Physicochemical and biocompatibility characterisation of lyophilised platelet-rich fibrin for bone tissue engineering

The preservation and functional stability of platelet-rich fibrin (PRF) are critical for its translational use in craniofacial bone tissue engineering. Preservation methods for frozen PRF (FPRF) remain controversial; freeze-thaw cycles can compromise fibrin structure and diminish bioactivity. Lyophilisation presents an alternative strategy that may enhance storage stability, handling and clinical shelf life while maintaining microarchitecture and biological efficacy. This study aimed to evaluate and compare the physicochemical and biological properties of LyPRF and FPRF. PRF samples were prepared from venous blood obtained from healthy donors and processed into LyPRF and FPRF. Comprehensive physicochemical characterisation was conducted using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). The kinetic release pattern of Growth factor (PDGF-AB, PDGF-BB, and TGF-β1) were quantified via enzyme-linked immunosorbent assay (ELISA), while biocompatibility was assessed using the MTT assay on human gingival fibroblasts (HGFs). SEM analysis demonstrated a more porous and interconnected fibrin network in LyPRF compared to FPRF. EDX analysis revealed higher mineral retention in LyPRF, and both FTIR and XRD confirmed the preservation of fibrin-associated protein structures in both groups. Notably, LyPRF exhibited sustained release of growth factors and enhanced fibroblast metabolic activity over 72 hours relative to FPRF. The aforementioned results demonstrates LyPRF exceptional physicochemical and biocompatibility characteristics, establishing it as a promising biomaterial for craniofacial bone tissue engineering.