Enhanced osteo–angiogenic coupling by a bioactive cell-free fat extract (CEFFE) delivered through electrospun fibers

Regeneration of functional bone tissue relies heavily on achieving adequate vascularization in engineered bone constructs following implantation. This process requires the close integration of osteogenesis and angiogenesis. Cell-free fat extract (CEFFE or FE), a recently emerging acellular fat extract containing abundant growth factors, holds significant potential for regulating osteo-angiogenic coupling and promoting regeneration of vascularized bone tissue. However, its specific role in modulating the coupling between angiogenesis and osteogenesis remains unclear. Our previous research demonstrated that FE-decorated electrospun fibers of polycaprolactone/gelatin (named FE-PDA@PCL/GT) exhibited pro-vasculogenic capabilities both <i>in vitro</i> and <i>in vivo</i> (D. Li, Q. Li, T. Xu, X. Guo, H. Tang, W. Wang, W. Zhang and Y. Zhang, Pro-vasculogenic fibers by PDA-mediated surface functionalization using cell-free fat extract (CEFFE), <i>Biomacromolecules</i> 2024, <b>25</b>, 1550-1562). Herein, we firstly demonstrated that the FE-PDA@PCL/GT fibers also significantly stimulated osteogenesis in a mouse calvaria osteoblast-like cell line MC3T3-E1 cells, as evidenced by the increased production of alkaline phosphatase (ALP), mineral deposits, and collagen I, as well as the upregulated expression of osteogenic marker genes in the osteoblasts. Using a transwell co-culture system, we further demonstrated that the release of FE from the FE-PDA@PCL/GT fibers not only promoted osteogenesis and angiogenesis but also markedly enhanced the paracrine functions and reciprocal communications between endothelial cells and osteoblasts. This dynamic interaction played a key role in the observed enhancement of osteo-angiogenic coupling. With the confirmed pro-osteogenic and pro-angiogenic properties of FE-PDA@PCL/GT, it is envisaged that these newly engineered bioactive fibers can be used to develop highly biomimicking bone constructs. These constructs are designed to promote native-like cell-scaffold and cell-cell interactions, which are essential for the effective regeneration of defected bone tissue with adequate vasculature.