Editorial

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3 pages, 177 KiB

Open AccessEditorial

Biocomposite Inks for 3D Printing

by

Gary Chinga-Carrasco

Bioengineering 2021, 8(8), 102; https://doi.org/10.3390/bioengineering8080102 - 22 Jul 2021

Viewed by 2040

Abstract

Three-dimensional (3D) printing has evolved massively during the last years and is demonstrating its potential in tissue engineering, wound dressings, cell culture models for drug testing, and prosthesis, to name a few [...] Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

Research

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14 pages, 3027 KiB

Open AccessArticle

3D Printed Nanocellulose Scaffolds as a Cancer Cell Culture Model System

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Bioengineering 2021, 8(7), 97; https://doi.org/10.3390/bioengineering8070097 - 10 Jul 2021

Cited by 11 | Viewed by 3298

Abstract

Current conventional cancer drug screening models based on two-dimensional (2D) cell culture have several flaws and there is a large need of more in vivo mimicking preclinical drug screening platforms. The microenvironment is crucial for the cells to adapt relevant in vivo characteristics [...] Read more.

Current conventional cancer drug screening models based on two-dimensional (2D) cell culture have several flaws and there is a large need of more in vivo mimicking preclinical drug screening platforms. The microenvironment is crucial for the cells to adapt relevant in vivo characteristics and here we introduce a new cell culture system based on three-dimensional (3D) printed scaffolds using cellulose nanofibrils (CNF) pre-treated with 2,2,6,6-tetramethylpyperidine-1-oxyl (TEMPO) as the structural material component. Breast cancer cell lines, MCF7 and MDA-MB-231, were cultured in 3D TEMPO-CNF scaffolds and were shown by scanning electron microscopy (SEM) and histochemistry to grow in multiple layers as a heterogenous cell population with different morphologies, contrasting 2D cultured mono-layered cells with a morphologically homogenous cell population. Gene expression analysis demonstrated that 3D TEMPO-CNF scaffolds induced elevation of the stemness marker CD44 and the migration markers VIM and SNAI1 in MCF7 cells relative to 2D control. T47D cells confirmed the increased level of the stemness marker CD44 and migration marker VIM which was further supported by increased capacity of holoclone formation for 3D cultured cells. Therefore, TEMPO-CNF was shown to represent a promising material for 3D cell culture model systems for cancer cell applications such as drug screening. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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15 pages, 3625 KiB

Open AccessArticle

Advanced Bioink for 3D Bioprinting of Complex Free-Standing Structures with High Stiffness

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Bioengineering 2020, 7(4), 141; https://doi.org/10.3390/bioengineering7040141 - 07 Nov 2020

Cited by 26 | Viewed by 4970

Abstract

One of the challenges in 3D-bioprinting is the realization of complex, volumetrically defined structures, that are also anatomically accurate and relevant. Towards this end, in this study we report the development and validation of a carboxylated agarose (CA)-based bioink that is amenable to [...] Read more.

One of the challenges in 3D-bioprinting is the realization of complex, volumetrically defined structures, that are also anatomically accurate and relevant. Towards this end, in this study we report the development and validation of a carboxylated agarose (CA)-based bioink that is amenable to 3D printing of free-standing structures with high stiffness at physiological temperature using microextrusion printing without the need for a fugitive phase or post-processing or support material (FRESH). By blending CA with negligible amounts of native agarose (NA) a bioink formulation (CANA) which is suitable for printing with nozzles of varying internal diameters under ideal pneumatic pressure was developed. The ability of the CANA ink to exhibit reproducible sol-gel transition at physiological temperature of 37 °C was established through rigorous characterization of the thermal behavior, and rheological properties. Using a customized bioprinter equipped with temperature-controlled nozzle and print bed, high-aspect ratio objects possessing anatomically-relevant curvature and architecture have been printed with high print reproducibility and dimension fidelity. Objects printed with CANA bioink were found to be structurally stable over a wide temperature range of 4 °C to 37 °C, and exhibited robust layer-to-layer bonding and integration, with evenly stratified structures, and a porous interior that is conducive to fluid transport. This exceptional layer-to-layer fusion (bonding) afforded by the CANA bioink during the print obviated the need for post-processing to stabilize printed structures. As a result, this novel CANA bioink is capable of yielding large (5–10 mm tall) free-standing objects ranging from simple tall cylinders, hemispheres, bifurcated ‘Y’-shaped and ‘S’-shaped hollow tubes, and cylinders with compartments without the need for support and/or a fugitive phase. Studies with human nasal chondrocytes showed that the CANA bioink is amenable to the incorporation of high density of cells (30 million/mL) without impact on printability. Furthermore, printed cells showed high viability and underwent mitosis which is necessary for promoting remodeling processes. The ability to print complex structures with high cell densities, combined with excellent cell and tissue biocompatibility of CA bodes well for the exploitation of CANA bioinks as a versatile 3D-bioprinting platform for the clinical translation of regenerative paradigms. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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12 pages, 2100 KiB

