Hydrogels were rendered 3-D printable by increasing the concentration of pNVP (4, 6, 8, and 12 mol%). For initial studies, only 0.1 wt% PAn-PAAMPSA was incorporated into the printable, electroconductive formulation. Electroconductive hydrogel inks were placed in 30 cc syringes with a 200-micron needle inner diameter. 3-D printing was done using an EnvisionTEC 3-D Bioplotter and accompanying software, Perfactory RP, and Bioplotter RP (EnvisionTEC GmbH, Gladbeck, Germany). The EnvisionTEC 3-D Bioplotter was housed within a custom-fabricated biosafety cabinet for BSL-2 printing of human cells. Hydrogels were printed at a z-offset of 0.12 mm, a printing speed of 8.0 mm/s, and an extrusion pressure of 0.8 bar. The process of fabrication is shown in Figure 1 B. The 3-D constructs were printed layer-by-layer, with UV light exposures for 30 s between layers. Following crosslinking, hydrogels were hydrated using the same protocol used for photopatterned hydrogels.
3d bioplotter
Manufactured by EnvisionTEC
Sourced in Germany
The 3D Bioplotter is a lab equipment product designed for additive manufacturing. It is capable of processing a variety of materials, including biomaterials, polymers, and ceramics, to create complex three-dimensional structures. The 3D Bioplotter is equipped with multiple extrusion heads and a temperature-controlled build platform, allowing for the precise deposition of materials.
Automatically generated - may contain errors
Lab products found in correlation
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Bioplotter rp, by EnvisionTEC (2 mentions)
Lsm 710, by Zeiss (2 mentions)
Perfactory rp, by EnvisionTEC (2 mentions)
Fisherbrand model 50 sonic dismembrator, by Thermo Fisher Scientific (1 mentions)
Tc treated, by Corning (1 mentions)
Poly ε caprolactone pcl, by Merck Group (1 mentions)
Multi well plates, by Greiner (1 mentions)
Uv box, by Avantor (1 mentions)
Irgacure 2959, by BASF (1 mentions)
Magics software, by EnvisionTEC (1 mentions)
Mw 45 000, by Merck Group (1 mentions)
Discovery v8 stereomicroscope, by Zeiss (1 mentions)
Omnicure s2000 uv lamp, by Excelitas (1 mentions)
T150 utm, by Agilent Technologies (1 mentions)
Quanta 200, by Thermo Fisher Scientific (1 mentions)
Maestro in vivo imaging system, by PerkinElmer (1 mentions)
Fibrinogen, by Merck Group (1 mentions)
Gelatin type 1, by Merck Group (1 mentions)
Inventor 2019, by Autodesk (1 mentions)
60 protocols using 3d bioplotter
1
3D Printable Electroconductive Hydrogel Fabrication
2
Bioprinting Endothelial Cell Adhesion Scaffolds
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Discs with the material used for the testing of adhesion and growth of endothelial cells were bioprinted. The procedure for printing was performed essentially as recently described [27 (link)]. The respective hydrogel preparation was filled into sterile 30 ml printing cartridges (Nordson EFD, Pforzheim; Germany) and centrifuged for 3 min at 1500 rpm to remove air bubbles. After connecting a 0.25 mm tapered polyethylene printing tip (Nordson EFD) the cartridge was placed into the preheated (25°C) printing head of the 3D-Bioplotter (Envisiontec, Gladbeck; Germany). At 25°C, using a pressure of 1.5 bar and a printing speed of 16 mm/s cylindrical scaffolds measuring 7.5 x 0.4 mm were printed as described [27 (link)]. The strand distance between the printed cylinders was set to 1 mm resulting in a pore size of the printed layers of approximately 0.5 x 0.5 mm. Those scaffolds were printed directly into sterile 94 mm Petri dishes (Greiner Bio-One, Frickenhausen; Germany), supplemented with 2.5% [w/v] CaCl2 as crosslinking solution. The printed discs with a diameter 9.5 mm had been hardened in 2.5% [w/v] CaCl2 for crosslinking for 5 min. During this treatment the discs shrunk to 7.5 mm.
