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The biochemical and physical properties of a scaffold can be tailored to elicit specific cellular responses. However, it is challenging to decouple their individual effects on cell-material interactions. Here, we solvent-cast 3D printed different ratios of high and low molecular weight (MW) poly(caprolactone) (PCL) to fabricate scaffolds with significantly different stiffnesses without affecting other properties. Ink viscosity was used to match processing conditions between inks and generate scaffolds with the same surface chemistry, crystallinity, filament diameter, and architecture. Increasing the ratio of low MW PCL resulted in a significant decrease in modulus. Scaffold modulus did not affect human mesenchymal stromal cell (hMSC) differentiation under osteogenic conditions. However, hMSC response was significantly affected by scaffold stiffness in chondrogenic media. Low stiffness promoted more stable chondrogenesis whereas high stiffness drove hMSC progression toward hypertrophy. These data illustrate how this versatile platform can be used to independently modify biochemical and physical cues in a single scaffold to synergistically enhance desired cellular response., (© 2024 The Authors. Journal of Biomedical Materials Research Part A published by Wiley Periodicals LLC.)

Recent advances in extrusion-based filament 3D printing technology enable the processability of high-performance polymers. Poly(ether ether ketone) (PEEK) is an important group of high-performance polymer that has been widely used in aerospace, automotive, and biomedical applications. The interlayer bonding strength of 3D printed PEEK is crucial for load-bearing applications, yet studies on 3D printed PEEK are sparse due to processing challenges. In this study, the three-point flexural test is used to study the interlayer bonding strength of 3D-printed PEEK specimens with respect to the printing process parameters, including nozzle temperature, print speed, layer height, and wait-time. A design of experiment (DOE) approach is developed to study correlations between printing parameters and the end-use properties, including flexural stress (σf) and strain at break (εf), flexural modulus (Ef), and crystallinity (χ). Our results show that the nozzle temperature, layer height, and wait-time significantly affect the interlayer bonding strength, with nozzle temperature being the most influential parameter to enhance interlayer bonding strength indicated by a significant increase in σf, εf, and χ. Thermal annealing post-printing is shown to increase the degree of χ and Ef, yet its effect on interlayer bonding strength is minimal, indicating that the interlayer bonding strength is primarily determined during the printing process. This study demonstrates the use of a three-point flexural test integrated with a versatile and robust DOE approach to study the interlayer bonding strength of PEEK to reduce product development time while improving mechanical properties.

Three-dimensional (3D) printing of biodegradable polymers has rapidly become a popular approach to create scaffolds for tissue engineering. This technique enables fabrication of complex architectures and layer-by-layer spatial control of multiple components with high resolution. The resulting scaffolds can also present distinct chemical groups or bioactive cues on the surface to guide cell behavior. However, surface functionalization often includes one or more post-fabrication processing steps, which typically produce biomaterials with homogeneously distributed chemistries that fail to mimic the biochemical organization found in native tissues. As an alternative, our laboratory developed a novel method that combines solvent-cast 3D printing with peptide-polymer conjugates to spatially present multiple biochemical cues in a single scaffold without requiring post-fabrication modification. Here, we describe a detailed, stepwise protocol to fabricate peptide-functionalized scaffolds and characterize their physical architecture and biochemical spatial organization. We used these 3D-printed scaffolds to direct human mesenchymal stem cell differentiation and osteochondral tissue formation by controlling the spatial presentation of cartilage-promoting and bone-promoting peptides. This protocol also describes how to seed scaffolds and evaluate matrix deposition driven by peptide organization., (© 2013-2021 The Journal of Biological Methods, All rights reserved.)