Paracrine activation of extracellular signal-regulated kinase in a simple in vitro model of wounded osteoblasts.

The immediate signal-transduction response of osteoblasts to acute trauma is poorly characterized. We have developed a simple in vitro model for osteoblast trauma to investigate aspects of the molecular mechanisms of wound healing in bone. Herein we report the specific, rapid, and transient phosphorylation of extracellular signal-regulated kinase (ERK) 1 and 2 in osteoblasts as a response to disruption ("wounding") of a confluent monolayer. The mitogen-activated protein kinase (MAPK) cascades of p38 and stress-activated protein kinase/c-jun N-terminal kinase (SAPK/JNK) were not activated by this perturbation. The response to wounding was equivalent to the activation of ERK by the addition of exogenous growth factors, and the perturbation-dependent phosphorylation of ERK can be suppressed by an inhibitor of heparin-binding growth factors. Conditioned media from wounded monolayers can induce the phosphorylation of ERK in unperturbed monolayers. Using immunohistochemistry, it was demonstrated that the cells with increased levels of phosphorylated ERK were not localized to the wound edges. These results indicate that ERK activation is the result of an autocrine/paracrine response by osteoblasts to trauma. We speculate that osteoblasts respond to trauma with the release of soluble factors as part of an autocrine/paracrine modulation of the wound-healing process in bone.