Hybrid 3D bioprinting of functionalized structures for tissue engineering

Official URL: https://risc01.sabanciuniv.edu/record=b2553611 _(Table of contents)

Tissue engineering is an interdisciplinary field of research aiming at developing methods and technologies for regenerating damaged tissues. It relies on a combinatory platform of biomaterials with cells and bioactive molecules to resemble the human microenvironment to stimulate tissue constructs. Hence, numerous factors, including biochemical, biophysical, and mechanical aspects of the host tissue, have to be taken into account for developing a successful tissue replacement. Skin replacements caused by traumas, injuries, and burns are a burden to the healthcare system globally. The human body cannot fully regenerate the tissue with all the functionalities and features in severe wounds or skin loss. Poor mechanical properties, scarring, delayed cell and biomolecules infiltration, and non/poor vascularization are the main challenges yet to be addressed. Three-dimensional (3D) bioprinting, also known as additive manufacturing (AM), a layer-by-layer fabrication method, is regarded as a gold standard technique with the ability of controlled deposition of biomaterials in the desired geometry by using computer-aided design (CAD) models. Together with the development of biomaterials and architecture design, 3D bioprinting could ease the long and complicated journey towards functional tissue regeneration. In this context, the fabrication of small fibers mimicking natural extracellular matrix (ECM), selection of functional material with good mechanical and biochemical properties, the inclusion of bioactive molecules to enhance functionality, and printability are prerequisite factors of successful scaffold fabrication. In this work, novel hybrid 3D bioprinting approaches have been developed for functionalized structures, mainly for skin tissue engineering. Within this framework, we first optimized the effect of printing parameters on fiber diameter for Melt Electrospinning Writing (MEW), a special 3D printing process, using response surface methodology (RSM) as a predictive tool. Then we copolymerized polycaprolactone (PCL) with polypropylene succinate to improve its degradation rate and hydrophilicity and functionalized it with silver nitrate to induce antibacterial properties, and finally, it was 3Dprinted using an extrusion-based printer. For preparing hybrid 3D bioprinting, we used a composite support-bath system based on Pluronic PF127 was formulated with the inclusion of Laponite RDS and calcium chloride as rheological modifier and stabilizer, respectively. The rheological characterization of support-bath showed thixotropic behavior with a high degree of recoverability which facilitated bioprinting of complex hydrogel structures within the support-bath through an extrusion system. Then, we fabricated a polymer-hydrogel construction using MEW-casting for skin tissue substitute. In this context, we first investigated the geometrical effect of melt electrowritten scaffolds on cord-like structure formation for pre-vascularization. Mesh scaffolds with 0-90and 60-120 degree orientations and honeycomb shape were explored and cell-laden gelatin hydrogels were infiltrated inside those PCL scaffolds, and the results suggested the potential of honeycomb structure for better mechanical and invitro properties. In the final stage, a functionalized hybrid MEW-hydrogel scaffold for wound healing was fabricated. A functionalized mesh structure of PCL-bioactive glass was created via MEW, and a gelatin hydrogel comprising basic fibroblast and vascular endothelial growth factors was cast within the mesh scaffold. In vivo implantation of hybrid scaffolds showed promising results for accelerating and functionality of the healed parts according to wound closure and histological evaluation.