Novel 3D printing technology for CT phantom coronary arteries with high geometrical accuracy for biomedical imaging applications
Cellularized scaffolds fabricated with hydrogel do not possess sufficient strength to act as stand-alone implant devices for hard tissue repair and regeneration. A thermoplastic polymer support structure typically provides the structural integrity to scaffolds while cells and growth factors in hydrogel provide biological stimulation for tissue formation. In this research, we investigated the viability of human adipose-derived mesenchymal stem cells (ASCs) mixed in an alginate-gelatin (1:1) hydrogel (bioink) that is deposited between polylactic acid (PLA)-borate glass composite filaments. Bioactive borate glass (13–93B3, also called B3 glass) is a rapidly dissolving biomaterial, and the physiologically relevant ionic dissolution products are known to stimulate cells in vitro and endogenous tissues in vivo. B3 glass was added to PLA in two different weight ratios (50% and 33%) to form PLA ?+ ?B3 glass composites, and the weight loss over time of the 3D printed composite scaffold, pH change of the surrounding media, and mechanical properties were investigated. Physical assessment of the composite scaffolds indicated improved mechanical properties and complete glass dissolution within two weeks. Cellularized scaffolds were bioprinted in three configurations: Bioink only, PLA ?+ ?Bioink, and PLA ?+ ?B3 glass ?+ ?Bioink, and cultured in dynamic conditions to investigate ASC viability. The results indicated a non-uniform cell viability along the scaffold thickness, with hypoxic-like conditions and lower viability at the bottom region to higher viability in the top layers of the scaffold.
https://www.sciencedirect.com/science/article/pii/S2405886620300026?dgcid=rss_sd_allhttps://www.sciencedirect.com/science/article/pii/S2405886620300026Design and characterisation of multi-functional strontium-gelatin nanocomposite bioinks with improved print fidelity and osteogenic capacityPublication date: June 2020
Source: Bioprinting, Volume 18
Author(s): Cesar R. Alcala-Orozco, Isha Mutreja, Xiaolin Cui, Dhiraj Kumar, Gary J. Hooper, Khoon S. Lim, Tim B.F. Woodfield
3D bioprinting of constructs for tissue engineering and regenerative medicine has steadily gained attention due to its potential to fabricate anatomically-precise living constructs, localise specific cell types and enable the regeneration of functional tissues in a clinical setting. However, the limited availability of bioinks that can be successfully 3D bioprinted with high fidelity and simultaneously provide encapsulated cells with a tailored, low-stiffness microenvironment supporting functional tissue formation remains an unmet challenge. To address both the physical and biological limitations of available bioinks, this study aimed to develop a nanocomposite bioink (Sr-GelMA) comprising of strontium-carbonate (Sr) nanoparticles and low concentration (5 w/v%) gelatin-methacryloyl (GelMA) hydrogel for extrusion-based 3D bioprinting of low-stiffness cell-laden scaffolds with high shape fidelity and bone-specific cell signalling factors. We systematically investigated the effect of Sr incorporation on hydrogel physico-chemical properties, print fidelity, scaffold shape retention, as well as cell viability, osteogenic differentiation and in vitro bone formation. Nanocomposite Sr-GelMA hydrogels retained their physical and mechanical properties, while rheological studies revealed a significant increase in viscosity profiles that led to notably enhanced printability compared to GelMA alone. Moreover, bioprinted Sr-GelMA scaffolds exhibited excellent shape fidelity evidenced by a defined pore geometry on the x-y-z axis, resulting in an interconnected bioink filament and pore network that was maintained even after long-term culture and osteogenic differentiation (28 days) of human mesenchymal stromal cells (hMSCs). The presence of clustered Sr nanoparticles in the cell-laden bioink allowed high quality bioprinting combined with high hMSC viability (>95%) post-fabrication. Furthermore, Sr addition resulted in enhanced osteogenic differentiation of hMSCs as revealed by higher alkaline phosphatase (ALP) levels, osteocalcin (OCN) and collagen type-I (Col I) expression, with mineralised nodule formation distributed homogenously throughout the bioprinted construct. This study demonstrated that strontium-based nanocomposite bioinks optimised for extrusion-based 3D bioprinting of osteoconductive scaffolds support long-term shape retention with promising potential for bone tissue regeneration.
