Improvements in measurement of epithelial tissue optical properties (OPs) in the ultraviolet and visible (UV–Vis) may lead to enhanced understanding of optical techniques for neoplasia detection. In this study, we investigated an approach based on fiber-optic measurement of reflectance to determine absorption and reduced scattering coefficients ( ${\mu }_a$ and ${\mu }_s^{\prime }$ ) in two-layer turbid media. Neural network inverse models were trained on simulation data for a wide variety of OP combinations ( ${\mu }_a=1–22.5$ , ${\mu }_s^{\prime }=5–42.5{\mathrm{cm}}^{-1}$ ). Experimental measurements of phantoms with top-layer thicknesses (D) ranging from 0.22 to $0.66\mathrm{mm}$ were performed at three UV–Vis wavelengths. OP estimation accuracy was calculated and compared to theoretical results. Mean prediction errors were strongly correlated with D and ranged widely, from 1.5 to $12.1{\mathrm{cm}}^{-1}$ . Theoretical analyses indicated the potential for improving accuracy with alternate probe geometries. Although numerous challenges remain, this initial experimental study of an unconstrained approach for fiber-optic-based OP determination in two-layer epithelial tissue indicates the potential to provide useful measurements.