Based on its biocompatibility, silk fibroin has been applied in the fabrication of biomaterials in various forms such as gels, powders, fibers, and membranes. Poly-methyl-methacrylate (PMMA), which has been successfully used as a filler material in vertebroplasty, has the disadvantage of excessive inherent stiffness, which may damage adjacent vertebral bodies in elderly patients with osteoporosis. A novel potential filler material, consisting of silk fibroin and PMMA (silk/PMMA), was tested to assess its cytotoxicity, genotoxicity, and intramuscular implantation experiment according to ISO10993. The MTT assay, Ames test, mouse lymphoma assay, and intramuscular implantation were performed using extracts from solidified silk/PMMA, according to the protocol in ISO10993. Extracts of silk/PMMA did not show cytotoxicity, genotoxicity or toxic reaction in intramuscular implantation. These findings suggest that the silk/PMMA composite materials may potentially be used as a nontoxic biomaterial.