Cultivation of 3D dermal tissue by application of autologous matrix

Abstract

The most common reasons for major skin loss are thermal trauma — burns and scalds that can result in rapid, extensive, deep wounds as well as chronic non-healing wounds. Treatment using common techniques is poor and depending on the trauma level can result in death. There is a substantial need for skin integrity restoration. The main goal of this study was to develop an autologous 3D skin model that could eventually be translated into clinical applications. The study examined a variety of factors — extracellular matrix components, cell count, culture medium modification and role of structurally and functionally high-quality 3D skin dermis layer tissue culture production. The results of this study are an essential prerequisite to standardise the use of both clinical, as well as in vitro test systems. Dermal cell lines applied in the study were isolated form patient biopsies obtained at Pauls Stradiòð Clinical University Hospital. Blood plasma type AB was used for fibrin matrix formation. As catalysts, CaCl2 or calcium gluconate, and tranexamic acid were applied. 3D tissue functionality was assessed by evaluation of gene expression and changes in growth factor secretion. Fibrin matrix formulations with 1% and 1.5% CaCl2 and 5 mg, 7 mg and 10 mg tranexamic acid concentration were tested. Better matrix properties were observed with higher concentration of CaCl2 and tranexamic acid. Differences in levels of collagen gene expression and growth factor secretion were observed. Changes in levels of fibroblast growth factor and gene expression were observed in fibrin matrix samples and the surface-cultivated cell culture monolayer, but structural protein synthesis was not detected.