Recent advances on enhancing 3D printing quality of protein-based inks: A review.

3D printing is an additive manufacturing technology that locates constructed models with computer-controlled printing equipment. To achieve high-quality printing, the requirements on rheological properties of raw materials are extremely restrictive. Given the special structure and high modifiability under external physicochemical factors, the rheological properties of proteins can be easily adjusted to suitable properties for 3D printing. Although protein has great potential as a printing material, there are many challenges in the actual printing process. This review summarizes the technical considerations for protein-based ink 3D printing. The physicochemical factors used to enhance the printing adaptability of protein inks are discussed. The post-processing methods for improving the quality of 3D structures are described, and the application and problems of fourth dimension (4D) printing are illustrated. The prospects of 3D printing in protein manufacturing are presented to support its application in food and cultured meat. The native structure and physicochemical factors of proteins are closely related to their rheological properties, which directly link with their adaptability for 3D printing. Printing parameters include extrusion pressure, printing speed, printing temperature, nozzle diameter, filling mode, and density, which significantly affect the precision and stability of the 3D structure. Post-processing can improve the stability and quality of 3D structures. 4D design can enrich the sensory quality of the structure. 3D-printed protein products can meet consumer needs for nutritional or cultured meat alternatives.

The correlation between rheological properties and extrusion-based printability in bioink artifact quantification.

Bioinks for cell-based bioprinting face availability limitations. Furthermore, the bioink development process needs comprehensive printability assessment methods and a thorough understanding of rheological factors' influence on printing outcomes. To bridge this gap, our study aimed to investigate the relationship between rheological properties and printing outcomes. We developed a specialized bioink artifact specifically designed to improve the quantification of printability assessment. This bioink artifact adhered to established criteria from extrusion-based bioprinting approaches. Seven hydrogel-based bioinks were selected and tested using the bioink artifact and rheological measurement. Rheological analysis revealed that the high-performing bioinks exhibited notable characteristics such as high storage modulus, low tan(δ), high shear-thinning capabilities, high yield stress, and fast, near-complete recovery abilities. Although rheological data alone cannot fully explain printing outcomes, certain metrics like storage modulus and tan(δ) correlated well (R2 > 0.9) with specific printing outcomes, such as gap-spanning capability and turn accuracy. This study provides a comprehensive examination of bioink shape fidelity across a wide range of bioinks, rheological measures, and printing outcomes. The results highlight the importance of considering the holistic view of bioink's rheological properties and directly measuring printing outcomes. These findings underscore the need to enhance bioink availability and establish standardized methods for assessing printability.

Advancements in 3D food printing: a comprehensive overview of properties and opportunities.

3D printing has numerous applications in the food industry that may enhance diversity, quality, healthiness, and sustainability. This innovative additive manufacturing technology has the ability to specifically tailor food properties for individuals. Nevertheless, several challenges still need to be overcome before 3D printing can be utilized more widely in the food industry. This article focuses on the development and characterization of "food inks" suitable for 3D printing of foods. Specifically, the main factors impacting successfully printed foods are highlighted, including material properties and printing parameters. The creation of a 3D printed food with the appropriate quality and functional attributes requires understanding and control of these factors. Food ink printability is an especially important factor that depends on their composition, structure, and physicochemical properties. Previous studies do not sufficiently describe the precise design and operation of 3D printers in sufficient detail, which makes comparing results challenging. Additionally, important physicochemical characteristics utilized in traditional food are not consistently reported in 3D inks, such as moisture content, water activity, and microbial contamination, which limits the practical application of the results. For this reason, we highlight important factors impacting 3D ink formulation and performance, then provide suggestions for standardizing and optimizing 3D printed foods.

3D Printing of Monolithic Proteinaceous Cantilevers Using Regenerated Silk Fibroin.

Silk fibroin, regenerated from Bombyx mori, has shown considerable promise as a printable, aqueous-based ink using a bioinspired salt-bath system in our previous work. Here, we further developed and characterized silk fibroin inks that exhibit concentration-dependent fluorescence spectra at the molecular level. These insights supported extrusion-based 3D printing using concentrated silk fibroin solutions as printing inks. 3D monolithic proteinaceous structures with high aspect ratios were successfully printed using these approaches, including cantilevers only supported at one end. This work provides further insight and broadens the utility of 3D printing with silk fibroin inks for the microfabrication of proteinaceous structures.

Void-free 3D Bioprinting for In-situ Endothelialization and Microfluidic Perfusion.

