3D Bioplotter Research Papers

In this work, we describe a new 3D printing methodology for the fabrication of multimaterial scaffolds involving the combination of thermoplastic extrusion and low temperature extrusion of bioinks. A fiber engraving technique was used to create a groove on the surface of a thermoplastic printed fiber using a commercial 3D printer and a low viscosity bioink was deposited into this groove. In contrast to traditional extrusion bioinks that rely on increased viscosity to prevent lateral spreading, this groove creates a defined space for bioink deposition. By physically constraining bioink spreading, a broader range of viscosities can be used. As proof-of-concept,…

Background The anatomical properties of the enthesis of the rotator cuff are hardly regained during the process of healing. The tendon-to-bone interface is normally replaced by fibrovascular tissue instead of interposition fibrocartilage, which impairs biomechanics in the shoulder and causes dysfunction. Tissue engineering offers a promising strategy to regenerate a biomimetic interface. Here, we report heterogeneous tendon-to-bone interface engineering based on a 3D-printed multiphasic scaffold. Methods A multiphasic poly(ε-caprolactone) (PCL)–PCL/tricalcium phosphate–PCL/tricalcium phosphate porous scaffold was manufactured using 3D printing technology. The three phases of the scaffold were designed to mimic the graded tissue regions in the tendon-to-bone interface—tendon, fibrocartilage, and…

The classic cell culture involves the use of support in two dimensions, such as a well plate or a Petri dish, that allows the culture of different types of cells. However, this technique does not mimic the natural microenvironment where the cells are exposed to. To solve that, three-dimensional bioprinting techniques were implemented, which involves the use of biopolymers and/or synthetic materials and cells. Because of a lack of information between data sources, the objective of this review paper is, to sum up, all the available information on the topic of bioprinting and to help researchers with the problematics with…

Hyaluronic acid (HA)-based hydrogels are widely used in biomedical applications due to their excellent biocompatibility. HA can be Ultraviolet (UV)-crosslinked by modification with methacrylic anhydride (HA-MA) and crosslinked by modification with 3,3′-dithiobis(propionylhydrazide) (DTP) (HA-SH) via click reaction. In the study presented in this paper, a 3D-bioprinted, double-crosslinked, hyaluronic-acid-based hydrogel for wound dressing was proposed. The hydrogel was produced by mixing HA-MA and HA-SH at different weight ratios. The rheological test showed that the storage modulus (G’) of the HA-SH/HA-MA hydrogel increased with the increase in the HA-MA content. The hydrogel had a high swelling ratio and a high controlled degradation…

The field of 3D bioprinting has rapidly grown, yet the fundamental ability to manipulate material properties has been challenging with current bioink methods. Here, we change bioink properties using our PEG cross-linking (PEGX) bioink method with the objective of optimizing cell viability while retaining control of mechanical properties of the final bioprinted construct. First, we investigate cytocompatible, covalent cross-linking chemistries for bioink synthesis (e.g. Thiol Michael type addition and bioorthogonal inverse electron demand Diels-Alder reaction). We demonstrate these reactions are compatible with the bioink method, which results in high cell viability. The PEGX method is then exploited to optimize extruded…

Silk, with highly crystalline structure and well-documented biocompatibility, is promising to be used as reinforcing material and build functionalized composite scaffolds. In the present study, we developed chitosan/silk composite scaffolds using silk particles, silk microfibres and nanofibres via 3D printing method. The three forms of silk fillers with varied shapes and dimensions were obtained via different processing methods and evaluated of their morphology, crystalline structure and thermal property. All silk fillers showed different degrees of improvement on printability in terms of ink rheology and printing shape fidelity. Different silk fillers led to different scaffold surface morphology and different roughness, while…

