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AUTHOR Asulin, Masha and Michael, Idan and Shapira, Assaf and Dvir, Tal
Title One-Step 3D Printing of Heart Patches with Built-In Electronics for Performance Regulation [Abstract]
Year 2021
Journal/Proceedings Advanced Science
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Abstract Three dimensional (3D) printing of heart patches usually provides the ability to precisely control cell location in 3D space. Here, one-step 3D printing of cardiac patches with built-in soft and stretchable electronics is reported. The tissue is simultaneously printed using three distinct bioinks for the cells, for the conducting parts of the electronics and for the dielectric components. It is shown that the hybrid system can withstand continuous physical deformations as those taking place in the contracting myocardium. The electronic patch is flexible, stretchable, and soft, and the electrodes within the printed patch are able to monitor the function of the engineered tissue by providing extracellular potentials. Furthermore, the system allowed controlling tissue function by providing electrical stimulation for pacing. It is envisioned that such transplantable patches may regain heart contractility and allow the physician to monitor the implant function as well as to efficiently intervene from afar when needed.
AUTHOR Afanasenkau, Dzmitry and Kalinina, Daria and Lyakhovetskii, Vsevolod and Tondera, Christoph and Gorsky, Oleg and Moosavi, Seyyed and Pavlova, Natalia and Merkulyeva, Natalia and Kalueff, Allan V. and Minev, Ivan R. and Musienko, Pavel
Title Rapid prototyping of soft bioelectronic implants for use as neuromuscular interfaces [Abstract]
Year 2020
Journal/Proceedings Nature Biomedical Engineering
Reftype Afanasenkau2020
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Neuromuscular interfaces are required to translate bioelectronic technologies for application in clinical medicine. Here, by leveraging the robotically controlled ink-jet deposition of low-viscosity conductive inks, extrusion of insulating silicone pastes and in situ activation of electrode surfaces via cold-air plasma, we show that soft biocompatible materials can be rapidly printed for the on-demand prototyping of customized electrode arrays well adjusted to specific anatomical environments, functions and experimental models. We also show, with the monitoring and activation of neuronal pathways in the brain, spinal cord and neuromuscular system of cats, rats and zebrafish, that the printed bioelectronic interfaces allow for long-term integration and functional stability. This technology might enable personalized bioelectronics for neuroprosthetic applications.
AUTHOR Kleger, Nicole and Cihova, Martina and Masania, Kunal and Studart, André R. and Löffler, Jörg F.
Title 3d printing of salt as a template for magnesium with structured porosity [Abstract]
Year 2019
Journal/Proceedings advanced materials
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Abstract Porosity is an essential feature in a wide range of applications that combine light weight with high surface area and tunable density. Porous materials can be easily prepared with a vast variety of chemistries using the salt-leaching technique. However, this templating approach has so far been limited to the fabrication of structures with random porosity and relatively simple macroscopic shapes. Here, a technique is reported that combines the ease of salt leaching with the complex shaping possibilities given by additive manufacturing (AM). By tuning the composition of surfactant and solvent, the salt-based paste is rheologically engineered and printed via direct ink writing into grid-like structures displaying structured pores that span from the sub-millimeter to the macroscopic scale. As a proof of concept, dried and sintered NaCl templates are infiltrated with magnesium (Mg), which is typically highly challenging to process by conventional AM techniques due to its highly oxidative nature and high vapor pressure. Mg scaffolds with well-controlled, ordered porosity are obtained after salt removal. The tunable mechanical properties and the potential to be predictably bioresorbed by the human body make these Mg scaffolds attractive for biomedical implants and demonstrate the great potential of this additive technique.
AUTHOR Daly, Andrew C. and Kelly, Daniel J.
Title Biofabrication of spatially organised tissues by directing the growth of cellular spheroids within 3D printed polymeric microchambers [Abstract]
Year 2019
Journal/Proceedings Biomaterials
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Successful tissue engineering requires the generation of human scale implants that mimic the structure, composition and mechanical properties of native tissues. Here, we report a novel biofabrication strategy that enables the engineering of structurally organised tissues by guiding the growth of cellular spheroids within arrays of 3D printed polymeric microchambers. With the goal of engineering stratified articular cartilage, inkjet bioprinting was used to deposit defined numbers of mesenchymal stromal cells (MSCs) and chondrocytes into pre-printed microchambers. These jetted cell suspensions rapidly underwent condensation within the hydrophobic microchambers, leading to the formation of organised arrays of cellular spheroids. The microchambers were also designed to provide boundary conditions to these spheroids, guiding their growth and eventual fusion, leading to the development of stratified cartilage tissue with a depth-dependant collagen fiber architecture that mimicked the structure of native articular cartilage. Furthermore, the composition and biomechanical properties of the bioprinted cartilage was also comparable to the native tissue. Using multi-tool biofabrication, we were also able to engineer anatomically accurate, human scale, osteochondral templates by printing this microchamber system on top of a hypertrophic cartilage region designed to support endochondral bone formation and then maintaining the entire construct in long-term bioreactor culture to enhance tissue development. This bioprinting strategy provides a versatile and scalable approach to engineer structurally organised cartilage tissues for joint resurfacing applications.
AUTHOR Khaled, Shaban A. and Alexander, Morgan R. and Irvine, Derek J. and Wildman, Ricky D. and Wallace, Martin J. and Sharpe, Sonja and Yoo, Jae and Roberts, Clive J.
Title Extrusion 3D Printing of Paracetamol Tablets from a Single Formulation with Tunable Release Profiles Through Control of Tablet Geometry [Abstract]
Year 2018
Journal/Proceedings AAPS PharmSciTech
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An extrusion-based 3D printer was used to fabricate paracetamol tablets with different geometries (mesh, ring and solid) from a single paste-based formulation formed from standard pharmaceutical ingredients. The tablets demonstrate that tunable drug release profiles can be achieved from this single formulation even with high drug loading (>{thinspace}80{%} w/w). The tablets were evaluated for drug release using a USP dissolution testing type I apparatus. The tablets showed well-defined release profiles (from immediate to sustained release) controlled by their different geometries. The dissolution results showed dependency of drug release on the surface area/volume (SA/V) ratio and the SA of the different tablets. The tablets with larger SA/V ratios and SA had faster drug release. The 3D printed tablets were also evaluated for physical and mechanical properties including tablet dimension, drug content, weight variation and breaking force and were within acceptable range as defined by the international standards stated in the US Pharmacopoeia. X-ray powder diffraction, differential scanning calorimetry and attenuated total reflectance Fourier transform infrared spectroscopy were used to identify the physical form of the active and to assess possible drug-excipient interactions. These data again showed that the tablets meet USP requirement. These results clearly demonstrate the potential of 3D printing to create unique pharmaceutical manufacturing, and potentially clinical, opportunities. The ability to use a single unmodified formulation to achieve defined release profiles could allow, for example, relatively straightforward personalization of medicines for individuals with different metabolism rates for certain drugs and hence could offer significant development and clinical opportunities.
AUTHOR Khaled, Shaban A. and Burley, Jonathan C. and Alexander, Morgan R. and Yang, Jing and Roberts, Clive J.
Title 3D printing of five-in-one dose combination polypill with defined immediate and sustained release profiles [Abstract]
Year 2015
Journal/Proceedings Journal of Controlled Release
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Abstract We have used three dimensional (3D) extrusion printing to manufacture a multi-active solid dosage form or so called polypill. This contains five compartmentalised drugs with two independently controlled and well-defined release profiles. This polypill demonstrates that complex medication regimes can be combined in a single personalised tablet. This could potentially improve adherence for those patients currently taking many separate tablets and also allow ready tailoring of a particular drug combination/drug release for the needs of an individual. The polypill here represents a cardiovascular treatment regime with the incorporation of an immediate release compartment with aspirin and hydrochlorothiazide and three sustained release compartments containing pravastatin, atenolol, and ramipril. X-ray powder diffraction (XRPD) and Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR) were used to assess drug-excipient interaction. The printed polypills were evaluated for drug release using {USP} dissolution testing. We found that the polypill showed the intended immediate and sustained release profiles based upon the active/excipient ratio used.
AUTHOR Zhang, Danwei and Jonhson, Win and Herng, Tun Seng and Ang, Yong Quan and Yang, Lin and Tan, Swee Ching and Peng, Erwin and He, Hui and Ding, Jun
Title A 3D-printing method of fabrication for metals{,} ceramics{,} and multi-materials using a universal self-curable technique for robocasting [Abstract]
Year 2019
Journal/Proceedings Materials Horizons
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Ceramics and metals are important materials that modern technologies are constructed from. The capability to produce such materials in a complex geometry with good mechanical properties can revolutionize the way we engineer our devices. Current curing techniques pose challenges such as high energy requirements{,} limitations of materials with high refractive index{,} tedious post-processing heat treatment processes{,} uneven drying shrinkages{,} and brittleness of green bodies. In this paper{,} a novel modified self-curable epoxide–amine 3D printing system is proposed to print a wide range of ceramics (metal oxides{,} nitrides{,} and carbides) and metals without the need for an external curing source. Through this technique{,} complex multi-material structures (with metal–ceramic and ceramic–ceramic combinations) can also be realized. Tailoring and matching the sintering temperatures of different materials through sintering additives and dopants{,} combined with a structural design providing maximum adhesion between interfaces{,} allow us to successfully obtain superior quality sintered multi-material structures. High-quality ceramic and metallic materials have been achieved (e.g.{,} zirconia with >98% theoretical density). Also{,} highly conductive metals and magnetic ceramics were printed and shaped uniquely without the need for a sacrificial support. With the addition of low molecular weight plasticizers and a multi-stage heat treatment process{,} crack-free and dense high-quality integrated multi-material structures fabricated by 3D printing can thus be a reality in the near future.
AUTHOR Khaled, Shaban A. and Alexander, Morgan R. and Wildman, Ricky D. and Wallace, Martin J. and Sharpe, Sonja and Yoo, Jae and Roberts, Clive J.
Title 3D extrusion printing of high drug loading immediate release paracetamol tablets [Abstract]
Year 2018
Journal/Proceedings International Journal of Pharmaceutics
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The manufacture of immediate release high drug loading paracetamol oral tablets was achieved using an extrusion based 3D printer from a premixed water based paste formulation. The 3D printed tablets demonstrate that a very high drug (paracetamol) loading formulation (80% w/w) can be printed as an acceptable tablet using a method suitable for personalisation and distributed manufacture. Paracetamol is an example of a drug whose physical form can present challenges to traditional powder compression tableting. Printing avoids these issues and facilitates the relatively high drug loading. The 3D printed tablets were evaluated for physical and mechanical properties including weight variation, friability, breaking force, disintegration time, and dimensions and were within acceptable range as defined by the international standards stated in the United States Pharmacopoeia (USP). X-ray Powder Diffraction (XRPD) was used to identify the physical form of the active. Additionally, XRPD, Attenuated Total Reflectance Fourier Transform Infrared spectroscopy (ATR-FTIR) and differential scanning calorimetry (DSC) were used to assess possible drug-excipient interactions. The 3D printed tablets were evaluated for drug release using a USP dissolution testing type I apparatus. The tablets showed a profile characteristic of the immediate release profile as intended based upon the active/excipient ratio used with disintegration in less than 60 s and release of most of the drug within 5 min. The results demonstrate the capability of 3D extrusion based printing to produce acceptable high-drug loading tablets from approved materials that comply with current USP standards.
