Characterization of a new full thickness in vitro skin model.

Abstract

Since 30 years, bioengineering allowed to reconstruct human tissues using normal human cells. Skin has been one of the first organ to be reconstructed thanks to the development of specific cell culture media and supports favoring the culture of human skin cells such as fibroblasts, keratinocytes or melanocytes. Skin models have evolved from epidermis to complex models including a dermis. The purpose of the present study was to design a reconstructed full thickness skin suitable to perform efficacy testing of molecules and plant extracts in vitro. First, we reconstructed epidermis with normal human keratinocytes displaying the expected multi layered morphology and expressing specific epidermal proteins (e-cadherin, claudin-1, p63, Ki67, Keratin 10, filaggrin and loricrin). Then, a dermal equivalent was developed using a collagen matrix allowing the growth of fibroblasts. The functionality of the dermis was demonstrated by the measurement of skin parameters such as rigidity or elasticity with Ballistometer® and other parameters such as the contraction over time and the expression of dermal proteins. The combination of these two compartments (dermis and epidermis) allowed to reconstruct a full thickness model. This study model allowed to study the communication between compartments and with the establishment of a dermo epidermal junction showing the expression of specific proteins (collagen XVII, laminin, collagen IV).