Analysis of the expression of FAP-α protein in 2D-keloid fibroblast cultures and in 3D models of keloid.

OBJECTIVE

Keloids arise most often as a result of abnormal wound healing. The lesion characterizes inflammation and fibroproliferation. Fibroblast activation protein alpha (FAP-α) is one of enzymes engaged in keloid formation. It is serine protease that facilitates cells invasion and growth. FAP-α possess also non-enzymatic activity and plays role in activation of cell signaling. The aim of the study was to assess the impact of culture conditions on the proliferation of keloid fibroblasts and the expression of FAP-α. Analysis of utility of 3D models of keloid in in vitro study of pathogenesis of keloids was also made.

METHODS

NHDF and KEL FIB cells were cultured in vitro in 2D cultures and 3D Matrigel models. The viability of cells was assayed spectrophotometrically with WST-1 test. FAP-α protein amount in cell cultures and 3D models was evaluated with the use of ELISA test.

RESULTS

KEL FIB fibroblasts exhibited higher viability than NHDF fibroblasts in all three models of keloid. The expression of FAP-α is different in normal and keloid fibroblasts. In vitro conditions influence the expression of FAP-α in NHDF cells but not in KEL FIB cells.

CONCLUSIONS

This preliminary study has shown that the expression of FAP-α, similarly to other enzymes engaged in keloid formation, is different in keloids in vivo and in in vitro models. FAP-α expression is modulated by in vitro conditions in normal fibroblasts but not in keloid fibroblasts.