Abstract
The discovery of induced pluripotent stem cells (iPSCs) rendered the reprogramming of terminally differentiated cells to primary stem cells with pluripotency possible and provided potential for the regeneration and restoration of cartilage defect. Chondrogenic differentiation of iPSCs is crucial for their application in cartilage tissue engineering. In this study we investigated the effect of 3D nanofibrous scaffolds on the chondrogenesis of iPSCs and articular cartilage defect restoration. Super-hydrophilic and durable mechanic polycaprolactone (PCL)/gelatin scaffolds were fabricated using two separate electrospinning processes. The morphological structure and mechanical properties of the scaffolds were characterized. The chondrogenesis of the iPSCs in vitro and the restoration of the cartilage defect was investigated using scanning electron microscopy (SEM), the Cell Counting Kit-8 (CCK-8), histological observation, RT-qPCR, and western blot analysis. iPSCs on the scaffolds expressed higher levels of chondrogenic markers than the control group. In an animal model, cartilage defects implanted with the scaffold-cell complex exhibited an enhanced gross appearance and histological improvements, higher cartilage-specific gene expression and protein levels, as well as subchondral bone regeneration. Therefore, we showed scaffolds with a 3D nanofibrous structure enhanced the chondrogenesis of iPSCs and that iPSC-containing scaffolds improved the restoration of cartilage defects to a greater degree than did scaffolds alone in vivo.
Highlights
Cartilage loss and the healing of arthritis is difficult and an optimal solution remains unavailable due to the poor blood supply and regenerative capability of cartilage
Since Yamanaka and colleagues retrodifferentiated somatic cells to an ESC-like state, namely induced pluripotent stem cells (iPSCs), numerous reports regarding safer and more efficient methods for generating iPSCs for clinical applications have been published [5,6,7]. iPSCs can be induced into a variety of cell lineages, including the osteochondral lineage [8,9], and studies using scaffolds or gel carriers to enhance the chondrogenesis of iPSCs have been performed
Unlike the PCL fibrous membranes obtained from the solution, the prepared PCL scaffolds generated from the emulsion were hydrophilic due to the amphiphilic properties of the remnant surfactant
Summary
Cartilage loss and the healing of arthritis is difficult and an optimal solution remains unavailable due to the poor blood supply and regenerative capability of cartilage. A competent scaffold for cartilage reconstruction should provide the necessary mechanical strength, directed and controlled degradation, as well as the appropriate porosity to allow the nutrients and waste to diffuse, promoting cell proliferation [10,11]. Rigid scaffolds, such as poly(lactic-co- glycolic acid; PLGA) and PLA can provide more support under load, which may be important after the initial surgery, but may affect the properties of maturing chondrocytes and hyaline cartilage if not broken down appropriately; their application requires more invasive surgical implantation procedures [12,13]. Gel scaffolds generally lack sufficient mechanical strength; maintaining the original spatial structure and original site of implantation is difficult
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