AUTHOR Daly, Andrew C. and Pitacco, Pierluca and Nulty, Jessica and Cunniffe, Gráinne M. and Kelly, Daniel J.
Title 3D printed microchannel networks to direct vascularisation during endochondral bone repair [Abstract]
Year 2018
Journal/Proceedings Biomaterials
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Bone tissue engineering strategies that recapitulate the developmental process of endochondral ossification offer a promising route to bone repair. Clinical translation of such endochondral tissue engineering strategies will require overcoming a number of challenges, including the engineering of large and often anatomically complex cartilage grafts, as well as the persistence of core regions of avascular cartilage following their implantation into large bone defects. Here 3D printing technology is utilized to develop a versatile and scalable approach to guide vascularisation during endochondral bone repair. First, a sacrificial pluronic ink was used to 3D print interconnected microchannel networks in a mesenchymal stem cell (MSC) laden gelatin-methacryloyl (GelMA) hydrogel. These constructs (with and without microchannels) were next chondrogenically primed in vitro and then implanted into critically sized femoral bone defects in rats. The solid and microchanneled cartilage templates enhanced bone repair compared to untreated controls, with the solid cartilage templates (without microchannels) supporting the highest levels of total bone formation. However, the inclusion of 3D printed microchannels was found to promote osteoclast/immune cell invasion, hydrogel degradation, and vascularisation following implantation. In addition, the endochondral bone tissue engineering strategy was found to support comparable levels of bone healing to BMP-2 delivery, whilst promoting lower levels of heterotopic bone formation, with the microchanneled templates supporting the lowest levels of heterotopic bone formation. Taken together, these results demonstrate that 3D printed hypertrophic cartilage grafts represent a promising approach for the repair of complex bone fractures, particularly for larger defects where vascularisation will be a key challenge.
AUTHOR Ng, Wei Long and Qi, Jovina Tan Zhi and Yeong, Wai Yee and Naing, May Win
Title Proof-of-concept: 3D bioprinting of pigmented human skin constructs [Abstract]
Year 2018
Journal/Proceedings Biofabrication
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Three-dimensional (3D) pigmented human skin constructs have been fabricated using a 3D bioprinting approach. The 3D pigmented human skin constructs are obtained from using three different types of skin cells (keratinocytes, melanocytes and fibroblasts from three different skin donors) and they exhibit similar constitutive pigmentation (pale pigmentation) as the skin donors. A two-step drop-on-demand bioprinting strategy facilitates the deposition of cell droplets to emulate the epidermal melanin units (pre-defined patterning of keratinocytes and melanocytes at the desired positions) and manipulation of the microenvironment to fabricate 3D biomimetic hierarchical porous structures found in native skin tissue. The 3D bioprinted pigmented skin constructs are compared to the pigmented skin constructs fabricated by conventional a manual-casting approach; in-depth characterization of both the 3D pigmented skin constructs has indicated that the 3D bioprinted skin constructs have a higher degree of resemblance to native skin tissue in term of the presence of well-developed stratified epidermal layers and the presence of a continuous layer of basement membrane proteins as compared to the manually-cast samples. The 3D bioprinting approach facilitates the development of 3D in vitro pigmented human skin constructs for potential toxicology testing and fundamental cell biology research.
AUTHOR Khaled, Shaban A. and Burley, Jonathan C. and Alexander, Morgan R. and Yang, Jing and Roberts, Clive J.
Title 3D printing of tablets containing multiple drugs with defined release profiles [Abstract]
Year 2015
Journal/Proceedings International Journal of Pharmaceutics
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Abstract We have employed three-dimensional (3D) extrusion-based printing as a medicine manufacturing technique for the production of multi-active tablets with well-defined and separate controlled release profiles for three different drugs. This ‘polypill’ made by a 3D additive manufacture technique demonstrates that complex medication regimes can be combined in a single tablet and that it is viable to formulate and ‘dial up’ this single tablet for the particular needs of an individual. The tablets used to illustrate this concept incorporate an osmotic pump with the drug captopril and sustained release compartments with the drugs nifedipine and glipizide. This combination of medicines could potentially be used to treat diabetics suffering from hypertension. The room temperature extrusion process used to print the formulations used excipients commonly employed in the pharmaceutical industry. Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR) and X-ray powder diffraction (XRPD) were used to assess drug–excipient interaction. The printed formulations were evaluated for drug release using {USP} dissolution testing. We found that the captopril portion showed the intended zero order drug release of an osmotic pump and noted that the nifedipine and glipizide portions showed either first order release or Korsmeyer–Peppas release kinetics dependent upon the active/excipient ratio used.
AUTHOR Schaffner, Manuel and Faber, Jakob A. and Pianegonda, Lucas and Rühs, Patrick A. and Coulter, Fergal and Studart, André R.
Title 3D printing of robotic soft actuators with programmable bioinspired architectures [Abstract]
Year 2018
Journal/Proceedings Nature Communications
Reftype Schaffner2018
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Soft actuation allows robots to interact safely with humans, other machines, and their surroundings. Full exploitation of the potential of soft actuators has, however, been hindered by the lack of simple manufacturing routes to generate multimaterial parts with intricate shapes and architectures. Here, we report a 3D printing platform for the seamless digital fabrication of pneumatic silicone actuators exhibiting programmable bioinspired architectures and motions. The actuators comprise an elastomeric body whose surface is decorated with reinforcing stripes at a well-defined lead angle. Similar to the fibrous architectures found in muscular hydrostats, the lead angle can be altered to achieve elongation, contraction, or twisting motions. Using a quantitative model based on lamination theory, we establish design principles for the digital fabrication of silicone-based soft actuators whose functional response is programmed within the material's properties and architecture. Exploring such programmability enables 3D printing of a broad range of soft morphing structures.
AUTHOR Schroeder, Thomas B. H. and Guha, Anirvan and Lamoureux, Aaron and VanRenterghem, Gloria and Sept, David and Shtein, Max and Yang, Jerry and Mayer, Michael
Title An electric-eel-inspired soft power source from stacked hydrogels [Abstract]
Year 2017
Journal/Proceedings Nature
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Progress towards the integration of technology into livingo ganisms requires electrical power sources that are biocompatible, mechanically flexible, and able to harness the chemical energy available inside biological systems. Conventional batteries were not designed with these criteria in mind. The electric organ of the knifefish Electrophorus electricus (commonly known as the electric eel) is, however, an example of an electrical power source that operates within biological constraints while featuring power characteristics that include peak potential differences of 600 volts and currents of 1 ampere1,2. Here we introduce an electric eel-inspired power concept that uses gradients of ions between miniature polyacrylamide hydrogel compartments bounded by a repeating sequence of cation- and anion-selective hydrogel membranes. The system uses a scalable stacking or folding geometry that generates 110 volts at open circuit or 27 milliwatts per square metre per gel cell upon simultaneous, self-registered mechanical contact activation of thousands of gel compartments in series while circumventing power dissipation before contact. Unlike typical batteries, these systems are soft, flexible, transparent, and potentially biocompatible. These characteristics suggest that artificial electric organs could be used to power next-generation implant materials such as pacemakers, implantable sensors, or prosthetic devices in hybrids of living and non-living systems3–6.�
AUTHOR Shin, Crystal S. and Cabrera, Fernando J. and Lee, Richard and Kim, John and Ammassam Veettil, Remya and Zaheer, Mahira and Adumbumkulath, Aparna and Mhatre, Kirti and Ajayan, Pulickel M. and Curley, Steven A. and Scott, Bradford G. and Acharya, Ghanashyam
Title 3D-Bioprinted Inflammation Modulating Polymer Scaffolds for Soft Tissue Repair [Abstract]
Year 2021
Journal/Proceedings Advanced Materials
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Abstract Development of inflammation modulating polymer scaffolds for soft tissue repair with minimal postsurgical complications is a compelling clinical need. However, the current standard of care soft tissue repair meshes for hernia repair is highly inflammatory and initiates a dysregulated inflammatory process causing visceral adhesions and postsurgical complications. Herein, the development of an inflammation modulating biomaterial scaffold (bioscaffold) for soft tissue repair is presented. The bioscaffold design is based on the idea that, if the excess proinflammatory cytokines are sequestered from the site of injury by the surgical implantation of a bioscaffold, the inflammatory response can be modulated, and the visceral adhesion formations and postsurgical complications can be minimized. The bioscaffold is fabricated by 3D-bioprinting of an in situ phosphate crosslinked poly(vinyl alcohol) polymer. In vivo efficacy of the bioscaffold is evaluated in a rat ventral hernia model. In vivo proinflammatory cytokine expression analysis and histopathological analysis of the tissues have confirmed that the bioscaffold acts as an inflammation trap and captures the proinflammatory cytokines secreted at the implant site and effectively modulates the local inflammation without the need for exogenous anti-inflammatory agents. The bioscaffold is very effective in inhibiting visceral adhesions formation and minimizing postsurgical complications.
AUTHOR Kwak, Chaesu and Young Ryu, Seoung and Park, Hyunsu and Lim, Sehyeong and Yang, Jeewon and Kim, Jieun and Hyung Kim, Jin and Lee, Joohyung
Title A pickering emulsion stabilized by chlorella microalgae as an eco-friendly extrusion-based 3D printing ink processable under ambient conditions [Abstract]
Year 2021
Journal/Proceedings Journal of Colloid and Interface Science
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Three-dimensional (3D) printing technology is actively utilized in various industrial fields because it facilitates effective and customizable fabrication of complex structures. An important processing route for 3D printing is the extrusion of inks in the form of colloidal suspensions or emulsions, which has recently attracted considerable attention because it allows for selection of a wide range of printing materials and is operable under ambient processing conditions. Herein, we investigate the 3D printability of complex fluids containing chlorella microalgae as an eco-friendly material for 3D printing. Two possible ink types are considered: aqueous chlorella suspensions and emulsions of oil and water mixtures. While the aqueous chlorella suspensions at high particle loading display the 3D-printable rheological properties such as high yield stress and good shape retention, the final structures after extruding and drying the suspensions under ambient conditions show a significant number of macroscopic defects, limiting their practical application. In contrast, the 3D structures produced from the oil-in-water Pickering emulsions stabilized by chlorella microalgae, which are amphiphilic and active at the oil–water interface, show significantly reduced defect formation. Addition of a fast-evaporable oil phase, hexane, is crucial in the mechanisms of enhanced cementation between the individual microalgae via increased inter-particle packing, capillary attraction, and hydrophobic interaction. Furthermore, addition of solid paraffin wax, which is crystalline but well-soluble in the hydrocarbon oil phase under ambient conditions, completely eliminates the undesirable defect formation via enhanced inter-particle binding, while maintaining the overall rheological properties of the emulsion. The optimal formulation of the Pickering emulsion is finally employed to produce a 3D scaffold of satisfactory structural integrity, suggesting that the chlorella-based ink, in the form of an emulsion, has potential as an eco-friendly 3D printing ink processable under ambient conditions.
AUTHOR Tan, Edgar Y. S. and Suntornnond, Ratima and Yeong, Wai Yee
Title High-Resolution Novel Indirect Bioprinting of Low-Viscosity Cell-Laden Hydrogels via Model-Support Bioink Interaction [Abstract]
Year 2021
Journal/Proceedings 3D Printing and Additive Manufacturing
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Abstract Bioprinting of unmodified soft extracellular matrix into complex 3D structures has remained challenging to fabricate. Herein, we established a novel process for the printing of low-viscosity hydrogel by using a unique support technique to retain the structural integrity of the support structure. We demonstrated that this process of printing could be used for different types of hydrogel, ranging from fast crosslinking gelatin methacrylate to slow crosslinking collagen type I. In addition, we evaluated the biocompatibility of the process by observing the effects of the cytotoxicity of L929 and the functionality of the human umbilical vein endothelium primary cells after printing. The results show that the bioprinted construct provided excellent biocompatibility as well as supported cell growth and differentiation. Thus, this is a novel technique that can be potentially used to enhance the resolution of the extrusion-based bioprinter.
