Angiogenesis is a physiological process of blood vessel formation. Despite its natural occurrence, excess of cutaneous vascularization can be associated with a variety of skin conditions, like circles under the eyes or rosacea in the face. While dark circles are explained by an excess in melanin production, purple rings are linked to an impaired blood circulation in the thin epidermis under the eyes. Likewise, red patches of rosacea in the face are due to abnormal dilatation of the blood vessels. In rosacea-affected patients, changes of the external temperature condition or an inappropriate food consumption trigger inflammation that leads to dilation of skin small blood vessels causing facial erythema.

Despite being considered medically harmless, circles and rosacea remain aesthetic concerns affecting the self-esteem of million people worldwide.

Several topical treatments are available to reduce visible circles and minimize rosacea flares. Vasoconstrictors combined to anti-inflammatory or anti-oxidative compounds are commonly adopted solutions. With new active ingredients arriving on the market, in vitro assays are required to demonstrate their efficacy to reduce the vascularization of affected skins.

In vitro, angiogenesis can be replicated using the tube formation assay. This assay relies on the property of cultured endothelial cells to replicates microvascular capillary in vitro. Typically, HUVEC (Human Umbilical Vascular Endothelial cells) quickly forms a network of tube-like structures when seeded on top of a reconstituted basement membrane (FIG 1).

This newly formed structure can be visualized using bright light microscopy to allow measurement of the length of the capillary tubes as well as the area within the mesh networking (FIG 2). Treatment with angiogenic growth factors increases the number of microvascular tubes and mesh areas. In the opposite, anti-angiogenic factor reduces capillary tube formation and therefore the networking.

Based on bright filed images, tube length and mesh area are directly measured to discriminate between angiogenic and anti-angiogenic factors: angiogenic factors increase the length of the formed tubes and the areas of each meshes, compared to anti-angiogenic factors that reduce capillary formation and therefore calculated mesh areas.

In conclusion, the endothelial cells tube formation assay is a simple and cost-effective method for assessing angiogenic regulators in vitro. Knowing that the quality of the basement membrane is highly influencing the performance of the test, StratiCELL has selected a basement membrane close to the in vivo condition to improve cell response, and free from growth factors to reduce the background.

Given the importance on reducing the epidermal microcirculation in case of hypervascularization, this assay is an important tool to define the anti-angiogenic properties of innovative cosmetic compounds. In combination with assays addressing the anti-inflammatory properties or the anti-oxidative properties of cosmetic actives, this assay available at StratiCELL allows to screen for new potential compounds reducing blood vessel dilatation and offers alternative solution to reduce facial skin conditions such as purple rings or rosacea redness.

To get more information on StratiCELL’ s latest testing services, register on www.StratiCELL.com or contact directly StratiCELL at info@StratiCELL.com

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CONTACT

Christel Boudry

Business Developer

cboudry@straticell.com

www.straticell.com

info@straticell.com