide
depending on the source; in France alone, 122 physicians
are registered as mesotherapists, but many more medical
doctors are using the technique on a daily basis.4–7 Com-
pounds classically used in skin rejuvenation mesotherapy/
biorevitalization include noncross-linked hyaluronic acid
(HA) or organic silicium as the principal ingredient and
amino acids, vitamins, flavonoids, plant extracts, and others
as secondary ingredients.8,9 Although the procedure is widely
used by aesthetic physicians, there is a considerable lack of
scientific information related to the clinical efficiency of such
skin rejuvenation mesotherapeutic products.
In European countries, injectable mesotherapeutic prod-
ucts are classified as CE (European Conformity) class III
medical devices to ensure the safety of the final user and the
subsequent follow-up of the product once on the market. Most
of the companies proposing mesotherapy/biorevitalization
products for skin rejuvenation claim temporal filler effect due
to noncross-linked HA, and subsidiary claims may include
biorevitalization, dermal moisturizing, dermal regeneration,
extracellular matrix (ECM) strengthening, or collagen booster
effect. Nevertheless, very little in vitro data supporting these
subsidiary claims can be found in the scientific literature.
In this study, we present the in vitro results that were
generated by an independent, external, certified laboratory
regarding the effect of RRS® (Repairs, Refills, Stimulates)
HA injectable mesotherapy/biorevitalization product from the
company SKIN TECH PHARMA GROUP. The experiments
were carried out in order to determine the effect of RRS HA
injectable on human fibroblast viability and expression of
type I collagen, and elastin genes in vitro.
Materials and methods
MTT assay
Human skin fibroblasts (BJ cells) were seeded at 7,000 cells/
well in 24-well cell culture plates and allowed to grow in
Dulbecco’s Modified Eagle’s Medium supplemented with
10% fetal bovine serum (FBS) for 24 hours at 37°C with 5%
CO2. The cells underwent starvation in serum-free medium
for 6 hours before treatment. Next, the cells were exposed to
the tested sample at the specified concentrations (0.2 mg/mL
and 1 mg/mL) in fresh medium without FBS and incubated
for 24 hours or 48 hours. Cells treated with the medium
alone were used as negative control, and cells treated with
100 µg/mL of human insulin were used as positive control.
All dilutions were tested in triplicate. No ethics statement
was required from the institutional review board for the use
of these cell lines.
After cell treatment, cell survival was determined using
the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazo-
lium bromide) assay. Briefly, 300 µL of MTT solution was
added into each well and incubated for 3 hours at 37°C. The
medium was removed, the precipitated blue formazan dye was
dissolved in isopropanol or dimethyl sulfoxide (300 µL per
well), and the absorbance was quantified spectrophotometri-
cally at 550 nm. The plate was shaken on a gyratory plate
shaker to ensure that all the crystals dissolved and formed
a homogeneous solution. The absorbance was measured on
a microplate reader (Tecan modello Sunrise remote), with
background clearing. Results were expressed as follows:
% ellviability
[OD 550 of tested product
of the c
nm 690 nm)
=-(
OOD 550 of negativecontronm 690 nm)l]( -×100
RNA extraction, real-time and
quantitative polymerase chain reaction
Human skin-derived normal fibroblasts (American Type Cul-
ture Collection CRL-2522) were seeded at 10,000 cells/well
in 24-well cell culture plates and allowed to grow in minimum
essential medium supplemented with 10% FBS for 24 hours
at 37°C with 5% CO2. The cells underwent starvation in
serum-free medium for 6 hours before treatment. Next, the
cells were exposed to scalar dilutions of the tested products
(0.2 mg/mL and 1 mg/mL) in fresh serum-free medium. The
culture medium containing the sample was replaced daily.
Untreated cells were used as negative control; cells treated
with human insulin were used as positive control, and all
experiments were performed in triplicate.
After 24 hours and 48 hours of exposure, total RNA
was extracted using a guanidine thiocyanate-based reagent
according to the manufacturer’s instructions (TRIZOL). After
precipitation and centrifugation (30 min, 12,000 rpm, 4°C),
RNA was resuspended in 20 µL of sterile water and its concen-
tration was determined spectrophotometrically. Then, 300 ng
of total RNA was retrotranscribed into complementary DNA
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In vitro study of RRS HA injectable mesotherapy
using random primers at 37°C for 2 hours in a thermal cycler
following the manufacturer’s instructions (Thermo Fisher
Scientific, Waltham, MA, USA). Changes in gene expres-
sion profile were analyzed by “real-time polymerase chain
reaction” technology using a SYBR green-based chemistry.
Primer pair sequences used for type I collagen and elastin gene
amplification as well as the normalizer actin were designed
by Eurofins (Eurofins BioPharma Product Testing Spain SLU,
Barcelona, Spain). Calculation of the fold change was done in
a two-step calculation according to the following formulas:
1. CT = CT (target) - CT (normalizer)
2. Fold change = 2 - CT.
Statistical analysis
GraphPad software was used for graphical representation
of the data and statistical analysis. Unpaired t-test was
used to determine the P-value and statistical significance.
Normality testing was done using Anderson–Darling nor-
mality test. P-values #0.05 were considered as statistically
significant.
Bibliographic revision
and search algorithm
A search in PubMed database from The National Library
of Medicine (NLM) using specific MeSH headings was
done. MeSH term was retrieved for mesotherapy but nei-
ther for biorevitalization nor for bio-revitalization terms.
The search algorithm used was as follows: “Mesotherapy”
[Mesh], 47 abstracts retrieved. A simple PubMed search with
mesotherapy as a term allowed retrieving 186 abstracts that
contained all previously identified abstracts. Further search
in PubMed using biorevitalization (n=1), biorevitalization
(n=4), mesotherapic (n=2), mesoterapica (n=2), and meso-
therapy (n=2) allowed the identification of 12 more abstracts.
Finally, nine more abstracts were manually added. Then, the
pool of 216 abstracts was reviewed and analyzed, of which
85 abstracts were related to medical indications such as pain
treatment, 61 to adverse events occurring with mesotherapy
procedures, and 70 were directly related to the use of meso-
therapy for aesthetic purpose. Among the abstracts describing
the use of mesotherapy for aesthetic purpose, 29 described
procedures related to fat reduction, four related to alopecia,
20 related to skin rejuvenation, ten related to adverse events,
and ten related to other types of aesthetic procedures. In
addition, among the 29 abstracts related to skin rejuvena-
tion procedures, two in vitro studies, one animal model,
four expert opinion, and 13 clinical studies were identified
(Supplementary material).
Results
RRS HA injectable medical device,
indication, and clinical survey
RRS HA injectable is a resorbable medical device using a
long-chain (average 2 .8 million Da), medical-grade, noncross-
linked HA at a final concentration of 6.66 mg/mL for filling
superficial dermis together with a dermal biorevitalization.
This medical device is indicated for skin antiage regenera-
tion, tired atonic skin, and fine wrinkles. The HA employed
is from nonanimal origin. The biorevitalization solution is
composed of vitamins, fatty acids, coenzymes, antioxidants,
polyphenols, amino acids, and trace elements.