AUTHOR King, William E. and Bowlin, Gary L.
Title Near-Field Electrospinning and Melt Electrowriting of Biomedical Polymers—Progress and Limitations [Abstract]
Year 2021
Journal/Proceedings Polymers
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Near-field electrospinning (NFES) and melt electrowriting (MEW) are the process of extruding a fiber due to the force exerted by an electric field and collecting the fiber before bending instabilities occur. When paired with precise relative motion between the polymer source and the collector, a fiber can be directly written as dictated by preprogrammed geometry. As a result, this precise fiber control results in another dimension of scaffold tailorability for biomedical applications. In this review, biomedically relevant polymers that to date have manufactured fibers by NFES/MEW are explored and the present limitations in direct fiber writing of standardization in published setup details, fiber write throughput, and increased ease in the creation of complex scaffold geometries are discussed.
AUTHOR Lagatuz, M. and Vyas, R. J. and Predovic, M. and Lim, S. and Jacobs, N. and Martinho, M. and Valizadegan, H. and Kao, D. and Oza, N. and Theriot, C. A. and Zanello, S. B. and Taibbi, G. and Vizzeri, G. and Dupont, M. and Grant, M. B. and Lindner, D. J. and Reinecker, H.-C. and Pinhas, A. and Chui, T. Y. and Rosen, R. B. and Moldovan, N. and Vickerman, M. B. and Radhakrishnan, K. and Parsons-Wingerter, P.
Title Vascular Patterning as Integrative Readout of Complex Molecular and Physiological Signaling by VESsel GENeration Analysis [Abstract]
Year 2021
Journal/Proceedings J Vasc Res
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The molecular signaling cascades that regulate angiogenesis and microvascular remodeling are fundamental to normal development, healthy physiology, and pathologies such as inflammation and cancer. Yet quantifying such complex, fractally branching vascular patterns remains difficult. We review application of NASA’s globally available, freely downloadable VESsel GENeration (VESGEN) Analysis software to numerous examples of 2D vascular trees, networks, and tree-network composites. Upon input of a binary vascular image, automated output includes informative vascular maps and quantification of parameters such as tortuosity, fractal dimension, vessel diameter, area, length, number, and branch point. Previous research has demonstrated that cytokines and therapeutics such as vascular endothelial growth factor, basic fibroblast growth factor (fibroblast growth factor-2), transforming growth factor-beta-1, and steroid triamcinolone acetonide specify unique “fingerprint” or “biomarker” vascular patterns that integrate dominant signaling with physiological response. In vivo experimental examples described here include vascular response to keratinocyte growth factor, a novel vessel tortuosity factor; angiogenic inhibition in humanized tumor xenografts by the anti-angiogenesis drug leronlimab; intestinal vascular inflammation with probiotic protection by Saccharomyces boulardii, and a workflow programming of vascular architecture for 3D bioprinting of regenerative tissues from 2D images. Microvascular remodeling in the human retina is described for astronaut risks in microgravity, vessel tortuosity in diabetic retinopathy, and venous occlusive disease.
AUTHOR Lin, Che-Wei and Su, Yu-Feng and Lee, Chih-Yun and Kang, Lin and Wang, Yan-Hsiung and Lin, Sung-Yen and Wang, Chih-Kuang
Title 3D printed bioceramics fabricated using negative thermoresponsive hydrogels and silicone oil sealing to promote bone formation in calvarial defects [Abstract]
Year 2020
Journal/Proceedings Ceramics International
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The purpose of the present work was to investigate the potential for application and the effectiveness of osteoconductive scaffolds with bicontinuous phases of 3D printed bioceramics (3DP-BCs) based on reverse negative thermoresponsive hydrogels (poly[(N-isopropylacrylamide)-co-(methacrylic acid)]; p(NiPAAm-MAA)). 3DP-BCs have bioceramic objects and microchannel pores when created using robotic deposition additive manufacturing. We evaluated the benefits of silicone oil sealing on the 3DP-BC green body during the sintering process in terms of densification and structural stability. The shrinkage, density, porosity, element composition, phase structure and microstructural analyses and compression strength measurements of sintered 3DP-BC objects are presented and discussed in this study. In addition, the results of cell viability assays and bone healing analyses of the calvarial bone defects in a rabbit model were used to evaluate 3DP-BC performance. The main results indicated that these 3DP-BC scaffolds have optimal continuous pores and adequate compressive strength, which can enable the protection of calvarial defects and provide an environment for cell growth. Therefore, 3DP-BC scaffolds have better new bone regeneration efficiency in rabbit calvarial bone defect models than empty scaffolds and mold-forming bioceramic scaffolds (MF-BCs).
AUTHOR Rupp, Harald and Binder, Wolfgang H.
Title 3D Printing of Core–Shell Capsule Composites for Post-Reactive and Damage Sensing Applications [Abstract]
Year 2020
Journal/Proceedings Advanced Materials Technologies
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Abstract 3D printing of multicomponent materials as an advantageous method over traditional mold casting methods is demonstrated, developing small core–shell capsule composites fabricated by a two-step 3D printing process. Using a two-print-head system (fused deposition modeling extruder and a liquid inkjet print head), micro-sized capsules are manufactured in sizes ranging from 100 to 800 µm. The thermoplastic polymer poly(ε-caprolactone) (PCL) is chosen as matrix/shell material due to its optimal interaction with the embedded hydrophobic liquids. First, the core–shell capsules are printed with model liquids and pure PCL to optimize the printing parameters and to ensure fully enclosed capsules inside the polymer. As a proof of concept, novel “click” reaction systems, used in self-healing and stress-detection applications, are manufactured in which PCL composites with nano- and micro-fillers are combined with reactive, encapsulated liquids. The so generated 3D printed core–shell capsule composite can be used for post-printing reactions and damage sensing when combined with a fluorogenic dye.
AUTHOR Jung, Harry and Lee, Ji Seung and Lee, Jun Ho and Park, Ki Joon and Lee, Jae Jun and Park, Hae Sang
Title A Feasibility Study for 3D-printed Poly(methyl methacrylate)-resin Tracheostomy Tube Using a Hamster Cheek Pouch Model
Year 2020
Journal/Proceedings In Vivo
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AUTHOR Wang, Zehao and Hui, Aiping and Zhao, Hongbin and Ye, Xiaohan and Zhang, Chao and Wang, Aiqin and Zhang, Changqing
Title A Novel 3D-bioprinted Porous Nano Attapulgite Scaffolds with Good Performance for Bone Regeneration [Abstract]
Year 2020
Journal/Proceedings International Journal of Nanomedicine
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BACKGROUND: Natural clay nanomaterials are an emerging class of biomaterial with great potential for tissue engineering and regenerative medicine applications, most notably for osteogenesis. MATERIALS AND METHODS: Herein, for the first time, novel tissue engineering scaffolds were prepared by 3D bioprinter using nontoxic and bioactive natural attapulgite (ATP) nanorods as starting materials, with polyvinyl alcohol as binder, and then sintered to obtain final scaffolds. The microscopic morphology and structure of ATP particles and scaffolds were observed by transmission electron microscope and scanning electron microscope. In vitro biocompatibility and osteogenesis with osteogenic precursor cell (hBMSCs) were assayed using MTT method, Live/Dead cell staining, alizarin red staining and RT-PCR. In vivo bone regeneration was evaluated with micro-CT and histology analysis in rat cranium defect model. RESULTS: We successfully printed a novel porous nano-ATP scaffold designed with inner channels with a dimension of 500 µm and wall structures with a thickness of 330 µm. The porosity of current 3D-printed scaffolds ranges from 75% to 82% and the longitudinal compressive strength was up to 4.32±0.52 MPa. We found firstly that nano-ATP scaffolds with excellent biocompatibility for hBMSCscould upregulate the expression of osteogenesis-related genes bmp2 and runx2 and calcium deposits in vitro. Interestingly, micro-CT and histology analysis revealed abundant newly formed bone was observed along the defect margin, even above and within the 3D bioprinted porous ATP scaffolds in a rat cranial defect model. Furthermore, histology analysis demonstrated that bone was formed directly following a process similar to membranous ossification without any intermediate cartilage formation and that many newly formed blood vessels are within the pores of 3D-printed scaffolds at four and eight weeks. CONCLUSION: These results suggest that the 3D-printed porous nano-ATP scaffolds are promising candidates for bone tissue engineering by osteogenesis and angiogenesis.
AUTHOR Lee, J. M. and Sing, S. L. and Yeong, W. Y.
Title Bioprinting of Multimaterials with Computer-aided Design/Computer -aided Manufacturing [Abstract]
Year 2020
Journal/Proceedings International Journal of Bioprinting; Vol 6, No 1 (2020)
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Multimaterials deposition, a distinct advantage in bioprinting, overcomes material’s limitation in hydrogel-based bioprinting. Multimaterials are deposited in a build/support configuration to improve the structural integrity of three-dimensional bioprinted construct. A combination of rapid cross-linking hydrogel has been chosen for the build/support setup. The bioprinted construct was further chemically cross-linked to ensure a stable construct after print. This paper also proposes a file segmentation and preparation technique to be used in bioprinting for printing freeform structures.
AUTHOR Diloksumpan, Paweena and de Ruijter, Myl{`{e}}ne and Castilho, Miguel and Gbureck, Uwe and Vermonden, Tina and van Weeren, P. Ren{'{e}} and Malda, Jos and Levato, Riccardo
Title Combining multi-scale 3D printing technologies to engineer reinforced hydrogel-ceramic interfaces [Abstract]
Year 2020
Journal/Proceedings Biofabrication
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Abstract
Multi-material 3D printing technologies that resolve features at different lengths down to the microscale open new avenues for regenerative medicine, particularly in the engineering of tissue interfaces. Herein, extrusion printing of a bone-biomimetic ceramic ink and melt electrowriting (MEW) of spatially organized polymeric microfibres are integrated for the biofabrication of an osteochondral plug, with a mechanically reinforced bone-to-cartilage interface. A printable physiological temperature-setting bioceramic, based on α-tricalcium phosphate, nanohydroxyapatite and a custom-synthesized biodegradable and crosslinkable poloxamer, was developed as bone support. The mild setting reaction of the bone ink enabled us to print directly within melt electrowritten polycaprolactone meshes, preserving their micro-architecture. Ceramic-integrated MEW meshes protruded into the cartilage region of the composite plug, and were embedded with mechanically soft gelatin-based hydrogels, laden with articular cartilage chondroprogenitor cells. Such interlocking design enhanced the hydrogel-to-ceramic adhesion strength >6.5-fold, compared with non-interlocking fibre architectures, enabling structural stability during handling and surgical implantation in osteochondral defects ex vivo. Furthermore, the MEW meshes endowed the chondral compartment with compressive properties approaching those of native cartilage (20-fold reinforcement versus pristine hydrogel). The osteal and chondral compartment supported osteogenesis and cartilage matrix deposition in vitro, and the neo-synthesized cartilage matrix further contributed to the mechanical reinforcement at the ceramic-hydrogel interface. This multi-material, multi-scale 3D printing approach provides a promising strategy for engineering advanced composite constructs for the regeneration of musculoskeletal and connective tissue interfaces.
