3D Bioplotter Research Papers
Optimization of Biomanufacturing process for Tissue Engineering applications
In recent years, tissue engineering has experienced significant advancements, mostly driven by the emergence of additive manufacturing technologies and the integration of biomaterials and cells. This advanced technique enables the creation of intricate structures with diverse components and properties, specifically designed for use in biomedical applications. The primary benefit of this technology is its ability to be customised, which helps minimise post-operative difficulties for patients with orthopaedic diseases and those undergoing tissue transplants. For this purpose, the essential components can be synthesised by the patient’s own cells. However, there are still other obstacles that need to be addressed in order…
Bioprinting process optimization: case study on PVA (Polyvinyl Alcohol) and Graphene Oxide biocompatible hydrogels
Scaffolds manufacturing for tissue engineering is an elaborate process since to fabricate a functional tissue, the engineered structures have to mimic the extracellular matrix. The key goal is to produce 3D scaffolds composed of macro- and micro- scale structures. The combination of different production technologies, as 3D bioprinting (BP) and electrospinning (ES), enables the fabrication of multiscale structures but, above all, the biomaterial choice is crucial to apply these technologies. Hydrogels based of Polyvinyl alcohol (PVA), a water soluble and biodegradable polymer, are able to create a highly hydrated environment that promotes cell attachment and proliferation, with limited mechanical properties.…
3D printing of multi-scale porous β-tricalcium phosphate scaffolds: Mechanical properties and degradation
Processing-structure-property relationships of 3D-printed multi-scale porous ceramics were investigated. Direct ink writing (DIW) of oil-templated colloidal pastes produced hierarchically porous beta-tricalcium phosphate (TCP) scaffolds. Print architecture and microporosity within filaments were varied, mimicking bone structure. The scaffolds exhibited 60–70 % porosity with interconnected macropores 300–700 μm and microporosity within the filaments at the 10 micron-scale. Varying surfactant and oil concentrations created two micro-pore morphologies – bubble-like pores (emulsion) and channel-like pores (capillary suspension). Emulsion scaffolds were stronger, stiffer and more reliable than capillary suspension scaffolds under both compression and bending. Reducing nozzle diameter and inter-filament distance improved strength and stiffness,…
Dynamic Alginate Hydrogel as an Antioxidative Bioink for Bioprinting
3D bioprinting holds great potential for use in tissue engineering to treat degenerative joint disorders, such as osteoarthritis. However, there is a lack of multifunctional bioinks that can not only support cell growth and differentiation, but also offer protection to cells against injuries caused by the elevated oxidative stress; this conditions is a common characteristic of the microenvironment of the osteoarthritis disease. To mitigate oxidative stress-induced cellular phenotype change and malfunction, an anti-oxidative bioink derived from an alginate dynamic hydrogel was developed in this study. The alginate dynamic hydrogel gelated quickly via the dynamic covalent bond between the phenylboronic acid…
Effect of internal lattice structure on the flexural strength of 3D printed hierarchical porous ultra-high temperature ceramic (ZrB2)
3D printing of technical ceramics using direct ink writing (DIW) of multiphase colloidal inks has the unique ability to create structures with hierarchical features. To facilitate the application of 3D printed hierarchical porous ultra-high temperature ceramics (UHTCs), additional limiting factors such as strength and the effect of 3D printed internal lattice structure need to be better understood. This study reports on the strength dependence of common DIW print parameters including internal lattice structure shape, nozzle diameter and spacings between adjacent filaments. The present study applies Weibull statistics to the experimental array that considers macro features introduced through print parameters as…
Self-Adaptive MoO3−x Subnanometric Wires Incorporated Scaffolds for Osteosarcoma Therapy and Bone Regeneration