RRS HA injectable is normally used to treat face/neck and
décolletage using not only a microdermal papule injection
technique (needle 30G1/2 or 32G) but also, more recently,
blunt cannulas that allow reduction of entry points and
ecchymoses. Classically, 5 mL of the product is injected
per area per session. The total number of sessions may vary
from four to six with a frequency of one session per week.
Figure 1 presents one example of application of RRS HA
injectable on the face and neck.
Market survey (claim procedure according to ISO 13485)
did not register any claim for this product since 2013.
Face
Neck
Before treatment Following four sessions
Figure 1 Example of application of RRS® (Repairs, Rells, Stimulates) hyaluronic
acid injectable on the face and neck.
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Deglesne et al
RRS HA injectable promotes
human broblast viability
Dermal fibroblasts ensure the production of the ECM
components composing the dermal connective tissue
responsible for the skin’s tensile strength and mechanical
properties.10 Therefore, all the in vitro experiments were
carried out on primary human fibroblasts as they repre-
sent the main cellular population of human dermis skin
compartment.
We first evaluated the effect of RRS HA injectable on
human fibroblast viability using MTT assay on resting
synchronized serum-starved human fibroblasts. The assay
was done in the presence of two different concentrations
of the RRS HA injectable (0.2 mg/mL and 1 mg/mL) and
compared to two control conditions, one lacking RRS HA
injectable (unstimulated control) and the other activated with
100 µm/mL of human insulin (stimulated control). Two dif-
ferent time points were evaluated, at 24 hours and 48 hours,
posttreatment with RRS HA injectable.
The results demonstrated that RRS HA injectable was
not toxic to human fibroblasts in vitro at none of the doses
(0.2 mg/mL and 1 mg/mL) and at none of the time points
tested (24 hours and 48 hours). Moreover, we found that
RRS HA injectable supported cell viability of human fibro-
blasts up to 15.8%±3.9% as measured by the MTT assay at
48 hours in the presence of 1 mg/mL of RRS HA injectable
(Figure 2).
RRS HA injectable promotes the
synthesis of type I collagen and elastin
gene transcripts by human broblasts
The dermis is largely composed of dense collagen-rich ECM.
Dermal collagen represents by far the most abundant ECM
protein and constitutes the bulk of skin (90% dry weight).11,12
Among the five types of collagen present in the skin, type I
collagen represents 80% of the total content and is therefore
the main structural collagen type in the skin.13 Another 1 5% is
represented by type III collagen and was shown to participate
in skin healing.14 Moreover, a network of elastic fibers in the
ECM of these tissues gives them the required resilience to
recoil after stretching. The main component of elastic fibers
is the elastin molecule that creates a natural cross-linking to
adjacent elastin molecules.
Collagen dermis content is believed to represent dermis
thickness, while elastin content represents the elastic proper-
ties of the skin. Therefore, we evaluated the expression of type
I collagen and elastin genes on synchronized serum-starved
human fibroblasts in the presence or absence of different
concentrations of RRS HA injectable at two different time
points (24 hours and 48 hours). Gene expression was assessed
by quantitative polymerase chain reaction assay as described
in the “Materials and methods” section. As for MTT assay,
human fibroblasts were synchronized using serum starvation.
Two control conditions were used for subsequent comparison
with RRS HA injectable culture conditions, one negative
control (lacking RRS HA injectable) and one positive control
(culture in the presence of 50 µg/mL of human insulin).
Results were expressed as a fold increase in gene transcript
expression among the activated condition and control condi-
tion. RRS HA injectable was able to effectively increase the
expression of messenger RNA codifying for type I collagen
and elastin as early as 24 hours in the presence of 1 mg/mL
of the product (4.37±0.64 and 5.1±1.39, respectively). The
maximum increase of expression was observed at 48 hours
for both type I collagen (9.7±2.44) and elastin (14.3±2.55)
in the presence of 1 mg/mL of the product. Interestingly,
even low concentrations of RRS HA injectable (0.2 mg/mL)
were able to increase the expression of both type I collagen
and elastin (4.5±1.18 and 3.6±0.84, respectively) at 48 hours
posttreatment (Figure 3).
Discussion
Our results demonstrated that RRS HA injectable promotes
human skin fibroblast viability and enhances the expression
of type I collagen and elastin genes in vitro. The relevance
140
24 hours 127.5 126.0
115.8
102.0
100 100
Control RRS HA
--
103.3 102.4
100 µg/mL
hu-insulin
0.2 mg/mL 1 mg/mL
48 hours
130
% cell viability
120
110
100
Figure 2 Cell viability of human broblasts cultured in vitro in the presence of two
concentrations (0.2 mg/mL and 1 mg/mL) of RRS® HA injectable evaluated using
MTT assay at 24 hours (gray color) and 48 hours (black color) posttreatment.
Notes: As positive control, human broblasts were activated with 100 µg/mL of
human insulin. Results are expressed as the percentage of viability compared to
negative control.
Abbreviation: HA, hyaluronic acid; RRS®, Repairs, Rells, Stimulates.
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In vitro study of RRS HA injectable mesotherapy
of using in vitro studies to characterize the action of meso-
therapy/biorevitalization products, classified as CE class III
medical devices, is justified, as the product is directly injected
into the dermis compartment where it will be in contact with
dermal fibroblasts. Viability results from an in vitro experi-
ment on human fibroblasts with RRS HA injectable are in line
with preclinical tests that were performed with the product
for the delivery of the CE class III certificate. These tests,
described in Table 1, demonstrate the biocompatibility and
lack of toxicity of the RRS HA injectable medical device.
They are part of the CE class III medical device evaluation
and are done to ensure further patient safety and have to be
passed for the product to have a chance to be approved.
Histological and ultrastructural studies have revealed that
the major alterations in aged skin are encountered in the der-
mal ECM.15 In young skin (20–30 years), intact collagen fibrils
are abundant, tightly packed, and well organized, whereas in
aged skin (.80 years), the collagen fibrils are fragmented and
disorganized. Investigations have revealed that the character-
istic features of aging of the skin connective tissue are largely
due to aberrant collagen homeostasis.16 Two interrelated
mechanisms are involved: reduced collagen biosynthesis and
increased collagen fibril fragmentation. Moreover, degenera-
tion in the elastic fiber network has also been reported as a
characteristic feature of aging skin. Networks of elastic fibers,
which are composed of elastin molecules, in the ECM of these
tissues give them the required resilience to recoil after stretch-
ing and are thus responsible for skin elasticity. In this in vitro
study, we found that RRS HA injectable could promote a
nearly tenfold increase in type I collagen gene expression and
a 14-fold increase in elastin gene expression demonstrating
that in vitro RRS HA injectable promotes ECM production
of key proteins by acting on skin fibroblasts. Nevertheless,
the mechanisms leading to the induction of expression of
collagen or elastin synthesis by-products containing HA are
poorly understood. Studies using nonanimal cross-linked HA
suggest that HA could promote fibroblast tension, resulting
in their activation.17
Our bibliographic review on mesotherapy/biorevital-
ization identified ten clinical studies out of 13, reporting
beneficial effects on skin rejuvenation,18–26 which include
improvement of skin hydration, skin elasticity, skin
anisotropy, skin roughness, and skin complexion radiance.
