Development of biomimetic mouldable osseous scaffold engineered from bioactive decellularized extracellular matrix-polymer-gel composite: a potential bone substitute.

Critical-sized bone defects remain a major clinical challenge in veterinary orthopaedics due to limited intrinsic regenerative capacity and the drawbacks of conventional grafting options. This study reports the development and systematic evaluation of a novel bioinspired osseous scaffold engineered from decellularized bubaline cancellous bone granules, collagen gel, and polycaprolactone, with optional enrichment using autologous adipose-derived stromal vascular fraction. Scaffold physicochemical characteristics-including porosity, material density, and ultrastructure-were assessed through liquid displacement assays, histomorphometry, and scanning electron microscopy, confirming complete decellularization, interconnected porosity, and favourable matrix architecture. Mechanical testing demonstrated significantly enhanced tensile strength in the composite scaffold compared with plain acellular bone-collagen constructs. In vitro biocompatibility was validated by adipose derived mesenchymal stem cell adhesion, proliferation, and migration, with DAPI staining confirming uniform cellular distribution and robust scaffold-cell interactions, along with Alizarin Red S Staining for assessment of osteogenic mineralization.