One of the latest trends in the regenerative medicine is the development of 3D-printing hydrogel scaffolds with biomimetic structures for tissue regeneration and organ reconstruction. However, it has been practically difficult to achieve a highly biomimetic hydrogel scaffolds with proper mechanical properties matching the natural tissue. Here, bacterial cellulose nanofibers (BCNFs) were applied to improve the structural resolution and enhance mechanical properties of silk fibroin (SF)/gelatin composite hydrogel scaffolds. The SF-based hydrogel scaffolds with hierarchical pores were fabricated via 3D-printing followed by lyophilization. Results showed that the tensile strength of printed sample increased significantly with the addition of BCNFs in…

In the present work we have revisited the application of quantitative ultrasound imaging (QUI) to cellular hydrogels, by using the reference phantom method (RPM) in combination with a local attenuation compensation algorithm. The investigated biological samples consisted of cell-laden collagen hydrogels with PC12 neural cells. These cell-laden hydrogels were used to calibrate the integrated backscattering coefficient (IBC) as a function of cell density, which was then used to generate parametric images of local cell density. The image resolution used for QUI and its impact on the relative IBC error was also investigated. Another important contribution of our work was the…

Lignin is a low-cost, natural polymer with abundant polar sites on its backbone that can be utilized for physical cross-linking of polymers. Here, we use lignin for additional cross-linking of hydrophilic polyether-based polyurethane (HPU) hydrogels, aiming to improve their mechanical properties and processability. Without reducing the swelling, simple addition of 2.5 wt % lignin increases the fracture energy and Young’s modulus of HPU hydrogels from, respectively, 1540 ± 40 to 2050 ± 50 J m–2 and 1.29 ± 0.06 to 2.62 ± 0.84 MPa. Lignin also increases the lap shear adhesiveness of hydrogels and induces an immediate load recovery of…

Three-dimensional bioprinting is an emerging technology for fabricating living 3D constructs, and it has shown great promise in tissue engineering. Bioinks are scaffold materials mixed with cells used by 3D bioprinting to form a required cell-laden structure. In this paper, a novel bioink made of gelatin methacrylamide (GelMA) and collagen (Col) doped with tyrosinase (Ty) is presented for the 3D bioprinting of living skin tissues. Ty has the dual function of being an essential bioactive compound in the skin regeneration process and also as an enzyme to facilitate the crosslink of Col and GelMA. Further, enzyme crosslinking together with photocrosslinking…

Soft tissue fixation of implant and bioelectrodes relies on mechanical means (e.g., sutures, staples, and screws), with associated complications of tissue perforation, scarring, and interfacial stress concentrations. Adhesive bioelectrodes address these shortcomings with voltage cured carbene‐based bioadhesives, locally energized through graphene interdigitated electrodes. Electrorheometry and adhesion structure activity relationships are explored with respect to voltage and electrolyte on bioelectrodes synthesized from graphene 3D‐printed onto resorbable polyester substrates. Adhesive leachates effects on in vitro metabolism and human‐derived platelet‐rich plasma response serves to qualitatively assess biological response. The voltage activated bioadhesives are found to have gelation times of 60 s or less…

Hydrogel bioprinting is a major area of focus in the field of tissue engineering. However, 3D printed hydrogel scaffolds often suffer from low printing accuracy and poor mechanical properties because of their soft nature and tendency to shrink. This makes it challenging to process them into structural materials. In this study, natural chitosan hydrogel scaffolds were, for the first time, reinforced with milled silk particles and fabricated by 3D printing. Compared with pure chitosan scaffolds, the addition of silk particles resulted in up to a 5-fold increase in compressive modulus as well as significantly better printing accuracy and improved scaffold…

Regenerative medicine has emerged as an important discipline that aims to repair injury or replace damaged tissues or organs by introducing living cells or functioning tissues. Successful regenerative medicine strategies will likely depend upon a simultaneous optimization strategy for the design of biomaterials, cell-seeding methods, cell-biomaterial interactions and molecular signaling within the engineered tissues. It remains a challenge to image three-dimensional (3-D) structures and functions of the cell-seeded scaffold in mesoscopic scale (>2∼3 mm). In this study, we utilized angled fluorescence laminar optical tomography (aFLOT), which allows depth-resolved molecular characterization of engineered tissues in 3-D to investigate cell viability, migration and…

