This study presents a novel trilayer eccentric Janus nanofiber structure fabricated via multiple-fluid electrospinning for the treatment of periodontitis. The nanofibers consist of three layers: an outer hydrophilic layer containing ketoprofen for rapid drug release and anti-inflammatory effects, a middle layer with metronidazole for sustained release, and an inner layer with nano-hydroxyapatite to promote alveolar bone growth. The fibers exhibit good wettability, mechanical properties, biodegradability, and antibacterial properties. They reduce inflammatory responses and promote osteoblast formation by modulating interleukin 6 and osteoprotegerin expression. The composite nanostructures also enhance fibroblast attachment, infiltration, proliferation, and differentiation. The developed fibrous implant films show strong potential for combined treatment of periodontitis. The study highlights the potential of multi-chamber nanofibers for advanced medical applications through a robust process-structure-performance relationship. The research demonstrates that electrospun fibrous membranes can serve as effective implantable barrier membranes for periodontal tissue regeneration. These membranes can prevent epithelial and connective tissue ingrowth, minimize bacterial migration, and guide bone regeneration. However, conventional materials have limited penetration into periodontal pockets and are ineffective for tissue regeneration. Electrospun fibrous membranes, with tunable porosity, controllable morphology, and high flexibility, are promising for periodontal treatment. The study suggests that multi-chamber nanostructures can be tailored to deliver multiple active ingredients for synergistic therapy. The research also highlights the importance of biodegradable, environmentally friendly materials for periodontal implants. Polymers such as PCL, PLGA, PLA, chitosan, and gelatin have been explored as candidates for electrospun scaffolds. However, natural polymers have limitations in mechanical properties and spinnability. Synthetic polymers like PCL offer advantages in biocompatibility and mechanical properties. The study demonstrates the potential of electrospun fibrous membranes for periodontal treatment, paving the way for the development of advanced fiber materials for medical applications.This study presents a novel trilayer eccentric Janus nanofiber structure fabricated via multiple-fluid electrospinning for the treatment of periodontitis. The nanofibers consist of three layers: an outer hydrophilic layer containing ketoprofen for rapid drug release and anti-inflammatory effects, a middle layer with metronidazole for sustained release, and an inner layer with nano-hydroxyapatite to promote alveolar bone growth. The fibers exhibit good wettability, mechanical properties, biodegradability, and antibacterial properties. They reduce inflammatory responses and promote osteoblast formation by modulating interleukin 6 and osteoprotegerin expression. The composite nanostructures also enhance fibroblast attachment, infiltration, proliferation, and differentiation. The developed fibrous implant films show strong potential for combined treatment of periodontitis. The study highlights the potential of multi-chamber nanofibers for advanced medical applications through a robust process-structure-performance relationship. The research demonstrates that electrospun fibrous membranes can serve as effective implantable barrier membranes for periodontal tissue regeneration. These membranes can prevent epithelial and connective tissue ingrowth, minimize bacterial migration, and guide bone regeneration. However, conventional materials have limited penetration into periodontal pockets and are ineffective for tissue regeneration. Electrospun fibrous membranes, with tunable porosity, controllable morphology, and high flexibility, are promising for periodontal treatment. The study suggests that multi-chamber nanostructures can be tailored to deliver multiple active ingredients for synergistic therapy. The research also highlights the importance of biodegradable, environmentally friendly materials for periodontal implants. Polymers such as PCL, PLGA, PLA, chitosan, and gelatin have been explored as candidates for electrospun scaffolds. However, natural polymers have limitations in mechanical properties and spinnability. Synthetic polymers like PCL offer advantages in biocompatibility and mechanical properties. The study demonstrates the potential of electrospun fibrous membranes for periodontal treatment, paving the way for the development of advanced fiber materials for medical applications.