https://www.sciencedirect.com/science/article/pii/S2405886619300417?dgcid=rss_sd_allhttps://www.sciencedirect.com/science/article/pii/S2405886619300417Cell death persists in rapid extrusion of lysis-resistant coated cardiac myoblastsPublication date: June 2020
Source: Bioprinting, Volume 18
Author(s): Calvin F. Cahall, Aman Preet Kaur, Kara A. Davis, Jonathan T. Pham, Hainsworth Y. Shin, Brad J. Berron
As the demand for organ transplants continues to grow faster than the supply of available donor organs, a new source of functional organs is needed. High resolution high throughput 3D bioprinting is one approach towards generating functional organs for transplantation. For high throughput printing, the need for increased print resolutions (by decreasing printing nozzle diameter) has a consequence: it increases the forces that cause cell damage during the printing process. Here, a novel cell encapsulation method provides mechanical protection from complete lysis of individual living cells during extrusion-based bioprinting. Cells coated in polymers possessing the mechanical properties finely-tuned to maintain size and shape following extrusion, and these encapsulated cells are protected from mechanical lysis. However, the shear forces imposed on the cells during extrusion still cause sufficient damage to compromise the cell membrane integrity and adversely impact normal cellular function. Cellular damage occurred during the extrusion process independent of the rapid depressurization.
https://www.sciencedirect.com/science/article/pii/S2405886619300132?dgcid=rss_sd_allhttps://www.sciencedirect.com/science/article/pii/S24058866193001323D bioprinting: A powerful tool to leverage tissue engineering and microbial systemsPublication date: June 2020
Source: Bioprinting, Volume 18
Author(s): Ecem Saygili, Asli Aybike Dogan-Gurbuz, Ozlem Yesil-Celiktas, Mohamed S. Draz
Bioprinting covers the precise deposition of cells, biological scaffolds and growth factors to produce desired tissue models. The main focus of bioprinting is the creation of functional three-dimensional (3D) biomimetic composites for various application areas. Successful creations of model tissues depend on certain parameters such as determination of optimum microenvironment conditions, selection of appropriate scaffold, and cell source. As the cell culture-based assays have vital roles in the biomedical field, bioprinted tissue analogs would provide unprecedented chances to study, screen, and treat diseases. Today’s 3D bioprinting technology is able to print cells and scaffolds simultaneously, which provides the opportunity for disease modeling. This paper presents a general overview of the current state of the art in bioprinting technologies and potential 3D cell culture systems now being developed to model microbial infections, host-pathogen interactions, niches for microbiota, biofilm formation, and assess microbial resistance to antibiotics.
https://www.sciencedirect.com/science/article/pii/S2405886619300405?dgcid=rss_sd_allhttps://www.sciencedirect.com/science/article/pii/S2405886619300405Introduction to the state-of-the-art 3D bioprinting methods, design, and applications in orthopedicsPublication date: June 2020
Source: Bioprinting, Volume 18
Author(s): Julia Anna Semba, Adam Aron Mieloch, Jakub Dalibor Rybka
Cartilage injuries and bone loss become increasingly prevalent in modern societies. Articular cartilage and menisci have low or no capacity for self-repair and none of the available treatments provide satisfactory, long-term outcomes. Additionally, despite self-regenerating capabilities of bone tissue, the mechanism may fail or become insufficient, creating the need for surgical bone replacement, which is restrained by natural graft accessibility. 3D bioprinting is a rapidly developing technology emerging as a promising remedial therapy in orthopedics. The extensive and ongoing studies in this field are focused on such topics as cartilage and bone biology, standardization of cell culture protocols, bioink formulation, and 3D bioprinting technology. Recent results of these examinations, focused on applications in orthopedics, are presented in this review.
https://www.sciencedirect.com/science/article/pii/S2405886619300399?dgcid=rss_sd_allhttps://www.sciencedirect.com/science/article/pii/S2405886619300399Fiber engraving for bioink bioprinting within 3D printed tissue engineering scaffoldsPublication date: June 2020
Source: Bioprinting, Volume 18
Author(s): Luis Diaz-Gomez, Maryam E. Elizondo, Gerry L. Koons, Mani Diba, Letitia K. Chim, Elizabeth Cosgriff-Hernandez, Anthony J. Melchiorri, Antonios G. Mikos
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, we fabricated and characterized a multimaterial scaffold containing poly(?-caprolactone) (PCL) as the thermoplastic polymer and a gelatin-based bioink. A 7.5 w/v% gelatin methacryloyl (GelMA) bioink loaded with either 5 w/v% poly(lactic-co-glycolic acid) (PLGA) microparticles containing fluorescent albumin or mouse fibroblasts (1 ?× ?106 ?cell/mL) was printed at 24 ?°C. The structure of the composite scaffolds had no significant decrease in porosity or mechanical properties as compared to the PCL control scaffolds, demonstrating the engraving technique did not significantly compromise the mechanical or structural integrity of the scaffold. The encapsulated PLGA microparticles were homogeneously distributed in the GelMA and remained in the scaffolds after incubation in PBS for 24 ?h ?at 37 ?°C. In addition, the viability of the fibroblasts encapsulated in the GelMA bioink and printed in the grooves of the PCL scaffolds was confirmed after 24 ?h of incubation. Overall, this work provides a new methodology for the preparation of 3D printed scaffolds containing a robust thermoplastic structure in combination with low viscosity bioinks.