AUTHOR Tan, Wen See and Juhari, Muhammad Aidil Bin and Shi, Qian and Chen, Shengyang and Campolo, Domenico and Song, Juha
Title Development of a new additive manufacturing platform for direct freeform 3D printing of intrinsically curved flexible membranes [Abstract]
Year 2020
Journal/Proceedings Additive Manufacturing
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DOI/URL URL DOI
Abstract
The wearable technology market has been expanding from wearable medical devices for non-invasive continuous monitoring of patient vital signs to wearable devices for tracking fitness activities that any person can access. Regardless of their form or function, desirable characteristics of wearable devices are the ability to be flexible, conformal, and easily attachable to the human body. However, as the human body is intrinsically curved and irregular, flat devices often have poor interfacial adhesion with the human body. This often leads to interfacial delamination and eventual detachment of the device. Therefore, a new additive manufacturing (AM) platform, a direct freeform 3D printing process (DF3DP), is proposed to allow direct construction of intrinsically curved 3D surfaces during the material deposition phase without the need for any pre-shaped supporting molds or templates. This 3D freeform printing process involves a supporting matrix made up of calcium alginate microgels, printing material made from silicone ink, and freeform printing paths derived from customized G-codes that conform exactly to the scanned human surface profile. Curved meshes mimicking the human elbow were used as a demonstration. A static contact stability test showed that the printed 3D silicone mesh was highly conformal to the model elbow surface as compared to a 2D flat mesh. A dynamic contact stability test was also conducted by subjecting both meshes to 100 cycles of mechanical flexion and extension, proving that intrinsically curved surfaces can provide better contact stability for complex human body surfaces undergoing motion than can flat surfaces. These results have proven that intrinsically curved membranes or structures fabricated by DF3DP can reduce the interfacial shear stress and occurrence of cracks and delamination while maintaining structural integrity and stability during use without compromising the comfort of the users. Our approach can resolve interfacial issues in flexible substrates and has great potential for epidermal devices or soft robotics via its long-term sustainable performance.
AUTHOR Zhang, Hua and Cong, Yang and Osi, Amarachi Rosemary and Zhou, Yang and Huang, Fangcheng and Zaccaria, Remo P. and Chen, Jing and Wang, Rong and Fu, Jun
Title Direct 3D Printed Biomimetic Scaffolds Based on Hydrogel Microparticles for Cell Spheroid Growth [Abstract]
Year 2020
Journal/Proceedings Advanced Functional Materials
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DOI/URL DOI
Abstract
Abstract Biocompatible hydrogel inks with shear-thinning, appropriate yield strength, and fast self-healing are desired for 3D bioprinting. However, the lack of ideal 3D bioprinting inks with outstanding printability and high structural fidelity, as well as cell-compatibility, has hindered the progress of extrusion-based 3D bioprinting for tissue engineering. In this study, novel self-healable pre-cross-linked hydrogel microparticles (pcHμPs) of chitosan methacrylate (CHMA) and polyvinyl alcohol (PVA) hybrid hydrogels are developed and used as bioinks for extrusion-based 3D printing of scaffolds with high fidelity and biocompatibility. The pcHμPs display excellent shear thinning when injected through a syringe and subsequently self-heal into gels as shear forces are removed. Numerical simulations indicate that the pcHμPs experience a plug flow in the nozzle with minimal disturbance, which favors a steady and continuous printing. Moreover, the pcHμPs show a self-supportive yield strength (540 Pa), which is critical for the fidelity of printed constructs. A series of biomimetic constructs with very high aspect ratio and delicate fine structures are directly printed by using the pcHμP ink. The 3D printed scaffolds support the growth of bone-marrow-derived mesenchymal stem cells and formation of cell spheroids, which are most important for tissue engineering.
AUTHOR Lee, Jia Min and Yeong, Wai Yee
Title Engineering macroscale cell alignment through coordinated toolpath design using support-assisted 3D bioprinting [Abstract]
Year 2020
Journal/Proceedings Journal of The Royal Society Interface
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DOI/URL DOI
Abstract
Aligned cells provide direction-dependent mechanical properties that influence biological and mechanical function in native tissues. Alignment techniques such as casting and uniaxial stretching cannot fully replicate the complex fibre orientation of native tissue such as the heart. In this study, bioprinting is used to direct the orientation of cell alignment. A 0°–90° grid structure was printed to assess the robustness of the support-assisted bioprinting technique. The variation in the angles of the grid pattern is designed to mimic the differences in fibril orientation of native tissues, where angles of cell alignment vary across the different layers. Through bioprinting of a cell–hydrogel mixture, C2C12 cells displayed directed alignment along the longitudinal axis of printed struts. Cell alignment is induced through firstly establishing structurally stable constructs (i.e. distinct 0°–90° structures) and secondly, allowing cells to dynamically remodel the bioprinted construct. Herein reports a method of inducing a macroscale level of controlled cell alignment with angle variation. This was not achievable both in terms of methods (i.e. conventional alignment techniques such as stretching and electrical stimulation) and magnitude (i.e. hydrogel features with less than 100 µm features).
AUTHOR Abu Awwad, Hosam Al-Deen M. and Thiagarajan, Lalitha and Kanczler, Janos M. and Amer, Mahetab H. and Bruce, Gordon and Lanham, Stuart and Rumney, Robin M. H. and Oreffo, Richard O. C. and Dixon, James E.
Title Genetically-programmed, mesenchymal stromal cell-laden & mechanically strong 3D bioprinted scaffolds for bone repair [Abstract]
Year 2020
Journal/Proceedings Journal of Controlled Release
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DOI/URL URL DOI
Abstract
Additive manufacturing processes used to create regenerative bone tissue engineered implants are not biocompatible, thereby restricting direct use with stem cells and usually require cell seeding post-fabrication. Combined delivery of stem cells with the controlled release of osteogenic factors, within a mechanically-strong biomaterial combined during manufacturing would replace injectable defect fillers (cements) and allow personalized implants to be rapidly prototyped by 3D bioprinting. Through the use of direct genetic programming via the sustained release of an exogenously delivered transcription factor RUNX2 (delivered as recombinant GET-RUNX2 protein) encapsulated in PLGA microparticles (MPs), we demonstrate that human mesenchymal stromal (stem) cells (hMSCs) can be directly fabricated into a thermo-sintered 3D bioprintable material and achieve effective osteogenic differentiation. Importantly we observed osteogenic programming of gene expression by released GET-RUNX2 (8.2-, 3.3- and 3.9-fold increases in OSX, RUNX2 and OPN expression, respectively) and calcification (von Kossa staining) in our scaffolds. The developed biodegradable PLGA/PEG paste formulation augments high-density bone development in a defect model (~2.4-fold increase in high density bone volume) and can be used to rapidly prototype clinically-sized hMSC-laden implants within minutes using mild, cytocompatible extrusion bioprinting. The ability to create mechanically strong 'cancellous bone-like’ printable implants for tissue repair that contain stem cells and controlled-release of programming factors is innovative, and will facilitate the development of novel localized delivery approaches to direct cellular behaviour for many regenerative medicine applications including those for personalized bone repair.
AUTHOR Eltaher, Hoda M. and Abukunna, Fatima E. and Ruiz-Cantu, Laura and Stone, Zack and Yang, Jing and Dixon, James E.
Title Human-scale tissues with patterned vascular networks by additive manufacturing of sacrificial sugar-protein composites [Abstract]
Year 2020
Journal/Proceedings Acta Biomaterialia
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DOI/URL URL DOI
Abstract
Combating necrosis, by supplying nutrients and removing waste, presents the major challenge for engineering large three-dimensional (3D) tissues. Previous elegant work used 3D printing with carbohydrate glass as a cytocompatible sacrificial template to create complex engineered tissues with vascular networks (Miller et al. 2012, Nature Materials). The fragile nature of this material compounded with the technical complexity needed to create high-resolution structures led us to create a flexible sugar-protein composite, termed Gelatin-sucrose matrix (GSM), to achieve a more robust and applicable material. Here we developed a low-range (25–37˚C) temperature sensitive formulation that can be moulded with micron-resolution features or cast during 3D printing to produce complex flexible filament networks forming sacrificial vessels. Using the temperature-sensitivity, we could control filament degeneration meaning GSM can be used with a variety of matrices and crosslinking strategies. Furthermore by incorporation of biocompatible crosslinkers into GSM directly, we could create thin endothelialized vessel walls and generate patterned tissues containing multiple matrices and cell-types. We also demonstrated that perfused vascular channels sustain metabolic function of a variety of cell-types including primary human cells. Importantly, we were able to construct vascularized human noses which otherwise would have been necrotic. Our material can now be exploited to create human-scale tissues for regenerative medicine applications. Statement of Significance Authentic and engineered tissues have demands for mass transport, exchanging nutrients and oxygen, and therefore require vascularization to retain viability and inhibit necrosis. Basic vascular networks must be included within engineered tissues intrinsically. Yet, this has been unachievable in physiologically-sized constructs with tissue-like cell densities until recently. Sacrificial moulding is an alternative in which networks of rigid lattices of filaments are created to prevent subsequent matrix ingress. Our study describes a biocompatible sacrificial sugar-protein formulation; GSM, made from mixtures of inexpensive and readily available bio-grade materials. GSM can be cast/moulded or bioprinted as sacrificial filaments that can rapidly dissolve in an aqueous environment temperature-sensitively. GSM material can be used to engineer viable and vascularized human-scale tissues for regenerative medicine applications.
AUTHOR Strauß, Svenja and Meutelet, Rafaela and Radosevic, Luka and Gretzinger, Sarah and Hubbuch, Jürgen
Title Image analysis as PAT-Tool for use in extrusion-based bioprinting [Abstract]
Year 2020
Journal/Proceedings Bioprinting
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DOI/URL URL DOI
Abstract
The technology of bioprinting is arousing a growing interest in biopharmaceutical research and industry. In order to accelerate process development in the field of bioprinting, image-based analysis methods are non-invasive, time- and cost-saving tools which are usable for printer characterization, bioink printability evaluation, and process optimization. Image processing can also be used for the study of reproducibility, since reliable production is important in the transition from research to industrial application, and more precisely to clinical studies. This study revolves around the establishment of an automated and image-based line analysis method for bioprinting applications which enables an easy comparison of 3D-printed lines. Diverse rheological properties of bioinks and the printing process affect the geometry of the resulting object. The line represents a simple geometry, where the influence of the rheological properties and printing parameters is directly apparent. Therefore, a method for line analysis was developed on the basis of image recognition. At first, the method is tested for several substances such as Nivea®, pure and colored Kolliphor solutions, and two commercially available hydrogel formulations which can be used as bioinks. These are Biogelx™-ink-RGD by Biogelx and Cellink® Bioink by Cellink. The examination of limitations showed that transparent materials such as Kolliphor-based solutions cannot be analyzed with the developed method whereas opaque materials such as Nivea® and both bioinks can be analyzed. In the course of process characterization, the method was used to investigate the shrinkage behavior for both bionks. With the help of the line analysis tool, a shrinkage behavior of both bioinks was demonstrated and thus, process time could be identified as a critical process parameter.
AUTHOR Rupp, Harald and Binder, Wolfgang H.