Open AccessArticle

Formulation and Characterization of Alginate Dialdehyde, Gelatin, and Platelet-Rich Plasma-Based Bioink for Bioprinting Applications

by

Lakshmi T. Somasekharan

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Naresh Kasoju

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Riya Raju

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Anugya Bhatt

Bioengineering 2020, 7(3), 108; https://doi.org/10.3390/bioengineering7030108 - 09 Sep 2020

Cited by 24 | Viewed by 6173

Abstract

Layer-by-layer additive manufacturing process has evolved into three-dimensional (3D) “bio-printing” as a means of constructing cell-laden functional tissue equivalents. The process typically involves the mixing of cells of interest with an appropriate hydrogel, termed as “bioink”, followed by printing and tissue maturation. An [...] Read more.

Layer-by-layer additive manufacturing process has evolved into three-dimensional (3D) “bio-printing” as a means of constructing cell-laden functional tissue equivalents. The process typically involves the mixing of cells of interest with an appropriate hydrogel, termed as “bioink”, followed by printing and tissue maturation. An ideal bioink should have adequate mechanical, rheological, and biological features of the target tissues. However, native extracellular matrix (ECM) is made of an intricate milieu of soluble and non-soluble extracellular factors, and mimicking such a composition is challenging. To this end, here we report the formulation of a multi-component bioink composed of gelatin and alginate -based scaffolding material, as well as a platelet-rich plasma (PRP) suspension, which mimics the insoluble and soluble factors of native ECM respectively. Briefly, sodium alginate was subjected to controlled oxidation to yield alginate dialdehyde (ADA), and was mixed with gelatin and PRP in various volume ratios in the presence of borax. The formulation was systematically characterized for its gelation time, swelling, and water uptake, as well as its morphological, chemical, and rheological properties; furthermore, blood- and cytocompatibility were assessed as per ISO 10993 (International Organization for Standardization). Printability, shape fidelity, and cell-laden printing was evaluated using the RegenHU 3D Discovery bioprinter. The results indicated the successful development of ADA–gelatin–PRP based bioink for 3D bioprinting and biofabrication applications. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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18 pages, 6803 KiB

Open AccessArticle

Additive Manufacturing of Prostheses Using Forest-Based Composites

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Bioengineering 2020, 7(3), 103; https://doi.org/10.3390/bioengineering7030103 - 01 Sep 2020

Cited by 19 | Viewed by 4640

Abstract

A custom-made prosthetic product is unique for each patient. Fossil-based thermoplastics are the dominant raw materials in both prosthetic and industrial applications; there is a general demand for reducing their use and replacing them with renewable, biobased materials. A transtibial prosthesis sets strict [...] Read more.

A custom-made prosthetic product is unique for each patient. Fossil-based thermoplastics are the dominant raw materials in both prosthetic and industrial applications; there is a general demand for reducing their use and replacing them with renewable, biobased materials. A transtibial prosthesis sets strict demands on mechanical strength, durability, reliability, etc., which depend on the biocomposite used and also the additive manufacturing (AM) process. The aim of this project was to develop systematic solutions for prosthetic products and services by combining biocomposites using forestry-based derivatives with AM techniques. Composite materials made of polypropylene (PP) reinforced with microfibrillated cellulose (MFC) were developed. The MFC contents (20, 30 and 40 wt%) were uniformly dispersed in the polymer PP matrix, and the MFC addition significantly enhanced the mechanical performance of the materials. With 30 wt% MFC, the tensile strength and Young´s modulus was about twice that of the PP when injection molding was performed. The composite material was successfully applied with an AM process, i.e., fused deposition modeling (FDM), and a transtibial prosthesis was created based on the end-user’s data. A clinical trial of the prosthesis was conducted with successful outcomes in terms of wearing experience, appearance (color), and acceptance towards the materials and the technique. Given the layer-by-layer nature of AM processes, structural and process optimizations are needed to maximize the reinforcement effects of MFC to eliminate variations in the binding area between adjacent layers and to improve the adhesion between layers. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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12 pages, 1660 KiB

