Objective To investigate the biomechanical effects of tropoelastin on self-assembly and degradation of the collagen in vitro. Methods The real-time dynamic scanning with atomic force microscopy was utilized to observe and analyze the changes in microstructure, orientation, area fraction, and average diameter of collagen fibrils during the self-assembly process. The D-band structure of collagen fibrils was characterized using transmission electron microscopy. Finally, data analysis was performed using Gwyddion and Matlab software to assess the structural characteristics and degradation resistance of collagen fibrils. Results Compared to the control group, the addition of tropoelastin not only promoted the collagen self-assembly and significantly increased the area occupied by the assemblies, but also caused a shift in their orientation from unidirectional to multidirectional. Upon the addition of collagenase, the assemblies underwent degradation; however, the degradation half-period decreased with the inclusion of tropoelastin, indicating that tropoelastin accelerated the degradation of collagen assemblies. Conclusions The addition of tropoelastin promotes the collagen self-assembly, and increases the area occupied by the self-assembled structures. However, the consistency of orientation in the assemblies is weakened, resulting in a reduced resistance of collagen fibrils to degradation by collagenase. This study lays a theoretical foundation for the preparation of collagen-based biomimetic functional materials.