Title Multicomponent Stress-Sensing Composites Fabricated by 3D-Printing Methodologies [Abstract]
Year 2020
Journal/Proceedings Macromolecular Rapid Communications
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DOI/URL DOI
Abstract
Abstract The preparation and characterization of mechanoresponsive, 3D-printed composites are reported using a dual-printing setup for both, liquid dispensing and fused-deposition-modeling. The here reported stress-sensing materials are based on high- and low molecular weight mechanophores, including poly(ε-caprolactone)-, polyurethane-, and alkyl(C11)-based latent copper(I)bis(N-heterocyclic carbenes), which can be activated by compression to trigger a fluorogenic, copper(I)-catalyzed azide/alkyne “click”-reaction of an azide-functionalized fluorescent dye inside a bulk polymeric material. Focus is placed on the printability and postprinting activity of the latent mechanophores and the fluorogenic “click”-components. The multicomponent specimen containing both, azide and alkyne, are manufactured via a 3D-printer to place the components separately inside the specimen into void spaces generated during the FDM-process, which subsequently are filled with liquids using a separate liquid dispenser, located within the same 3D-printing system. The low-molecular weight mechanophores bearing the alkyl-C11 chains display the best printability, yielding a mechanochemical response after the 3D-printing process.
AUTHOR Plou, Javier and García, Isabel and Charconnet, Mathias and Astobiza, Ianire and García-Astrain, Clara and Matricardi, Cristiano and Mihi, Agustín and Carracedo, Arkaitz and Liz-Marzán, Luis M.
Title Multiplex SERS Detection of Metabolic Alterations in Tumor Extracellular Media [Abstract]
Year 2020
Journal/Proceedings Advanced Functional Materials
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DOI/URL DOI
Abstract
Abstract The composition and intercellular interactions of tumor cells in the tissues dictate the biochemical and metabolic properties of the tumor microenvironment. The metabolic rewiring has a profound impact on the properties of the microenvironment, to an extent that monitoring such perturbations could harbor diagnostic and therapeutic relevance. A growing interest in these phenomena has inspired the development of novel technologies with sufficient sensitivity and resolution to monitor metabolic alterations in the tumor microenvironment. In this context, surface-enhanced Raman scattering (SERS) can be used for the label-free detection and imaging of diverse molecules of interest among extracellular components. Herein, the application of nanostructured plasmonic substrates comprising Au nanoparticles, self-assembled as ordered superlattices, to the precise SERS detection of selected tumor metabolites, is presented. The potential of this technology is first demonstrated through the analysis of kynurenine, a secreted immunomodulatory derivative of the tumor metabolism and the related molecules tryptophan and purine derivatives. SERS facilitates the unambiguous identification of trace metabolites and allows the multiplex detection of their characteristic fingerprints under different conditions. Finally, the effective plasmonic SERS substrate is combined with a hydrogel-based three-dimensional cancer model, which recreates the tumor microenvironment, for the real-time imaging of metabolite alterations and cytotoxic effects on tumor cells.
AUTHOR Diloksumpan, Paweena and Bolaños, Rafael Vindas and Cokelaere, Stefan and Pouran, Behdad and de Grauw, Janny and van Rijen, Mattie and van Weeren, René and Levato, Riccardo and Malda, Jos
Title Orthotopic Bone Regeneration within 3D Printed Bioceramic Scaffolds with Region-Dependent Porosity Gradients in an Equine Model [Abstract]
Year 2020
Journal/Proceedings Advanced Healthcare Materials
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DOI/URL DOI
Abstract
Abstract The clinical translation of three-dimensionally printed bioceramic scaffolds with tailored architectures holds great promise toward the regeneration of bone to heal critical-size defects. Herein, the long-term in vivo performance of printed hydrogel-ceramic composites made of methacrylated-oligocaprolactone-poloxamer and low-temperature self-setting calcium-phosphates is assessed in a large animal model. Scaffolds printed with different internal architectures, displaying either a designed porosity gradient or a constant pore distribution, are implanted in equine tuber coxae critical size defects. Bone ingrowth is challenged and facilitated only from one direction via encasing the bioceramic in a polycaprolactone shell. After 7 months, total new bone volume and scaffold degradation are significantly greater in structures with constant porosity. Interestingly, gradient scaffolds show lower extent of remodeling and regeneration even in areas having the same porosity as the constant scaffolds. Low regeneration in distal regions from the interface with native bone impairs ossification in proximal regions of the construct, suggesting that anisotropic architectures modulate the cross-talk between distant cells within critical-size defects. The study provides key information on how engineered architectural patterns impact osteoregeneration in vivo, and also indicates the equine tuber coxae as promising orthotopic model for studying materials stimulating bone formation.
AUTHOR Athanasiadis, Markos and Afanasenkau, Dzmitry and Derks, Wouter and Tondera, Christoph and Murganti, Francesca and Busskamp, Volker and Bergmann, Olaf and Minev, Ivan R.
Title Printed elastic membranes for multimodal pacing and recording of human stem-cell-derived cardiomyocytes [Abstract]
Year 2020
Journal/Proceedings npj Flexible Electronics
Reftype Athanasiadis2020
DOI/URL DOI
Abstract
Bioelectronic interfaces employing arrays of sensors and bioactuators are promising tools for the study, repair and engineering of cardiac tissues. They are typically constructed from rigid and brittle materials processed in a cleanroom environment. An outstanding technological challenge is the integration of soft materials enabling a closer match to the mechanical properties of biological cells and tissues. Here we present an algorithm for direct writing of elastic membranes with embedded electrodes, optical waveguides and microfluidics using a commercial 3D printing system and a palette of silicone elastomers. As proof of principle, we demonstrate interfacing of cardiomyocytes derived from human induced pluripotent stem cells (hiPSCs), which are engineered to express Channelrhodopsin-2. We demonstrate electrical recording of cardiomyocyte field potentials and their concomitant modulation by optical and pharmacological stimulation delivered via the membrane. Our work contributes a simple prototyping strategy with potential applications in organ-on-chip or implantable systems that are multi-modal and mechanically soft.
AUTHOR Tan, Wen See and Shi, Qian and Chen, Shengyang and Bin Juhari, Muhammad Aidil and Song, Juha
Title Recyclable and biocompatible microgel-based supporting system for positive 3D freeform printing of silicone rubber [Abstract]
Year 2020
Journal/Proceedings Biomedical Engineering Letters
Reftype Tan2020
DOI/URL DOI
Abstract
Additive manufacturing (AM) of biomaterials has evolved from a rapid prototyping tool into a viable approach for the manufacturing of patient-specific implants over the past decade. It can tailor to the unique physiological and anatomical criteria of the patient’s organs or bones through precise controlling of the structure during the 3D printing. Silicone elastomers, which is a major group of materials in many biomedical implants, have low viscosities and can be printed with a special AM platform, known as freeform 3D printing systems. The freeform 3D printing systems are composed of a supporting bath and a printing material. Current supporting matrices that are either commercially purchased or synthesized were usually disposed of after retrieval of the printed part. In this work, we proposed a new and improved supporting matrix comprises of synthesized calcium alginate microgels produced via encapsulation which can be recycled, reused, and recovered for multiple prints, hence minimizing wastage and cost of materials. The dehydration tolerance of the calcium alginate microgels was improved through physical means by the addition of glycerol and chemical means by developing new calcium alginate microgels encapsulated with glycerol. The recyclability of the heated calcium alginate microgels was also enhanced by a rehydration step with sodium chloride solution and a recovery step with calcium chloride solution via the ion exchange process. We envisaged that our reusable and recyclable biocompatible calcium alginate microgels can save material costs, time, and can be applied in various freeform 3D printing systems.
AUTHOR Sanz-Fraile, Hector and Amorós, Susana and Mendizabal, Irene Isabel and Gálvez-Montón, Carolina and Prat-Vidal, Cristina and Bayés-Genís, Antoni and Navajas, Daniel and Farre, Ramon and Otero, Jorge
Title Silk-reinforced Collagen Hydrogels with Raised Multiscale Stiffness for Mesenchymal Cells 3D Culture [Abstract]
Year 2020
Journal/Proceedings Tissue Engineering Part A
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DOI/URL DOI
Abstract
Type I collagen hydrogels are of high interest in tissue engineering. With the evolution of 3D bioprinting technologies, a high number of collagen-based scaffolds have been reported for the development of 3D cell cultures. A recent proposal was to mix collagen with silk fibroin derived from Bombyx Mori silkworm. Nevertheless, due to the difficulties in the preparation and the characteristics of the protein, several problems like phase separation and collagen denaturation appears during the procedure. Therefore, the common solution is to diminish the concentration of collagen although in that way the most biologically relevant component is reduced. In the present work, we present a new, simple and effective method to develop a collagen-silk hybrid hydrogel with high collagen concentration and with increased stiffness approaching that of natural tissues, which could be of high interest for the development of cardiac patches for myocardial regeneration and for preconditioning of mesenchymal stem cells to improve their therapeutic potential. Sericin in the silk was preserved by using a physical solubilizing procedure which results in a preserved fibrous structure of type I collagen, as shown by ultrastructural imaging. The macro- and micromechanical properties of the hybrid hydrogels measured by tensile stretch and Atomic Force Microscopy respectively, showed a more than two-fold stiffening as compared with collagen-only hydrogels. Rheological measurements showed improved printability properties for the developed biomaterial. The suitability of the hydrogels for 3D cell culture was assessed by 3D bioprinting bone marrow-derived mesenchymal stem cells cultured within the scaffolds. The result was a biomaterial with improved printability characteristics that better resembled the mechanical properties of natural soft tissues while preserving biocompatibility owing to the high concentration of collagen.
AUTHOR Döhler, Diana and Kang, Jiheong and Cooper, Chris Brittain and Tok, Jeffrey B.-H. and Rupp, Harald and Binder, Wolfgang H. and Bao, Zhenan
Title Tuning the Self-Healing Response of Poly(dimethylsiloxane)-Based Elastomers [Abstract]
Year 2020
Journal/Proceedings ACS Appl. Polym. Mater.
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DOI/URL DOI
Abstract
We present a comprehensive investigation of mechanical properties of supramolecular polymer networks with rationally developed multistrength hydrogen-bonding interactions. Self-healing poly(dimethylsiloxane) (PDMS)-based elastomers with varying elasticity, fracture toughness, and the ability to dissipate strain energy through the reversible breakage and re-formation of the supramolecular interactions were obtained. By changing the ratio between isophorone diisocyanate (IU), 4,4′-methylenebis(cyclohexyl isocyanate) (MCU), and 4,4′-methylenebis(phenyl isocyanate) (MPU) and by varying the molecular weight of the PDMS precursor, we obtained a library of poly(urea)s to study the interplay of mechanical performance and self-healability. The Young’s moduli of the presented materials ranged between 0.4 and 13 MPa and increased with decreasing molecular weight of the PDMS precursor and increasing content of MCU or MPU units related to the formation of stronger hydrogen-bonding interactions. By exchanging MPU against MCU units, we achieved an optimum balance between mechanical properties and self-healing performance, and by the additional reduction of the molecular weight of the precursor polymer, a minimum recovery of 80% in stress within 12 h at room temperature was observed. Selected poly(urea)s could be processed via 3D printing by the conventional extrusion method, obtaining dimensionally stable and freestanding objects. We present a comprehensive investigation of mechanical properties of supramolecular polymer networks with rationally developed multistrength hydrogen-bonding interactions. Self-healing poly(dimethylsiloxane) (PDMS)-based elastomers with varying elasticity, fracture toughness, and the ability to dissipate strain energy through the reversible breakage and re-formation of the supramolecular interactions were obtained. By changing the ratio between isophorone diisocyanate (IU), 4,4′-methylenebis(cyclohexyl isocyanate) (MCU), and 4,4′-methylenebis(phenyl isocyanate) (MPU) and by varying the molecular weight of the PDMS precursor, we obtained a library of poly(urea)s to study the interplay of mechanical performance and self-healability. The Young’s moduli of the presented materials ranged between 0.4 and 13 MPa and increased with decreasing molecular weight of the PDMS precursor and increasing content of MCU or MPU units related to the formation of stronger hydrogen-bonding interactions. By exchanging MPU against MCU units, we achieved an optimum balance between mechanical properties and self-healing performance, and by the additional reduction of the molecular weight of the precursor polymer, a minimum recovery of 80% in stress within 12 h at room temperature was observed. Selected poly(urea)s could be processed via 3D printing by the conventional extrusion method, obtaining dimensionally stable and freestanding objects.