3
3D-Printed PLA/HA Scaffolds for MSC Culture
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All chemicals were obtained from Sigma Aldrich (Shanghai, China). A 3D-Bioplotter™ (Envision TEC GmbH, Germany) was used to construct the 3D-printed scaffolds using a computer. First, PLA particles (mean molecular weight ~20.0×104 Da) were dissolved in 1, 4-dioxane. HA powders (particle size ~50 m) were then added to the PLA solution and stirred for 12 h to form a uniform PLA/HA paste using a ratio of 7:3. Next, the cylinder models were loaded into 3D bioplotter software. The 3D-printed scaffolds were fabricated to have an outer diameter of 5 mm, an inner diameter of 2 mm, and a height of 15 mm for the in-vivo experiments.
High-resolution micro-computed tomography (Scanco Medical, μCT; Switzerland) was used to assess the porosity and average pore size of the scaffolds. The data were analyzed by μCT. Scanning electron microscopy (SEM; USA) was performed to observe the microstructure and morphology of the 3D-printed scaffolds.
Mesenchymal stem cells (MSCs) isolated from the eBM were used. Cell suspension was seeded at a density of 1×105 cells on the 3D-printed scaffolds in 24-well plates. Cell spreading and focal adhesion formation were assessed after 24 hours of culture (37 °C and 5% carbon dioxide in a humidified incubator). SEM was used to observe the attachment of cells onto the scaffolds.
High-resolution micro-computed tomography (Scanco Medical, μCT; Switzerland) was used to assess the porosity and average pore size of the scaffolds. The data were analyzed by μCT. Scanning electron microscopy (SEM; USA) was performed to observe the microstructure and morphology of the 3D-printed scaffolds.
Mesenchymal stem cells (MSCs) isolated from the eBM were used. Cell suspension was seeded at a density of 1×105 cells on the 3D-printed scaffolds in 24-well plates. Cell spreading and focal adhesion formation were assessed after 24 hours of culture (37 °C and 5% carbon dioxide in a humidified incubator). SEM was used to observe the attachment of cells onto the scaffolds.
4
Calcium Phosphate Cement 3D Printing
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A 3D Bioplotter (EnvisionTec, Gladbeck, Germany) with a low-temperature printing head as well as conical needles made of polypropylene with inner diameters of 0.2 and 0.25 mm were used to print the different geometries. The CPC paste used was made by Innotere (Innotere, Radebeul, Germany). It consisted of synthetic calcium and phosphate salts finely dispersed in a biocompatible oil phase of short-chain triglycerides (caprycil/capric triglycerides) together with two further emulsifiers (polyoxyl-35-castor oil/cetyl phosphate). The triglycerides and the polyoxyl-35-castor oil (castor oil) were both based on pure vegetable raw materials.
5
Bioprinting of GelMA and GelMA-AuNPs Scaffolds
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GelMA pre-polymer solution was prepared as explained in Section 2.1 , sterile filtered (PES membrane, 0.22 µm), and transferred into the sterile plotting cartridge (Nordson EFD, Westlake, OH). The plotting cartridge inserted into the printing head of a 3D Bioplotter (Envisiontec, Gladbeck, Germany) and the GelMA and GelMA-AuNPs solutions were cooled down to 20 °C. The pre-polymer solutions were extruded through a 23G needle with an inner diameter of 330 µm and a length of 3.81 cm (general purpose dispersing tips, Nordson EFD, East Providence, RI, USA) onto the printing plate, which was cooled down to 4 °C. Layer-by-layer printing, a speed of 18 mm/s, a 2.3 bar extrusion pressure, and a 200 µm layer thickness were used to form 0°/90° oriented strands with a 1.2 mm distance. CAD files specifying the cylindrical geometry (diameter: 30 mm, thickness: 3 mm) of the scaffolds were used as the input to produce the physical model for the 3D Bioplotter (EnvisionTech GmbH, Gladbeck, Germany) and converted into G-code via Visual Machines software (EnvisionTech GmbH, Gladbeck, Germany) for the printing process. 3D constructs were printed and then exposed to 12.5 mW/cm2 UV light (BlueWave® 75 UV Light Curing Spot Lamp, 365 nm, Torrington, CT, USA) for 60 s for permanent crosslinking.
