Multifunctional Three-Layer Collagen-Based Membrane for Periodontal Guided Tissue Regeneration

Guided tissue regeneration (GTR) and guided bone regeneration (GBR) are pivotal surgical techniques for addressing periodontal defects arising from periodontitis. These methods utilize barrier membranes to isolate soft tissue from bone defects, preventing premature fibroblast infiltration and promoting osteoblast colonization. However, infections caused by periodontitis-associated pathogens significantly elevate the risk of membrane failure, underscoring the need for advanced membrane designs. This study introduces a multifunctional membrane, known as guided tissue/bone regeneration membrane (GTBRM), engineered to optimize composition, structure, and bioactivity. The GTBRM comprises two porous collagen layers for soft and hard tissue regeneration and 2% silver nanoparticles (AgNPs) and β-tricalcium phosphate nanoparticles (β-TCP NPs) for antibacterial and osteogenic properties, respectively, and a dense poly(vinyl alcohol) (PVA)/cellulose nanocrystal (CNC) core for mechanical strength and cell occlusion. Physicochemical properties of the membrane were characterized using scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDAX), X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, tensile strength testing, and biodegradation assessments. In vitro studies with MG-63 cells, human gingival fibroblasts (HGF), and bone marrow mesenchymal stem cells (BMSCs) demonstrated biocompatibility, cell adhesion, proliferation, and osteogenic differentiation. The incorporation of AgNPs conferred significant antibacterial efficacy against Enterococcus faecalis (E. faecalis) and Fusobacterium nucleatum (F. nucleatum), and β-TCP NPs enhanced osteoblast activity. In vivo studies in rat radial bone and oral defect models revealed that GTBRM markedly improved osteogenesis and buccal soft tissue regeneration. These preclinical findings suggest that the GTBRM exhibits potential for periodontal regeneration by providing biocompatibility, space maintenance, antibacterial properties, and osteoconductivity. However, further studies are needed to validate its clinical efficacy and translational potential.