Boosting angiogenesis in ovo by biofunctionalizing collagen membranes with platelet-rich fibrin and hyaluronic acid: Implications for regenerative oral surgery?

Background Biocompatible collagen membranes (CM) are widely used in regenerative dentistry, particularly in guided tissue regeneration (GTR) and guided bone regeneration (GBR). While resorbable CMs offer advantages such as reduced patient morbidity and enhanced wound healing, their barrier function can impede vascularization, potentially compromising graft survival. Biofunctionalization of CMs with platelet-rich fibrin (PRF) and hyaluronic acid (HA) has shown promise in enhancing angiogenesis. Objectives This study evaluates the pro-angiogenic effects of biofunctionalizing collagen membranes with advanced PRF (A-PRF), injectable PRF (i-PRF), and HA using the chorioallantoic membrane (CAM) assay in ovo. Methods Three porcine-derived collagen membranes (Mucoderm®, Bio-Gide®, and Smartbrane®) were biofunctionalized with A-PRF, i-PRF, or HA and applied to the CAM assay on day 7 of incubation. To assess impact on angiogenesis, vascularization was evaluated at multiple time points (24 h, 48 h, 72 h, 96 h, and 120 h) using immunohistochemical staining (hematoxylin-eosin, α-smooth muscle actin, CD105) and AI-assisted image analysis (IKOSA® software). Results Biofunctionalization of Mucoderm® with A-PRF significantly enhanced angiogenesis up to 96 hours, as evidenced by increased vessel area, length, and branching points (p < 0.05). I-PRF biofunctionalization of Bio-Gide® also promoted angiogenesis between 24 and 72 hours, outperforming the native membrane. While HA biofunctionalization of Bio-Gide® and Smartbrane® resulted in significantly increased angiogenesis at 48 and 72 hours (p < 0.05), its effects were less pronounced than those achieved with PRF variants. Native Bio-Gide® exhibited greater pro-angiogenic potential than native Mucoderm®; however, biofunctionalized membranes generally outperformed native variants. Conclusions Biofunctionalization of collagen membranes with A-PRF and i-PRF significantly enhances angiogenesis in ovo, with A-PRF showing sustained effects up to 96 hours. HA also promotes angiogenesis and represents a viable, cost-effective alternative that does not require blood collection. Both PRF and HA biofunctionalization are hold promise for improving vascularization in GBR/GTR applications, and clinical randomized studies are warranted to fully evaluate their pro-angiogenic potential and clinical efficacy.