The goal of this work is to develop an innovative method that combines bioprinting and endoscopic imaging to repair tympanic membrane perforations (TMPs). TMPs are a serious health issue because they can lead to both conductive hearing loss and repeated otitis media. TMPs occur in 3 to 5% of cases after ear tube placement as well as in cases of acute otitis media (the second most common infection in pediatrics), chronic otitis media with or without cholesteatoma, or as a result of barotrauma to the ear. About 55,000 tympanoplasties, the surgery performed to reconstruct TMPs, are performed every year and…

Collagen is the most abundant protein found in mammals and it exhibits a low immunogenicity, high biocompatibility and biodegradability when compared with others natural polymers. For this reason, it has been explored for the development of biologically instructive biomaterials with applications for tissue substitution and regeneration. Marine origin collagen has been pursued as an alternative to the more common bovine and porcine origins. This study focused on squid (Teuthoidea: Cephalopoda), particularly the Antarctic squid Kondakovia longimana and the Sub-Antarctic squid Illex argentinus as potential collagen sources. In this study, collagen has been isolated from the skins of the squids using…

Continuous composite fibres composed of polypyrrole (PPy) nanoparticles and reduced graphene oxide (rGO) at different mass ratios were fabricated using a single step wet-spinning approach. The electrical conductivity of the composite fibres increased significantly with the addition of rGO. The mechanical properties of the composite fibres also improved by the addition of rGO sheets compared to fibres containing only PPy. The ultimate tensile strength of the fibres increased with the proportion of rGO mass present. The elongation at break was greatest for the composite fibre containing equal mass ratios of PPy nanoparticles and rGO sheets. L929 fibroblasts seeded onto fibres…

The application of magnetic nanoparticles (MNPs) in tissue engineering (TE) approaches opens several new research possibilities in this field, enabling a new generation of multifunctional constructs for tissue regeneration. This study describes the development of sophisticated magnetic polymer scaffolds with aligned structural features aimed at applications in tendon tissue engineering (TTE). Tissue engineering magnetic scaffolds are prepared by incorporating iron oxide MNPs into a 3D structure of aligned SPCL (starch and polycaprolactone) fibers fabricated by rapid prototyping (RP) technology. The 3D architecture, composition, and magnetic properties are characterized. Furthermore, the effect of an externally applied magnetic field is investigated on…

An ideal behavior of a tissue engineering scaffold is that it degrades and reshapes at a rate that matches the formation of new tissues. However, this ideal situation may not occur as the scaffold often undergoes too slow or too fast degradation. To test the promise of the active control of scaffold degradation, in this work, a photo/water dual-degradable porous scaffold was designed and fabricated using a 3D fiber deposition (3DF) system from a linear biopolymer (named PLANB) that combined the o-nitrobenzyl linkages and hydrolysable ester bone in the polymer chains. The chemical structure, molecular weight and polydispersity of PLANB…

A multimaterial bio-ink method using polyethylene glycol crosslinking is presented for expanding the biomaterial palette required for 3D bioprinting of more mimetic and customizable tissue and organ constructs. Lightly crosslinked, soft hydrogels are produced from precursor solutions of various materials and 3D printed. Rheological and biological characterizations are presented, and the promise of this new bio-ink synthesis strategy is discussed.

Graphene-based polymer composites are a very promising class of compounds for tissue engineering scaffolds. However, in general the methods of synthesis are environmentally hazardous and residual toxic materials can affect the biocompatibility significantly. In this paper a simple, scalable, environmentally-friendly, microwave-assisted synthesis is described that results in conducting graphene/polycaprolactone composites that retain the processability and biocompatibility of the pristine polymer without introducing possibly hazardous reducing agents. Composites of polycaprolactone and graphene oxide were synthesised in a single step by the ring-opening polymerisation of ε-caprolactone in the presence of dispersed graphene oxide nanosheets under microwave irradiation. The graphene oxide provides a…

