To address this issue, Syntivia has validated an ex-vivo model close to sensitive skin. It will enable us to test ingredients and formulas under development for fragile skin.

A human skin model to target sensitive skin

We have chosen the skin explant as our favoured model. As the explant is obtained from surgically derived human skin, it is the most representative of the complexity of human skin.

We are working on a ex-vivo skin model with a barrier defect, dehydration and expressing the main markers of sensitive skin. The skin is first stripped and then exposed to a pro-inflammatory treatment, thus presenting a phenotype similar to that of sensitive skin. (figure 1).

Figure 1: Human skin explant after stripping and exposition to irritant molecules to mimic fragile skin phenotype.

Damage to the barrier function and loss of moisture are recurrent cutaneous phenomena in fragile or sensitive skin, and must be prevented.

 Impaired skin barrier:

After 7 days of treatment, we measure the thickness and quality of the reconstructed stratum corneum.
The stratum corneum gradually re-forms but shows an immature appearance or may show parakeratosis after 7 days.

In addition, main markers of keratinocyte differenciation in the stratum corneum after immunolabelling (figure 2):

• Filaggrin: crucial pour l’agrégation des filaments de kératine dans les cellules de l’épiderme pour la maturation
  
• Loricrin: major protein of the horny envelope
  
• Ceramides: essential lipids in the stratum corneum
• Involucrin (not shown): structural protein involved in the formation of the horny envelope by binding to cell membrane lipids.

The expression of these markers decrease in our model and can be reactivated.

Figure 2: Stratum corneum markers detected by immunofluorescence in healthy and artificially irritated skin after 7 days of culture.

Dehydration :

Transepidermal water loss (TEWL) significantly increases while glycerol skin content decreases (figure 3). Indeed, a low concentration of glycerol may lead to decrease the water content of both epidermal and dermal compartments. The structural changes to the skin that could result would lead to a change in cutaneous micro-relief and an increased TEWL.

Figure 3: Singinficant decrease in Transepidermal Water Loss (TEWL) (A) and Glycerol content (B) in healthy and artificially irritated skin during 7 days of culture.

Inflammation and Neuroinflammation mediators:

The model is characterized by the production of inflammation mediators associated with non-specific inflammation but also TRPV1 associated to neuro-inflammation (figure 4):

• Tumor Necrosis Factor α (TNFα)
• Prostaglandin E2 (PGE2) : particularly associated with sensitive skin.
• Transient Receptor Potential, Vanilloid Family 1 (TRPV1)

TRPV1 expression increases in the skin model and may contribute to activation of inflammation mediators. This ion receptor plays an important role in pain perception and thermoregulation. In people with sensitive skin, TRPV1 may be more reactive or expressed in greater quantities, leading to increased sensitivity to external stimuli.

Figure 5: Activation of the expression of inflammation mediators TNFα and PGE2 (A) and activation of the expression of TRPV1(B) in healthy and artificially irritated skin during 7 days of culture.

EVALUATION OF SOOTHING PRODUCTS

To conclude, this ex vivo human skin models are ideal to study the effect of soothing compounds and formula dedicated to sensitive skins. This will not only optimize the chances of success in clinical trials, but also identify the product’s biological pathways. Other readouts and markers can be analyzed on this model.

Don’t hesitate to contact us  for more information on our human skin models and product testing. You can also share your projects with us so that we can set up a study adapted to your needs.

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Human skin has excellent regenerative capacities that ageing or external stress can alter by ageing or external stress. In order to test ingredients or products capable of improving this tissue regeneration; it is necessary to develop adapted ex vivo human skin models. These skin models not only allow a better understanding of the biological phenomena involved, but also better prediction of the in vivo effects.

Alteration of the barrier function and loss of hydration are recurrent cutaneous phenomena in fragile skin. Therefore it is necessary to prevent these phenomena to preserve the health of the skin.

We offer two ex vivo skin models that show the different characteristic alterations of skin dryness:
– Alteration of the stratum corneum
– Decrease in the expression of essential stratum corneum markers
– Increased transepidermal water loss (TEWL)

Tissue regeneration in skin model after stripping

The skin explants were successively stripped until almost all the stratum corneum was removed. We then treated the skin models for 7 days with or without a formulated cosmetic active ingredient under development.