Two major challenges of 3D bioprinting are the retention of structural fidelity and efficient endothelialization for tissue vascularization. We address both of these issues by introducing a versatile 3D bioprinting strategy, in which a templating bioink is deposited layer-by-layer alongside a matrix bioink to establish void-free multimaterial structures. After crosslinking the matrix phase, the templating phase is sacrificed to create a well-defined 3D network of interconnected tubular channels. This void-free 3D printing (VF-3DP) approach circumvents the traditional concerns of structural collapse, deformation and oxygen inhibition, moreover, it can be readily used to print materials that are widely considered "unprintable". By pre-loading endothelial cells into the templating bioink, the inner surface of the channels can be efficiently cellularized with a confluent endothelial layer. This in-situ endothelialization method can be used to produce endothelium with a far greater uniformity than can be achieved using the conventional post-seeding approach. This VF-3DP approach can also be extended beyond tissue fabrication and towards customized hydrogel-based microfluidics and self-supported perfusable hydrogel constructs.

Void-free 3D Bioprinting for In-situ Endothelialization and Microfluidic Perfusion.

Two major challenges of 3D bioprinting are the retention of structural fidelity and efficient endothelialization for tissue vascularization. We address both of these issues by introducing a versatile 3D bioprinting strategy, in which a templating bioink is deposited layer-by-layer alongside a matrix bioink to establish void-free multimaterial structures. After crosslinking the matrix phase, the templating phase is sacrificed to create a well-defined 3D network of interconnected tubular channels. This void-free 3D printing (VF-3DP) approach circumvents the traditional concerns of structural collapse, deformation and oxygen inhibition, moreover, it can be readily used to print materials that are widely considered "unprintable". By pre-loading endothelial cells into the templating bioink, the inner surface of the channels can be efficiently cellularized with a confluent endothelial layer. This in-situ endothelialization method can be used to produce endothelium with a far greater uniformity than can be achieved using the conventional post-seeding approach. This VF-3DP approach can also be extended beyond tissue fabrication and towards customized hydrogel-based microfluidics and self-supported perfusable hydrogel constructs.

Low-temperature solvent-based 3D printing of PLGA: a parametric printability study.

In this paper, a novel low-temperature 3 D printing technique is introduced and characterized through a parametric printability study to fabricate poly-lactic-co-glycolic acid (PLGA) constructs using methyl ethyl ketone (MEK) as a solvent. The effects of varying concentrations of PLGA in MEK solvent, lactic to glycolic ratio of PLGA, the molecular weight of PLGA, and the scaling of PLGA constructs on the printability are investigated. PLGA concentrations of higher than 80% w/v, lactic to glycolic ratio more than 75%, molecular weight more than 100 kDa, and printing through nozzles smaller than 0.96 mm internal diameter are recommended for 3 D printing of PLGA constructs with high shape fidelity. Ultimately, a vacuum drying solvent removal process is implemented, and Proton Nuclear Magnetic Resonance (1H-NMR) spectroscopy is used to confirm complete removal of the solvent from PLGA constructs. The results of this study can be used for the development of drug-eluting implants.

Using Synergistic Multiple Dynamic Bonds to Construct Polymers with Engineered Properties.

Dynamic bonds have achieved significant attention for their ability to impart fascinating properties to polymeric materials, such as high mechanical strength, self-healing, shape memory, 3D printability, and conductivity. Incorporating multiple dynamic bonds into polymer systems affords an attractive and efficient approach to endow multiple functionalities. This mini-review focuses on the use of complementary dynamic interactions to control the properties of soft materials. Owing to the diversity in dynamic chemistries that can be explored, the scope of this article is restricted to polymers and does not include colloids, amphiphiles, liquid crystals, or biological soft matter.

Development of a Simple Mechanical Screening Method for Predicting the Feedability of a Pharmaceutical FDM 3D Printing Filament.

PURPOSE: The filament-based feeding mechanism employed by the majority of fused deposition modelling (FDM) 3D printers dictates that the materials must have very specific mechanical characteristics. Without a suitable mechanical profile, the filament can cause blockages in the printer. The purpose of this study was to develop a method to screen the mechanical properties of pharmaceutically-relevant, hot-melt extruded filaments to predetermine their suitability for FDM. METHODS: A texture analyzer was used to simulate the forces a filament is subjected to inside the printer. The texture analyzer produced a force-distance curve referred to as the flexibility profile. Principal Component Analysis and Correlation Analysis statistical methods were then used to compare the flexibility profiles of commercial filaments to in-house made filaments. RESULTS: Principal component analysis showed clearly separated clustering of filaments that suffer from mechanical defects versus filaments which are suitable for printing. Correlation scores likewise showed significantly greater values with feedable filaments than their mechanically deficient counterparts. CONCLUSION: The screening method developed in this study showed, with statistical significance and reproducibility, the ability to predetermine the feedability of extruded filaments into an FDM printer.