Despite various studies to minimize host reaction following a biomaterial implantation, an appealing strategy in regenerative medicine is to actively use such an immune response to trigger and control tissue regeneration. We have developed an in vitro model to modulate the host response by tuning biomaterials‘ surface properties through surface modifications techniques as a new strategy for tissue regeneration applications. Results showed tunable surface topography, roughness, wettability, and chemistry by varying treatment type and exposure, allowing for the first time to correlate the effect of these surface properties on cell attachment, morphology, strength and proliferation, as well as proinflammatory (IL-1β,…

In a versatile modular scaffold system, gradient nonwovens of in situ crosslinked gelatin nanofibers (CGN), fabricated by reactive electrospinning, are laminated with perforated layers and nonwovens of thermoplastic non-crosslinked biodegradable polyesters. The addition of glyoxal to a gelatin solution in a non-toxic solvent mixture consisting of acetic acid, ethyl acetate, and water (5:3:2 w/w/w) enables the in situ crosslinking of gelatin nanofibers during electrospinning. The use of this fluorine-free crosslinking system eliminates the need of post-treatment crosslinking and purification steps typical for conventional CGN scaffolds. The slowly progressing crosslinking of the dissolved gelatin in the presence of glyoxal increases the…

The costs associated with the treatment of medical device and surgical site infections are a major cause of concern in the global healthcare system. To prevent transmission of such infections, a prophylactic surface system that provides protracted release of antibacterial silver ions using low intensity direct electric current (LIDC; 28 μA system current at 6 V) activation has been recently developed. To ensure the safety for future in vivo studies and potential clinical applications, this study assessed the biocompatibility of the LIDC-activated interdigitated silver electrodes-based surface system; in vitro toxicity to human epidermal keratinocytes, human dermal fibroblasts, and normal human…

The requirement of a multifunctional scaffold for tissue engineering capable to offer at the same time tunable structural properties and bioactive interface is still unpaired. Here we present three-dimensional (3D) biodegradable polymeric (PCL) scaffolds with controlled morphology, macro-, micro-, and nano-mechanical performances endowed with bioactive moieties (RGD peptides) at the surface. Such result was obtained by a combination of rapid prototyping (e.g., 3D fiber deposition) and surface treatment approach (aminolysis followed by peptide coupling). By properly designing process conditions, a control over the mechanical and biological performances of the structure was achieved with a capability to tune the value of…

Spinal cord injury (SCI) represents a significant health and social problem, and therefore it is vital to develop novel strategies that can specifically target it. In this context, the objective of the present work was to develop a new range of three-dimensional (3D) tubular structures aimed at inducing the regeneration within SCI sites. Up to six different 3D tubular structures were initially developed by rapid prototyping: 3D bioplotting–based on a biodegradable blend of starch. These structures were then further complemented by injecting Gellan Gum, a polysaccharide-based hydrogel, in the central area of structures. The mechanical properties of these structures were…

Scaffolds are of great importance for tissue engineering because they enable the production of functional living implants out of cells obtained from cell culture. These scaffolds require individual external shape and well defined internal structure with interconnected porosity. The problem of the fabrication of prototypes from computer assisted design (CAD) data is well known in automotive industry. Rapid prototyping (RP) techniques are able to produce such parts. Some RP techniques exist for hard tissue implants. Soft tissue scaffolds need a hydrogel material. No biofunctional and cell compatible processing for hydrogels exists in the area of RP. Therefore, a new rapid…

In the year 2000 a new rapid prototyping (RP) technology was developed at the Freiburg Materials Research Center to meet the demands for desktop fabrication of scaffolds useful in tissue engineering. A key feature of this RP technology is the three-dimensional (3D) dispensing of liquids and pastes in liquid media. In contrast to conventional RP systems, mainly focused on melt processing, the 3D dispensing RP process (3D plotting) can apply a much larger variety of synthetic as well as natural materials, including aqueous solutions and pastes, to fabricate scaffolds for application in tissue engineering. For the first time, hydrogel scaffolds…