Purpose: Bone tissue engineering is a promising approach for bone reconstruction in oral and maxillofacial surgery. The aim of this study was to investigate the microstructure and biocompatibility of a novel albumin scaffold developed from human serum on human alveolar osteoblasts. Materials and Methods: Samples of mandibular bone were obtained during routine oral surgery. Osteoblast cells were cultured and plated in a spongy, noncalcified protein scaffold prepared with plasmatic albumin crossed with a glutaraldehyde-type agent (study group) and in a large-particle mineralized cancellous allograft (control group). Measurement of the differentiation marker alkaline phosphatase and histologic examination were performed after 30 days of incubation. The cultures were examined for cell growth patterns and morphology by scanning electron microscopy and histomorphometry. Results: Cultured osteoblasts showed comparable phenotypic profiles and expressed alkaline phosphatase in albumin scaffold. Hematoxylin-eosin staining revealed a bonelike extracellular matrix in study scaffold and mineralization of osteoblasts cultured in the albumin scaffold was confirmed by von Kossa staining. Conclusion: Osteoblasts were able to proliferate in vitro and synthesize a bonelike extracellular matrix and mineralized tissue. The results indicate that this novel albumin scaffold is a favorable substrate for the growth and differentiation of osteoblasts and a promising material for bone tissue engineering and repair of bone defects. Keywords: bone culture, osteoblasts, scaffold, tissue engineering