A new ex vivo skin model to mimic pollen allergens exposition and evaluate preventive or cleansing effects of skin care products
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Presented by: Laurent PENO-MAZZARINO
Introduction
By their form and function, infundibula are natural site of accumulation of many exogenous elements, such as microorganisms, chemicals, pollutant matters or allergens such as pollen allergens.
This accumulation is a source of discomfort in most cases but may lead to severe allergic reactions in sensitive subjects. In order to evaluate the preventive or cleansing efficacy of care or hygiene skin products, a new study model using hairy skin explants on Perfex vivo supports was developed to mimic the deposit of pollen allergen in the hair follicle infundibula.
Methods
From a hairy abdominoplasty (obtained from aesthetic surgery), large explants were prepared and placed on Perfex vivo culture supports (BIO-EC culture device). After few hours of stabilization in open air under controlled temperature and with culture medium nutrition, stratum corneum reaches its normal physiology and environment.
Then, Recombinant Pollen allergen Phl p 5b protein was applied on the whole surface of the explant to mimic a pollen exposure. After 6 hours of contact, skin explants comprising several infundibula were collected and fixed with formalin solution, then impregnated and embedded with paraffin for immunohistochemistry analysis. The presence of pollen proteins was revealed by immunostaining of the Phl p 5b protein on serial paraffin or frozen skin sections, using several antibody references.
In a second step, two types of product application (preventive or curative) were carried out to define if pollen allergen accumulation in infundibulum can be reduced or removed.
In order to evaluate if a product is able to reduce the accumulation of pollen protein in the infundibula, products were applied prior to the application of the pollen. To investigate the performance of cleansing product, explants were washed after 6hrs of allergen contact.
Results
Several trials with different skin types, mode of application, antibodies allowing to visualize the allergen and tissue treatments (paraffin vs frozen / antigen retrieval) were carried out, to determine the optimum conditions to detect pollen protein in the infundibulum.
The best method to identify and reveal the accumulation of pollen in the infundibula was immunostaining of Phl p 5b pollen protein on formalin- fixed paraffin- embedded skin sections revealed by immunohistochemistry (peroxidase coupled secondary antibody).
A film forming product applied prior to pollen exposure significantly reduced pollen deposit in infundibula, offering a preventive benefit effect.
Cleansing with a cosmetic product after pollen exposure managed to significantly remove accumulated pollen from infundibula.
Conclusion discussion
The model based on hairy skin explants on Perfex vivo support allows to mimic skin exposure to allergens like pollen and their deposit in infundibula. This model can highlight protective effects of film forming products by reducing the accumulation of pollen on the skin. In addition, this model allows to evaluate cleansing effects of product applied to remove pollen from the skin.
By their form and function, infundibula are natural site of accumulation of many exogenous elements, such as microorganisms, chemicals, pollutant matters or allergens such as pollen allergens.
This accumulation is a source of discomfort in most cases but may lead to severe allergic reactions in sensitive subjects. In order to evaluate the preventive or cleansing efficacy of care or hygiene skin products, a new study model using hairy skin explants on Perfex vivo supports was developed to mimic the deposit of pollen allergen in the hair follicle infundibula.
Methods
From a hairy abdominoplasty (obtained from aesthetic surgery), large explants were prepared and placed on Perfex vivo culture supports (BIO-EC culture device). After few hours of stabilization in open air under controlled temperature and with culture medium nutrition, stratum corneum reaches its normal physiology and environment.
Then, Recombinant Pollen allergen Phl p 5b protein was applied on the whole surface of the explant to mimic a pollen exposure. After 6 hours of contact, skin explants comprising several infundibula were collected and fixed with formalin solution, then impregnated and embedded with paraffin for immunohistochemistry analysis. The presence of pollen proteins was revealed by immunostaining of the Phl p 5b protein on serial paraffin or frozen skin sections, using several antibody references.
In a second step, two types of product application (preventive or curative) were carried out to define if pollen allergen accumulation in infundibulum can be reduced or removed.
In order to evaluate if a product is able to reduce the accumulation of pollen protein in the infundibula, products were applied prior to the application of the pollen. To investigate the performance of cleansing product, explants were washed after 6hrs of allergen contact.
Results
Several trials with different skin types, mode of application, antibodies allowing to visualize the allergen and tissue treatments (paraffin vs frozen / antigen retrieval) were carried out, to determine the optimum conditions to detect pollen protein in the infundibulum.
The best method to identify and reveal the accumulation of pollen in the infundibula was immunostaining of Phl p 5b pollen protein on formalin- fixed paraffin- embedded skin sections revealed by immunohistochemistry (peroxidase coupled secondary antibody).
A film forming product applied prior to pollen exposure significantly reduced pollen deposit in infundibula, offering a preventive benefit effect.
Cleansing with a cosmetic product after pollen exposure managed to significantly remove accumulated pollen from infundibula.
Conclusion discussion
The model based on hairy skin explants on Perfex vivo support allows to mimic skin exposure to allergens like pollen and their deposit in infundibula. This model can highlight protective effects of film forming products by reducing the accumulation of pollen on the skin. In addition, this model allows to evaluate cleansing effects of product applied to remove pollen from the skin.