Osteogenic culture of human adipose stem cells on electrospun multispiral poly(ε-caprolactone)/β-tricalcium phosphate composite scaffold in a perfusion bioreactor.

The present study investigated the effect of a dynamic perfusion system on the osteogenic differentiation of human adipose-derived mesenchymal stem cells (hASCs) cultured within a 3D multispiral scaffold architecture. The scaffolds were constructed by winding electrospun poly(ε-caprolactone)/β-tricalcium phosphate (PCL/β-TCP) composite membranes into hierarchical multispiral structures, thereby providing a biomimetic microenvironment for cell growth and tissue formation. To evaluate the role of mechanical stimuli on mineralization, cell-laden constructs were comparatively assessed in two different environments over a 28-day period: (i) conventional static culture and (ii) dynamic perfusion culture in a custom-designed bioreactor operated at a constant flow rate of 4.375 mL/min per channel. The in vitro biological performance and structural development of the engineered constructs were characterized using scanning electron microscopy, histological analyses (hematoxylin and eosin, Von Kossa, and Alizarin Red S stainings), and quantitative biochemical assays, including the MTT assay for metabolic activity and the o-cresolphthalein complexone assay for calcium deposition. The results revealed that while both environments facilitated robust cellular viability, the dynamic perfusion system significantly enhanced mineralized matrix production and mechanical performance compared to the static group. Overall, these findings indicate that the synergistic combination of perfusion-induced mechanical stimulation and hierarchical multispiral PCL/β-TCP scaffolds create a favorable osteogenic microenvironment that significantly enhances mineralized tissue formation. This integrated strategy establishes a promising platform for bone tissue engineering and offers potential for the regeneration of complex skeletal disorders.