The blood-brain barrier (BBB) maintains the homeostasis of the brain microenvironment, which is crucial for neuronal activity and function. Under pathological conditions, the BBB may fail due to yet unknown mechanisms. BBB failure is accompanied by an increase in the transendothelial permeability to substances such as sucrose that are normally extruded. Furthermore, altered BBB function may also lead to development of abnormal drug extrusion mechanisms including expression of multiple drug resistance proteins. Therefore, it is not surprising that strategies have been developed to "repair" the BBB in order to restore normal brain homeostasis and penetration/extrusion of pharmacologically active (noxious) substances. To this end, steroidal hormones and synthetic analogues such as dexamethasone (DEX) have been used to counteract BBB failure. However, several side effects limit the usefulness of steroid treatment in humans leading to the quest for developing novel strategies for BBB repair. We here show that, in an in vitro model of the BBB based on a co-culture of endothelial cells (EC) and glia, the natural compound glycerophosphoinositol (GPI) may replicate the effects of DEX. Thus, GPI in concentrations ranging from 3 to 100 microM promoted both BBB formation and repair in a dose dependent fashion. Similar effects were obtained with an elevated dose of DEX (10 microM); at higher concentrations (100 microM), DEX was cytotoxic. We conclude that the endogenous anti-inflammatory agent GPI may ameliorate BBB function with efficacy comparable to that of steroids, but with significantly fewer side effects. Further experiments will confirm the efficacy of this treatment in vivo and elucidate the pathways that lead to BBB repair after exposure to GPI.