PENETRATION OF SCHISTOSOMA-MANSONI CERCARIAE INTO A LIVING SKIN EQUIVALENT

We evaluated the use of a living skin equivalent (LSE) as a suitable membrane for Schistosoma mansoni cercarial penetration. LSE is a living artificial skin composed of a dermal layer containing human dermal fibroblasts embedded in a collagen lattice and an epidermal layer consisting of differentiated human keratinocytes. The keratinocytes differentiate into a stratum corneumlike layer, whereas the dermal-epidermal junction forms a layer similar, but not identical to, the basement membrane. We exposed LSE to 50 cercariae for 0, 3, 6, 20, and 30 hr at 37 C, and the percentage of penetration was evaluated by counting cercariae remaining on the LSE surface. No cercarial penetration was observed in the first 15 min of exposure; however, penetration was detected at all other times. Maximum penetration rates were observed at 20 hr (80%). In other experiments LSE was pretreated topically with 0 or 4 mug/cm2 linoleic acid, then exposed to between 800 and 1,000 cercariae for 18-20 hr at 37 C. LSE pretreated with linoleate had significantly higher penetration rates than untreated membranes (81% +/- 2.51% vs. 65.9% +/- 6.97%, P = 0.03). Increasing linoleate concentrations from 10 to 40 mug/CM2 gradually decreased the ability of cercariae to penetrate the membrane. Some LSE membranes also were processed for light microscopy, and we present photomicrographs showing schistosomulae within the epidermal and dermal layers of the LSE. We conclude that despite the time it takes for cercariae to penetrate LSE, these membranes may allow investigators to examine, in vitro, host-parasite interactions at the level of the skin.