Laser micro-machining is a rapidly growing technique to create, manufacture and fabricate microstructures on different materials ranging from metals and ceramics to polymers. Micro- and nano-machining on different materials has been helpful and useful for various biomedical applications. This study focuses on the micro-machining of innovative barbed sutures using an ultrashort pulse laser, specifically a femtosecond (fs) laser system. Two bioresorbable polymeric materials, namely, catgut and poly (4-hydroxybutyrate) (P4HB), were studied and micro-machined using the femtosecond (fs) laser system. The optimized laser parameter was used to fabricate two different barb geometries, namely, straight and curved barbs. The mechanical properties were evaluated via tensile testing, and the anchoring performance was studied by means of a suture-tissue pull-out protocol using porcine dermis tissue which was harvested from the medial dorsal site. Along with the evaluation of the mechanical and anchoring properties, the thermal characteristics and degradation profiles were assessed and compared against mechanically cut barbed sutures using a flat blade. The mechanical properties of laser-fabricated barbed sutures were significantly improved when compared to the mechanical properties of the traditionally/mechanically cut barbed sutures, while there was not any significant difference in the anchoring properties of the barbed sutures fabricated through either of the fabrication techniques. Based on the differential scanning calorimetry (DSC) results for thermal transitions, there was no major impact on the inherent material properties due to the laser treatment. This was also observed in the degradation results, where both the mechanically cut and laser-fabricated barbed sutures exhibited similar profiles throughout the evaluation time period. It was concluded that switching the fabrication technique from mechanical cutting to laser fabrication would be beneficial in producing a more reproducible and consistent barb geometry with more precision and accuracy.