Current Status of Bioinks for Micro-Extrusion-Based 3D Bioprinting.

Recent developments in 3D printing technologies and design have been nothing short of spectacular. Parallel to this, development of bioinks has also emerged as an active research area with almost unlimited possibilities. Many bioinks have been developed for various cells types, but bioinks currently used for 3D printing still have challenges and limitations. Bioink development is significant due to two major objectives. The first objective is to provide growth- and function-supportive bioinks to the cells for their proper organization and eventual function and the second objective is to minimize the effect of printing on cell viability, without compromising the resolution shape and stability of the construct. Here, we will address the current status and challenges of bioinks for 3D printing of tissue constructs for in vitro and in vivo applications.

Three-Dimensional Bioprinting of GelMA Hydrogels with Culture Medium: Balancing Printability, Rheology and Cell Viability for Tissue Regeneration.

Three-dimensional extrusion bioprinting technology aims to become a fundamental tool for tissue regeneration using cell-loaded hydrogels. These biomaterials must have highly specific mechanical and biological properties that allow them to generate biosimilar structures by successive layering of material while maintaining cell viability. The rheological properties of hydrogels used as bioinks are critical to their printability. Correct printability of hydrogels allows the replication of biomimetic structures, which are of great use in medicine, tissue engineering and other fields of study that require the three-dimensional replication of different tissues. When bioprinting cell-loaded hydrogels, a small amount of culture medium can be added to ensure adequate survival, which can modify the rheological properties of the hydrogels. GelMA is a hydrogel used in bioprinting, with very interesting properties and rheological parameters that have been studied and defined for its basic formulation. However, the changes that occur in its rheological parameters and therefore in its printability, when it is mixed with the culture medium necessary to house the cells inside, are unknown. Therefore, in this work, a comparative study of GelMA 100% and GelMA in the proportions 3:1 (GelMA 75%) and 1:1 (GelMA 50%) with culture medium was carried out to determine the printability of the gel (using a device of our own invention), its main rheological parameters and its toxicity after the addition of the medium and to observe whether significant differences in cell viability occur. This raises the possibility of its use in regenerative medicine using a 3D extrusion bioprinter.

Boosting the Performance of PEDOT:PSS Based Electronics Via Ionic Liquids.

The conducting polymer poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) offers superior advantages in electronics due to its remarkable combination of high electrical conductivity, excellent biocompatibility, and mechanical flexibility, making it an ideal material among electronic skin, health monitoring, and energy harvesting and storage. Nevertheless, pristine PEDOT:PSS films exhibit limitations in terms of both low conductivity and stretchability; while, conventional processing techniques cannot enhance these properties simultaneously, facing the dilemma that highly conductive interconnected PEDOT:PSS domains are susceptible to tensile strain. Via modifying PEDOT:PSS with ionic liquids (ILs), not only a synergistic enhancement of the electrical and mechanical properties can be achieved but also the requirements for the printable bioelectronic are satisfied. In this comprehensive review, the task of providing a thorough examination of the mechanisms and applications of ILs as modifiers for PEDOT:PSS is undertaken. First, the theoretical mechanisms governing the interactions between ILs and PEDOT:PSS are discussed in detail. Then, the enhanced properties and the elucidation of the underlying mechanisms achieved through the incorporation of ILs are reviewed. Next, specific applications of ILs-modified PEDOT:PSS relevant to bioelectronic devices are presented. Last, there is a concise summary and a discussion regarding the opportunities and challenges in this exciting field.

Shrinkage and Cracking Properties of Cellulose Fiber-Concrete Composites for 3D Printing by Leveraging Internal Curing.

Compared with conventional formwork casting materials, 3D printed concrete (3DPC) is characterized by large amounts of cementitious materials, a low aggregate-binder ratio, and a large water evaporation area, which make the printed materials and structures highly prone to plastic shrinkage and cracking. In this study, cellulose fibers were incorporated into concrete to improve its moisture distribution and increase its early-age strength. The effects of both dry and prewet cellulose fibers on properties of 3DPC were experimentally investigated. To ensure consistency in the amounts of dry fibers used, 0.5-2% dry cellulose fibers and 1-4% prewet cellulose fibers were adopted. The effects of the added cellulose fibers on printability, mechanical strength, shrinkage, and cracking performance of the 3DPC were experimentally studied. Particularly, a constraint method was developed to access the cracking behavior of 3DPC. Favorable shrinkage resistance was achieved, and the 120-day shrinkage decreased by 17.9% and 23.3% by addition of 2% dry fibers and 4% prewet fibers, respectively. Cracking was eliminated with addition of 4% prewet fibers, without influencing the printability and mechanical properties.

