This work investigates the effect of a bio-based plasticizer derived from used sunflower oil on the crystallization behavior of poly (lactic acid) (PLA), comparing it with that of the conventional plasticizer tributyrin. This study aims to explore biodegradable alternatives to petroleum-based materials and to evaluate their potential in modulating PLA crystallization kinetics without altering the crystalline structure of the resulting sustainable material solutions with tailored performance. PLA-based films containing 5%, 10%, and 15% plasticizer were prepared and characterized by differential scanning calorimetry (DSC), polarized optical microscopy (POM), and X-Ray diffraction (XRD). DSC analysis showed a decrease in the glass transition temperatures upon plasticization, with tributyrin producing a more pronounced effect than the recycled sunflower oil plasticizer. XRD patterns confirmed that the crystalline form of PLA remained unchanged regardless of plasticizer type or content. POM revealed that both plasticizers influenced crystallization kinetics, with the bio-plasticizer promoting larger and more sparsely distributed spherulites than tributyrin, indicating differences in nucleation efficiency and crystal growth. Crystallization kinetics were further analyzed using the Avrami model, the Lauritzen-Hoffman theory, and the isoconversional method. Avrami analysis indicated that nucleation mechanisms were largely unaffected, although the overall crystallization rate increased upon plasticization. Lauritzen-Hoffman analysis confirmed crystallization in Regime III, controlled by nucleation, while isoconversional analysis showed reduced activation energy in plasticized PLA. These findings highlight the ability of bio-derived plasticizers to modulate PLA crystallization, promoting the valorization of a food industry residue as a sustainable plasticizer. This study hopes to contribute relevant knowledge to emerging areas of polymer processing, such as 3D printing, to develop sustainable and high-performance PLA-based materials.