Open AccessArticle

3D Printing of Gelled and Cross-Linked Cellulose Solutions; an Exploration of Printing Parameters and Gel Behaviour

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Bioengineering 2020, 7(2), 30; https://doi.org/10.3390/bioengineering7020030 - 27 Mar 2020

Cited by 13 | Viewed by 6195

Abstract

In recent years, 3D printing has enabled the fabrication of complex designs, with low-cost customization and an ever-increasing range of materials. Yet, these abilities have also created an enormous challenge in optimizing a large number of process parameters, especially in the 3D printing [...] Read more.

In recent years, 3D printing has enabled the fabrication of complex designs, with low-cost customization and an ever-increasing range of materials. Yet, these abilities have also created an enormous challenge in optimizing a large number of process parameters, especially in the 3D printing of swellable, non-toxic, biocompatible and biodegradable materials, so-called bio-ink materials. In this work, a cellulose gel, made out of aqueous solutions of cellulose, sodium hydroxide and urea, was used to demonstrate the formation of a shear thinning bio-ink material necessary for an extrusion-based 3D printing. After analysing the shear thinning behaviour of the cellulose gel by rheometry a Design of Experiments (DoE) was applied to optimize the 3D bioprinter settings for printing the cellulose gel. The optimum print settings were then used to print a human ear shape, without a need for support material. The results clearly indicate that the found settings allow the printing of more complex parts with high-fidelity. This confirms the capability of the applied method to 3D print a newly developed bio-ink material. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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10 pages, 2528 KiB

Open AccessCommunication

Nanocellulose-Based Inks—Effect of Alginate Content on the Water Absorption of 3D Printed Constructs

by Eduardo Espinosa,

Daniel Filgueira

, Alejandro Rodríguez and

Gary Chinga-Carrasco

Bioengineering 2019, 6(3), 65; https://doi.org/10.3390/bioengineering6030065 - 30 Jul 2019

Cited by 38 | Viewed by 7427

Abstract

2,2,6,6-tetramethylpyperidine-1-oxyl (TEMPO) oxidized cellulose nanofibrils (CNF) were used as ink for three-dimensional (3D) printing of porous structures with potential as wound dressings. Alginate (10, 20, 30 and 40 wt%) was incorporated into the formulation to facilitate the ionic cross-linking with calcium chloride (CaCl [...] Read more.

2,2,6,6-tetramethylpyperidine-1-oxyl (TEMPO) oxidized cellulose nanofibrils (CNF) were used as ink for three-dimensional (3D) printing of porous structures with potential as wound dressings. Alginate (10, 20, 30 and 40 wt%) was incorporated into the formulation to facilitate the ionic cross-linking with calcium chloride (CaCl₂). The effect of two different concentrations of CaCl₂ (50 and 100 mM) was studied. The 3D printed hydrogels were freeze-dried to produce aerogels which were tested for water absorption. Scanning Electronic Microscopy (SEM) pictures demonstrated that the higher the concentration of the cross-linker the higher the definition of the printed tracks. CNF-based aerogels showed a remarkable water absorption capability. Although the incorporation of alginate and the cross-linking with CaCl₂ led to shrinkage of the 3D printed constructs, the approach yielded suitable porous structures for water and moisture absorption. It is concluded that the 3D printed biocomposite structures developed in this study have characteristics that are promising for wound dressings devices. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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12 pages, 3131 KiB

Open AccessArticle

3D Printing High-Consistency Enzymatic Nanocellulose Obtained from a Soda-Ethanol-O₂ Pine Sawdust Pulp

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Bioengineering 2019, 6(3), 60; https://doi.org/10.3390/bioengineering6030060 - 16 Jul 2019

Cited by 12 | Viewed by 7178

Abstract

Soda-ethanol pulps, prepared from a forestry residue pine sawdust, were treated according to high-consistency enzymatic fibrillation technology to manufacture nanocellulose. The obtained nanocellulose was characterized and used as ink for three-dimensional (3D) printing of various structures. It was also tested for its moisture [...] Read more.

