Three-Dimensional Printing of Hydrogel as Skin Substitute and Comparative Evaluation of Melanin Production

Cell culture in two dimensions has been the main instrument in cellular and molecular biology. But there are limitations to two-dimensional culture when it comes to tissue engineering and in vivo reproduction. Tissue engineering technology enabled the creation of biomedical scaffolds, which are mostly utilized to biofabricate different artificial human organs. Tissue architecture that encourage cell proliferation can be produced using direct bioprinting technology. The development of bioinks for 3D bioprinting is consistently seen as a problem in the domains of biofabrication and tissue engineering. This study aimed to determine if Fibroblasts and Keratinocytes could grow on hydrogel scaffolds as efficiently as they can in the culture plates. Melanocytes were co-cultured, and the production of melanin was assessed in a two- and three-dimensional culture system. Scaffolds were fabricated using 8% alginate and 6% gelatin and 3D-printed using a cell link printer. FTIR was used to determine the precise composition of the gels. SEM analysis was performed for the cells present in gel and the topology of the cells. In addition, 8% alginate and 6% alginate gel scaffolds were analyzed for swelling and degradation over time in the cell growth medium and PBS. Furthermore, a gene expression study of cell cultures on scaffolds was performed through qPCR. A live/dead assay was performed to determine cell viability for cells grown on scaffolds for 7, 14, and 21 days. Most of the cells were shown to be viable, similar to the control cells grown on a plate. The findings from the SEM showed that cells were grown on the gel surface, remained viable even after 21 days, and displayed circular cells stacked three-dimensionally on the gel surface in the 3D scaffold. The MTT assay was performed to check the viability of cells cultured on a 3D-printed scaffold for 1, 5, and 15 days. We observed about 40% viable cells after 15 days, as shown by the MTT assay. Furthermore, a co-culture study with Melanocyte showed an increased production of melanin in a 3D culture as compared to a 2D culture. Our findings suggest that an alginate and gelatin polymer can be used as a cellular matrix for epithelial cell culture. Further, in vivo and ex vivo experiments are needed to validate the results for future applications in tissue engineering for wound healing and other tissue engineering domains.