Residual tumor cells and bone tissue defects are two critical challenges in clinical osteosarcoma treatment. Herein, a subnanomedicine concept is proposed by developing a self-adaptive functional tissue engineering scaffold constructed by integrating MoO3−x subnanometric wires onto 3D printing bioactive glass scaffolds. The MoO3−x subnanometric wires are synthesized by a one-pot hydrothermal method, which aggregate in an acidic tumor microenvironment and react with hydrogen peroxide to produce reactive oxygen species for specific chemodynamic therapy. However, they can degrade rapidly under physiological conditions without causing toxicity. Moreover, self-adaptively enhanced photothermal conversion enables tumor-targeting photothermal therapy while enhancing chemodynamic therapy. Additionally, the Mo5+-Mo6+…
Utilisation of waste wool through fabrication of 3D water-resistant polyvinyl alcohol composite: Impact of micro-sized wool powders
Polyvinyl alcohol (PVA) is a biodegradable polymer having applications in several industries, such as textiles and paper manufacturing. Due to the water solubility, it is also a useful solvent in 3D dispensing to extrude printable solutions at a low temperature. However, its hydrophilicity is a drawback when considering the property of the end-use material if water contact is required. Using the water solubility of PVA as a benefit during 3D dispensing but avoiding the hydrophilicity in the printed material is tricky and unresolved. This study proposes a technique to achieve this phenomenon by taking advantage of the natural hydrophobicity of…
Rapid Customization and Manipulation Mechanism of Micro-Droplet Chip for 3D Cell Culture
A full PDMS micro-droplet chip for 3D cell culture was prepared by using SLA light-curing 3D printing technology. This technology can quickly customize various chips required for experiments, saving time and capital costs for experiments. Moreover, an injection molding method was used to prepare the full PDMS chip, and the convex mold was prepared by light-curing 3D printing technology. Compared with the traditional preparation process of micro-droplet chips, the use of 3D printing technology to prepare micro-droplet chips can save manufacturing and time costs. The different ratios of PDMS substrate and cover sheet and the material for making the convex…
In-situ 4-point flexural testing and synchrotron micro X-ray computed tomography of 3D printed hierarchical-porous ultra-high temperature ceramic
3D printed ceramics have received much attention of late due to the ability to manufacture complex near net shapes with a range of structures across multiple length scales. The introduction of hierarchical features offers a wider array of properties, yet with this comes additional unknowns as to their limits including the mechanisms behind failures. The present work applies in-situ Synchrotron micro X-ray computed tomography (μXCT) with 4-point flexural testing to study and further understand the failure pattern of 3D printed hierarchical porous ultra-high temperature ceramics. Samples were imaged at incremental load steps to observe the propagation of defects until final…
3D Printing and Performance Study of Porous Artificial Bone Based on HA-ZrO2-PVA Composites
An ideal artificial bone implant should have similar mechanical properties and biocompatibility to natural bone, as well as an internal structure that facilitates stomatal penetration. In this work, 3D printing was used to fabricate and investigate artificial bone composites based on HA-ZrO2-PVA. The composites were proportionally configured using zirconia (ZrO2), hydroxyapatite (HA) and polyvinyl alcohol (PVA), where the ZrO2 played a toughening role and PVA solution served as a binder. In order to obtain the optimal 3D printing process parameters for the composites, a theoretical model of the extrusion process of the composites was first established, followed by the optimization…
Dynamic hyaluronic acid hydrogel with covalent linked gelatin as an anti-oxidative bioink for cartilage tissue engineering
In the past decade, cartilage tissue engineering has arisen as a promising therapeutic option for degenerative joint diseases, such as osteoarthritis, in the hope of restoring the structure and physiological functions. Hydrogels are promising biomaterials for developing engineered scaffolds for cartilage regeneration. However, hydrogel-delivered mesenchymal stem cells or chondrocytes could be exposed to elevated levels of reactive oxygen species (ROS) in the inflammatory microenvironment after being implanted into injured joints, which may affect their phenotype and normal functions and thereby hinder the regeneration efficacy. To attenuate ROS induced side effects, a multifunctional hydrogel with an innate anti-oxidative ability was produced…
Direct ink writing of hierarchical porous alumina-stabilized emulsions: Rheology and printability