Soda-ethanol pulps, prepared from a forestry residue pine sawdust, were treated according to high-consistency enzymatic fibrillation technology to manufacture nanocellulose. The obtained nanocellulose was characterized and used as ink for three-dimensional (3D) printing of various structures. It was also tested for its moisture sorption capacity and cytotoxicity, as preliminary tests for evaluating its suitability for wound dressing and similar applications. During the high-consistency enzymatic treatment it was found that only the treatment of the O₂-delignified pine pulp resulted in fibrillation into nano-scale. For 3D printing trials, the material needed to be fluidized further. By 3D printing, it was possible to fabricate various structures from the high-consistency enzymatic nanocellulose. However, the water sorption capacity of the structures was lower than previously seen with porous nanocellulose structures, indicating that further optimization of the material is needed. The material was found not to be cytotoxic, thus showing potential as material, e.g., for wound dressings and for printing tissue models. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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Review

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31 pages, 7647 KiB

Open AccessEditor’s ChoiceReview

Advantages of Additive Manufacturing for Biomedical Applications of Polyhydroxyalkanoates

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Bioengineering 2021, 8(2), 29; https://doi.org/10.3390/bioengineering8020029 - 23 Feb 2021

Cited by 21 | Viewed by 5738

Abstract

In recent years, biopolymers have been attracting the attention of researchers and specialists from different fields, including biotechnology, material science, engineering, and medicine. The reason is the possibility of combining sustainability with scientific and technological progress. This is an extremely broad research topic, [...] Read more.

In recent years, biopolymers have been attracting the attention of researchers and specialists from different fields, including biotechnology, material science, engineering, and medicine. The reason is the possibility of combining sustainability with scientific and technological progress. This is an extremely broad research topic, and a distinction has to be made among different classes and types of biopolymers. Polyhydroxyalkanoate (PHA) is a particular family of polyesters, synthetized by microorganisms under unbalanced growth conditions, making them both bio-based and biodegradable polymers with a thermoplastic behavior. Recently, PHAs were used more intensively in biomedical applications because of their tunable mechanical properties, cytocompatibility, adhesion for cells, and controllable biodegradability. Similarly, the 3D-printing technologies show increasing potential in this particular field of application, due to their advantages in tailor-made design, rapid prototyping, and manufacturing of complex structures. In this review, first, the synthesis and the production of PHAs are described, and different production techniques of medical implants are compared. Then, an overview is given on the most recent and relevant medical applications of PHA for drug delivery, vessel stenting, and tissue engineering. A special focus is reserved for the innovations brought by the introduction of additive manufacturing in this field, as compared to the traditional techniques. All of these advances are expected to have important scientific and commercial applications in the near future. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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23 pages, 1313 KiB

Open AccessReview

Additive Biomanufacturing with Collagen Inks

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Bioengineering 2020, 7(3), 66; https://doi.org/10.3390/bioengineering7030066 - 01 Jul 2020

Cited by 32 | Viewed by 5978

Abstract

Collagen is a natural polymer found abundantly in the extracellular matrix (ECM). It is easily extracted from a variety of sources and exhibits excellent biological properties such as biocompatibility and weak antigenicity. Additionally, different processes allow control of physical and chemical properties such [...] Read more.

Collagen is a natural polymer found abundantly in the extracellular matrix (ECM). It is easily extracted from a variety of sources and exhibits excellent biological properties such as biocompatibility and weak antigenicity. Additionally, different processes allow control of physical and chemical properties such as mechanical stiffness, viscosity and biodegradability. Moreover, various additive biomanufacturing technology has enabled layer-by-layer construction of complex structures to support biological function. Additive biomanufacturing has expanded the use of collagen biomaterial in various regenerative medicine and disease modelling application (e.g., skin, bone and cornea). Currently, regulatory hurdles in translating collagen biomaterials still remain. Additive biomanufacturing may help to overcome such hurdles commercializing collagen biomaterials and fulfill its potential for biomedicine. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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20 pages, 4522 KiB

Open AccessReview

Nanocellulose-Based Inks for 3D Bioprinting: Key Aspects in Research Development and Challenging Perspectives in Applications—A Mini Review

by

Xiaoju Wang

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Qingbo Wang

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Chunlin Xu

Bioengineering 2020, 7(2), 40; https://doi.org/10.3390/bioengineering7020040 - 29 Apr 2020

Cited by 68 | Viewed by 11723

Abstract

Nanocelluloses have emerged as a catalogue of renewable nanomaterials for bioink formulation in service of 3D bioprinting, thanks to their structural similarity to extracellular matrices and excellent biocompatibility of supporting crucial cellular activities. From a material scientist’s viewpoint, this mini-review presents the key [...] Read more.

