A disclosed preparation method for a polylactic acid fiber three-dimensional bionic porous ordered scaffold comprises the following steps: slicing polylactic acid (PLA) with good biodegradability and biocompatibility, and successively performing spinning and winding to prepare fiber assemblies in parallel arrangement; using a medical binder to bond the loose and parallel-arranged PLA fiber assemblies to form a stable structure; and then curing and naturally cooling to room temperature. The polylactic acid fiber three-dimensional bionic porous ordered scaffold prepared by using the method has relatively small thickness and weight, relatively large pore diameter and porosity, and relatively good order degree, pore communication property, hydrophily, hydroscopicity, mechanical properties and adsorption capability on gelatin. The ordered scaffold has relatively good application prospect, is applicable to aspects of cell vectors of blood vessel engineering and nerve tissue engineering, ligament base materials, medicine loading and release, nerve regeneration of nerve tissue engineering, implantation type functional muscle, restoration of knee damage and bone arthritis, bone tissue engineering, and the like.