Bio-inspired multi-scaled (hierarchical) porous structures have remarkable strength and stiffness-to-density properties. Direct ink writing (DIW) or robocasting, an additive manufacturing (or also commonly known as 3D printing) material extrusion technique is able to create near-net-shaped complex geometries. A new approach of combining DIW, colloidal particle-stabilized emulsion paste inks and partial densification to create tailored architectures of hierarchical porosity on three scales has been demonstrated. The printed and sintered ceramic lattice structures possess relatively high overall porosity of 78.7% (on average), comprising mainly (64.7%) open porosity. The effects of formulation (surfactant and oil concentrations, solids particle size, and mixing speed) on…
3D Printable and Biocompatible Iongels for Body Sensor Applications
Soft-ionic materials with biocompatibility and 3D printability are needed to develop next-generation devices to interface between electronic and biological signals. Herein, thermoreversible and biocompatible ionic liquid gels or iongels, which can be processed by direct ink writing are reported. The iongels are designed by taking advantage of polyvinyl alcohol/phenol interactions to gelify biocompatible cholinium carboxylate ionic liquids. The obtained iongels are stable, soft, and flexible materials (Young modulus between 14 and 70 kPa) with high ionic conductivity (1.8 × 10–2 S cm–1). Interestingly, they presented thermoreversible properties with gel–sol transitions ranging from 85 and 110 °C, which allows the iongel…
Mesoporous calcium silicate and titanium composite scaffolds via 3D-printing for improved properties in bone repair
Calcium silicate (CS) composite bone tissue engineering scaffolds were three-dimensionally printed using titanium metallic powders as the second strengthening phase for overcoming the inherent brittleness and fast degradability. In order to promote the sintering process of all composite scaffolds, mesoporous structure was further introduced into sol-gel-derived CS powders obtaining mesoporous CS (MCS) with larger surface area. The influences of mesoporous structure, sintering temperature and Ti content have been investigated through comparisons of the final scaffold composition, microstructure, compressive strength and in vitro stability. Results showed that CS matrix materials reacted with Ti could form less degradable CaTiO3 and TiC ceramic…
Cellular, Mineralized, and Programmable Cellulose Composites Fabricated by 3D Printing of Aqueous Pastes Derived from Paper Wastes and Microfibrillated Cellulose
Combining recycling of paper wastes (WPs) with extrusion‐based additive manufacturing represents a sustainable route to cellular cellulose composites tailored for lightweight construction. Particularly, shear mixing of shredded WPs with an aqueous solution of a polymer binder like polyvinyl alcohol (PVA) yields aqueous pastes suitable for 3D printing. As a shear thinning additive, both WP and microfibrillated cellulose account for enhanced shear thinning and dimensional stability. Opposite to the formation of dense WP/PVA composites by melt extrusion, 3D printing of aqueous pastes produces cellular cellulose/PVA composites exhibiting hierarchical pore architectures. In spite of low densities around 0.8 g cm−3, high Young’s…
Silicon substituted hydroxyapatite/VEGF scaffolds stimulate bone regeneration in osteoporotic sheep
Silicon-substituted hydroxyapatite (SiHA) macroporous scaffolds have been prepared by robocasting. In order to optimize their bone regeneration properties, we have manufactured these scaffolds presenting different microstructures: nanocrystalline and crystalline. Moreover, their surfaces have been decorated with vascular endothelial growth factor (VEGF) to evaluate the potential coupling between vascularization and bone regeneration. In vitro cell culture tests evidence that nanocrystalline SiHA hinders pre-osteblast proliferation, whereas the presence of VEGF enhances the biological functions of both endothelial cells and pre-osteoblasts. The bone regeneration capability has been evaluated using an osteoporotic sheep model. In vivo observations strongly correlate with in vitro cell culture…
3D-printed ternary SiO2CaOP2O5 bioglass-ceramic scaffolds with tunable compositions and properties for bone regeneration
