Wound healing research has advanced with biodegradable polymers as a promising treatment, especially for chronic wounds. Zebrafish offer a cost-effective, regenerative, and human-relevant model for real-time, reproducible skin repair studies. The primary objective of this study is to investigate the efficacy of biodegradable polymers, PLLA (poly-l-lactic acid) and PDLLA (poly-D, L lactic acid), in promoting skin wound healing in zebrafish, while understanding their mechanisms of action at the cellular and molecular levels. Using a laser-based ablation method, which provides precision and reproducibility, skin wounds are created in zebrafish and treated with these polymers to test their wound closure-promoting efficacy. The healing process was monitored by capturing the dynamic change of skin wound size and further analyzed through molecular techniques to evaluate key biomarker genes that are important for collagen deposition, angiogenesis, and inflammation. Additionally, a comprehensive biosafety assessment of PDLLA and PLLA is conducted to ensure their biocompatibility and safety for therapeutic applications. Results showed that usage of PLLA and PDLLA, especially when they were combined, significantly enhanced the wound healing process in zebrafish. Overall, this study highlighted the effectiveness of PLLA and PDLLA in enhancing skin wound healing, providing valuable insights into their mechanisms and paving the way for future research and clinical applications in human wound care.