6
Bioprinting of Alginate-based Hydrogel
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The printing process was carried out in a support bath. The fully transparent and biocompatible support bath was prepared as previously described using a microparticulate formulation method [28 (link)]. Briefly, 0.1% (w/v) microparticulate (composed of 0.32% (w/v) sodium alginate, 0.25% (w/v) xanthan gum, 9.56 × 10−3 M calcium carbonate and 19.15 × 10−3 M D-(+)-gluconic acid δ-lacton) was resuspended in 1% (w/v) xanthan gum to form the support bath. The bioink at a 3% (w/v) concentration was generated by dissolving lyophilized SpGel powder in PBS solution, which was supplemented with 0.04% (v/v) blue food dye to visualize the printing process and the final pattern. A 10-ml syringe fitted with a 30G needle was loaded with freshly prepared bioink (5 ml) and then mounted onto a 3D extrusion printer (3D-bioplotter, Envisiontec). The extrusion pressure and printing speed were kept constant at 1 bar and 25 mm/s, respectively. After printing, the SpGel hydrogel in the support bath solidified at 37 °C to form the designed pattern.
7
Anatomically Engineered Meniscal Scaffolds
8
3D Scaffolds Fabrication with PLGA-nHA-Collagen
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A set of 3D scaffolds were fabricated using a 3D Bioplotter (EnvisionTEC, Gladbeck, NRW, Germany). Briefly, 2.1 g of ground PLGA was mixed with 0.9 g of nHA and 3.6 mL of HFP in a 25 mL beaker for 120 s. In addition, 0.225 g of collagen and 3 mL of HFP were mixed for 90 s in another 25 mL beaker. Both beakers were then sealed and kept under a fume hood for 23 h. Then, 0.42 g of ground PEG (20% of PLGA mass) was added to the PLGA-nHA beaker. Finally, the collagen solution was poured into the beaker, and the solution was mixed for 5 min, followed by sonication at an amplitude of 40 μm for 3 min with a Fisherbrand Model 50 Sonic Dismembrator (Fisher Scientific, Portsmouth, NH, USA). The 3D Bioplotter was calibrated prior to scaffold fabrication. The 3D geometry used for 3D plotting was a 20 mm × 20 mm × 3 mm block partitioned into 300 μm layers, which enabled some overlap between the successive layers to prevent delamination [20 (link)]. The 3D Bioplotter was set to a pressure of 1.5 bar, a dispensing speed of 2.4 mm/s, and a temperature of 20 °C. The edge-to-edge distance between the plotted strands was set to 1000 μm. The scaffolds were dried under the fume hood for 14 days. The plasma-treated scaffold samples used for the BCA assay were immersed in PBS immediately after the plasma treatment.
9
3D-Printed Silk Fibroin/Gelatin Hydrogel Scaffolds
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The 3D bioplotter (Envision TEC, Germany) was used to prepare 3D-printed SF/GT hydrogel scaffolds. HRP with a concentration of 60 U/mL was added to the three pregel solutions prepared in section 2.2.3 , and the mixtures were then transferred into printer cartridges. The dosing pressure of the syringe pump was set between 1 and 1.5 bar, and the moving speed of the dispensing unit was set to 10–15 mm/s. The nozzle size was selected to be 200 μm, and the fiber spacing was set to 0.6 mm. The temperature of the cartridge and printing platform were set to 20–30 °C and 4 °C, respectively. Printed scaffolds were immediately transferred into 5 mM H2O2 solution and left to rest for 30 min to obtain enzymatically crosslinked hydrogel scaffolds (denoted as E-SF0GT15, E-SF2.5GT15, and E-SF5GT15). The scaffolds were then transferred into 75% methanol solution for 12 h, after which they were washed with deionized water three times to remove the methanol and obtain the final hydrogels (denoted as EM-SF0GT15, EM-SF2.5GT15, and EM-SF5GT15).
10
Bioprinting Microgel-Based Constructs
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Bioinks were made fresh and loaded to 50 cc cartridge, which was then placed in the printer (3D Bioplotter, EnvisionTec) nozzle set at a certain temperature. The nozzle temperature was determined when the bioinks could hang a smooth filament at the nozzle during manual extrusion. The pneumatic pressure was determined when the bioinks was extruded at a volume equal to an ≈1 mm diameter spheroid during 1 s purge. Printing speed was set at 2.5 mm s–1 for tubular and thin lattice structures. For centimeter-sized structure printing, printing speed was set at 10 mm s–1 with a higher pneumatic pressure accordingly. Unless otherwise stated, 25-gauge needle (inner diameter of 260 μm) was used for all the tested groups. Due to the presence of a continuous matrix phase and the smaller size of microgels than the needle, the printing process was smoothly conducted without observing undesired aggregating or jamming of microgels within the bioinks. After printing, the constructs were treated with UV light (10 mW cm–2, 2.5 min in the air plus 2.5 min in photoinitiator solution), followed by 37 °C incubation.
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