Development of agarose–gelatin bioinks for extrusion-based bioprinting and cell encapsulation

Abstract

Three-dimensional bioprinting continues to advance as an attractive biofabrication technique to employ cell-laden hydrogel scaffolds in the creation of precise, user-defined constructs that can recapitulate the native tissue environment. Development and characterisation of new bioinks to expand the existing library helps to open avenues that can support a diversity of tissue engineering purposes and fulfil requirements in terms of both printability and supporting cell attachment. In this paper, we report the development and characterisation of agarose-gelatin hydrogel blends as a bioink for extrusion-based bioprinting. Four different agarose-gelatin hydrogel blend formulations with varying gelatin concentration were systematically characterised to evaluate suitability as a potential bioink for extrusion-based bioprinting. Additionally, autoclave and filter sterilisation methods were compared to evaluate their effect on bioink properties. Finally, the ability of the agarose-gelatin bioink to support cell viability and culture after printing was evaluated using SH-SY5Y cells encapsulated in bioprinted droplets of the agarose-gelatin. All bioink formulations demonstrate rheological, mechanical and swelling properties suitable for bioprinting and cell encapsulation. Autoclave sterilisation significantly affected the rheological properties of the agarose-gelatin bioinks compared to filter sterilisation. SH-SY5Y cells printed and differentiated into neuronal-like cells using the developed agarose-gelatin bioinks demonstrated high viability (>90%) after 23 days in culture. This study demonstrates the properties of agarose-gelatin as a printable and biocompatible material applicable for use as a bioink.

Effect of Flow Rate Modulation on Alginate Emulsification in Multistage Microfluidics

“…The phenomenon of spheroidal droplets produced from the plug form as shown in Figure 10 also confirmed the results of a previous study by Saeki [46]. Meanwhile, the outer double emulsion drop diameter in the current study is relatively larger compared to other studies, which makes the results of the current study applicable for bioink production in bioprinting technology, where bulk encapsulation of cells is preferred [47]. Comparison of droplet size from previous studies and current result is presented in Table 5.…”

Advancements of 3D bioprinting in regenerative medicine: Exploring cell sources for organ fabrication

“…The utilization of natural materials in 3D bioprinting has garnered significant attention due to their exceptional biocompatibility, biodegradability, and potential to enhance cell survival, function, adhesion, and self-organization [ [73] , [74] , [75] ]. Extensive research and optimization have been conducted on several natural materials, including collagen, gelatin, alginate, hyaluronic acid, and chitosan, to develop them as bioink materials for 3D bioprinting [ [76] , [77] , [78] , [79] , [80] , [81] , [82] , [83] , [84] ]. Acellular tissue matrix, which contains mixed tissue-specific ECM components, is often incorporated into the bioink materials to provide a tissue-specific environment with signaling and mechanical properties [ 85 , 86 ].…”

3D Bioprinting Using Hydrogels: Cell Inks and Tissue Engineering Applications

“…The addition of agarose to the cell ink allowed for greater viscosity, smaller drop size, and improved printer accuracy [24]. In a very recent extrusion-based bioprinting application, Dravid et al (2022) characterize agarose-gelatin hydrogel blends and describe desirable mechanical and rheological properties for bioprinting [25]. In addition, this group was able to print SH-SY5Y cells, which differentiated into neuronal-like cells using the described agarose-gelatin cell ink.…”