https://www.sciencedirect.com/science/article/pii/S2405886620300038?dgcid=rss_sd_allhttps://www.sciencedirect.com/science/article/pii/S2405886620300038Multilayered microcasting of agarose–collagen composites for neurovascular modelingPublication date: March 2020
Source: Bioprinting, Volume 17
Author(s): Hossein Heidari, Hayden Taylor
The in vitro fabrication of vascular networks is one of the most complex challenges currently faced in tissue engineering. We describe a method to create multi-layered, cell-laden hydrogel microstructures with coaxial geometries and heterogeneous elastic moduli. The technique can be used to build in vitro vascular structures that are fully embedded in physiologically realistic hydrogels. Our technique eliminates rigid polymeric surfaces from the vicinity of the cells—overcoming a limitation of many microfluidic models—and allows layers of multiple cell types to be defined with tailored ECM composition and stiffness, and in direct contact with each other. We demonstrate channels with internal diameters as small as 175 ??m, and agarose–collagen (AC) gels whose Young’s moduli range from 1.4–8.3 ?kPa. We also show co-axial geometries with layer thicknesses as small as 125 ??m. One potential application of such structures is to simulate brain microvasculature. Towards this goal, the composition and mechanical properties of the composite AC hydrogels are optimized for cell viability and biological performance in both 2D and 3D culture. Seven-day viability of human microvascular endothelial cells (HMECs) and SY5Y glial cells is found to be maximized with a collagen content of 0.05% (w/v) when agarose content ranges between 0.25% and 1% (w/v). Additionally, we quantify the roles of type I bovine and rat-tail collagen, Matrigel, and poly-d-lysine–collagen–Matrigel coatings in promoting HMEC spreading, proliferation and confluence. 3D triple-layer vascular constructs have been fabricated, composed of a cannular monolayer of HMECs surrounded by two regions of SY5Ys with differing spatial densities. The endothelia are confluent and maintain trans-endothelial electrical resistance (TEER) values around 300 ?? ?cm2 over 11.5 days. This prototype opens the way for intricate multi-luminal blood vessels to be fabricated in vitro.
https://www.sciencedirect.com/science/article/pii/S2405886619300387?dgcid=rss_sd_allhttps://www.sciencedirect.com/science/article/pii/S2405886619300387Chondroprotective and osteogenic effects of silk-based bioinks in developing 3D bioprinted osteochondral interfacePublication date: March 2020
Source: Bioprinting, Volume 17
Author(s): Joseph Christakiran Moses, Triya Saha, Biman B. Mandal
Attributing cell instructive features and multifunctionality to biological inks (bioinks) employed for three-dimensional (3D) printing strategies is very much essential to bring about a paradigm shift in developing next generation smart intuitive 3D bioprinted constructs. Giving perspective to this notion, we explore here the feasibilities in developing multifunctional silk-based cartilage and bone bioinks for recreating heterogeneous complicated tissue constructs such as the osteochondral interface. In this regard, the developed silk based bioinks exhibit shear thinning behaviour with quick thixotropic recovery (~90% recovery) aiding in printing self-standing structures with decent print fidelity. The hydrogel network within the 3D bioprinted constructs present good permeability enabling in forming an undulating demarcation region at the bioprinted osteochondral interface. Furthermore, the cartilage and bone inks used for the microextrusion based bioprinting of osteochondral constructs facilitate the spatial maturation and differentiation of encapsulated stem cells towards osteogenic and chondrogenic lineages. The incorporation of strontium doped nano-apatites activates hypoxia inducible factor (HIF-1?) related genes, conferring proangiogenic and chondroprotective traits to the bioinks. Involvement of strontium in down regulating cyclooxygenase-2 via inhibiting prostaglandins (PGE2) pathway enabled anti-osteoclastic activity while favouring M2 macrophage biasness. Altogether, these findings corroborate the potential of the developed nanocomposite bioinks for fabricating clinically viable grafts for osteochondral defect repair associated with osteoporosis.