AUTHOR Rupp, Harald and Döhler, Diana and Hilgeroth, Philipp and Mahmood, Nasir and Beiner, Mario and Binder, Wolfgang H.
Title 3D Printing of Supramolecular Polymers: Impact of Nanoparticles and Phase Separation on Printability [Abstract]
Year 2019
Journal/Proceedings Macromolecular Rapid Communications
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Abstract
Abstract 3D printing of linear and three-arm star supramolecular polymers with attached hydrogen bonds and their nanocomposites is reported. The concept is based on hydrogen-bonded supramolecular polymers, known to form nano-sized micellar clusters. Printability is based on reversible thermal- and shear-induced dissociation of a supramolecular polymer network, which generates stable and self-supported structures after printing, as checked via melt-rheology and X-ray scattering. The linear and three-arm star poly(isobutylene)s PIB-B2 (Mn = 8500 g mol −1), PIB-B3 (Mn = 16 000 g mol −1), and linear poly(ethylene glycol)s PEG-B2 (Mn = 900 g mol−1, 8500 g mol −1) are prepared and then probed by melt-rheology to adjust the viscosity to address the proper printing window. The supramolecular PIB polymers show a rubber-like behavior and are able to form self-supported 3D printed objects at room temperature and below, reaching polymer strand diameters down to 200–300 µm. Nanocomposites of PIB-B2 with silica nanoparticles (12 nm, 5–15 wt%) are generated, in turn leading to an improvement of their shape persistence. A blend of the linear polymer PIB-B2 and the three-arm star polymer PIB-B3 (ratio ≈ 3/1 mol) reaches an even higher structural stability, able to build free-standing structures.
AUTHOR Creusen, Guido and Roshanasan, Ardeshir and Garcia Lopez, Javier and Peneva, Kalina and Walther, Andreas
Title Bottom-up design of model network elastomers and hydrogels from precise star polymers [Abstract]
Year 2019
Journal/Proceedings Polymer Chemistry
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Abstract
We introduce a platform for the simultaneous design of model network hydrogels and bulk elastomers based on well-defined water-soluble star polymers with a low glass transition temperature (Tg). This platform is enabled via the development of a synthetic route to a new family of 4-arm star polymers based on water-soluble poly(triethylene glycol methyl ether acrylate) (p(mTEGA)){,} which after quantitative introduction of functional end-groups can serve as suitable building blocks for model network formation. We first describe in detail the synthesis of highly defined star polymers using light and Cu-wire mediated Cu-based reversible deactivation radical polymerization. The resulting polymers exhibit narrow dispersities and controlled arm length at very high molecular weights{,} and feature a desirable low Tg of −55 °C. Subsequently{,} we elucidate the rational design of the stiffness and elasticity in covalent model network elastomers and hydrogels formed by fast photo-crosslinking for different arm lengths{,} and construct thermally reversible model network hydrogels based on dynamic supramolecular bonds. In addition{,} we describe preliminary 3D-printing applications. This work provides a key alternative to commonly used star-poly(ethylene glycol) (PEG) for model hydrogel networks{,} and demonstrates access to new main and side chain chemistries{,} thus chain stiffnesses and entanglement molecular weight{,} and{,} critically{,} enables the simultaneous study of the mechanical behavior of bulk network elastomers and swollen hydrogels with the same network topology. In a wider perspective{,} this work also highlights the need for advancing precision polymer chemistry to allow for an understanding of architectural control for the rational design of functional mechanical network materials.
AUTHOR Athanasiadis, Markos and Pak, Anna and Afanasenkau, Dzmitry and Minev, Ivan R.
Title Direct Writing of Elastic Fibers with Optical, Electrical, and Microfluidic Functionality [Abstract]
Year 2019
Journal/Proceedings Advanced Materials Technologies
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DOI/URL DOI
Abstract
Abstract Direct Ink Writing is an additive fabrication technology that allows the integration of a diverse range of functional materials into soft and bioinspired devices such as robots and human-machine interfaces. Typically, a viscoelastic ink is extruded from a nozzle as a continuous filament of circular cross section. Here it is shown that a careful selection of printing parameters such as nozzle height and speed can produce filaments with a range of cross-sectional geometries. Thus, elliptic cylinder-, ribbon-, or groove-shaped filaments can be printed. By using the nozzle as a stylus for postprint filament modification, even filaments with an embedded microfluidic channel can be produced. This strategy is applied to directly write freeform and elastic optical fibers, electrical interconnects, and microfluidics. The integration of these components into simple sensor-actuator systems is demonstrated. Prototypes of an optical fiber with steerable tip and a thermal actuation system for soft tissues are presented.
AUTHOR Mestre, Rafael and Patiño, Tania and Barceló, Xavier and Anand, Shivesh and Pérez-Jiménez, Ariadna and Sánchez, Samuel
Title Force Modulation and Adaptability of 3D-Bioprinted Biological Actuators Based on Skeletal Muscle Tissue [Abstract]
Year 2019
Journal/Proceedings Advanced Materials Technologies
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DOI/URL DOI
Abstract
Abstract The integration of biological systems into robotic devices might provide them with capabilities acquired from natural systems and significantly boost their performance. These abilities include real-time bio-sensing, self-organization, adaptability, or self-healing. As many muscle-based bio-hybrid robots and bio-actuators arise in the literature, the question of whether these features can live up to their expectations becomes increasingly substantial. Herein, the force generation and adaptability of skeletal-muscle-based bio-actuators undergoing long-term training protocols are analyzed. The 3D-bioprinting technique is used to fabricate bio-actuators that are functional, responsive, and have highly aligned myotubes. The bio-actuators are 3D-bioprinted together with two artificial posts, allowing to use it as a force measuring platform. In addition, the force output evolution and dynamic gene expression of the bio-actuators are studied to evaluate their degree of adaptability according to training protocols of different frequencies and mechanical stiffness, finding that their force generation could be modulated to different requirements. These results shed some light into the fundamental mechanisms behind the adaptability of muscle-based bio-actuators and highlight the potential of using 3D bioprinting as a rapid and cost-effective tool for the fabrication of custom-designed soft bio-robots.
AUTHOR Tondera, Christoph and Akbar, Teuku Fawzul and Thomas, Alvin Kuriakose and Lin, Weilin and Werner, Carsten and Busskamp, Volker and Zhang, Yixin and Minev, Ivan R.
Title Highly Conductive, Stretchable, and Cell-Adhesive Hydrogel by Nanoclay Doping [Abstract]
Year 2019
Journal/Proceedings Small
Reftype
DOI/URL DOI
Abstract
Abstract Electrically conductive materials that mimic physical and biological properties of tissues are urgently required for seamless brain–machine interfaces. Here, a multinetwork hydrogel combining electrical conductivity of 26 S m−1, stretchability of 800%, and tissue-like elastic modulus of 15 kPa with mimicry of the extracellular matrix is reported. Engineering this unique set of properties is enabled by a novel in-scaffold polymerization approach. Colloidal hydrogels of the nanoclay Laponite are employed as supports for the assembly of secondary polymer networks. Laponite dramatically increases the conductivity of in-scaffold polymerized poly(ethylene-3,4-diethoxy thiophene) in the absence of other dopants, while preserving excellent stretchability. The scaffold is coated with a layer containing adhesive peptide and polysaccharide dextran sulfate supporting the attachment, proliferation, and neuronal differentiation of human induced pluripotent stem cells directly on the surface of conductive hydrogels. Due to its compatibility with simple extrusion printing, this material promises to enable tissue-mimetic neurostimulating electrodes.
AUTHOR Fortunato, Gabriele Maria and Maria, Carmelo De and Eglin, David and Serra, Tiziano and Vozzi, Giovanni
Title An ink-jet printed electrical stimulation platform for muscle tissue regeneration [Abstract]
Year 2018
Journal/Proceedings Bioprinting
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DOI/URL URL DOI
Abstract
Conducting polymeric materials have been used to modulate response of cells seeded on their surfaces. However, there is still major improvement to be made related to their biocompatibility, conductivity, stability in biological milieu, and processability toward truly tissue engineered functional device. In this work, conductive polymer, poly(3,4-ethylene-dioxythiophene):polystyrene-sulfonate (PEDOT:PSS), and its possible applications in tissue engineering were explored. In particular PEDOT:PSS solution was inkjet printed onto a gelatin substrate for obtaining a conductive structure. Mechanical and electrical characterizations, structural stability by swelling and degradation tests were carried out on different PEDOT-based samples obtained by varying the number of printed PEDOT layers from 5 to 50 on gelatin substrate. Biocompatibility of substrates was investigated on C2C12 myoblasts, through metabolic activity assay and imaging analysis during a 7-days culture period, to assess cell morphology, differentiation and alignment. The results of this first part allowed to proceed with the second part of the study in which these substrates were used for the design of an electrical stimulation device, with the aim of providing the external stimulus (3 V amplitude square wave at 1 and 2 Hz frequency) to guide myotubes alignment and enhance differentiation, having in this way promising applications in the field of muscle tissue engineering.
AUTHOR Ng, Wei Long and Goh, Min Hao and Yeong, Wai Yee and Naing, May Win
Title Applying macromolecular crowding to 3D bioprinting: fabrication of 3D hierarchical porous collagen-based hydrogel constructs [Abstract]
Year 2018
Journal/Proceedings Biomaterials Science
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DOI/URL DOI
Abstract
Native tissues and/or organs possess complex hierarchical porous structures that confer highly-specific cellular functions. Despite advances in fabrication processes{,} it is still very challenging to emulate the hierarchical porous collagen architecture found in most native tissues. Hence{,} the ability to recreate such hierarchical porous structures would result in biomimetic tissue-engineered constructs. Here{,} a single-step drop-on-demand (DOD) bioprinting strategy is proposed to fabricate hierarchical porous collagen-based hydrogels. Printable macromolecule-based bio-inks (polyvinylpyrrolidone{,} PVP) have been developed and printed in a DOD manner to manipulate the porosity within the multi-layered collagen-based hydrogels by altering the collagen fibrillogenesis process. The experimental results have indicated that hierarchical porous collagen structures could be achieved by controlling the number of macromolecule-based bio-ink droplets printed on each printed collagen layer. This facile single-step bioprinting process could be useful for the structural design of collagen-based hydrogels for various tissue engineering applications.
AUTHOR Bastola, A. K. and Paudel, M. and Li, L.
