The emerging technique of three-dimensional (3D) printing provides a simple, fast, and flexible way to fabricate structures with arbitrary spatial features and may prove useful in the development of standardized, phantom-based performance test methods for biophotonic imaging. Acrylonitrile Butadiene Styrene (ABS) is commonly used in the printing process, given its low cost and strength. In this study, we evaluate 3D printing as an approach for fabricating biologically-relevant optical phantoms for hyperspectral reflectance imaging (HRI). The initial phase of this work involved characterization of absorption and scattering coefficients using spectrophotometry. The morphology of phantoms incorporating vessel-like channels with diameters on the order of hundreds of microns was examined by microscopy and OCT. A near-infrared absorbing dye was injected into channels located at a range of depths within the phantom and imaged with a near-infrared HRI system (650-1100 nm). ABS was found to have scattering coefficients comparable to biological tissue and low absorption throughout much of the visible and infrared range. Channels with dimensions on the order of the resolution limit of the 3D printer (~0.2 mm) exhibited pixelation effects as well as a degree of distortion along their edges. Furthermore, phantom porosity sometimes resulted in leakage from channel regions. Contrast-enhanced channel visualization with HRI was possible to a depth of nearly 1 mm – a level similar to that seen previously in biological tissue. Overall, our ABS phantoms demonstrated a high level of optical similarity to biological tissue. While limitations in printer resolution, matrix homogeneity and optical property tunability remain challenging, 3D printed phantoms have significant promise as samples for objective, quantitative evaluation of performance for biophotonic imaging modalities such as HRI.