Skin barrier function impairment and recovery

Haematoxylin/eosin staining and specific labelling of Filaggrin in yellow and Loricrin in blue in untreated, striped skin, exposed or unexposed to a test product.

After 7 days, the untreated skin shows a very immature looking stratum corneum which contains a parakeratosis (incomplete maturation of epidermal keratinocytes, resulting in abnormal retention of nuclei in the stratum corneum).

In contrast, treatment with the test product allows a reconstruction of the upper layers of the epidermis and reactivation of the expression of essential markers of the stratum corneum.

Tissue regeneration in skin model after a detergent application

The skin explants were exposed to a detergent for several hours. We then treated the skin models for 2 or 3 days with or without a formulated cosmetic active ingredient under development.

Skin barrier function impairment and recovery

Haematoxylin/eosin staining and specific labelling of Loricrin in blue and Ceramids in orange in untreated or skin exposed to a detergent, treated or not with to a test product.

After 2 days, the untreated skin shows a strong deterioration of the stratum corneum and a marked decrease of its biomarkers.

On the other hand, skin that has received the test product shows a slight improvement in the reconstruction of the stratum corneum and in the expression of its essential biomarkers.

Increased dehydration

Measurement of the Trans epidermal water loss with Aquaflux on human skin explants

Detergent-exposed skin explants show a significant increase in TEWL. This effect may be reversed by a moisturizer or repairing products.

To conclude

These ex vivo human skin models are ideal to study the effect of topical treatments on:

– Tissue regeneration
– Barrier function recovery
– Moisturizing properties

Don’t hesitate to contact us  for more information on our human skin models and product testing. You can also share your projects with us so that we can set up a study adapted to your needs.

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From cell culture to skin explant, the research in cosmetic and dermatology research has a constant need for innovation.

The aim of pre-clinical studies is to identify the active ingredients and cosmetic products with an activity on the skin, to find their biological mode of action but also to optimise and anticipate results during clinical studies.

Our aim has been to develop the most suitable skin models to be able to demonstrate anti-ageing claims in order to identify new active compounds and formulas.

Two new models of ageing

From cell culture to skin explant, the research in cosmetic and dermatology research has a constant need for innovation.

Skin explants obtained from plastic surgery biopsies are exposed to proteases. These will specifically target the main collagen of the DEJ or Dermis, respectively Collagen IV and Collagen I. The skin explants are then kept alive to allow treatments in the presence of cosmetic or dermatological formulas.

ALTERED DEJ MODEL

Hematoxylin and eosin coloration and specific labelling of Collagen IV in untreated skin (left) and skin expose to protease (right)

A destructuring of the basal lamina is visible which leads to a weakening of the DEJ and a partial separation of the epidermis and dermis.

The marking of Collagen IV significantly decreases in the DEJ. The aim is to obtain a partial digestion which will allow us to quantify the reverse effect.

ALTERED DERMIS MODEL

Masson trichrome coloration and specific IHC labelling of Collagen I in untreated skin (left) and skin expose to protease (right)

The coloration and the specific labelling demonstrate a decrease of Collagen I density in the skin. We obtained a premature ageing that can be used to test new anti-ageing compounds.

To anticipate clinical studies

These new models of ageing allow better design of clinical studies and increase the chances of positive results.They also provide valuable clues about the biological functioning of the products tested in the skin in depth.

Several products under development have been tested on these models and allow the induced ageing to be reversed.

We are currently pursuing our research on these models. If you want additional information or if you want to test your products in development, please do not hesitate to contact us.

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EVIDENCE BASED COSMETICS

Dr Karl Lintner, independent consultant for the cosmetic industry (KAL’IDEES-Beauty Ideas), and Dr Claire Leduc, scientific business developer and marketing manager of Syntivia, will present their approach of the development of cosmetic active ingredients and formulas based on scientific evidence.

“A better definition of pre-clinical studies, through judicious choices of skin models and study conditions, will enable clinical tests to be anticipated more reliably in order to optimise the obtaining of convincing results”.

