Bioprinting and Preliminary Testing of Highly Reproducible Novel Bioink for Potential Skin Regeneration

Abstract

Three-dimensional (3D) bioprinting is considered as a novel approach in biofabricating cell-laden constructs that could potentially be used to promote skin regeneration following injury. In this study, a novel crosslinked chitosan (CH)–genipin (GE) bioink laden with keratinocyte and human dermal fibroblast cells was developed and printed successfully using an extruder-based bioprinter. By altering the composition and degree of CH–GE crosslinking, bioink printability was further assessed and compared with a commercial bioink. Rheological analysis showed that the viscosity of the optimised bioink was in a suitable range that facilitated reproducible and reliable printing by applying low pressures ranging from 20–40 kPa. The application of low printing pressures proved vital for viability of cells loaded within the bioinks. Further characterisation using MTT assay showed that cells were still viable within the printed construct at 93% despite the crosslinking, processing and after subjecting to physiological conditions for seven days. The morphological study of the printed cells showed that they were mobile within the bioink. Furthermore, the multi-layered 3D printed constructs demonstrated excellent self-supportive structures in a consistent manner.

Current Insight of Printability Quality Improvement Strategies in Natural-Based Bioinks for Skin Regeneration and Wound Healing

“…Bioinks are known as biological materials fluids loaded into the 3D-bioprinter system to construct scaffolds with a specific design using a layer-by-layer printing technique [ 20 ]. The main components involved to create the bioinks are biomaterials, cells, and suitable crosslinkers.…”

“…Human dermal fibroblasts (HDFs) are differentiated from highly proliferative progenitor fibroblasts and responsible for maintaining skin tissues’ structural support [ 20 ]. During the formation of the dermal and wound repair process, HDFs are essential in response to stimuli and ECM proteins’ formation [ 21 ].…”

Current Advances in 3D Bioprinting Technology and Its Applications for Tissue Engineering

“…Admane et al proved that 3D bioprinted skin fabricated using a mixture of silk and gelatin was dimensionally stable relative to the collagen-based skin structure [ 179 ]. More recently, Hafezi et al designed an alginate and chitosan–genipin–PEG-based three-layer skin tissue [ 180 ]. They also optimized the crosslinking ratio of chitosan and genipin to alter printability, comparable to that of commercial bio-ink.…”

State of the Art on Biomaterials for Soft Tissue Augmentation in the Oral Cavity. Part II: Synthetic Polymers-Based Biomaterials

“…The 3D construct is assembled in an additive manner by printing layer by layer [ 34 ] (arrow). Figure reproduced and adapted from Hafezi et al (2020) [ 36 ]. Copyright MDPI, 2020.…”