Nanocelluloses have emerged as a catalogue of renewable nanomaterials for bioink formulation in service of 3D bioprinting, thanks to their structural similarity to extracellular matrices and excellent biocompatibility of supporting crucial cellular activities. From a material scientist’s viewpoint, this mini-review presents the key research aspects of the development of the nanocellulose-based bioinks in 3D (bio)printing. The nanomaterial properties of various types of nanocelluloses, including bacterial nanocellulose, cellulose nanofibers, and cellulose nanocrystals, are reviewed with respect to their origins and preparation methods. Different cross-linking strategies to integrate into multicomponent nanocellulose-based bioinks are discussed in terms of regulating ink fidelity in direct ink writing as well as tuning the mechanical stiffness as a bioactive cue in the printed hydrogel construct. Furthermore, the impact of surface charge and functional groups on nanocellulose surface on the crucial cellular activities (e.g., cell survival, attachment, and proliferation) is discussed with the cell–matrix interactions in focus. Aiming at a sustainable and cost-effective alternative for end-users in biomedical and pharmaceutical fields, challenging aspects such as biodegradability and potential nanotoxicity of nanocelluloses call for more fundamental comprehension of the cell–matrix interactions and further validation in in vivo models. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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24 pages, 2361 KiB

Open AccessReview

3D Bioprinting Strategies for the Regeneration of Functional Tubular Tissues and Organs

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Bioengineering 2020, 7(2), 32; https://doi.org/10.3390/bioengineering7020032 - 31 Mar 2020

Cited by 74 | Viewed by 10261

Abstract

It is difficult to fabricate tubular-shaped tissues and organs (e.g., trachea, blood vessel, and esophagus tissue) with traditional biofabrication techniques (e.g., electrospinning, cell-sheet engineering, and mold-casting) because these have complicated multiple processes. In addition, the tubular-shaped tissues and organs have their own design [...] Read more.

It is difficult to fabricate tubular-shaped tissues and organs (e.g., trachea, blood vessel, and esophagus tissue) with traditional biofabrication techniques (e.g., electrospinning, cell-sheet engineering, and mold-casting) because these have complicated multiple processes. In addition, the tubular-shaped tissues and organs have their own design with target-specific mechanical and biological properties. Therefore, the customized geometrical and physiological environment is required as one of the most critical factors for functional tissue regeneration. 3D bioprinting technology has been receiving attention for the fabrication of patient-tailored and complex-shaped free-form architecture with high reproducibility and versatility. Printable biocomposite inks that can facilitate to build tissue constructs with polymeric frameworks and biochemical microenvironmental cues are also being actively developed for the reconstruction of functional tissue. In this review, we delineated the state-of-the-art of 3D bioprinting techniques specifically for tubular tissue and organ regeneration. In addition, this review described biocomposite inks, such as natural and synthetic polymers. Several described engineering approaches using 3D bioprinting techniques and biocomposite inks may offer beneficial characteristics for the physiological mimicry of human tubular tissues and organs. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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Other

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6 pages, 2777 KiB

Open AccessPerspective

Biofabrication of Bacterial Constructs: New Three-Dimensional Biomaterials

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Amin Shavandi

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Esmat Jalalvandi

Bioengineering 2019, 6(2), 44; https://doi.org/10.3390/bioengineering6020044 - 14 May 2019

Cited by 8 | Viewed by 6747

Abstract

An enormous number of bacteria live in almost every environment; from deep oceans to below the surface of the earth or in our gastrointestinal tract. Although biofabrication is growing and maturing very quickly, the involvement of bacteria in this process has not been [...] Read more.

An enormous number of bacteria live in almost every environment; from deep oceans to below the surface of the earth or in our gastrointestinal tract. Although biofabrication is growing and maturing very quickly, the involvement of bacteria in this process has not been developed at a similar pace. From the development of a new generation of biomaterials to green bioremediation for the removal of hazardous environmental pollutants or to develop innovative food products in a recent trend, researchers have used cutting-edge biofabrication techniques to reveal the great potential of 3D structured bacterial constructs. These 3D bacterial workhouses may fundamentally change our approach toward biomaterials. Full article

(This article belongs to the Special Issue Biocomposite Inks for 3D Printing)

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