A reduction in the number and depth of wrinkles was also
demonstrated. Moreover, skin biopsy after 3 months showed
a decrease in interleukin-1β, interleukin-6, and matrix metal-
loproteinase-1 (MMP1) and an increase in type I collagen and
elastic fibers.23 Nevertheless, two clinical studies evaluating
both skin properties and histological biopsies did not find any
significant benefit of mesotherapy/biorevitalization for skin
Figure 3 Effect on the expression of type I collagen and elastin genes.
Notes: Evaluation of type I collagen and elastin transcript expression of human broblasts cultured in vitro in the presence of two concentrations (0.2 mg/mL and 1 mg/mL)
of RRS® HA injectable evaluated using quantitative PCR at 24 (gray color) and 48 (black color) hours posttreatment. Results are expressed as a fold increase of the gene
transcript expression against the control condition (without treatment). As positive control, human broblasts were activated with 100 µg/mL of human insulin. (A) Collagen
type I transcript expression in the different culture conditions. (B) Elastin transcript expression in the different culture conditions.
Abbreviations: PCR, polymerase chain reaction; HA, hyaluronic acid; RRS®, Repairs, Rells, Stimulates.
Table 1 Preclinical tests performed with RRS HA injectable
Test Guideline
Tests for in vitro cytotoxicity on mammal broblast ISO 10993-5: 2009
Tests for systemic toxicity ISO 10993-11: 2006
Intracutaneous reactivity test ISO 10993-10: 2010
Tests for irritation and delayed-type hypersensitivity ISO 10993-10: 2010
Genotoxicity, carcinogenicity toxicity assays ISO 10993-3: 2014
Note: List of biocompatibility studies performed for the RRS® product on cell lines,
animals, and bacteria.
Abbreviations: RRS®, Repairs, Rells, Stimulates; HA, hyaluronic acid.
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Deglesne et al
rejuvenation.27,28 These discordant results may be explained
by the fact that formulations used in all those eleven studies
were all very different; some combining noncross-linked HA
with a biorevitalization solution, whereas others using only
HA or organic silicon. Moreover, biorevitalization solution
formulas may greatly vary between fabricants.
Diverse effects of mesotherapy/biorevitalization products
on skin rejuvenation have also been demonstrated in vitro by
Jäger et al.29 Indeed, they found that some products preserved
the viability of normal human skin fibroblasts while stimulating
type I collagen, MMP1, and tissue inhibitor of MMP1 produc-
tion, whereas other products led to apoptosis and/or necrosis
of human fibroblasts. More recently, these diverse effects of
mesotherapy/biorevitalization products on skin rejuvenation
were again emphasized by Avantaggiato et al.30 They found that
there was an association between the HA content of the products
and the expression of elastin and elastase genes. In that study,
higher HA concentrations were associated with lower elastin
gene activation but higher expression of elastase gene. However,
genes encoding for hyaluronan synthase 1, hyaluronidase 1,
and desmoplakin, although enhanced in culture in the pres-
ence of the different products, did not appear to be related to
the HA content of the different products. The negative effect
is eventually related to some of the other components of the
solution used for this study. They concluded that it is important
for the physician to choose which medical devices can be used
in relation to the effect expected.
Conclusion
In conclusion, we suggest that as the number of these products is
currently increasing, there is a real need to evaluate their safety
and efficiency. This study demonstrated that RRS HA inject-
able promotes human skin fibroblast viability and enhances the
expression of type I collagen and elastin genes in vitro.
Acknowledgments
All in vitro experiments and collection of data corresponding
to the experiments were performed by Eurofins BioPharma
Product Testing Spain SLU, an external independent certified
laboratory. This manuscript has been revised by Eurofins to
validate that all the described data conform to their results.
Patient pictures were kindly provided by Dr Jane Ranneva
after obtaining informed consent from the patients.
Author contributions
All authors made substantial contributions to conception
and design; took part in either drafting the article or revis-
ing it critically for important intellectual content; gave final
approval of the version to be published; and agree to be
accountable for all aspects of the work in ensuring that ques-
tions related to the accuracy or integrity of any part of the
work are appropriately investigated and resolved.
Disclosure
All four authors are working for SKIN TECH PHARMA
GROUP and elaborated the design of the experiments and
the redaction of the following manuscript. Nevertheless, all
in vitro experiments and collection of data corresponding
to the experiments were performed by Eurofins BioPharma
Product Testing Spain SLU, an external independent certi-
fied laboratory. This manuscript has been revised by Euro-
fins to validate that all the described data conform to their
results. The authors report no other conflicts of interest in
this work.
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12. Li Y, Lei D, Swindell WR, et al. Age-associated increase in skin fibro-
blast-derived prostaglandin E(2) contributes to reduced collagen levels
in elderly human skin. J Invest Dermatol. 2015;135(9):2181–2188.
13. Diegelmann RF. Collagen metabolism. Wounds. 2001;13:177–182.
14. Leung A, Crombleholme TM, Keswani SG. Fetal wound healing: impli-
cations for minimal scar formation. Curr Opin Pediatr. 2012;24(3):
371–378.
15. Quan T, Little E, Quan H, Qin Z, Voorhees JJ, Fisher GJ. Elevated matrix
metalloproteinases and collagen fragmentation in photodamaged human
skin: impact of altered extracellular matrix microenvironment on dermal
fibroblast function. J Invest Dermatol. 2013;133(5):1362–1366.
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16. Quan T, Qin Z, Robichaud P, Voorhees JJ, Fisher GJ. CCN1 contributes
to skin connective tissue aging by inducing age-associated secretory
phenotype in human skin dermal fibroblasts. J Cell Commun Signal.
2011;5(3):201–207.
17. Quan T, Wang F, Shao Y, et al. Enhancing structural support of the
dermal microenvironment activates fibroblasts, endothelial cells, and
keratinocytes in aged human skin in vivo. J Invest Dermatol. 2013;
133(3):658–667.
18. Herreros FO, Cintra ML, Adam RL, de Moraes AM, Metze K.
Remodeling of the human dermis after application of salicylate silanol.
Arch Dermatol Res. 2007;299(1):41–45.
19. Lacarrubba F, Tedeschi A, Nardone B, Micali G. Mesotherapy/
biorevitalization for skin rejuvenation: assessment of the subepidermal
low-echogenic band by ultrasound evaluation with cross-sectional
B-mode scanning. Dermatol Ther. 2008;21(Suppl 3):S1–S5.
20. Rozhanets AR, Turova EA, Kul’chitskaia DB. Combined application
of electric myo-stimulation and meso-therapy for the correction of
age-related changes of the facial skin. Vopr Kurortol Fizioter Lech Fiz
Kult. 2010;4:26–30.
