Technological watch

Fabrication and characterization of porous bone scaffold of bovine hydroxyapatite-glycerin by 3D printing technology

Clinically used tissue-engineered devices are few and far between. The search for biocompatible materials has been a great focus for the field for the past 30 years. The ones that exist are hard to work with when manufacturing patient-specific shapes with complex external and internal pore geometries because of their material and mechanical properties. Photo-cross-linkable polymers have arisen as a potential 3D printable solution for this problem. Here we present a new sugar-based polymer, isosorbide-derived polyester as a potential candidate to help fill this gap.

Isosorbide-derived polyesters containing different amount of double bonds were synthesized to allow the material to be crosslinked or functionalized. Three resin formulations with different amounts of double bonds available (5, 12 and 17%) were used. After initial thermal and photo-crosslinking studies, Isosorbide-derived unsaturated polyesters were then mixed with a photoinitiator, co-crosslinker, and solvent to form a 3D printable resin formulation. Thin films of that resin were then cured using a Digital Light Projection (DLP) printer and analyzed. An attempt was made to modulate the cured thickness by adding HMB as a photoattenuator. We assessed 3D printing success and tensile green strength for the 3 resin formulations.

We were able to 3D print unsaturated polyesters from isosorbide with a DLP printer when enough double bonds were present to facilitate crosslinking and curing of the material. Finally, thin films were then seeded with fibroblasts following the ISO 10993-5 protocol. These coupons were found safe (i.e., non-cytotoxic and biocompatible), potentially marking the beginning of a new approach to preparing resorbable implants for tissue engineering uses. 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. 3D printing technology for CT phantom coronary arteries with high geometrical accuracy for biomedical imaging applicationsPublication date: June 2020

Source: Bioprinting, Volume 18

Author(s): Karolina Stepniak, Ali Ursani, Narinder Paul, Hani Naguib

There is a growing interest in using Computed Tomography (CT) and Hounsfield Unit (HU) measurements in identifying and assessing non-calcified plaque; however, the complex geometry of the coronary arteries poses a challenge in achieving good image quality, which is crucial in providing patients with an accurate diagnosis. Minimizing artifacts associated with cardiac motion is also an important step in improving CT diagnostic accuracy of Coronary Artery Disease. Existing arterial phantoms are commonly straight, short tubular sections that are often rigid, and do not represent the geometry of the entire arterial network, with which image quality and Hounsfield Unit measurements vary and are dependent on. In this study, the process of manufacturing a plaque phantom with physiologically accurate geometry of the coronary arteries is demonstrated. A computer model is obtained by segmenting CTCA images, and several flexible commercially available materials are used to 3D print the model. The static and dynamic mechanical properties of the 3D printing materials are investigated under physiologically relevant loading and the CT numbers of contrast-enhanced tubular samples with 50%, 75%, and 90% concentric stenosis are characterized and compared with ranges for lipid-rich and fibrous plaque. The proposed plaque phantom design offers the possibility of investigating the effect of non-calcified plaque geometry and arterial motion on various parameters in CT optimization studies. with bioactive glass loaded polylactic acid composite and human adipose stem cellsPublication date: June 2020

Source: Bioprinting, Volume 18

Author(s): Krishna C.R. Kolan, Julie A. Semon, August T. Bindbeutel, Delbert E. Day, Ming C. Leu

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. 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. 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. 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. 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. 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. 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.

Graphical abstract

Publication date: 01/06/2020

Author: Joko Triyono, Raasyid Alfiansyah, Heru Sukanto, Dody Ariawan, Yohanes NugrohoAbstractThis research aims to know the mechanical properties of bone graft bio-composites of bovine hydroxyapatite (BHA)-Glycerin by deposition using three-dimensional (3D) Prin


This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 870292.