Kefir is a natural probiotic produced by kefir grains fermentation. Biofilms produced from fresh kefir grains in natura were studied for presenting structural characteristics that will be of great interest in the area of regenerative medicine. This work presents a study on the surface of kefir biofilms, obtained by the cultivation of kefir grains in commercial white sugar. Four different films were produced, varying the concentration of sugar. The crystallinity of the biofilms was analyzed and revealed that sugar concentration influences biofilm amorphousness. Morphology showed that the biofilms presented excellent superficial adhesiveness. Fractal parameters were studied and revealed that there was homogeneity in the biofilm microtexture. Both fractal succolarity and surface entropy showed that the degree of water penetration and topographic homogeneity of the biofilms was not influenced by sugar concentration. These results show that kefir biofilms have excellent structural and morphological properties to be used in the biomedical field.