Simple ternary SiO2CaOP2O5 bioglasses proved sufficient osteogenesis capacity. In this study, the bioglasses were 3D printed into porous scaffolds and SiO2/CaO molar ratio was altered (from 90/5 to 60/35) to achieve tunable glass-ceramic compositions after thermal treatment. Scaffolds possessed interconnected porous structure with controllable porosities via 3D printing technique. In addition, microstructure and properties of mechanical strength, degradation, ion dissolution and apatite formation were investigated. Characterization results showed that higher content of SiO2 produced more homogeneous crystalline particles and sintering compactness, thus led to higher strength. For scaffolds with higher CaO content, more glasses were maintained and faster degradation rate…
3D Printing Nanoscale Bioactive Glass Scaffolds Enhance Osteoblast Migration and Extramembranous Osteogenesis through Stimulating Immunomodulation
Bioactive glass (BG) can repair bone defects, however, it is not clear whether BG has the ability for bone augmentation without making any bone defect. Unlike the intramembranous osteogenesis in bone defect repair, the extramembranous osteogenesis occurs outside the cortical bone and the osteoprogenitor cells show the reversed migration. Herein, nanoscale bioactive glass scaffolds (BGSs) are fabricated, and their role and immunomodulation‐related mechanism in the extramembranous osteogenesis are investigated. The in vitro migration and differentiation of calvaria preosteoblasts are studied by culturing with peripheral macrophage‐conditioned medium after stimulating with BGSs. The results indicate that the proinflammatory environment significantly promotes preosteoblast…
3D Printed, PVA–PAA Hydrogel Loaded-Polycaprolactone Scaffold for the Delivery of Hydrophilic In-Situ Formed Sodium Indomethacin
3D printed polycaprolactone (PCL)-blended scaffolds have been designed, prepared, and evaluated in vitro in this study prior to the incorporation of a polyvinyl alcohol–polyacrylic acid (PVA–PAA) hydrogel for the delivery of in situ-formed sodium indomethacin. The prepared PCL–PVA–PAA scaffold is proposed as a potential structural support system for load-bearing tissue damage where inflammation is prevalent. Uniaxial strain testing of the PCL-blended scaffolds were undertaken to determine the scaffold’s resistance to strain in addition to its thermal, structural, and porosimetric properties. The viscoelastic properties of the incorporated PVA–PAA hydrogel has also been determined, as well as the drug release profile of…
3D-printed IFN-γ-loading calcium silicate-β-tricalcium phosphate scaffold sequentially activates M1 and M2 polarization of macrophages to promote vascularization of tissue engineering bone
To promote vascularization of tissue-engineered bone, IFN-γ polarizing macrophages to M1 was loaded on 5% calcium silicate/β-tricalcium phosphate (CaSiO3-β-TCP) scaffolds. IFN-γ and Si released from the scaffold were designed to polarize M1 and M2 macrophages, respectively. β-TCP, CaSiO3-β-TCP, and IFN-γ@CaSiO3-β-TCP were fabricated and biocompatibilities were evaluated. Polarizations of macrophages were detected by flow cytometry. Human umbilical vein endothelial cells with GFP were cultured and induced on Matrigel with conditioned culture medium extracted from culture of macrophages loaded on scaffolds for evaluating angiogenesis. Four weeks after the scaffolds were subcutaneously implanted into C57B1/6, vascularization was evaluated by visual observation, hematoxylin and…
3D Bio-Plotted Tricalcium Phosphate/Zirconia Composite Scaffolds to Heal Large Size Bone Defects
β-TCP-Zirconia scaffolds with different architectures were fabricated by means of 3D-Bioplotting in order to enhance the mechanical and in-vitro ability of the scaffold to heal large size bone defects. In the present study scaffold architecture with different strand orientations (0o-90o, 0o-45o-135o-180o, 0o-108o-216o and 0o-72o-144o-36o-108o) were fabricated, characterized and evaluated for mechanical strength and cell proliferation ability. β-TCP powder (25µm) and PVA (Polyvinyl Alcohol) was acquired from Fisher Scientific, India. Zirconia (18 to 32 µm) was procured from Lobachemie, India. In brief 7.5%, PVA in distilled water was used as a binder and was mixed with 10 grams of (70/30) TCP-Zirconia…
Bioprinted fibrin-factor XIII-hyaluronate hydrogel scaffolds with encapsulated Schwann cells and their in vitro characterization for use in nerve regeneration
The blood clotting protein fibrin contains cell-binding domains, providing potential advantage for the fabrication of tissue repair scaffolds and for live cell encapsulation. However, fabrication of fibrin scaffolds with encapsulated cells using three dimensional (3D) printing has proven challenging due to the mechanical difficulties of fabricating protein hydrogel scaffolds with defined microstructure. For example, extrusion based 3D printing of fibrin is generally unfeasible because of the low viscosity of precursor fibrinogen solution. Here we describe a novel technique for bioprinting of fibrin scaffolds by extruding fibrinogen solution into thrombin solution, utilizing hyaluronic acid (HA) and polyvinyl alcohol (PVA) to increase…