Two new ageing skin models

During their conference entitled From petri dish to pre-clinical for predicting clinical results, they will present two new ageing skin models. These models are based on human skin explants, explants that have been subjected to various alterations to mimic natural skin ageing. As different skin compartments are targeted, this also makes it possible to evaluate the activity of cosmetic active ingredients and products more precisely, to focus on a sigle aspect of ageing.

“It is not always easy to detect differences in activity on healthy tissue. We have chosen to target key proteins located in different compartments of the skin in order to unbalance the skin physiology.

The homeostasis of the tissue, thus altered, makes it easier to study a repairing effect of the active ingredients, formulas or cosmetic products.”

Come and discover how we have developed our models, and if you would like more information or would like to discuss with our research team to define your own skin models, please do not hesitate to contact us.

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A strong partnership

NeoVirTech is a biotechnology company developing autofluorescent viruses for imaging and antiviral discovery. The company has developed the ANCHOR^TM technology. It allows live cell imaging of virus infection, replication and propagation. The company has been highlighted by the PharmaTechOutlook magazine in the Top 10 drug discovery company in Europe in 2019.

Syntivia is specialized in the research & development of active ingredients or molecules in cosmetic and dermatology fields. We design and conducts preclinical evaluations to demonstrate the efficacy of active compounds.

A part of our expertise is based on our solid knowledge of skin physiology and our creativity in validating new study models.

The combination of our skills has enabled us to develop a model dedicated to the evaluation of products targeting viruses applied to the surface of the skin.

A proven study model

We have used living human skin samples. We artificially contaminated them with an autofluorescent virus. Following skin adsorption and treatment with antiviral product we can quantify the number of infectious particles. In parallel, the impact of the product applied is evaluated on the skin structure and viability. We provide here an unprecedented relationship between viral disinfection potential and impact on the skin.

Protocol of virus and product application: Evaluation of the antiviral effect and safety of the skin

This protocol gives a 24-hour answer on the effectiveness of your products on virus on the skin.

Quantification of infectious and replicating particles: Cells containing auto-fluorescent viral particles are quantified by automated microscopy (Thermo Scientific CellInsight CX7 high content screening microscope). More than 4000 cells are analysed by sample.

The hand sanitizer we tested reduced the percentage of infected cells down to 3.5%. Our results show that it is crucial not to dilute hydroalcoholic solutions in order to preserve their efficacy on viruses on the surface of the skin.

Structural analysis of the skin explants: after histological staining Masson trichrome (left) and immunohistochemistry Decorin in green, Elastin in red, Nuclei in blue (right).

Syntivia brings its expertise to analyse the effect of antiviral solutions on the human skin itself. Indeed, it is well known that the repeated daily use of sanitizers, de such as hydroalcoholic solutions, can deeply dehydrate the skin and create discomfort. Our histological and immunochemical analysis show that the hydroalcoholic solution does not have a deleterious effect on the skin, which structural integrity is preserved.

We are glad to be able to offer tailored studies to our clients for the evaluation of their products.  They are needed more than ever to ensure efficacy and user security. Do not hesitate to contact us for more information!

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Human skin model of accelerated aging

UV rays are the main cause of premature skin aging. UV radiations cause DNA damage, apoptosis and deleterious morphological changes to the skin characteristic of photoaging.

Experimental protocol to develop UVA exposed human skin model.

Human skin explants were chronically irradiated with UVA in order to mimic daily sunlight exposure. We used UVA for their ability to penetrate the skin deeply. Afterwards, effects on the epidermis were observed and quantified after histological staining and immunoflurescence. Skin morphology, as well as several markers involved in DNA repair and epidermal differentiation, were studied. Dermis deterioration was evaluated by transmission electronic microscopy and zymography in situ.

Epidermal disruption

The integrity of the epidermis is clearly affected by the UV treatment.

A disorganization of the layer is observed, characterized by a modification of cell cohesion and superposition. Moreover, the thickness of the epidermis decreased. The number of sunburn cells, that carry chromatin condensation within the nuclei, significantly increases.

Morphological modifications of skin after UVA irradiation and sunburn cells quantification

The histone H2AX becomes phosphorylated (ɣ-H2AX) in response to the formation of DNA breaks. On the irradiated skin, we detected a significant increase in positive ɣ-H2Ax stained nuclei representative of DNA damage.

