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In ligament reconstruction involving anterior cruciate ligament surgery, biological fixation between the transferred ligament and bone tissue is critical for achieving successful outcomes. Here, we administered chitin fabrics into the bone tunnels and evaluated their efficacy in promoting biological fixation. An animal model on the rat's patellar ligament was employed. First, bone tunnels were created in the lateral condyle of the femur. The ligament was then separated from the tibial tuberosity, and half was inserted into the tunnel and fixed with the use of end button. Animals in the experimental group were treated with microfiber nonwoven chitin fabric, whereas control animals received no treatment. Specimens were collected at 2, 4, and 6 weeks after surgery, and the fixation strength was measured by mechanical tests. Histological sections were prepared from samples prepared 4 weeks after surgery, and the diameter of bone tunnel and the width ratio of collagenous tissue in the bone tunnel were measured. Administration of chitin significantly increased the mean fixation strength at 4 and 6 weeks after surgery. Furthermore, chitin also promoted bone formation in the bone tunnel and increased the density of collagen fibers. Thus, microfiber nonwoven chitin fabric enhanced the biological fixation of the ligament to the bone tissue. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2355-2360, 2018., (© 2017 Wiley Periodicals, Inc.)