Innovative In Vivo Approaches to Bone Regeneration: Combining Chitosan‐Xanthan Gum Membranes, Electrical Stimulation, and Laser Therapy

Critical bone defects present significant clinical challenges due to their debilitating nature and associated costs. Tissue engineering offers a promising solution, employing biomaterials like porous chitosan (CH) and xanthan gum (XG) membranes as scaffolds for tissue regeneration. Electrical stimulation (ES) and laser (L) therapy, known for osteogenic and anti‐inflammatory effects, respectively, may synergistically enhance bone healing when combined with these membranes. This study evaluated CH/XG membranes and their association with ES and L for the treatment of critical bone defects (25 mm 2 ) created in 80 male Wistar rats. The animals were divided into four groups: untreated, CH/XG, CH/XG + ES, and CH/XG + L, assessed at days 30 and 90. Histomorphometric and real‐time PCR analyses revealed greater tissue and blood vessel formation in the CH/XG, CH/XG + ES, and CH/XG + L groups compared to the untreated group. At day 30, reduced Runx‐2 and Osx expression in the treated groups suggested an initial high inflammatory response. The CH/XG group showed pronounced inflammation (elevated Il‐1β and pro‐inflammatory markers) alongside an anti‐inflammatory response. Conversely, compared with the CH/XG‐only group, the CH/XG + ES and CH/XG + L groups generally showed lower expression of the evaluated pro‐inflammatory markers after 30 days, together with treatment‐ and time‐dependent changes in anti‐inflammatory markers. Although inflammation limited the initial effectiveness of CH/XG membranes, significant positive impacts were observed after 90 days. Combining membranes to cover bone defects and adjunctive therapies, as demonstrated in this study, emerges as a beneficial approach for enhancing bone repair outcomes. Adding osteoconductive or osteoinductive fillers may bring further healing improvements.