DNA damage observed and quantified after ɣ-H2AX labelling

The alteration of the intermediate keratinocyte differentiation state as well as the appearance of apoptotic cells confirm the deleterious and pro-aging effect on the epidermis.

Dermis disorganization

UV exposition causes a desassembly of the dermal fibers and a disruption of their organization.

The papillary and reticular dermis is formally altered by the treatment. The damage is characterized by a fragmentation of the fibers and a decrease in their density. Observed by transmission electronic microscopy, fibers are dispersed and noncohesive.

Extracellular matrix strucure observed by Transmission Electronic Microscopy (TEM)

Activation of proteases is responsible for the alteration of dermal fibers. UVA rays penetrating into the dermis are involved in this process of activation.

In irradiated skin, we have detected a strong increase in protease activity in both the epidermis and dermis. It was detected by in situ zymography.

In situ zymography of global skin proteases carried out on a cryosection of human skin explants

Evaluation of protective or repairing effects

Our human skin model is representative of premature aging and photoaging, characterized by epidermis alteration and accumulation of dermis malfunctions.

We have shown a strong alteration of the epidermis. The cells of the upper layers lose their cohesion, DNA lesions appear while keratinocyte differentiation is affected. We also observe disorganized dermis through the elastosis and the degradation of collagen I by matrix metalloproteases.

The life span of the skin explant is between 7 and 10 days. It allows skin markers to be measured only in the short term. Tested ingredient or product can be applied before, during or after UV exposition. The method of application chosen makes it possible to support a protective and/or repairing effect.

Don’t hesitate to contact us  for more information on our other human skin models and product testing. You can share your projects with us so that we can set up a study adapted to your needs.

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How we developed this new cosmetic antipollution ingredient

Syntivia covers the entire process of cosmetic ingredient developmen; from the design and conception of an active compound over the demonstration of biological activity to clinical evaluation. Our experts guide you through the entire process. The story of VITALISIS® perfectly illustrates our way of working. Thus, to develop this new antipollution ingredient, we first identified unexplored active ingredients using genomic screening. On the other hand, we validated its metabolic pathways by screening protein expression. Moreover, targeted tests helped us increase the ingredient’s action range, after which we studied use on human skin using skin biomarkers and structural analysis. All those tests allowed us to set up the ingredient’s clinical evaluation.

Steps of cosmetic active ingredient development

The importance of thorough biological analysis

To reveal the significance of the effects VITALISIS® has on human skin and demonstrate its preventative and curative action; we used a varied selection of human skin models, i.e. cells, explants and in-vivo models. These models were submitted to a wide range of biological evaluation techniques. Indeed, VITALISIS® was tested for gene / protein expression and for effects on skin appearance. In order to reproduce the dermatological effects of an urban environment, the models were also exposed to a range of stress factors : UV, heavy metals, cigarette smoke and urban pollution.

Illustrating the biological activity of VITALISIS ®

PROTECTION FROM OXYDATIVE STRESS

The detoxifying propreties of VITALISIS®  were tested on several skin models:
– Keratinocytes exposed to UVA by quantifying H₂O₂
– Keratinocytes exposed to UVA by quantifying H₂O₂
– Volunteers who lead ab urban life by quantifying carbonylated proteins

SOOTHING EFFECT

We tested the soothing properties of VITALISIS® on several skin models:
– Keratinocytes and dermal fibroblasts by quantifying genes expression of inflammation mediators
– Keratinocytes exposed to IL1β by quantifying IL8 production
– Volunteers who lead an urban life by assessing redness

DNA PROTECTION

The protective effect of VITALISIS® on DNA was demonstrated:
– On keratinocytes and fibroblasts exposed to an extract of cigarette smoke by quantifying DNA damage and a DNA repair protein.
– On skin explants exposed to UVA rays by quantifying DNA damage

PREVENTION FROM SENESCENCE

VITALISIS® also maintains cell longevity, which was demonstrated on several skin models
– Keratinocytes and fibroblasts exposed to cigarette smoke extract and heavy metal by quantifying cell viability
– Skin explants exposed to UVA rays by studying the expression of a senescence biomarker
– Volunteers under urban life by assessing skin texture

PRESERVATION OF DERMAL FIBERS

We demonstrated the protective effects VITALISIS® has on dermal fibers, using:
– skin explants that were exposed to UVA by quantifying elastic fibers and studying protease expression.