21. Taieb M, Gay C, Sebban S, Secnazi P. Hyaluronic acid plus mannitol
treatment for improved skin hydration and elasticity. J Cosmet Dermatol.
2012;11(2):87–92.
22. Baspeyras M, Rouvrais C, Liégard L, et al. Clinical and biometrologi-
cal efficacy of a hyaluronic acid-based mesotherapy/biorevitalization
product: a randomised controlled study. Arch Dermatol Res. 2013;
305(8):673–682.
23. Savoia A, Landi S, Baldi A. A new minimally invasive mesotherapy/
biorevitalization technique for facial rejuvenation. Dermatol Ther
(Heidelb). 2013;3(1):83–93.
24. Liu S, Chen Z, Cai X, et al. Effects of dimethylaminoethanol and
compound amino acid on D-galactose induced skin aging model of rat.
Scientific World Journal. 2014;2014:507351.
25. Tedeschi A, Lacarrubba F, Micali G. Mesotherapy/biorevitalization with
an intradermal hyaluronic acid formulation for skin rejuvenation: an
intrapatient, placebo-controlled, long-term trial using high-frequency
ultrasound. Aesthetic Plast Surg. 2015;39:129–133.
26. Sparavigna A, Tenconi B, De Ponti I. Antiaging, photoprotective, and
brightening activity in biorevitalization: a new solution for aging skin.
Clin Cosmet Investig Dermatol. 2015;8:57–65.
27. Amin SP, Phelps RG, Goldberg DJ. Mesotherapy for facial skin reju-
venation: a clinical, histologic, and electron microscopic evaluation.
Dermatol Surg. 2006;32(12):1467–1472.
28. El-Domyati M, El-Ammawi TS, Moawad O, et al. Efficacy of
mesotherapy/biorevitalization in facial rejuvenation: a histological
and immunohistochemical evaluation. Int J Dermatol. 2012;51(8):
913–919.
29. Jäger C, Brenner C, Habicht J, Wallich R. Bioactive reagents used in
mesotherapy/biorevitalization for skin rejuvenation in vivo induce
diverse physiological processes in human skin fibroblasts in vitro-
a pilot study. Exp Dermatol. 2012;21(1):72–75.
30. Avantaggiato A, Girardi A, Palmieri A, Pascali M, Carinci F. Comparison
of bio-revitalizing injective products: a study on skin fibroblast cultures.
Rejuvenation Res. 2015;8(3):270–276.
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Deglesne et al
Table S1 Review details about the abstracts related to skin rejuvenation and mesotherapy
Authors PMID Year Study type Product tested Patient
number
Interventional/
observational
Efciency analysis Side effects Results
Amin et al117199654 2006 Open label
clinical trial
Multivitamin and hyaluronic acid (HA) solution 10 Interventional Photographs evaluation/histology/electron
microscopic analysis of collagen bers
measurements
No side effects Mesotherapy (MT) does not appear to provide any signicant
benet
Iorizzo et al218472058 2008 Expert opinion
Herreros
et al3
17333223 2007 Open label
clinical trial
0.1% salicylate silanol 7 Interventional The histological features of punch biopsies
of the treated area and the nontreated
contralateral arm were compared and the
collagen and elastic bers quantied
No side effects The application of silicon may stimulate the production of
collagen and elastic bers
Lacarrubba
et al4
19076625 2008 Open label
clinical trial
HA salts of biotechnological origin (1,000 kDa) 20 Interventional Ultrasound evaluation with cross-sectional
B-mode scanning
No side effects A statistically signicant (P,0.001) increase of SLEB
echogenicity (with a mean increase of pixel numbers equal to
31.3%) was observed in 15 of 19 subjects who completed the
study
Rozhanets
et al5
21086592 2010 Randomized
clinical trial
MT and electrostimulation (EMS) 60 Interventional Well-being-Activity-Mood test/Skin XP Pro
system and skin microcirculation by laser
Doppler owmetry (LDF)
No side effects The results of the study indicate that combined MT + EMS
therapy signicantly improves the state of facial skin, decreases
its pigmentation, reduces the number and depth of wrinkles,
enhances skin moisture, improves its elasticity, and decreases
porosity
Braccini et al621284223 2010 Expert opinion
Sturm et al721299793 2011 Review
Jäger et al822151394 2012 Experimental
in vitro
NCTF135® and NCTF135HA®, Soluvit® N and
Meso-BK
Collagen expression, cell proliferation, and
morphological changes using normal human
skin broblast cultures in vitro
Cell proliferation was comparable to control cultures in the
presence of hyaluronic acid, NCTF135® and NCTF135HA®;
moreover, a higher expression of collagen type-1,
matrix metalloproteinase-1 and tissue inhibitor of matrix
metalloproteinase-1 was noticed in the presence of those
products. In comparison, addition of Soluvit® N and Meso-BK
led to apoptosis and/or necrosis of human broblasts.
Taieb et al922672272 2012 Open label
clinical trial
13.5 mg/g uncross-linked HA +0.9% mannitol
(HA + mannitol)
34 Interventional Evaluation form/physician skin evaluation No side effects HA + mannitol is effective for skin hydration, anisotropy, and
roughness when treated using a depot technique but not with
picotage technique
El-Domyati
et al10
22788806 2012 Clinical Revitacare®, Bio-Revitalization 6 Interventional Photographs/skin biopsies No side effects The clinical evaluation of volunteers at baseline, end of
treatment, and 3 months posttreatment revealed no signicant
differences. Histological and immunostaining analysis of types I,
III, and VII collagen, newly synthesized collagen, total elastin,
and tropoelastin showed no statistically signicant changes
(P.0.05) after MT injection
Turova et al11 22994060 2012 Clinical Procaine 95 Interventional Biological age No side effects The study revealed the most signicant decrease of the
biological age (calculated as described by LM Belozerova)
in the patients treated with procaine electrophoresis (91%
improvement). The mesotherapeutic administration of procaine
also produced the benecial effect (59% improvement). Neither
procaine galvanization nor placebo MT caused any signicant
change of the variables characterizing the biological age
Baspeyras
et al12
PMC3778226 2013 Randomized
clinical trial
Glytone® 1 professional, Merz Pharmaceutical GmbH 55 Interventional Photographs/skin elasticity was measured
using a Cutometer® MPA580/dermis
thickness was measured by echography using a
high-frequency (20 MHz) ultrasound scanner
87.7% experienced one or several
adverse effects after injection.
