• The polylactic acid/collagen bilayer membrane fabricated by electrospinning exhibits a dense layer with smaller fiber diameter and pore size, and a loose layer with larger fiber diameter and pore size, providing a suitable structure for guided tissue regeneration.
• Glutaraldehyde vapor cross-linking was selected as the optimal method, yielding a cross-linking degree of 17.42% and improved mechanical properties.
• The membrane shows asymmetric wettability: the loose layer is hydrophilic while the dense layer is hydrophobic, which may facilitate cell attachment and barrier function.
• Comprehensive biocompatibility tests confirmed that the membrane is non-cytotoxic, non-hemolytic, non-pyrogenic, non-toxic, non-irritating, and non-sensitizing, indicating excellent biocompatibility for clinical application.