Emerging studies have showed that silk fibroin might be as a naturally occurring degradable biomaterial for tissue engineering with unique mechanical properties, biocompatib ility and processability. To optimize the formation of the regenerated silk fibroin film, in thi s study, we examined the genotoxic potential of the regenerated silk fibroin films by differe nt cross-linking mode with micronucleus and the single cell gel electrophoresis (comet) assa y. The data showed that MNF in group PC and group SD showed significantly no differenc e with blank group (P>0.05), and no evident comet tails were observed. However, the resul t of EC group was significantly different from the control group(P<0.01). Taken together, R SFFs in group PC and group SD have no evident genotoxicity, which is better than group EC.