Adverse events were generally of
mild or moderate intensity and
expected (hematoma, edema,
papule, erythema, or other
transient inammatory reactions):
46.8% occurred in the HA-treated
hemiface and 25.7% in the control
Dermis thickness signicantly increased after HA treatment
at 1 M (+3.4%, P=0.028) and 3 M (+4%, P=0.008) and after
control treatment at 1 M only (+2.5%, P=0.015). The HA ller
signicantly improved complexion radiance at 3 M compared
with the control (P=0.012) and for 51% of subjects, their
skin status. Nonreticulated HA-based MT signicantly and
sustainably improves skin elasticity and complexion radiance
(Continued)
Supplementary material
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49
In vitro study of RRS HA injectable mesotherapy
Table S1 Review details about the abstracts related to skin rejuvenation and mesotherapy
Authors PMID Year Study type Product tested Patient
number
Interventional/
observational
Efciency analysis Side effects Results
Amin et al117199654 2006 Open label
clinical trial
Multivitamin and hyaluronic acid (HA) solution 10 Interventional Photographs evaluation/histology/electron
microscopic analysis of collagen bers
measurements
No side effects Mesotherapy (MT) does not appear to provide any signicant
benet
Iorizzo et al218472058 2008 Expert opinion
Herreros
et al3
17333223 2007 Open label
clinical trial
0.1% salicylate silanol 7 Interventional The histological features of punch biopsies
of the treated area and the nontreated
contralateral arm were compared and the
collagen and elastic bers quantied
No side effects The application of silicon may stimulate the production of
collagen and elastic bers
Lacarrubba
et al4
19076625 2008 Open label
clinical trial
HA salts of biotechnological origin (1,000 kDa) 20 Interventional Ultrasound evaluation with cross-sectional
B-mode scanning
No side effects A statistically signicant (P,0.001) increase of SLEB
echogenicity (with a mean increase of pixel numbers equal to
31.3%) was observed in 15 of 19 subjects who completed the
study
Rozhanets
et al5
21086592 2010 Randomized
clinical trial
MT and electrostimulation (EMS) 60 Interventional Well-being-Activity-Mood test/Skin XP Pro
system and skin microcirculation by laser
Doppler owmetry (LDF)
No side effects The results of the study indicate that combined MT + EMS
therapy signicantly improves the state of facial skin, decreases
its pigmentation, reduces the number and depth of wrinkles,
enhances skin moisture, improves its elasticity, and decreases
porosity
Braccini et al621284223 2010 Expert opinion
Sturm et al721299793 2011 Review
Jäger et al822151394 2012 Experimental
in vitro
NCTF135® and NCTF135HA®, Soluvit® N and
Meso-BK
Collagen expression, cell proliferation, and
morphological changes using normal human
skin broblast cultures in vitro
Cell proliferation was comparable to control cultures in the
presence of hyaluronic acid, NCTF135® and NCTF135HA®;
moreover, a higher expression of collagen type-1,
matrix metalloproteinase-1 and tissue inhibitor of matrix
metalloproteinase-1 was noticed in the presence of those
products. In comparison, addition of Soluvit® N and Meso-BK
led to apoptosis and/or necrosis of human broblasts.
Taieb et al922672272 2012 Open label
clinical trial
13.5 mg/g uncross-linked HA +0.9% mannitol
(HA + mannitol)
34 Interventional Evaluation form/physician skin evaluation No side effects HA + mannitol is effective for skin hydration, anisotropy, and
roughness when treated using a depot technique but not with
picotage technique
El-Domyati
et al10
22788806 2012 Clinical Revitacare®, Bio-Revitalization 6 Interventional Photographs/skin biopsies No side effects The clinical evaluation of volunteers at baseline, end of
treatment, and 3 months posttreatment revealed no signicant
differences. Histological and immunostaining analysis of types I,
III, and VII collagen, newly synthesized collagen, total elastin,
and tropoelastin showed no statistically signicant changes
(P.0.05) after MT injection
Turova et al11 22994060 2012 Clinical Procaine 95 Interventional Biological age No side effects The study revealed the most signicant decrease of the
biological age (calculated as described by LM Belozerova)
in the patients treated with procaine electrophoresis (91%
improvement). The mesotherapeutic administration of procaine
also produced the benecial effect (59% improvement). Neither
procaine galvanization nor placebo MT caused any signicant
change of the variables characterizing the biological age
Baspeyras
et al12
PMC3778226 2013 Randomized
clinical trial
Glytone® 1 professional, Merz Pharmaceutical GmbH 55 Interventional Photographs/skin elasticity was measured
using a Cutometer® MPA580/dermis
thickness was measured by echography using a
high-frequency (20 MHz) ultrasound scanner
87.7% experienced one or several
adverse effects after injection.
Adverse events were generally of
mild or moderate intensity and
expected (hematoma, edema,
papule, erythema, or other
transient inammatory reactions):
46.8% occurred in the HA-treated
hemiface and 25.7% in the control
Dermis thickness signicantly increased after HA treatment
at 1 M (+3.4%, P=0.028) and 3 M (+4%, P=0.008) and after
control treatment at 1 M only (+2.5%, P=0.015). The HA ller
signicantly improved complexion radiance at 3 M compared
with the control (P=0.012) and for 51% of subjects, their
skin status. Nonreticulated HA-based MT signicantly and
sustainably improves skin elasticity and complexion radiance
(Continued)
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50
Deglesne et al
Table S1 (Continued)
Authors PMID Year Study type Product tested Patient
number
Interventional/
observational
Efciency analysis Side effects Results
Savoia et al13 PMC3680640 2013 Clinical Formulation A, tocopherol, arginine, sodium chloride,
adenine, inositol, sodium hyaluronate, ubiquinone,
lysine, cytosine, acetyl cysteine, water, cyanocobalamin,
isoleucine, glutamine, polysorbate 20, folic acid, leucine,
guanine, riboavin, threonine
Pyridoxine HCL, valine, thiamine HCL, histidine
hydrochloride, nicotinamide, tyrosine, calcium
pantothenate, phenylalanine, ascorbic acid, glutamic acid,
aspartic acid, asparagine, cysteine, proline, serine, alanine,
methionine, glycine, tryptophan
Formulation B, which was composed of HA, idebenone,
polysorbate 20, water, acetyl cysteine, sodium chloride
50 Interventional Punch biopsies were taken from randomly
selected participants, baseline, and after
6 weeks, and stained for interleukin (IL)-6,
IL-1β, MMP1, and collagen type I. Clinical
evaluation was based on the Global Aesthetic
Scale (GAIS) and on the Wrinkle Severity
Rating Scale (WSRS)
No side effects The results produced were statistically analyzed and resulted
in a signicant and long-lasting effect on facial rejuvenation.
