Physical Modification of Hybrid Hydrogels to Fabricate Full-Scale Construct Using Three-Dimensional Bio-Printing Process.

Bioprinting for regenerative medicine has been gaining a lot of popularity in today's world. Despite being one of the rigorously studied fields, there are still several challenges yet to be solved. Geometric fidelity and mechanical complexities stand as roadblocks when it comes to the printability of the customized constructs. Exploring the rheological properties of the compositions helps us understand the physical and mechanical properties of the biomaterials which are closely tied to the printability of the filament and eventually, geometric fidelity of the constructs. To ensure the structural integrity of the constructs, viscosity enhancers such as carboxymethyl cellulose (CMC) and crosslinkers like CaCl2 and CaSO4 were used. These crosslinkers can be used before (precrosslinking) and after (postcrosslinking) the extrusion of considered compositions to investigate and compare the outcome. To do this, mixtures of CMC (viscosity enhancer), Alginate, and CaCl2 and CaSO4 (crosslinkers) were prepared at various concentrations maintaining minimum solid content (≤8%). Each composition was subjected to a set of rheological tests like flow curve for shear thinning behavior, three points thixotropic for recovery rate, and amplitude test for gelation point. Various geometric fidelity identification tests were conducted and correlated with their physical properties. Some compositions were used to fabricate large-scale constructs (in cm-scale) to demonstrate their capability. This research is a thorough investigation of compositions when they are introduced to crosslinkers and viscosity enhancers which can be crucial for the 3D printing world.

3D Bio-Printability of Hybrid Pre-Crosslinked Hydrogels.

Maintaining shape fidelity of 3D bio-printed scaffolds with soft biomaterials is an ongoing challenge. Here, a rheological investigation focusing on identifying useful physical and mechanical properties directly related to the geometric fidelity of 3D bio-printed scaffolds is presented. To ensure during- and post-printing shape fidelity of the scaffolds, various percentages of Carboxymethyl Cellulose (CMC) (viscosity enhancer) and different calcium salts (CaCl2 and CaSO4, physical cross-linkers) were mixed into alginate before extrusion to realize shape fidelity. The overall solid content of Alginate-Carboxymethyl Cellulose (CMC) was limited to 6%. A set of rheological tests, e.g., flow curves, amplitude tests, and three interval thixotropic tests, were performed to identify and compare the shear-thinning capacity, gelation points, and recovery rate of various compositions. The geometrical fidelity of the fabricated scaffolds was defined by printability and collapse tests. The effect of using multiple cross-linkers simultaneously was assessed. Various large-scale scaffolds were fabricated (up to 5.0 cm) using a pre-crosslinked hybrid. Scaffolds were assessed for the ability to support the growth of Escherichia coli using the Most Probable Number technique to quantify bacteria immediately after inoculation and 24 h later. This pre-crosslinking-based rheological property controlling technique can open a new avenue for 3D bio-fabrication of scaffolds, ensuring proper geometry.