Exploring Novel Applications for Hydrogels Derived from Modified Celluloses.

The valorization of lignocellulosic biomass by-products holds significant economic and ecological potential, considering their global overproduction. This paper introduces the fabrication of a novel wheat-straw-based hydrogel and a new microcellulose-based hydrogel through 2,2,6,6-tetramethylpiperidinyl-1-oxy (TEMPO) oxidation. In this study, Fourier transform infrared (FTIR) analysis was employed for the detection of carboxyl groups, neutralization titration was conducted using a conductivity meter, viscosity analysis was performed using a rheometer, and transmittance analysis was carried out using a spectrophotometer. Two novel hydrogels based on TEMPO oxidation have been developed. Among them, the bio-based hydrogel derived from oxidized wheat straw exhibited exceptional printability and injectability. We found that the oxidation degree of microcellulose reached 56-69%, and the oxidation degree of wheat straw reached 56-63%. The cross-linking of 4% oxidized wheat straw and calcium chloride was completed in 400 seconds, and the viscosity exceeded 100,000 Pa·s. In summary, we have successfully created low-cost hydrogels through the modification of wheat straw and microcellulose, transforming lignocellulosic biomass by-products into a sustainable source of polymers. This paper verifies the future applicability of biomass materials in 3D printing.

Printing GelMA bioinks: a strategy for buildingin vitromodel to study nanoparticle-based minocycline release and cellular protection under oxidative stress.

Owing to its thermoresponsive and photocrosslinking characteristics, gelatin methacryloyl (GelMA)-based biomaterials have gained widespread usage as a novel and promising bioink for three-dimensional bioprinting and diverse biomedical applications. However, the flow behaviors of GelMA during the sol-gel transition, which are dependent on time and temperature, present significant challenges in printing thick scaffolds while maintaining high printability and cell viability. Moreover, the tunable properties and photocrosslinking capabilities of GelMA underscore its potential for localized drug delivery applications. Previous research has demonstrated the successful incorporation of minocycline (MH) into GelMA scaffolds for therapeutic applications. However, achieving a prolonged and sustained release of concentrated MH remains a challenge, primarily due to its small molecular size. The primary aim of this study is to investigate an optimal extrusion printing method for GelMA bioink in extrusion bioprinting, emphasizing its flow behaviors that are influenced by time and temperature. Additionally, this research seeks to explore the potential of GelMA bioink as a carrier for the sustained release of MH, specifically targeting cellular protection against oxidative stress. The material properties of GelMA were assessed and further optimization of the printing process was conducted considering both printability and cell survival. To achieve sustained drug release within GelMA, the study employed a mechanism using metal ion mediation to facilitate the interaction between MH, dextran sulfate (DS), and magnesium, leading to the formation of nanoparticle complexes (MH-DS). Furthermore, a GelMA-basedin vitromodel was developed in order to investigate the cellular protective properties of MH against oxidative stress. The experimental results revealed that the printability and cell viability of GelMA are significantly influenced by the printing duration, nozzle temperature, and GelMA concentrations. Optimal printing conditions were identified based on a thorough assessment of both printability and cell viability. Scaffolds printed under these optimal conditions exhibited exceptional printability and sustained high cell viability. Notably, it was found that lower GelMA concentrations reduced the initial burst release of MH from the MH-dextran sulfate (MH-DS) complexes, thus favoring more controlled, sustained release profiles. Additionally, MH released under these conditions significantly enhanced fibroblast viability in anin vitromodel simulating oxidative stress.

Impact of starch amylose and amylopectin on the rheological and 3D printing properties of corn starch.