Clinical evaluation to demonstrate real-life efficacy of this new antipollution ingredient

At last, the solid results from the biological analysis helped the Syntivia team design and monitor the clinical study. The study included 42 women (21 VITALISIS® / 21 placebo), aged between 30 and 50, with an urban lifestyle and a dull complexion. These volunteers used the products twice a day after facial cleansing. As a result, after one month, volunteers were unanimous: their skin felt detoxified and purified with an added feeling of freshness.

The Syntivia team combined expertise in cosmetic ingredient development and high-level creativity to obtain an efficient antipollution ingredient. It perfectly meets the needs of urban populations. The cosmetic active ingredient was launched by SOLLICE BIOTECH at InCosmetic 2018 in Amsterdam.

More information on VITALISIS® is available on the Sollice Biotech website. To get in touch with the Syntivia team and discuss active ingredient development or study design, don’t hesitate to contact us.

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Syntivia uses a skin cell model to carry out solid cosmetic tests. We developed a blue light irradiation dose corresponding to 1h to 2h40 of screen exposure. It allows us to test and validate cosmetic product’s efficacy against blue light-induced skin damages.

Cellular model: primary cells in culture

Proper conditions allow cells in culture to live, grow and maintain their functions as if they were within the skin. Syntivia uses primary cells, Normal Human Epidermal Keratinocytes (NHEK) in 2D culture. Cells are isolated from skin epidermis ans kept growing under adequate conditions to reproduce the human skin configuration.
Our equipment allows us to obtain reliable results on this model by analyzing:

• Cell viability
• Biomarkers involved in skin water balance
• DNA damages signaling
• Senescence

Normal Human Epidermal Keratinocytes in 2D culture.

Effects of blue light on skin cells

We developed two conditions of blue light exposure with increasing doses: single or chronic irradiations. Based on this model and these conditions, we carried out various tests. Both models are of interest:

• Single irradiation rapidly generates lesions. This condition is interesting to test products that immediatly protect the skin.
• Chronic irradiation mimics closely the reality of skin exposed to blue light on a daily basis. The effects induced by this condition are more important than those of a single irradiation. This condition allows to test products that will have a long-term effect.

The various parameters analyzed allow to measure the increasing protection power of cosmetic active ingredients.

Effect of blue light on cell viability

A viability test was carried out on NHEK exposed in a single or chronic way to blue light. This allows to identify the cytotoxic dose in both conditions.

Viability test after a single exposure (left) and after chonic exposure (right) to blue light irradiation. Increasing irradiation doses were tested.

The cell viability is inversely proportional to the dose of blue light applied.

Effect of blue light on DNA in skin cells

Epidermal cells were exposed to various doses of blue light and γ-H2AX expression, a marker of DNA damage [1], was detected by immunofluorescence  and quantified.

γ-H2AX immunofluorescence detection by fluorescence microscopy on unirradiated NHEK (left) and irradiated NHEK (right). Cell nucleus are labelled in blue and γ-H2AX in red.

γ-H2AX labelling quantification after a single irradiation (left) and after chronic irradiations (right). Increading irradiation doses were tested.

Cells irradiated with increasing doses of blue light (once or three times) clearly show an increase of γ-H2AX expression, proof of DNA damages.

Effect of blue light on skin water balance

Epidermal cells were irradiated with blue light at various doses and Aquaporin 3 expression, hydration marker [2], was detected by immunofluorescence  and quantified.

Aquaporin 3 immunofluorescence detection by fluorescence microscopy on unirradiated NHEK (left) and irradiated NHEK (right). Cell nucleus are labelled in blue and Aquaporin 3 in red.

Aquaporine 3 quantification after a single irradiation (left) and after chronic irradiations (right). Increasing irradiation doses were tested.

Exposure to blue light reduces the expression of Aquaporin 3, demonstrating the impact of this stress on the skin’s water balance.

Effect of blue light on cell senescence

After exposure to blue light, Lipofuscin, a marker of cellular senescence [3], was labelled with Sudan Black B coloration and observed with an optical microscope with transmitted light coupled to a colour camera.

