Abstract
Acne is an inflammatory skin disease of the pilosebaceous unit, involving 4 essential factors: hyperseborrhea combined to a modification of sebum composition, colonization by Cutibacterium (formerly Propionibacterium) acnes, hyperkeratinization and secreted inflammation. Mimicking these compromised skins is a pre-requisite to understand the complex pathophysiology, and thus to screen potential therapeutic molecules. Most of published cutaneous 3D in vitro models mimicking acneic skin have focused either on the colonization by C. acnes or on the modification of sebum composition. In this context, the aim of this study was to set up a more complex model by combining these two features of acne physiopathology. To do so, normal human keratinocytes were used to generate reconstructed epidermis (RE). During emersion step, RE were either untreated (control) or treated with: - 5.104 CFU of a virulent C. acnes (CA) strain, in order to mimic the colonization by this opportunistic microorganism: CA RE; - peroxidized squalene (PS) in topic during 1 day, in order to mimic the altered sebum composition: PS RE; - a combination of C. acnes and peroxidized squalene, in order to mimic acneic skin in a more physiological way: CA-PS RE For each condition, hyperkeratinization and inflammation were checked at day 11 (D11) and 14 (D14). As expected, CA-PS RE harbors a significant increase of both ratio of SC/total ER thickness and cytokeratin-5 staining, thus revealing hyper-keratinization of the model. As evidence of inflammation, supernatant presents higher level of IL8 than CA RE and PS RE (P<0.005). Regarding antimicrobial defenses, expression of specific markers (CAMP, HBD2 and RNase 7) was significantly increased. In conclusion, by combining two main features of acne (i.e. C. acnes invasion and sebum alteration) this model succeeded in mimicking acne, as it harbors characteristics observed in acneic patients: hyper-keratinization and secreted inflammation. This relevant model is thus suitable for the screening of molecules dedicated to acne treatment.
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