Alb-PRF Hybrid Membranes Functionalized with Carbonated Hydroxyapatite and Doxycycline for Bone Regeneration and...

Bone tissue engineering requires biomaterials capable of simultaneously supporting regeneration and preventing infection. Platelet-rich fibrin (PRF) has been widely used due to its autologous origin and growth factor release, but its rapid resorption limits its clinical applications. Albumin-PRF (Alb-PRF) membranes were developed to improve stability, and their combination with carbonated nanostructured hydroxyapatite (nCHA) may further reinforce osteoconductive properties. In this proof-of-concept study, we fabricated Alb-PRF, Alb-nCHA-PRF, and Alb-nCHA-PRF + doxycycline (DOX) membranes and characterized their physicochemical, antimicrobial, and biological performance in vitro. Membrane stability was monitored for up to 14 days; DOX incorporation and release were evaluated by autofluorescence and spectrophotometry; antimicrobial activity was assessed against E. faecalis and S. aureus; and MG-63 osteoblast-like cells were used to test cytocompatibility, proliferation, mineralization, and alkaline phosphatase (ALP) activity. The release of 27 cytokines and growth factors was quantified by multiplex immunoassay. Alb-PRF exhibited morphological integrity and an enhanced trophic secretome, and supported proliferation and late mineralization. nCHA incorporation reduced cell proliferation and secretome output, while DOX conferred sustained antibacterial activity and enhanced early ALP expression even with attenuated cytokine release, positively impacting mineralization, when compared to nCHA alone. These preliminary results provide preliminary feasibility evidence that Alb-PRF can be engineered as a multifunctional scaffold combining antimicrobial and regenerative functions, though some trade-offs indicate the need for dose optimization and validation with in vivo models.