Evaluation of photographs at 0, 1, and 2 months revealed
signicant clinical improvement: brightness, texture, and
rmness of the skin. The analysis of the GAIS and WSRS scores
in the two groups demonstrated statistically signicant results
after 2 months. The biopsies taken from randomly selected
participants at baseline and after 3 months showed a decrease
in IL-1β, IL-6, and MMP1 and an increase in collagen 1
Liu et al14 PMC4123624 2014 Animal model Intradermal microinjection of saline, 3.48% amino acids
(AA), 0.1% DMAE, 0.2% DMAE, 0.1% DMAE + AA, or
0.2% DMAE + AA
Expression of types I and II collagen and MMP1 was highly
upregulated in both 0.1% DMAE + AA and 0.2% DMAE +
AA groups compared with aging control. In contrast, TIMP-1
expression levels of various aging groups were signicantly
reduced when compared to sham control
El-Domyati
et al15
25514823 2015 Clinical Different noninvasive modality for the treatment of skin
aging, including radiofrequency, Nd: YAG 1320-nm laser
and Er: YAG 2940-nm laser mini-peels, intense pulsed
light (IPL), MT injection, and electrooptical synergy
36 Histology TGF-βHowever, no signicant differences (P.0.05) were observed
in TGF-β level in response to IPL or MT treatments in
comparison with baseline
Tedeschi
et al16
25539986 2014 Clinical, placebo
controlled
MT with HA 22 Interventional High-frequency ultrasound (22 MHz) was
performed to evaluate SLEB echogenicity
changes during treatment
No side effects Eighteen of 22 patients completed the study. At the end of
4 weeks, an ultrasound increase of dermal echogenicity was
observed in 13 subjects (seven of group A and six of group B),
which we considered as “responders.” In these patients, the
Student’s t-test showed a signicant increase from baseline of
SLEB pixel numbers of +24% (P,0.01) versus +6% with placebo.
In the same subjects, after an additional 4 months of monthly
injections, the mean increase was +18% (P,0.05) versus +4%
with placebo. In patients from group B who completed
10 months of treatment, the increase from baseline of SLEB
pixel numbers was +18% (P,0.05) versus 0% with placebo
Sparavigna
et al17
PMC4330006 2015 Randomized
clinical trial
Viscoderm®Skinkò E 64 Interventional The induced erythema was graded 24±4 hours
after irradiation, according to the COLIPA
reference visual score: 0= no erythema, 1/2=
perceptible redness reaction (MED), 1=
moderate erythema, 2= severe erythema.
Visual scores of every response to MED-testing
were then compared with the ones obtained
by irradiated skin previously injected with the
study product and irradiated skin previously
injected with a placebo (physiological solution
for injection). UVB irradiation corresponding
to 1 MED was performed 24 hours after the
study product/placebo microinjection, and
clinical evaluation of the induced erythema
was scored 24±4 hours after irradiation. The
photoprotective efcacy was expressed as a
percentage of erythema visual score variation vs
placebo.
Only one adverse event occurred
during the trial: on the day after
the second injection procedure,
one subject showed edema on
the lower eyelids, more marked
on the right side, which resolved
a few days after the application of
an anti-inammatory cream
Instrumental assessment showed, as early as after the second
biorevitalizing treatment, the antiaging efcacy of the tested
product; there was a clinical and statistically signicant
improvement of prolometric parameters, skin brightness,
pigmentation, and deep skin hydration. The study product
induced a statistically signicant decrease of the visual score of
the UVB-induced erythema compared with baseline, which was
statistically different from placebo
Prikhnenko18 PMC4396578 2015 Review
(Continued)
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51
In vitro study of RRS HA injectable mesotherapy
Table S1 (Continued)
Authors PMID Year Study type Product tested Patient
number
Interventional/
observational
Efciency analysis Side effects Results
Savoia et al13 PMC3680640 2013 Clinical Formulation A, tocopherol, arginine, sodium chloride,
adenine, inositol, sodium hyaluronate, ubiquinone,
lysine, cytosine, acetyl cysteine, water, cyanocobalamin,
isoleucine, glutamine, polysorbate 20, folic acid, leucine,
guanine, riboavin, threonine
Pyridoxine HCL, valine, thiamine HCL, histidine
hydrochloride, nicotinamide, tyrosine, calcium
pantothenate, phenylalanine, ascorbic acid, glutamic acid,
aspartic acid, asparagine, cysteine, proline, serine, alanine,
methionine, glycine, tryptophan
Formulation B, which was composed of HA, idebenone,
polysorbate 20, water, acetyl cysteine, sodium chloride
50 Interventional Punch biopsies were taken from randomly
selected participants, baseline, and after
6 weeks, and stained for interleukin (IL)-6,
IL-1β, MMP1, and collagen type I. Clinical
evaluation was based on the Global Aesthetic
Scale (GAIS) and on the Wrinkle Severity
Rating Scale (WSRS)
No side effects The results produced were statistically analyzed and resulted
in a signicant and long-lasting effect on facial rejuvenation.
Evaluation of photographs at 0, 1, and 2 months revealed
signicant clinical improvement: brightness, texture, and
rmness of the skin. The analysis of the GAIS and WSRS scores
in the two groups demonstrated statistically signicant results
after 2 months. The biopsies taken from randomly selected
participants at baseline and after 3 months showed a decrease
in IL-1β, IL-6, and MMP1 and an increase in collagen 1
Liu et al14 PMC4123624 2014 Animal model Intradermal microinjection of saline, 3.48% amino acids
(AA), 0.1% DMAE, 0.2% DMAE, 0.1% DMAE + AA, or
0.2% DMAE + AA
Expression of types I and II collagen and MMP1 was highly
upregulated in both 0.1% DMAE + AA and 0.2% DMAE +
AA groups compared with aging control. In contrast, TIMP-1
expression levels of various aging groups were signicantly
reduced when compared to sham control
El-Domyati
et al15
25514823 2015 Clinical Different noninvasive modality for the treatment of skin
aging, including radiofrequency, Nd: YAG 1320-nm laser
and Er: YAG 2940-nm laser mini-peels, intense pulsed
light (IPL), MT injection, and electrooptical synergy
36 Histology TGF-βHowever, no signicant differences (P.0.05) were observed
in TGF-β level in response to IPL or MT treatments in
comparison with baseline
Tedeschi
et al16
25539986 2014 Clinical, placebo
controlled
MT with HA 22 Interventional High-frequency ultrasound (22 MHz) was
performed to evaluate SLEB echogenicity
changes during treatment
No side effects Eighteen of 22 patients completed the study. At the end of
4 weeks, an ultrasound increase of dermal echogenicity was
observed in 13 subjects (seven of group A and six of group B),
which we considered as “responders.” In these patients, the
Student’s t-test showed a signicant increase from baseline of
SLEB pixel numbers of +24% (P,0.01) versus +6% with placebo.
In the same subjects, after an additional 4 months of monthly
injections, the mean increase was +18% (P,0.05) versus +4%
with placebo. In patients from group B who completed
10 months of treatment, the increase from baseline of SLEB
pixel numbers was +18% (P,0.05) versus 0% with placebo
Sparavigna
et al17
PMC4330006 2015 Randomized
clinical trial
Viscoderm®Skinkò E 64 Interventional The induced erythema was graded 24±4 hours
after irradiation, according to the COLIPA
reference visual score: 0= no erythema, 1/2=
perceptible redness reaction (MED), 1=
moderate erythema, 2= severe erythema.
