Three-dimensional (3D) bioprinting, where cells, hydrogels, and structural polymers can be printed layer by layer into complex designs, holds great promise for advances in medicine and the biomedical sciences. In principle, this technique enables the creation of highly patient-specific disease models and biomedical implants. However, an ability to tailor surface biocompatibility and interfacial bonding between printed components, such as polymers and hydrogels, is currently lacking. Here we demonstrate that an atmospheric pressure plasma jet (APPJ) can locally activate polymeric surfaces for the reagent-free covalent attachment of proteins and hydrogel in a single-step process at desired locations. Polyethylene and poly-ε-caprolactone were used as example polymers. Covalent attachment of the proteins and hydrogel was demonstrated by resistance to removal by rigorous sodium dodecyl sulfate washing. The immobilized protein and hydrogel layers were analyzed using Fourier transform infrared and X-ray photoelectron spectroscopy. Importantly, the APPJ surface activation also rendered the polymer surfaces mildly hydrophilic as required for optimum biocompatibility. Water contact angles were observed to be stable within a range where the conformation of biomolecules is preserved. Single and double electrode designs of APPJs were compared in their characteristics relevant to localized surface functionalization, plume length, and shape. As a proof of efficacy in a biological context, APPJ-treated polyethylene functionalized with fibronectin was used to demonstrate improvements in cell adhesion and proliferation. These results have important implications for the development of a new generation of 3D bioprinters capable of spatially patterned and tailored surface functionalization performed during the 3D printing process in situ.
Three-dimensional porous scaffolds have the potential to revolutionize a number of fields, including stem cell research, biomedical implants, tissue engineering, and regeneration, as well as energy technologies, filtration, and sensing. The ability to precisely engineer their surface properties is paramount to successful and enduring application. Plasma treatments promise to deliver homogeneous surfaces with tailored characteristics while generating few by-products and not relying on the diffusion of liquids into a porous network. However, forming plasma inside complex interconnected pores is a significant challenge and requires much parameter fine-tuning. This study uses numerical modeling to investigate key parameters which affect plasma breakdown and makes practical recommendations for achieving treatment homogeneity. We consider a dielectric barrier discharge (DBD) system, where nitrogen gas flows through a glass tube containing a porous scaffold, which is surrounded by a cylindrical electrode, with ground electrodes at either end of the tube. The parameters investigated include the scaffolds' pore size and porosity as well as electrode size and position. The results showed that homogeneous treatment is achieved by avoiding alternative breakdown pathways such as a poor seal between the scaffold and the glass containment tube, using a narrow supply electrode centered at the scaffold, and positioning ground electrodes as close as possible to the supply electrode without arcing. This strategy maximizes the electric field strength at a given voltage, allowing for higher pressures to be used, which in turn give a more homogeneous mean free path inside the pores.
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