Collagen is a structural protein that is widely present in vertebrates, being usually distributed in tissues in the form of fibers. In living organisms, fibers are organized in different orientations in various tissues. As the structural base in connective tissue and load-bearing tissue, the orientation of collagen fibers plays an extremely important role in the mechanical properties and physiological and biochemical functions. The study on mechanics role in formation of oriented collagen fibers enables us to understand how discrete cells use limited molecular materials to create tissues with different structures, thereby promoting our understanding of the mechanism of tissue formation from scratch, from invisible to tangible. However, the current understanding of the mechanism of fiber orientation is still insufficient. In addition, existing fabrication methods of oriented fibers are varied and involve interdisciplinary study, and the achievements of each experiment are favorable to the construction and improvement of the fiber orientation theory. To this end, this review focuses on the preparation methods of oriented fibers and proposes a model explaining the formation process of oriented fibers in tendons based on the existing fiber theory. Impact statement As the structural base in connective tissue and load-bearing tissue, the orientation of collagen fibers plays an extremely important role in the mechanical properties and physiological and biochemical functions. However, the current understanding of the mechanism of fiber orientation is still insufficient, which is greatly responsible for the challenge of functional tissue repair and regeneration. Understanding the mechanism of fiber orientation can promote the successful application of fiber orientation scaffolds in tissue repair and regeneration, as well as providing an insight for the mechanism of tissue histomorphology.