Electrospun nanofibers have shown great potential as scaffolds for regenerative medicine because of its biomimicry. However, traditional two-dimensional electrospun nanofiber mats inhibit their applications due to the dense structure and lack of effective cell infiltration. Herein, we report a new method of expanding electrospun nanofiber mats in the third dimension using a modified gas-foaming technique. The resulted nanofiber scaffolds show layered structures with controllable gap widths and layer thicknesses on the order of microns. Expanded nanofiber scaffolds possess significantly higher porosity than traditional two-dimensional nanofiber membranes, while simultaneously maintaining nanotopographic cues. The distributions of gap widths and layer thicknesses are directly dependent on the processing time of nanofiber mats within the gas bubble forming solution. In vitro testing demonstrates robust cellular infiltration and proliferation within expanded nanofiber scaffolds as compared to limited cellular proliferation on the surface of traditional nanofiber mats. Importantly, cell alignment was observed throughout the expanded and aligned nanofiber scaffolds after incubation for 7 days. The presented method was further applied to fabricate tubular scaffolds composed of expanded nanofibers. Together, this novel class of scaffolds holds significant promise for applications in regenerative medicine and building 3D in vitro tissue models for drug screening and biological study.

Herein, we report a robust method to fabricate expanded nanofiber scaffolds with controlled size and thickness using a customized mold during the modified gas-foaming process. The expansion of nanofiber membranes was also simulated using a computational fluid model. Expanded nanofiber scaffolds implanted subcutaneously in rats showed cellular infiltration, whereas non-expanded scaffolds only had surface cellular attachment. Compared to unexpanded nanofiber scaffolds, more CD68+ and CD163+ cells were observed within expanded scaffolds at all tested time points post-implantation. More CCR7+ cells appeared within expanded scaffolds at week 8 post-implantation. In addition, new blood vessels were present within the expanded scaffolds at week 2. The formed multinucleated giant cells within expanded scaffolds were heterogeneous expressing CD68, CCR7, or CD163 markers. Together, the present study demonstrated that the expanded nanofiber scaffolds promoted cellular infiltration/tissue integration, a regenerative response, and neovascularization after subcutaneous implantation in rats. The use of expanded electrospun nanofiber scaffolds offers a promising method for in situ tissue repair/regeneration and generation of three-dimensional tissue models/constructs.

Electrospun bioactive glass fibers show great potential as scaffolds for bone tissue engineering due to their architectural biomimicry of the bone extracellular matrix and their composition capable of providing soluble bioactive cues for bone regeneration and remodeling. Trace elements can be doped to further promote osteogenesis and angiogenesis during bone regeneration. Cationic substitution of strontium for calcium in bioactive glass positively enhances osteoblast phenotype, while suppressing osteoclast activity. Further, the addition of copper spontaneously improves the vascularization during neobone formation. The objective of this study was to fabricate and characterize electrospun bioactive glass fibers doped with strontium and copper and evaluate their potential for bone repair/regeneration in vitro. Different ratios of strontium and copper were doped in electrospun bioactive glass fibers. The released strontium and copper from doped fibers could reach effective concentrations within 40 h and last for 4 weeks. These bioactive glass fibers demonstrate their bioactivity by promoting osteoblastic and endothelial cell activity and inhibiting the formation of osteoclasts or bone resorbing cells. Additionally, in vitro cell culture of different cell types in the presence of extraction solutions of the electrospun bioactive glass fibers showed that the dopants achieved their individual goals without causing significant cytotoxicity. Altogether, this novel class of bioactive glass fibers holds great promise for bone regeneration.

An ideal synthetic bone graft is a combination of the porous and nanofibrous structure presented by natural bone tissue as well as osteoinductive biochemical factors such as bone morphogenetic protein 2 (BMP-2). In this work, ultralight 3D hybrid nanofiber aerogels composed of electrospun PLGA-collagen-gelatin and Sr-Cu codoped bioactive glass fibers with incorporation of heptaglutamate E7 domain specific BMP-2 peptides have been developed and evaluated for their potential in cranial bone defect healing. The nanofiber aerogels are surgically implanted into 8 mm × 1 mm (diameter × thickness) critical-sized defects created in rat calvariae. A sustained release of E7-BMP-2 peptide from the degradable hybrid aerogels significantly enhances bone healing and defect closure over 8 weeks in comparison to unfilled defects. Histomorphometry and X-ray microcomputed tomography (µ-CT) analysis reveal greater bone volume and bone formation area in case of the E7-BMP-2 peptide loaded hybrid nanofiber aerogels. Further, histopathology data divulged a near complete nanofiber aerogel degradation along with enhanced vascularization of the regenerated tissue. Together, this study for the first time demonstrates the fabrication of 3D hybrid nanofiber aerogels from 2D electrospun fibers and their loading with therapeutic osteoinductive BMP-2 mimicking peptide for cranial bone tissue regeneration.

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