Lipofuscin staining and quantification on unirradiated NHEK and after chronic irradiations with the highest blue light dose (right). Cell nucleus are in pink and lipofuscin in blue.

Keratinocytes chronically exposed to blue light become senescent, as shown by the lipofuscin staining. Cell senescence induces skin’s premature aging.

NB: A same activation of senescence was measured on Normal Human Dermal Fibroblasts (NHDF) exposed to blue light.

Our cell model is hightly sensitive to blue light and perfectly adapted to evaluate cosmetic active ingredients

Blue light irradiation results in loss of viability, emergence of DNA damages,  unbalanced skin moisture, and senescent cells. These effects can be countered to precisely define your product’s cosmetic claims regarding protection against blue light damages.

What’s next?

Studies are currently running to validate ex vivo models to carry out blue light cosmetic tests on skin explants. Do not hesitate to share your projects with us so that we can set up a study adapted to your needs.

Sources :

[1] Oh et al. UV-induced histone H2AX phosphorylation and DNA damage related proteins accumulate and persist in nucleotide excision repair-deficient XP-B cells. DNA Repair (Amst). 2011 January 2. 10(1): 5–15. doi:10.1016/j.dnarep.2010.09.004.

[2] Avola et al. Blue light induces down regulation of aquaporin 1, 3 and 9 in human keratinocytes. Cells 2018, 7, 197. 

[3] Georgakopoulou et al. Specific lipofuscin staining as a novel biomarker to detect replicative and stress-induced senescence. A method applicable in cryo-preserved and arhival tissues. AGING, January 2013, Vol. 5 No 1.

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“Syntivia is a fast-growing company, most of the data is stored in Excel but we are reaching the limits of this solution. After 9 years of research our history is significant and the team is growing, which makes file sharing complicated and capitalization difficult. INEON came to propose us to co-develop a solution that meets these needs, knowing that with INEON Free we already have a solution to manage part of our work. »
Philippe Bedos, PhD, Managing Director of Syntivia.

“In line with our strategy, we wanted to benefit from the very specific know-how of Syntivia’s team to develop an offer for cosmetic laboratories. In this sector as in Life Science, the players have designed solutions that are very expensive and too complete to be easy to use. »
Christophe Pierrat, Chief Operating Officer of INEON Biotech.

About Syntivia
SYNTIVIA is a Toulouse-based company created in 2010 and located on the Oncopole site, at the heart of life science research. SYNTIVIA uses the latest scientific advances to develop innovative and effective cosmetic active ingredients. The company has extensive expertise in cosmetic biology, high-tech equipment and in-depth knowledge of the market. SYNTIVIA uses its expertise to meet the latest cosmetic trends and regulatory requirements in this sector. It develops products that meet the market’s needs in terms of innovation, efficiency and safety.

About INEON Biotech
INEON Biotech S.A.S was created in December 2017 thanks to the association of private investors and BNP Paribas Développement. As a start-up in software publishing, it offers laboratories innovative solutions in SAAS mode. Resolutely intuitive, these solutions allow to manage stocks of chemical compounds or reagents as well as biological molecules such as plasmids and proteins. Ineon S2 follows the analysis of the results, including those of screening projects for drug development. Other modules make it possible to ensure the safety of their handling, to monitor the budgetary impact of each project and soon the management of plants and plant extracts.

Don’t hesitate to contact us for more information on this collaboration.

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Skin melanocytes synthetize a pigment, the melanin, which is transferred via melanocytes dendrites to keratinocytes. Melanin transfer results in skin pigmentation darkening of the skin, but also creating age spots.

Although melanin plays an important role in photoprotection against UV-induced DNA damages, melanin production can be altered by external stresses (sun exposure, pollution, …) and internal factors (hormones, inflammation, aging). Such malfunction can lead to pigmentation defect that has to be treated with adequate skin care products. Improvement of the skin color homogeneity can be provided by cosmetic products and need to be demonstrated by solid research studies.

Syntivia as an expert company in the research and development of cosmetic active ingredients is developping various models to study both activation and down regulation of skin pigmentation.

Skin models

Primary cells in culture

Proper conditions allow the cells in culture to live, grow and maintain their functions as if they were within the skin.

