Fabric placed over human skin may change the stratum corneum (SC) hydration level, an important phenomenon because increases in normal levels of SC hydration pose potential health problems. The skin becomes more susceptible to abrasive damage, increases in permeability, and is more prone to microbial growth. The purpose of this study was to determine the effect of fiber type and fabric moisture content on SC hydration. Using three similarly constructed knit fabrics, six fabric type/moisture content combinations were selected for the tests. Using an occluded plastic dome, fabric samples were placed on both “normal” and “hydrated” volar forearm skin of subjects for a specified time period, then removed. Two minutes after removal, evaporative water loss and skin temperature were measured. The design of the study was a randomized complete block with all possible treatment combinations applied to each subject. Data were analyzed using analyses of variance on the linear and quadratic coefficients of best fit lines and Bonferroni t tests. For “normal” skin, SC hydration generally increased as fabric moisture content increased. The SC was significantly drier after being in contact with cotton fabric at regain than at the two moisture content levels above regain, and also under polyester fabric. For hydrated skin, the hydration state was significantly lower under the cotton fabric at regain than at 38.6% moisture content and at saturation, but was not significantly different under the polyester fabric at regain and at saturation.