Visual scores of every response to MED-testing
were then compared with the ones obtained
by irradiated skin previously injected with the
study product and irradiated skin previously
injected with a placebo (physiological solution
for injection). UVB irradiation corresponding
to 1 MED was performed 24 hours after the
study product/placebo microinjection, and
clinical evaluation of the induced erythema
was scored 24±4 hours after irradiation. The
photoprotective efcacy was expressed as a
percentage of erythema visual score variation vs
placebo.
Only one adverse event occurred
during the trial: on the day after
the second injection procedure,
one subject showed edema on
the lower eyelids, more marked
on the right side, which resolved
a few days after the application of
an anti-inammatory cream
Instrumental assessment showed, as early as after the second
biorevitalizing treatment, the antiaging efcacy of the tested
product; there was a clinical and statistically signicant
improvement of prolometric parameters, skin brightness,
pigmentation, and deep skin hydration. The study product
induced a statistically signicant decrease of the visual score of
the UVB-induced erythema compared with baseline, which was
statistically different from placebo
Prikhnenko18 PMC4396578 2015 Review
(Continued)
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52
Deglesne et al
Table S1 (Continued)
Authors PMID Year Study type Product tested Patient
number
Interventional/
observational
Efciency analysis Side effects Results
Avantaggiato
et al19
25640228 2015 Experimental
in vitro
Five different commercial medical devices containing
6.2 mg/mL, 10 mg/mL, 10 mg/mL, 13 mg/mL, and
20 mg/mL, respectively, of HA
RT-PCR and PCR HA concentration seems to be inversely correlated to elastin
gene activation. Regarding the neutrophil elastase gene, the two
medical devices with the higher concentration of HA displayed
the greater effect. Genes encoding for hyaluronan synthase 1,
hyaluronidase 1, and desmoplakin were enhanced, but the HA
content of the different products did not seem to be directly
related to gene activation. Therefore, the explanation for the
differences must be studied further with respect to elements
that are distinctive for each device
Rozhanets
et al5
21086592 2010 Open label
clinical trial
MT and EMS 60 Interventional Facial skin conditions were assessed using a
Skin XP Pro system and skin microcirculation
by LDF. The psychological status of the
patients was evaluated with the help of the
Well-being-Activity-Mood test
The results of the study indicate that combined MT + EMS
therapy signicantly improves the state of facial skin, decreases
its pigmentation, reduces the number and depth of wrinkles,
enhances skin moisture, improves its elasticity, and decreases
porosity
Abbreviations: SLEB, subepidermal low echogenicity band; TGF, transforming growth factor; PMID, PubMed identication; PMC, PubMed Central; IPL, intense pulsed light;
M, month; PCR, polymerase chain reaction; DMAE, dimethylethanolamine; TIMP-1, metallopeptidase inhibitor 1; RT-PCR, reverse transcription polymerase chain reaction;
LDF, laser doppler owmetry.
Total abstract
n=211
Medical purpose
(pain treatment and
others) n=80
Adverse events
n=71
n=60n=61 n=10
Aesthetic purpose
n=70
Fat reduction
n=26
Alopecia
n=4
Others
n=10
Skin rejuvenation
n=20
In vitro study
n=2
Animal model
n=1
Clinical studies
n=13
Expert opinion and
review
n=4
Figure S1 Analysis of the 211 selected abstracts.
Notes: Abstracts were individually revised for specic topics (medical use of the mesotherapy/biorevitalization, adverse events, aesthetic purpose, including fat reduction,
alopecia, and skin rejuvenation). Finally, abstracts on skin rejuvenation were separated into four groups, including in vitro studies, expert opinions/reviews, animal models,
and clinical studies. Original paper for each abstract within the skin rejuvenation group was retrieved and analyzed.
References
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53
In vitro study of RRS HA injectable mesotherapy
Table S1 (Continued)
Authors PMID Year Study type Product tested Patient
number
Interventional/
observational
Efciency analysis Side effects Results
Avantaggiato
et al19
25640228 2015 Experimental
in vitro
Five different commercial medical devices containing
6.2 mg/mL, 10 mg/mL, 10 mg/mL, 13 mg/mL, and
20 mg/mL, respectively, of HA
RT-PCR and PCR HA concentration seems to be inversely correlated to elastin
gene activation. Regarding the neutrophil elastase gene, the two
medical devices with the higher concentration of HA displayed
the greater effect. Genes encoding for hyaluronan synthase 1,
hyaluronidase 1, and desmoplakin were enhanced, but the HA
content of the different products did not seem to be directly
related to gene activation. Therefore, the explanation for the
differences must be studied further with respect to elements
that are distinctive for each device
Rozhanets
et al5
21086592 2010 Open label
clinical trial
MT and EMS 60 Interventional Facial skin conditions were assessed using a
Skin XP Pro system and skin microcirculation
by LDF. The psychological status of the
patients was evaluated with the help of the
Well-being-Activity-Mood test
The results of the study indicate that combined MT + EMS
therapy signicantly improves the state of facial skin, decreases
its pigmentation, reduces the number and depth of wrinkles,
enhances skin moisture, improves its elasticity, and decreases
porosity
Abbreviations: SLEB, subepidermal low echogenicity band; TGF, transforming growth factor; PMID, PubMed identication; PMC, PubMed Central; IPL, intense pulsed light;
M, month; PCR, polymerase chain reaction; DMAE, dimethylethanolamine; TIMP-1, metallopeptidase inhibitor 1; RT-PCR, reverse transcription polymerase chain reaction;
LDF, laser doppler owmetry.
5. Rozhanets AR, Turova EA, Kul’chitskaia DB. Combined application
of electric myo-stimulation and meso-therapy for the correction of
age-related changes of the facial skin. Vopr Kurortol Fizioter Lech Fiz
Kult. 2010;4:26–30.
6. Braccini F, Dohan Ehrenfest DM. [Advantages of combined therapies
in cosmetic medicine for the treatment of face aging: botulinum
toxin, fillers and mesotherapy]. Rev Laryngol Otol Rhinol (Bord).
2010;131(2):89–95. French.
7. Sturm LP, Cooter RD, Mutimer KL, Graham JC, Maddern GJ. A sys-
tematic review of dermal fillers for age-related lines and wrinkles. ANZ
J Surg. 2011;81(1–2):9–17.
8. Jäger C, Brenner C, Habicht J, Wallich R. Bioactive reagents used in
mesotherapy/biorevitalization for skin rejuvenation in vivo induce
diverse physiological processes in human skin fibroblasts in vitro-a
pilot study. Exp Dermatol. 2012;21(1):72–75.
9. Taieb M, Gay C, Sebban S, Secnazi P. Hyaluronic acid plus mannitol
treatment for improved skin hydration and elasticity. J Cosmet Dermatol.
2012;11(2):87–92.
10. El-Domyati M, El-Ammawi TS, Moawad O, et al. Efficacy of meso-
therapy/biorevitalization in facial rejuvenation: a histological and immu-
nohistochemical evaluation. Int J Dermatol. 2012;51(8):913–919.