3D scaffold with effective multidrug sequential release against bacteria biofilm
Bone infection is a feared complication following surgery or trauma that remains as an extremely difficult disease to deal with. So far, the outcome of therapy could be improved with the design of 3D implants, which combine the merits of osseous regeneration and local multidrug therapy so as to avoid bacterial growth, drug resistance and the feared side effects. Herein, hierarchical 3D multidrug scaffolds based on nanocomposite bioceramic and polyvinyl alcohol (PVA) prepared by rapid prototyping with an external coating of gelatin-glutaraldehyde (Gel-Glu) have been fabricated. These 3D scaffolds contain three antimicrobial agents (rifampin, levofloxacin and vancomycin), which have been…
Three-dimensional printed strontium-containing mesoporous bioactive glass scaffolds for repairing rat critical-sized calvarial defects
The development of a new generation of biomaterials with high osteogenic ability for fast osseointegration with host bone is being intensively investigated. In this study, we have fabricated three-dimensional (3-D) strontium-containing mesoporous bioactive glass (Sr-MBG) scaffolds by a 3-D printing technique. Sr-MBG scaffolds showed uniform interconnected macropores (∼400 μm), high porosity (∼70%) and enhanced compressive strength (8.67 ± 1.74 MPa). Using MBG scaffolds as a control, the biological properties of Sr-MBG scaffolds were evaluated by apatite-forming ability, adhesion, proliferation, alkaline phosphatase activity and osteogenic gene expression of osteoblast-like cells MC3T3-E1. Furthermore, Sr-MBG scaffolds were used to repair critical-sized rat calvarial…
From nanoparticles to fibres: effect of dispersion composition on fibre properties
A polyvinyl alcohol (PVA)-stabilized polypyrrole nanodispersion has been optimised for conductivity and processability by decreasing the quantity of PVA before and after synthesis. A reduction of PVA before synthesis leads to the formation of particles with a slight increase in dry particle diameter (51 ± 6 to 63 ± 3 nm), and conversely a reduced hydrodynamic diameter. Conductivity of the dried nanoparticle films was not measureable after a reduction of PVA prior to synthesis. Using filtration of particles after synthesis, PVA content was sufficiently reduced to achieve dried thin film conductivity of 2 S cm−1, while the electroactivity of the…
Three-dimensional printing of strontium-containing mesoporous bioactive glass scaffolds for bone regeneration
In this study, we fabricated strontium-containing mesoporous bioactive glass (Sr-MBG) scaffolds with controlled architecture and enhanced mechanical strength using a three-dimensional (3-D) printing technique. The study showed that Sr-MBG scaffolds had uniform interconnected macropores and high porosity, and their compressive strength was ∼170 times that of polyurethane foam templated MBG scaffolds. The physicochemical and biological properties of Sr-MBG scaffolds were evaluated by ion dissolution, apatite-forming ability and proliferation, alkaline phosphatase activity, osteogenic expression and extracelluar matrix mineralization of osteoblast-like cells MC3T3-E1. The results showed that Sr-MBG scaffolds exhibited a slower ion dissolution rate and more significant potential to stabilize the…
Bone repair by cell-seeded 3D-bioplotted composite scaffolds made of collagen treated tricalciumphosphate or tricalciumphosphate-chitosan-collagen hydrogel or PLGA in ovine critical-sized calvarial defects
The aim of this study was to investigate the osteogenic effect of three different cell-seeded 3D-bioplotted scaffolds in a ovine calvarial critical-size defect model. The choice of scaffold-materials was based on their applicability for 3D-bioplotting and respective possibility to produce tailor-made scaffolds for the use in cranio-facial surgery for the replacement of complex shaped boneparts. Scaffold raw-materials are known to be osteoinductive when being cell-seeded [poly(L-lactide-co-glycolide) (PLGA)] or having components with osteoinductive properties as tricalciumphosphate (TCP) or collagen (Col) or chitosan. The scaffold-materials PLGA, TCP/Col, and HYDR (TCP/Col/chitosan) were cell-seeded with osteoblast-like cells whether gained from bone (OLB) or from…