Andreia Feital da Costa Pereira, Renan Lage, Mamy Honda Igarashi, Sheila Gomes da Silva, Maurizio Mercuri, Elis Angela de Alcantara Viana, Lucia de Fatia Holanda Paiva de Araújo,Amanda Francielli Pereira, Suelma Natalie de Melo Oliveira, Michelle Sabrina Silva,Gustavo Facchini, Ana Lúcia Alves Tabarini Pinheiro, Samara Eberlin.
Atopic dermatitis (AD) is a chronic, recurrent and inflammatory skin disease, characterized by the presence of eczematous lesions and pruritus. Despite the large number of clinical and experimental studies, and subsequent advances in this field, the pathophysiology of AD remains unclear. The most recent hypotheses about the pathogenesis of AD involve numerous factors: genetic predisposition, environmental factors, altered immune function and skin barrier dysfunction. Innate and adaptive immunity has a definitive role in the development, maintenance and crisis of AD. This process involves a complex interaction between T cells, antimicrobial peptides, cytokines and keratinocytes. The alteration in the balance of the immune system directly influences the rupture of the skin barrier, characteristic of AD lesions, making individuals predisposed to skin infections. In the present study we evaluated the preclinical efficacy of an active complex (AC) in anti-inflammatory response in culture of human monocytes/macrophages (THP-1) stimulated with lipoteichoic acid (LTA) to induce the inflammatory microenvironment. Additionally, we evaluated the pre-clinical efficacy of a moisturizing formulation containing the AC (Hydra-AI) in the synthesis of fillagrin, involucrin and loricrin in ex vivo skin fragments submitted to epidermal barrier rupture. Cultures of THP-1 were differentiated into phagocytes with 12-myristate 13-acetate phorbol. After, the cultures were stimulated with LTA (10 µM) to induce the inflammatory microenvironment and treated with AC at 0.100; 0.0316 and 0.0100 mg/ml. Cell lysate and culture supernatants were collected for quantification of interleukins 4, 10, 13, 17, 31, and NF-?B using ELISA technique. In parallel, ex vivo skin fragments were treated with Hydra-AI for two consecutive days, with a 24-hour interval between each application. Epidermis was ruptured using 4% sodium lauryl sulfate (SDS) and after 24 hours the fragments were treated again with Hydra-AI. Fragments were collected, fixed and cryopreserved for evaluation by immunofluorescence of filaggrin, involucrin and loricrin. The results obtained demonstrated that the treatment with AC performed on the LTA-stimulated THP-1 cell cultures was able to significantly reduce the inflammatory mediators IL-4, 10, 13, 17, 31 and the signaling pathway NF-?B when compared to the only stimulated group. In addition, immunofluorescence evaluations demonstrated that HYDRA-AI stimulate the synthesis of filaggrin, involucrin and loricrin in fragments of ex vivo skin submitted to rupture of epidermal barrier with SDS. The results obtained allow us to infer that AC presents an anti-inflammatory activity reducing the exacerbated production of mediators associated with the development of AD, which contribute to the commitment of cutaneous barrier. In addition, HYDRA-AI upregulates the synthesis of filaggrin, loricrin and involucrin, suggesting a positive effect on the skin barrier protection, hydration, proliferation, differentiation, epidermal cohesion and synthesis of NMF. Taken together, these results demonstrate that AC and moisturizing formulation containing AC (HYDRA-AI) have a protective and restorative effect of sensitive skin, associated with atopic dermatitis.