The effect of high energy ball milling, HEBM, and the presence of kaolin on the structure, morphology, and biofilm development of polylactic acid, PLA, were studied. Biofilm development was evaluated in terms of structural and/or morphological variations so as the sole presence of kaolin. Composites based on PLA filled with kaolin were prepared by HEBM followed by hot pressing to obtain films. Structure was studied by X-ray diffraction and Fourier transformed infrared spectroscopy whereas morphology was inspected by scanning electron microscopy and atomic force microscopy. To study biofilm development on the surface of these materials, Pseudomonas fluorescens B52 were used. The shear forces from the milling process favor kaolin dispersion within the PLA. Longer milling times and cryogenic conditions improve clay dispersion. Subsequent hot pressing process enhances the most ordered structure of PLA (alpha-phase) which is also favored with previous milling at longer times and under cryogenic conditions. Changes in P. fluorescens biofilm development are mainly due to modifications of surface properties induced by structural variations, being the most ordered structures those which better support bacterial adhesion and proliferation.