Hyaluronic acid promotes biomineralization of osteoblast-like cells – observations on two different barrier membranes

Abstract Background Guided bone regeneration (GBR) relies on biocompatible membranes to support osteogenesis. 1,4-butanediol diglycidyl ether (BDDE)-crosslinked hyaluronic acid (xHyA) has shown promise in enhancing bone regeneration, yet its mechanisms remain unclear. Objective This study evaluates the osteogenic effects of xHyA-functionalized native pericardium collagen membrane (NPCM) and ribose-crosslinked collagen membrane (RCCM) using an airlift culture model with SaOS-2 cells. Transcriptomic changes following xHyA treatment were also investigated. Methods SaOS-2 cells were cultured on NPCM and RCCM, with or without xHyA functionalization. Cytocompatibility, alkaline phosphatase (ALP) activity, mineralization (Von Kossa staining), and RNA sequencing were assessed. Differential gene expression and pathway enrichment analyses were conducted on cells exposed to two xHyA concentrations. Results Both membrane types supported cell viability, though NPCM allowed cellular infiltration while RCCM maintained barrier integrity. xHyA significantly enhanced ALP activity and mineral deposition across both substrates. RNA sequencing revealed minimal upregulation of classical osteogenic genes but identified differential expression in pathways related to focal adhesion, VEGF signaling, and stem cell differentiation. IGF1R, FYN, and MAPK14 were consistently upregulated regardless of xHyA concentration. Conclusion xHyA functionalization enhances osteogenic activity, evidenced by increased ALP and mineralization, likely via modulation of cell-matrix interactions rather than classical osteogenic gene activation. These findings support using xHyA-functionalized membranes in GBR and warrant further investigation in vivo.