11. Turova EA, Konchugova TV, Balaban EI, Fadeeva NI, Golovach AV,
Teniaeva EA. [The influence of procaine electrophoresis and meso-
therapy on the characteristics of biological age]. Vopr Kurortol Fizioter
Lech Fiz Kult. 2012;4:19–22. Russian.
12. Baspeyras M, Rouvrais C, Liégard L, et al. Clinical and biometro-
logical efficacy of a hyaluronic acid-based mesotherapy/biorevital-
ization product: a randomised controlled study. Arch Dermatol Res.
2013;305(8):673–682.
13. Savoia A, Landi S, Baldi A. A new minimally invasive mesotherapy/
biorevitalization technique for facial rejuvenation. Dermatol Ther
(Heidelb). 2013;3(1):83–93.
14. Liu S, Chen Z, Cai X, et al. Effects of dimethylaminoethanol and
compound amino acid on D-galactose induced skin aging model of rat.
Scientific World Journal. 2014;2014:507351.
15. El-Domyati M, El-Ammawi TS, Medhat W, Moawad O,
Mahoney MG, Uitto J. Expression of transforming growth factor-β
after different non-invasive facial rejuvenation modalities. Int J Der-
matol. 2015;54(4):396–404.
16. Tedeschi A, Lacarrubba F, Micali G. Mesotherapy/biorevitalization with
an intradermal hyaluronic acid formulation for skin rejuvenation: an
intrapatient, placebo-controlled, long-term trial using high-frequency
ultrasound. Aesthetic Plast Surg. 2015;39:129–133.
17. Sparavigna A, Tenconi B, De Ponti I. Antiaging, photoprotective, and
brightening activity in biorevitalization: a new solution for aging skin.
Clin Cosmet Investig Dermatol. 2015;8:57–65.
18. Prikhnenko S. Polycomponent mesotherapy/biorevitalization formula-
tions for the treatment of skin aging and improvement of skin quality.
Clin Cosmet Investig Dermatol. 2015;8:151–157.
19. Avantaggiato A, Girardi A, Palmieri A, Pascali M, Carinci F. Comparison
of bio-revitalizing injective products: a study on skin fibroblast cultures.
Rejuvenation Res. 2015;8(3):270–276.
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... In androgenetic alopecia, hair loss is heritable and androgen-dependent: the genetically predisposed hair follicles are the target for hair follicle miniaturization, leading to a replacement of pigmented hairs by barely visible, depigmented hairs in affected areas up to a decline in visible scalp hair density [3]. Mesotherapy/biorevitalization is a minimally invasive technique that consists of local intradermal therapy (LIT) with medical devices or other substances [4]. This approach is commonly used as a treatment for the rejuvenation of the skin but also of the hair follicle subjected to intrinsic, but also extrinsic aging. ...
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The assessment of the biological age is needed to estimate the level of health and the efficacy of measures being undertaken to slow down the ageing processes and prolong the active life of the patients. Earlier studies have demonstrated the improvement of both physical health and psychic status of the elderly patients following intravenous administration of procaine. The objective of the present work was to evaluate effect of various methods of administration of a 2% procaine solution on the biological age. A total of 95 subjects at the age between 30 and 75 years were enrolled in the study. The procaine solutions were applied to the collar region. The patients of group 1 were treated with 2% procaine electrophoresis, those in group 2 with procaine galvanization with the same parameters. Mesotherapy using the 2% procaine solution was given to the patients of group 3. It was substituted by placebo (0.9% physiological solution) in the patients of group 4 (controls). The study revealed the most significant decrease of the biological age (calculated as described by L. M. Belozerova) in the patients treated with procaine electrophoresis (91% improvement). The mesotherapeutic administration of procaine also produced the beneficial effect (59% improvement). Neither procaine galvanization nor placebo mesotherapy caused any significant change of the variables characterizing the biological age.
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Production of type I collagen declines during aging, leading to skin thinning and impaired function. Prostaglandin E2 (PGE2) is a pleiotropic lipid mediator that is synthesized from arachidonic acid by the sequential actions of cyclooxygenases (COX) and PGE synthases (PTGES). PGE2 inhibits collagen production by fibroblasts in vitro. We report that PTGES1 and COX2 progressively increase with aging in sun-protected human skin. PTGES1 and COX2 mRNA was increased 3.4-fold and 2.7-fold, respectively, in the dermis of elderly (>80 years) versus young (21-30 years) individuals. Fibroblasts were the major cell source of both enzymes. PGE2 levels were increased 70% in elderly skin. Fibroblasts in aged skin display reduced spreading due to collagen fibril fragmentation. To investigate the relationship between spreading and PGE2 synthesis, fibroblasts were cultured on micropost arrays or hydrogels of varying mechanical compliance. Reduced spreading/mechanical force resulted in increased expression of both PTGES1 and COX2 and elevated levels of PGE2. Inhibition of PGE2 synthesis by diclofenac enhanced collagen production in skin organ cultures. These data suggest that reduced spreading/mechanical force of fibroblasts in aged skin elevates PGE2 production, contributing to reduced collagen production. Inhibition of PGE2 production may be therapeutically beneficial for combating age-associated collagen deficit in human skin.Journal of Investigative Dermatology accepted article preview online, 23 April 2015. doi:10.1038/jid.2015.157.
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Human skin is largely composed of a collagen-rich connective tissue, which provides structural and functional support. The collagen-rich connective tissue is produced, organized, and maintained by dermal fibroblasts. During aging, dermal collagen fibrils undergo progressive loss and fragmentation, leading to thin and structurally weakened skin. Age-related alterations of collagen fibrils impairs skin structure and function and creates a tissue microenvironment that promotes age-related skin diseases, such as delayed wound healing and skin cancer development. This mini-review describes cellular mechanisms that give rise to self-perpetuating, collagen fibril fragmentation that creates an age-associated dermal microenvironment, which contributes to decline of human skin function. © 2015 S. Karger AG, Basel.
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Bio-revitalization is a commonly used technique in aesthetic medicine to improve skin quality and appearance by intra dermal injection of hyaluronic acid (HA) containing compounds. The present research compares different HA containing injectable as regard their effects on cultured skin fibroblasts over time ( 24, 48, 72 hours) by using RT-PCR and a panel of genes involved in dermal integrity. Human dermal fibroblasts were seeded on a layer of five different commercial medical devices containing 6.2, 10, 10bis, 13 and 20 % respectively of HA. The products differ not only in the HA concentrations but in the content and quality of other ingredients, moreover one of these products contains cross-linked HA. Differences among medical devices were found. In particular HA concentration seems to be inversely correlated to the Elastin genes activation. As regard to Neutrophil Elastase gene, the two medical devices with the higher concentration in HA displayed the greater effect. Genes encoding for the HA synthase 1, Hyaluronidase 1 and Desmoplakin are enhanced but the HA content of the different products seems to be not directly related to genic activation so the explanation for the differences must be researched in further elements that are distinctive of the specific device. For the physician it is important to choose among drugs or medical devices which ones to use and in what protocols. The present study performed a comparison that can be useful to better address the skin improvement therapies in aging and in its prevention.