Title Development of hybrid magnetorheological elastomers by 3D printing [Abstract]
Year 2018
Journal/Proceedings Polymer
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Abstract
Intelligent or smart materials have one or more properties that can be significantly changed in a controlled fashion by external stimuli, such as temperature, pH, electric or magnetic fields, etc. Magnetorheological (MR) materials are a class of smart materials whose properties can be varied by applying an external magnetic field. In this work, the possibility of employing a suitable 3D printing technology for the development of one of the smart MR materials, the magnetorheological elastomer (MRE) has been explored. In order to achieve such 3D printing, a multi-material printing is implemented, where a controlled volume of MR fluid is encapsulated within an elastomer matrix in the layer-by-layer fashion. The choice of printing materials determines the final structure of the 3D printed hybrid MR elastomer. Printing with a vulcanizing MR suspension produces the solid MR structure inside the elastomer matrix while printing with a non-vulcanizing MR suspension (MR fluid) results in the structures that the MR fluid is encapsulated inside the elastomer matrix. The 3D printability of different materials has been studied by measuring their rheological properties and we found that the highly shear thinning and thixotropic properties are important for 3D printability. The quality of the printed filaments strongly depends on the key printing parameters such as extrusion pressure, initial height and feed rate. The experimental results from the forced vibration testing show that the 3D printed MR elastomers could change their elastic and damping properties when exposed to the external magnetic field. Furthermore, the 3D printed MR elastomer also exhibits the anisotropic behavior when the direction of the magnetic field is changed with respect to the orientation of the printed filaments. This study has demonstrated that the 3D printing is viable for fabrication of hybrid MR elastomers with controlled structures of magnetic particles or MR fluids.
AUTHOR Huang, Yun-An and Ho, Chris T. and Lin, Yu-Hsuan and Lee, Chen-Ju and Ho, Szu-Mo and Li, Ming-Chia and Hwang, Eric
Title Nanoimprinted Anisotropic Topography Preferentially Guides Axons and Enhances Nerve Regeneration [Abstract]
Year 2018
Journal/Proceedings Macromolecular Bioscience
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DOI/URL DOI
Abstract
Abstract Surface topography has a profound effect on the development of the nervous system, such as neuronal differentiation and morphogenesis. While the interaction of neurons and the surface topography of their local environment is well characterized, the neuron–topography interaction during the regeneration process remains largely unknown. To address this question, an anisotropic surface topography resembling linear grooves made from poly(ethylene-vinyl acetate) (EVA), a soft and biocompatible polymer, using nanoimprinting, is established. It is found that neurons from both the central and peripheral nervous system can survive and grow on this grooved surface. Additionally, it is observed that axons but not dendrites specifically align with these grooves. Furthermore, it is demonstrated that neurons on the grooved surface are capable of regeneration after an on-site injury. More importantly, these injured neurons have an accelerated and enhanced regeneration. Together, the data demonstrate that this anisotropic topography guides axon growth and improves axon regeneration. This opens up the possibility to study the effect of surface topography on regenerating axons and has the potential to be developed into a medical device for treating peripheral nerve injuries.
AUTHOR Suntornnond, R. and Tan, E. Y. S. and An, J. and Chua, C. K.
Title A highly printable and biocompatible hydrogel composite for direct printing of soft and perfusable vasculature-like structures [Abstract]
Year 2017
Journal/Proceedings Scientific Reports
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DOI/URL URL DOI
Abstract
Vascularization is one major obstacle in bioprinting and tissue engineering. In order to create thick tissues or organs that can function like original body parts, the presence of a perfusable vascular system is essential. However, it is challenging to bioprint a hydrogel-based three-dimensional vasculature-like structure in a single step. In this paper, we report a new hydrogel-based composite that offers impressive printability, shape integrity, and biocompatibility for 3D bioprinting of a perfusable complex vasculature-like structure. The hydrogel composite can be used on a non-liquid platform and is printable at human body temperature. Moreover, the hydrogel composite supports both cell proliferation and cell differentiation. Our results represent a potentially new vascularization strategy for 3D bioprinting and tissue engineering.
AUTHOR Bastola, A. K. and Hoang, V. T. and Li, L.
Title A novel hybrid magnetorheological elastomer developed by 3D printing [Abstract]
Year 2017
Journal/Proceedings Materials and Design
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DOI/URL URL
Abstract
Abstract In this study, a novel magnetorheological (MR) hybrid elastomer has been developed using a 3D printing method. In such an MR hybrid elastomer, a controlled volume of an MR fluid was encapsulated layer by layer into an elastomer matrix by means of a 3D printer and each layer was a composite structure consisting of an MR fluid and an elastomer. Similar to current MR fluids and MR elastomers, mechanical properties of 3D printed MR hybrid elastomers could be controlled via an externally applied magnetic field. The experimental results showed that the relative change in the damping capability of the new MR elastomer was more pronounced than the change in its stiffness when exposed to an external magnetic field. The study demonstrated that the 3D printing technique is feasible for fabrication of MR elastomers with controlled microstructures including magnetic particles or MR fluids. The 3D printed MR hybrid elastomer is also a potential material as a tunable spring-damper element.
AUTHOR Lorson, Thomas and Jaksch, Sebastian and Lübtow, Michael M. and Jüngst, Tomasz and Groll, Jürgen and Lühmann, Tessa and Luxenhofer, Robert
Title A Thermogelling Supramolecular Hydrogel with Sponge-Like Morphology as a Cytocompatible Bioink [Abstract]
Year 2017
Journal/Proceedings Biomacromolecules
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DOI/URL DOI
Abstract
Biocompatible polymers that form thermoreversible supramolecular hydrogels have gained great interest in biomaterials research and tissue engineering. When favorable rheological properties are achieved at the same time, they are particularly promising candidates as material that allow for the printing of cells, so-called bioinks. We synthesized a novel thermogelling block copolymer and investigated the rheological properties of its aqueous solution by viscosimetry and rheology. The polymers undergo thermogelation between room temperature and body temperature, form transparent hydrogels of surprisingly high strength (G′ > 1000 Pa) and show rapid and complete shear recovery after stress. Small angle neutron scattering suggests an unusual bicontinuous sponge-like gel network. Excellent cytocompatibility was demonstrated with NIH 3T3 fibroblasts, which were incorporated and bioplotted into predefined 3D hydrogel structures without significant loss of viability. The developed materials fulfill all criteria for future use as bioink for biofabrication.
AUTHOR Ribeiro, Alexandre and Blokzijl, Maarten Michiel and Levato, Riccardo and Visser, Claas Willem and Castilho, Miguel and Hennink, Wim E. and Vermonden, Tina and Malda, Jos
Title Assessing bioink shape fidelity to aid material development in 3D bioprinting [Abstract]
Year 2017
Journal/Proceedings Biofabrication
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Abstract
Abstract During extrusion-based bioprinting, the deposited bioink filaments are subjected to deformations, such as collapse of overhanging filaments, which compromises the ability to stack several layers of bioink, and fusion between adjacent filaments, which compromises the resolution and maintenance of a desired pore structure. When developing new bioinks, approaches to assess their shape fidelity after printing would be beneficial to evaluate the degree of deformation of the deposited filament and to estimate how similar the final printed construct would be to the design. However, shape fidelity has been prevalently assessed qualitatively through visual inspection after printing, hampering the direct comparison of the printability of different bioinks. In this technical note, we propose a quantitative evaluation for shape fidelity of bioinks based on testing the filament collapse on overhanging structures and the filament fusion of parallel printed strands. Both tests were applied on a hydrogel platform based on poloxamer 407 and poly(ethylene glycol) (PEG) blends, providing a library of hydrogels with different yield stresses. The presented approach is an easy way to assess bioink shape fidelity, applicable to any filament-based bioprinting system and able to quantitatively evaluate this aspect of printability , based on the degree of deformation of the printed filament. In addition, we built a simple theoretical model that relates filament collapse with bioink yield stress. The results of both shape fidelity tests underline the role of yield stress as one of the parameters influencing the printability of a bioink. The presented quantitative evaluation will allow for reproducible comparisons between different bioink platforms.
AUTHOR Siqueira, Gilberto and Kokkinis, Dimitri and Libanori, Rafael and Hausmann, Michael K. and Gladman, Amelia Sydney and Neels, Antonia and Tingaut, Philippe and Zimmermann, Tanja and Lewis, Jennifer A. and Studart, André R.
Title Cellulose Nanocrystal Inks for 3D Printing of Textured Cellular Architectures [Abstract]
Year 2017
Journal/Proceedings Advanced Functional Materials
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DOI/URL DOI
Abstract
3D printing of renewable building blocks like cellulose nanocrystals offers an attractive pathway for fabricating sustainable structures. Here, viscoelastic inks composed of anisotropic cellulose nanocrystals (CNC) that enable patterning of 3D objects by direct ink writing are designed and formulated. These concentrated inks are composed of CNC particles suspended in either water or a photopolymerizable monomer solution. The shear-induced alignment of these anisotropic building blocks during printing is quantified by atomic force microscopy, polarized light microscopy, and 2D wide-angle X-ray scattering measurements. Akin to the microreinforcing effect in plant cell walls, the alignment of CNC particles during direct writing yields textured composites with enhanced stiffness along the printing direction. The observations serve as an important step forward toward the development of sustainable materials for 3D printing of cellular architectures with tailored mechanical properties.
AUTHOR Paxton, Naomi Claire and Smolan, Willi and Böck, Thomas and Melchels, Ferry P. W. and Groll, Juergen and Juengst, Tomasz
Title Proposal to Assess Printability of Bioinks for Extrusion-Based Bioprinting and Evaluation of Rheological Properties Governing Bioprintability [Abstract]
Year 2017
Journal/Proceedings Biofabrication
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Abstract
Abstract The development and formulation of printable inks for extrusion-based 3D bioprinting has been a major challenge in the field of biofabrication. Inks, often polymer solutions with the addition of crosslinking to form hydrogels, must not only display adequate mechanical properties for the chosen application, but also show high biocompatibility as well as printability. Here we describe a reproducible two-step method for the assessment of the printability of inks for bioprinting, focussing firstly on screening ink formulations to assess fibre formation and the ability to form 3D constructs before presenting a method for the rheological evaluation of inks to characterise the yield point, shear thinning and recovery behaviour. In conjunction, a mathematical model was formulated to provide a theoretical understanding of the pressure-driven, shear thinning extrusion of inks through needles in a bioprinter. The assessment methods were trialled with a commercially-available crème, poloxamer 407, alginate-based inks and an alginate-gelatin composite material. Yield stress was investigated by applying a stress ramp to a number of inks, which demonstrated the necessity of high yield for printable materials. The shear thinning behaviour of the inks was then characterised by quantifying the degree of shear thinning and using the mathematical model to predict the window of printer operating parameters in which the materials could be printed. Furthermore, the model predicted high shear conditions and high residence times for cells at the walls of the needle and effects on cytocompatibility at different printing conditions. Finally, the ability of the materials to recover to their original viscosity after extrusion was examined using rotational recovery rheological measurements. Taken together, these assessment techniques revealed significant insights into the requirements for printable inks and shear conditions present during the extrusion process and allow the rapid and reproducible characterisation of a wide variety of inks for bioprinting.
AUTHOR Levato, Riccardo and Webb, William R. and Otto, Iris A. and Mensinga, Anneloes and Zhang, Yadan and van Rijen, Mattie and van Weeren, René and Khan, Ilyas M. and Malda, Jos
Title The bio in the ink: cartilage regeneration with bioprintable hydrogels and articular cartilage-derived progenitor cells
Year 2017
Journal/Proceedings Acta Biomaterialia
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DOI/URL URL
AUTHOR Minas, Clara and Carnelli, Davide and Tervoort, Elena and Studart, André R.