Syntivia uses primary cells, normal human melanocytes in 2D culture and in co-culture with normal human keratinocytes. Cells are isolated from skin of different phototypes to reproduce the human skin configuration.

Our equipment allows us to obtain reliable results on this model: gene/protein expression, melanin quantification, melanin transfer and melanocyte dendricity.

Normal Human Epidermal Melanocytes and Normal Human Epidermal Keratinocytes in 2D culture.

Epidermal microtissues

The epidermal microtissue is a mixed cellular model composed of the two main cell types of the epidermis: melanocytes and keratinocytes. This model is created from human primary cells grown as spheroids; we can choose the age and phototype of the donor. This model is produced and cultivated in 96-well plates, making it suitable for screening. This is an epidermis model that can be adapted to specific issues: pigmentation, inflammation, toxicity.

Epidermis microtissue composed of NHEM and NHEK.

Skin explants

Syntivia uses a skin explant model obtained by plastic surgery. The skin explant is kept alive for 7 days to allow ex vivo pigmentation studies to predict in vivo skin response.

Skin explant

Skin pigmentation studies

Study of pigmentation biological pathways

We use genomic chips on genes involved in each of the pigmentation processes. We can work on all our skin models. We use the Fluidigm’s Biomark^TM technology, this high-tech tool based on microfluidic technology allows to obtain:

• A large amount of data
• Flexible conditions
• Precise quantification of gene expression

The system’s technoloy is robust and yields reliable statistics on a wide range of targeted genes.

Cosmetogenomic chips have been especially designed to test active ingredients or cosmetic formulas.

Cosmetogenomic chip results: activation and inhibition of gene expression on skin explant.

Melanin synthesis: induction and inhibition of melanin production

We are able to reliably quantify melanin synthesis in human skin cells. It allows to identify compounds capable of activating or inhibiting melanin neosynthesis.
Models used: primary cells and epidermis microtissue.

Melanin content assay. Melanin production can be quantified after treatment with active compounds by spectroscopy.

Melanin transfer: Imaging and quantification

Melanin is transmitted from melanocytes to keratinocytes by cellular vesicles called melanosomes. Using fluorescence microscopy and flow cytometry, we can visualize and quantify the transfer of melanosomes from one cell type to another in 2D or 3D models.
Models used: primary cells in co-culture and epidermis microtissue.

Left: Melanosome transfer from melanocytes to keratinocytes visualized by fluorescent microscope in 2D after specific labelling (co-culture of melanocytes and keratinocytes). Right: Melanin transfer from melanocytes to keratinocytes can be reliably quantified by flow cytometry after specific labelling (dissociated microtissue).

Melanocyte dendricity: Imaging and quantification

Melanin transfer is directly connected to melanocyte dendricity. The melanosomes containing melanin are transported by a network of microtubules to the dendritic ends of melanocytes. In order to ensure the transfer of melanosomes to the cytoplasm of keratinocytes, melanocytes dendrites creep in between neighbouring keratinocytes. We can visualize and quantify melanocyte dendricity.

Co-culture of melanocytes and keratinocytes activated for melanin transfer, acquired with a confocal microscope. The Imaris software was used to proceed to the surfacing of melanocytes and 3D imaging.

The cell morphology can be visualized in 3D on epidermis microtissue by SPIM (Selective Plan Illumination Microscopy). The cells are labelled prior to spheroid formation with melanocytes in green and keratinocytes in red.

Observation of melanocyte dendricity in epidermal spheroid by 3D surfacing.

Pigmentation studies directly on human skin

Proteins implicated in melanin synthesis can be quantified by fluorescent labelling in skin explants and linked with melanin production.

Labelling of Tyrosinase in a skin section observed by fluorescent microscope.

Skin explants can be treated topically with any cosmetic compound. Melanin production is observed by histology and quantified by image processing.

Treatment with test product twice a day for 7 days and quantification of melanin in the skin after coloration.

Our expertise in skin pigmentation study has been built through the development of many active ingredients and cosmetic products. Do not hesitate to share your projects with us so that we can set up a study adapted to your needs.

The Syntivia team is completing these studies with more details, do not hesitate to contact us for more information on pigmentation studies.

