ABSTRACT
Tissue-engineered materials for clinical purposes have led to the
development of in vitro models as alternatives to animal testing.
The development of this work is to understand and apply the
basic principles of paracrine interactions arising between human
keratinocytes and fibroblasts for detecting an irritant-induced
inflammatory reaction for future testing purposes.We have used two
irritants, sodium dodecyl sulphate (SDS) and potassium diformate
(Formi R©) at subtoxic concentrations and studied interleukin-1 alpha (IL-1α) release from human keratinocytes as the primary
response to an irritant. Fibroblasts were cultured separately and
used to link the primary cytokine response to a series of secondary
responses using a keratinocyte-conditioned medium. Interleukin-
6 (IL-6) and interleukin-8 (IL-8) release from fibroblast cells was
measured and used to confirm the early amplification stage of
irritant-induced inflammatory signaling between keratinocytes and
fibroblasts. We describe herein the methods to conduct these
experiments in the context of paracrine models for in vitro testing
of irritant chemicals at subtoxic concentrations, as an alternative to
in vivo models.