This study evaluated the influence of the amylose and amylopectin on the physicochemical properties and printing performance of corn starch gels. Amylose in starch-based gels enhances their storage modulus and the support performance of printed products by promoting the formation of cross-linked gel structures and crystalline structures. However, the higher amylose content in starch gels makes extrusion difficult, resulting in intermittent extrusion in 3D printing. Despite the increased shear-thinning ability of high-amylose starch, its low water retention capacity leads to water loss and rough printed morphology. Additionally, starch with 72 % amylose content exhibits insufficient adhesive properties for effective layer bonding, negatively impacting structural integrity. While gels with 72 % and 56 % amylose content demonstrate higher viscosity and enhanced mechanical properties, their poor adhesion limits the quality of printed layers. Conversely, waxy starch gel demonstrates continuous extrusion and adhesion but lacks adequate support. The 27 % corn starch gel achieves the highest 3D printing accuracy at 88.12 %, suggesting an optimal amylose-amylopectin ratio for desired ink material performance. These findings enhance our understanding of the relationship between amylose content in starch and 3D printing performance, providing a theoretical basis for the development of starch-based printing products.

Improvement of 3D printing age-friendly brown rice food on rough texture, swallowability, and in vitro digestibility using fermentation properties of different probiotics.

Probiotics can promote the balance of the intestinal microbial community and enhance the biological activity of food. They are beneficial to the health of elderly people. Therefore, five different probiotics (4% of the total weight) were added to pasted brown rice to study the printability, swallowability, and digestibility of fermented inks (at 40 °C for 10 h). The results showed that probiotics reduced the apparent viscosity and resistance to deformation of brown rice inks. The inks with Lactobacillus bulgaricus (LB), Bifidobacterium longum (BL), and Lactiplantibacillus plantarum (LP) had better printing properties and finer appearances. Probiotics significantly reduced the adhesiveness, gumminess, and hardness of inks but had little effect on cohesiveness. LB, Streptococcus thermophilus (ST), and LP were categorized as having class 4 consistency with easy-to-swallow characteristics. The growth and multiplication of probiotics detached the internal structure of brown rice inks and reduced the relative crystallinity. They also modulated the nutrient content and flavor components by producing short-chain fatty acids, and improved the digestion of starch.

Preparation of shellac nanoparticles-chitosan complexes stabilized Pickering emulsion gels and its application in ß-carotene delivery.

Shellac nanoparticles (SNPs)-based Pickering emulsion gels show promise as delivery carriers but face challenges due to poor emulsifying properties. This study aimed to fabricate stable emulsion gels using SNPs and chitosan (CS) complexes, creating a ß-carotene delivery system. The effects of oil phase fractions, emulsifier concentrations and SNPs/CS ratios on rheological properties and the structural properties of emulsion were investigated. The formation of SNPs/CS complexes was through hydrogen bonding and electrostatic interactions. By adjusting the SNPs/CS ratio to 1/0.33, the contact angle of the complexes was optimized to approximately 90°. SNPs/CS complexes served dual roles as emulsifiers and gelling agents in the emulsion gels. Notably, the gel strength (storage modulus) of the emulsion gels remained unchanged after the encapsulation of ß-carotene. Emulsion gels with SNPs/CS (1/0.25) complexes showed the highest ß-carotene bioaccessibility at 80.4 %. Furthermore, this system could expand the use of shellac-based emulsion gels in food applications.

Recent research and applications in lipid-based food and lipid-incorporated bioink for 3D printing.

Three-dimensional (3D) printing, as an emerging digital production technology, has recently been receiving increasing attention in food processing. It is important to understand the effect of key ingredients of food materials on the printing, which makes it possible to achieve a wider range of structures using few nozzles and to provide tailored nutrition and personalization. This comprehensive review delves into the latest research on 3D-printed lipid-based foods, encompassing a variety of products such as chocolate, processed cheese, as well as meat. It also explores the development and application of food bioinks that incorporate lipids as a pivotal component, including those based on starch, protein, oleogels, bigels, and emulsions, as well as emulsion gels. Moreover, this review identifies the current challenges and presents an outlook on future research directions in the field of 3D food printing, especially the research and application of lipids in food 3D printing.

Application of xanthan and locust bean gum mix or sorbitol in the jelly formulation to improved jelly 3D printing using a fused deposition modeling printer.

This study examined the impacts of applying a xanthan and locust bean gum mix or sorbitol to a jelly formulation on the rheological parameters necessary for 3D printing a jelly applying the fused deposition modeling method. A jelly formulation was fortified with a gum mix (xanthan gum:locust bean gum = 0.625:0.375) at 1% (w/w), or added with sorbitol instead of sugar. Both treatments increased the values of storage modulus and yield stress, related to fidelity and shape retention, and adding the gum mix, in particular, increased the gel strength. Applying these treatments to the formulation that lacks the rheological parameters and gel strength required for 3D printing changed those values in a direction fulfilling the material requirements. This research confirmed that the application of xanthan and locust bean gum mix or sorbitol could adjust the properties of materials used in 3D printing for improved printability.