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NeoVirTech, an award winning innovative French company focusing its activity on antiviral discovery and virus imaging, recently acquired a Thermo Scientific CellInsight CX7 high content screening microscope. This integrated performant system allows for high quality in screening and analysis, rapid discovery of active compounds with highly reliable quantification and very high-definition imaging.

The CellInsight CX7 High-Content Screening Platform offers a wide choice of imaging modes to extract information from biological samples. The entire fluorescent spectrum can be used to optimize assays, and either widefield or confocal optics are available for any channel. This microscope also offers 4-color brightfield options for colorimetric analysis of tissue sections.

Thermo Scientific CellInsight CX7 high content screening microscope.

NeoVirTech provides its equipment and screening expertise in this collaboration for cosmetic active ingredients screening.

On the other hand, Syntivia provides its cellular models and biology expertise in cosmetic science. All skin cell types, alive or fixed, can be processed and analysed. Keratinocytes, fibroblasts, melanocytes, adipocytes, sebocytes, neural cells and more can be treated and analysed with your cosmetic compound under a wide range of conditions.

NeoVirTech’s screening expertise combined with Syntivia’s cellular culture and labelling techniques allows to obtain large amounts of reliable data, 2D high quality imaging precise quantification of visible or fluorescent markers to detect the activity of various cosmetic compounds on skin cells.

Proteins involved in skin biology, labelled by immunofluorescence in normal human epidermal keratinocytes (NHEK) – © Syntivia.

“We are delighted to start a collaboration with Syntivia, one of the experts in the discovery and development of dermo-cosmetic compounds. This collaboration will extend and diversify our screening activities, maximise our screening platform and provide rapid quantitative and high-quality data to our partners.” – Franck Gallardo, PhD, CEO and Founder of NeoVirTech.

Don’t hesitate to contact us for more information on cosmetic ingredient screening.

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The right model to study fully functional sebaceous glands

Syntivia’s hi-tech laboratory uses the skin explant model HairSkin^® (Genoskin), obtained from plastic surgery, which is kept in a state of survival for 7 days to allow ex vivo studies of sebaceous glands.

Analyzing sebaceous glands in situ for relevant results

Thanks to these skin explants, the sebaceous glands can be observed both in 2D-sections and using 3D microscopy. This approach allows visualizing both sebocyte differentiation through immunostaining and lipidic production with a specific staining method. The Syntivia team also quantifies the volume of sebaceous glands.

These analysis techniques are a promising new strategy to test active cosmetic ingredients and visualize their effects on appendages involved in sebum production.

Ex vivo sebaceous glands in 2D

Cross section of sebaceous gland after histological coloration. US : Undifferentiated Sebocytes, DS : Differentiated Sebocytes.

Immunochemistry labelling of sebaceous glands in skin explant to detect proteins implicated in sebaceous gland maturation: Marker associated with lipid production (A, B) Marker of undifferentiated sebocytes labelling (C, D). Nuclei are labelled with DAPI in blue.

The red labelling associated with lipid production is linked to differentiated sebocytes. Intense labeling in sebaceous gland lobules is observed. A gradient is visible as central cells are more intense than at the periphery, which highlights the sebocyte differentiation gradient in the glands.

Peripheral cell layers show green labelling where sebocytes are undifferentiated and do not produce lipids yet.

Ex vivo sebaceous glands in 3D

The sebaceous glands are analyzed directly inside the skin explants, which are made transparent using our transparization technique.

Volumic reconstruction of sebaceous glands in a skin explant using light-sheet fluorescence microscopy (LSFM) and analyzed with Imaris software

The volume of each gland can be quantified in the whole skin explant to compare sebaceous gland volume in samples treated with different compounds.

Volumic resconstruction of sebaceous glands in skin explants exposed to several topical treatments, images acquired using light-sheet fluorescence microscopy (LSFM) and analyzed with Imaris software

Quantification of the volume of each sebaceous gland in skin explants exposed to several topical treatments

The average of the volume values obtained reveals a visible trend: the sebaceous glands are slightly larger after treatment with linoleic acid while they atrophy with the test product.

The Syntivia team is still completing these studies with more details, but don’t hesitate to contact us for more information on these new evaluation techniques.

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