Chitosan has been widely studied for use in many areas, such as for its applications in the biomedical, engineering and pharmaceutical fields, as well as in industry, because of its unique properties, including biodegradability, antimicrobial activity, polycationic nature and biocompatibility. Thanks to the rapid development of materials science, chitosan applications are now possible in textiles. However, there are still many limitations of chitosan fibers in terms of their high electrostaticity, poor mechanical properties and high cost, which are obstacles that inhibit potential applications of chitosan fiber in the industry. Generally, in order to achieve the best performance with chitosan and enhance its commercial value, chitosan fibers are usually blended with long cotton fibers in the textile industry. Therefore, based on preliminary experiments and feedback from the industry, this study was carried out to further investigate the relationship between fiber length, fiber interaction and yarn performance. The results of this study would therefore help to reduce the production cost of yarns with the blending parameters used and also expand the utilization and applications beyond medical applications to fashion-based functional wear. The sliver-blending method offers better tensile properties of yarn samples, while the fiber-blending method offers higher uniformity of fiber distribution. This study would help to reduce the production cost of yarns by blending and also to expand the utilization and application not limited to fashion-based functional wear.