Title 3D Printing of Emulsions and Foams into Hierarchical Porous Ceramics [Abstract]
Year 2016
Journal/Proceedings Advanced Materials
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DOI/URL DOI
Abstract
Bulk hierarchical porous ceramics with unprecedented strength-to-weight ratio and tunable pore sizes across three different length scales are printed by direct ink writing. Such an extrusion-based process relies on the formulation of inks in the form of particle-stabilized emulsions and foams that are sufficiently stable to resist coalescence during printing.
AUTHOR Sommer, Marianne R. and Schaffner, Manuel and Carnelli, Davide and Studart, André R.
Title 3D Printing of Hierarchical Silk Fibroin Structures [Abstract]
Year 2016
Journal/Proceedings ACS Applied Materials and Interfaces
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DOI/URL DOI
Abstract
Like many other natural materials, silk is hierarchically structured from the amino acid level up to the cocoon or spider web macroscopic structures. Despite being used industrially in a number of applications, hierarchically structured silk fibroin objects with a similar degree of architectural control as in natural structures have not been produced yet due to limitations in fabrication processes. In a combined top-down and bottom-up approach, we exploit the freedom in macroscopic design offered by 3D printing and the template-guided assembly of ink building blocks at the meso- and nanolevel to fabricate hierarchical silk porous materials with unprecedented structural control. Pores with tunable sizes in the range 40–350 μm are generated by adding sacrificial organic microparticles as templates to a silk fibroin-based ink. Commercially available wax particles or monodisperse polycaprolactone made by microfluidics can be used as microparticle templates. Since closed pores are generated after template removal, an ultrasonication treatment can optionally be used to achieve open porosity. Such pore templating particles can be further modified with nanoparticles to create a hierarchical template that results in porous structures with a defined nanotopography on the pore walls. The hierarchically porous silk structures obtained with this processing technique can potentially be utilized in various application fields from structural materials to thermal insulation to tissue engineering scaffolds.
AUTHOR Suntornnond, Ratima and Tan, Edgar Yong Sheng and An, Jia and Chua, Chee Kai
Title A Mathematical Model on the Resolution of Extrusion Bioprinting for the Development of New Bioinks [Abstract]
Year 2016
Journal/Proceedings Materials
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DOI/URL URL DOI
Abstract
Pneumatic extrusion-based bioprinting is a recent and interesting technology that is very useful for biomedical applications. However, many process parameters in the bioprinter need to be fully understood in order to print at an adequate resolution. In this paper, a simple yet accurate mathematical model to predict the printed width of a continuous hydrogel line is proposed, in which the resolution is expressed as a function of nozzle size, pressure, and printing speed. A thermo-responsive hydrogel, pluronic F127, is used to validate the model predictions. This model could provide a platform for future correlation studies on pneumatic extrusion-based bioprinting as well as for developing new bioink formulations.
AUTHOR Melchels, Ferry P. W. and Blokzijl, Maarten M. and Levato, Riccardo and Peiffer, Quentin C. and de Ruijter, Myl{`{e}}ne and Hennink, Wim E. and Vermonden, Tina and Malda, Jos
Title Hydrogel-based reinforcement of 3D bioprinted constructs [Abstract]
Year 2016
Journal/Proceedings Biofabrication
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Abstract
Progress within the field of biofabrication is hindered by a lack of suitable hydrogel formulations. Here, we present a novel approach based on a hybrid printing technique to create cellularized 3D printed constructs. The hybrid bioprinting strategy combines a reinforcing gel for mechanical support with a bioink to provide a cytocompatible environment. In comparison with thermoplastics such as IMG [http://ej.iop.org/images/1758-5090/8/3/035004/bfaa2f97ieqn1.gif] {$epsilon $} -polycaprolactone, the hydrogel-based reinforcing gel platform enables printing at cell-friendly temperatures, targets the bioprinting of softer tissues and allows for improved control over degradation kinetics. We prepared amphiphilic macromonomers based on poloxamer that form hydrolysable, covalently cross-linked polymer networks. Dissolved at a concentration of 28.6%w/w in water, it functions as reinforcing gel, while a 5%w/w gelatin-methacryloyl based gel is utilized as bioink. This strategy allows for the creation of complex structures, where the bioink provides a cytocompatible environment for encapsulated cells. Cell viability of equine chondrocytes encapsulated within printed constructs remained largely unaffected by the printing process. The versatility of the system is further demonstrated by the ability to tune the stiffness of printed constructs between 138 and 263 kPa, as well as to tailor the degradation kinetics of the reinforcing gel from several weeks up to more than a year.
AUTHOR Geven, Mike A. and Sprecher, Christoph and Guillaume, Olivier and Eglin, David and Grijpma, Dirk W.
Title Micro-porous composite scaffolds of photo-crosslinked poly(trimethylene carbonate) and nano-hydroxyapatite prepared by low-temperature extrusion-based additive manufacturing [Abstract]
Year 2016
Journal/Proceedings Polymers for Advanced Technologies
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DOI/URL DOI
Abstract
Complex bony defects such as those of the orbital floor are challenging to repair. Additive manufacturing techniques open up possibilities for the fabrication of implants with a designed macro-porosity for the reconstruction of such defects. Apart from a designed macro-porosity for tissue ingrowth, a micro-porosity in the implant struts will be beneficial for nutrient diffusion, protein adsorption and drug loading and release. In this work, we report on a low-temperature extrusion-based additive manufacturing method for the preparation of composite photo-crosslinked structures of poly(trimethylene carbonate) with bone-forming nano-hydroxyapatite and noricaritin (derived from bone growth stimulating icariin). In this method, we extrude a dispersion of nano-hydroxyapatite and noricaritin particles in a solution of photo-crosslinkable poly(trimethylene carbonate) in ethylene carbonate into defined three-dimensional structures. The ethylene carbonate is subsequently crystallized and extracted after photo-crosslinking. We show that this results in designed macro-porous structures with micro-pores in the struts. The dispersion used to fabricate these structures shows favorable properties for extrusion-based processing, such as a sharp crystallization response and shear thinning. The formed photo-crosslinked materials have a micro-porosity of up to 48%, and the E modulus, ultimate tensile strength and toughness are in excess of 24 MPa, 2.0 N/mm2 and 113 N/mm2 respectively. A sustained release of noricaritin from these materials was also achieved. The results show that the technique described here is promising for the fabrication of micro-porous photo-crosslinked composite structures of poly(trimethylene carbonate) with nano-hydroxyapatite and that these may be applied in the reconstruction of orbital floor defects. Copyright © 2016 John Wiley & Sons, Ltd.
AUTHOR M{"{u}}ller, Michael and Becher, Jana and Schnabelrauch, Matthias and Zenobi-Wong, Marcy
Title Nanostructured Pluronic hydrogels as bioinks for 3D bioprinting [Abstract]
Year 2015
Journal/Proceedings Biofabrication
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Abstract
Bioprinting is an emerging technology in the field of tissue engineering as it allows the precise positioning of biologically relevant materials in 3D, which more resembles the native tissue in our body than current homogenous, bulk approaches. There is however a lack of materials to be used with this technology and materials such as the block copolymer Pluronic have good printing properties but do not allow long-term cell culture. Here we present an approach called nanostructuring to increase the biocompatibility of Pluronic gels at printable concentrations. By mixing acrylated with unmodified Pluronic F127 it was possible to maintain the excellent printing properties of Pluronic and to create stable gels via UV crosslinking. By subsequent elution of the unmodified Pluronic from the crosslinked network we were able to increase the cell viability of encapsulated chondrocytes at day 14 from 62% for a pure acrylated Pluronic hydrogel to 86% for a nanostructured hydrogel. The mixed Pluronic gels also showed good printability when cells where included in the bioink. The nanostructured gels were, with a compressive modulus of 1.42 kPa, mechanically weak, but we were able to increase the mechanical properties by the addition of methacrylated hyaluronic acid. Our nanostructuring approach enables Pluronic hydrogels to have the desired set of properties in all stages of the bioprinting process.
AUTHOR Kesti, Matti and M{"{u}}ller, Michael and Becher, Jana and Schnabelrauch, Matthias and D{textquoteright}Este, Matteo and Eglin, David and Zenobi-Wong, Marcy
Title A versatile bioink for three-dimensional printing of cellular scaffolds based on thermally and photo-triggered tandem gelation [Abstract]
Year 2014
Journal/Proceedings Acta Biomaterialia
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DOI/URL URL DOI
Abstract
Abstract Layer-by-layer bioprinting is a logical choice for the fabrication of stratified tissues like articular cartilage. Printing of viable organ replacements, however, is dependent on bioinks with appropriate rheological and cytocompatible properties. In cartilage engineering, photocrosslinkable glycosaminoglycan-based hydrogels are chondrogenic, but alone have generally poor printing properties. By blending the thermoresponsive polymer poly(N-isopropylacrylamide) grafted hyaluronan (HA-pNIPAAM) with methacrylated hyaluronan (HAMA), high-resolution scaffolds with good viability were printed. HA-pNIPAAM provided fast gelation and immediate post-printing structural fidelity, while {HAMA} ensured long-term mechanical stability upon photocrosslinking. The bioink was evaluated for rheological properties, swelling behavior, printability and biocompatibility of encapsulated bovine chondrocytes. Elution of HA-pNIPAAM from the scaffold was necessary to obtain good viability. HA-pNIPAAM can therefore be used to support extrusion of a range of biopolymers which undergo tandem gelation, thereby facilitating the printing of cell-laden, stratified cartilage constructs with zonally varying composition and stiffness.
AUTHOR M{"{u}}ller, Michael and Becher, Jana and Schnabelrauch, Matthias and Zenobi-Wong, Marcy
Title Printing thermoresponsive reverse molds for the creation of patterned two-component hydrogels for 3D cell culture. [Abstract]
Year 2013
Journal/Proceedings Journal of visualized experiments : JoVE
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DOI/URL URL
Abstract
Bioprinting is an emerging technology that has its origins in the rapid prototyping industry. The different printing processes can be divided into contact bioprinting(1-4) (extrusion, dip pen and soft lithography), contactless bioprinting(5-7) (laser forward transfer, ink-jet deposition) and laser based techniques such as two photon photopolymerization(8). It can be used for many applications such as tissue engineering(9-13), biosensor microfabrication(14-16) and as a tool to answer basic biological questions such as influences of co-culturing of different cell types(17). Unlike common photolithographic or soft-lithographic methods, extrusion bioprinting has the advantage that it does not require a separate mask or stamp. Using CAD software, the design of the structure can quickly be changed and adjusted according to the requirements of the operator. This makes bioprinting more flexible than lithography-based approaches. Here we demonstrate the printing of a sacrificial mold to create a multi-material 3D structure using an array of pillars within a hydrogel as an example. These pillars could represent hollow structures for a vascular network or the tubes within a nerve guide conduit. The material chosen for the sacrificial mold was poloxamer 407, a thermoresponsive polymer with excellent printing properties which is liquid at 4 degrees C and a solid above its gelation temperature ~20 degrees C for 24.5% w/v solutions(18). This property allows the poloxamer-based sacrificial mold to be eluted on demand and has advantages over the slow dissolution of a solid material especially for narrow geometries. Poloxamer was printed on microscope glass slides to create the sacrificial mold. Agarose was pipetted into the mold and cooled until gelation. After elution of the poloxamer in ice cold water, the voids in the agarose mold were filled with alginate methacrylate spiked with FITC labeled fibrinogen. The filled voids were then cross-linked with UV and the construct was imaged with an epi-fluorescence microscope.