It’s tough to fabricate advanced tissue constructs with the required mechanical properties and construction integrity while you’re with bioprinting in mushy tissue engineering. Usually, scientists will use supplies, like PCL, to strengthen the within of 3D bioprinting constructs, however the lengthy degradation interval just isn’t nice. However a crew of researchers – Pei Zhuang, Wei Lengthy Ng, Jia An, Chee Kai Chua, and Lay Poh Tan – from Singapore’s Nanyang Technological College have proposed a novel UV-assisted, extrusion-based (UAE) 3D bioprinting technique that might assist fabricate mushy tissue constructs with the specified structural integrity. The crew lays out their work in a paper titled “Layer-by-layer ultraviolet assisted extrusion-based (UAE) bioprinting of hydrogel constructs with excessive side ratio for mushy tissue engineering purposes.”
The summary reads, “Herein, we current a facile bioprinting technique that mixes the fast extrusion-based bioprinting method with an in-built ultraviolet (UV) curing system to facilitate the layer-by-layer UV curing of bioprinted photo-curable GelMA-based hydrogels to attain mushy but secure cell-laden constructs with excessive side ratio for mushy tissue engineering. GelMA is supplemented with a viscosity enhancer (gellan gum) to enhance the bio-ink printability and form constancy whereas sustaining the biocompatibility earlier than crosslinking through a layer-by-layer UV curing course of. This strategy might finally fabricate mushy tissue constructs with excessive side ratio (size to diameter) of ≥ 5. The consequences of UV supply on printing decision and cell viability have been additionally studied. As a proof-of-concept, small constructing models (3D lattice and tubular constructs) with excessive side ratio are fabricated. Moreover, we have now additionally demonstrated the power to carry out multi-material printing of tissue constructs with excessive side ratio alongside each the longitudinal and transverse instructions for potential purposes in tissue engineering of soppy tissues.”
Utilizing 3D bioprinting to exactly deposit cells and biomaterials can actually assist facilitate interactions between them, which permits tissue maturation. A very good microenvironment is important for regulation, however as beforehand talked about, in relation to mushy tissue engineering, it’s tough to attain this. Whereas there are various bioprinting approaches, extrusion-based appears greatest due to its compatibility with bioinks, ease of operation, and fast fabrication pace.
“A perfect bio-ink ought to exhibit good printability, biocompatibility and compliant tissue stiffness,” the researchers defined. “Many of the present bio-inks are modified from pure biomaterials similar to gelatin ([30–34]) and collagen ([35–37]) to kind new composite bio-inks with tunable properties. Significantly, gelatin methacryloyl (GelMA) has been recognized as a promising bio-ink owing to its wonderful organic properties and tunable bodily properties ([38, 39]).”
Bioinks based mostly in GelMA are sometimes utilized in regenerative medication and tissue engineering, but when there are excessive concentrations of this materials, restricted cell exercise can happen due to excessive crosslinking density, in addition to stiffness of the photo-crosslinked constructs; low concentrations may cause poor form constancy and low printing decision.
“Therefore, additional optimization is required to enhance the steadiness and printability of GelMA bio-inks. A plethora of strategies have been explored to enhance the rheological conduct of GelMA, such because the addition of varied supplies like nanosilicates (), partial crosslinking GelMA with enzymes (), or by means of cooling course of,” said the researchers. “Amongst these strategies, gellan gum, which is a non-toxic polysaccharide, has been found as a promising rheological modifier to enhance the rheological property of the bio-ink.”
The crew selected to make use of the minimal obligatory quantity of gellan gum (GG) with a view to stability biocompatibility and “endow the improved printability of GelMA-GG,” whereas they used layer-by-layer UAE bioprinting when making thick cell-laden tissue constructs with a view to reinforce decision and construction stability.
“As such, we have now demonstrated the power to manufacture bioprinted constructs with excessive side ratio through a layer-by-layer UAE bioprinting technique,” the researchers defined.
“The examine affords a brand new bioprinting technique to generate secure 3D buildings with compliant mechanical property and excessive side ratio utilizing GelMA-based (GelMA-GG) bio-inks for engineering of soppy tissue constructs.”
They investigated 30 totally different combos of GelMA-GG bio-inks in three phases:
Bioink preparation phaseBioprinting phasePost-printing section
The crew loaded the cells into the composite bioinks to guage their potential sedimentation and ease of cell encapsulation. The bioinks have been then printed utilizing the UAE technique with a view to consider accuracy and printability of the constructs. The researchers adjusted the print parameters with a view to “obtain buildings with excessive side ratio,” after which investigated the “UV results on printing decision and cell conduct,” earlier than lastly evaluating the 3D printed constructs for his or her corresponding cell conduct and materials properties.
The crew used a regenHU bioprinter, with a temperature of 25 ± 1°C and 27G needle, for its experiments, and fabricated GelMA-GG constructs in an oblong form.
“To optimize the printing course of, 2D buildings have been first printed to find out the bio-ink printability after which additional checks have been performed to find out the optimum printing strain and feed fee. Moreover, UV scanning pace has proven essential results on printing decision and cell viability,” the researchers wrote. “To check the affect of UV scanning pace on printing decision, C2C12 encapsulated GelMA-GG constructs with 1, three, 5, 7, 9 and 11 layers have been printed into grid sample underneath fastened printing pace and strain. The width of the printed filaments have been measured to find out the change of printing decision extra time. In the meantime, Stay/lifeless staining and ImageJ was used to analyse the cell viability within the printed constructs. Cells within the backside layer of the constructs have been by imaged by fluorescent microscope to find out the UV affect on cell viability by cell counting evaluation. After which, 3D grid and tubular constructs of various side ratio at various diameter have been printed with layer-by-layer UV curing strategy utilizing the recognized optimum printing parameters with all of the printable bio-inks.”
C2C12 cells have been additionally printed, in order that the crew might examine how materials stiffness and microstructure influenced cell conduct. They have been capable of simply select composite bio-inks for particular tissue engineering purposes by performing a guide casting strategy.
“The GelMA-based bio-inks have exhibited nice biocompatibility for cells because of the presence of RGD peptides,” the researchers concluded.
“To strike a stability between printability and biocompatibility, minimal ideally suited quantity of gellan gum was added to implement the printability of the bioinks with out compromising the biocompatibility. In-depth characterization and analysis on the totally different composite GelMA-GG bio-inks have been carried out to pick out an acceptable vary of GelMA-GG bio-inks by means of our proposed parametric examine.
“From our work, an acceptable vary of bio-ink viscosity decrease than Zero.124 Pa·s at 37°C was discovered to be appropriate for cell encapsulation and to attain a homogeneous cell-laden bio-inks. Materials viscosity of Zero.2-1.Zero Pa·s at a printing temperature of 25°C is advisable for printing of advanced 3D cell-laden constructs with excessive side ratio utilizing our layer-by-layer UV-assisted bioprinting technique. The essential function of UV supply in printing course of has been investigated, particularly, the UV affect on printing decision and cell survival fee. As well as, a powerful correlation between materials microstructure and stiffness has been proven in our examine and their synergistic affect on cell conduct has been investigated.”
The researchers mentioned their technique may very well be tailored for all light-curable supplies.
Focus on this and different 3D printing subjects at 3DPrintBoard.com or share your ideas beneath.
Credit score : Supply Hyperlink