May 29, 2020 by admin 0 Comments

3D-Bioprinted Aptamer-Functionalized Bio-inks for Spatiotemporally Controlled Growth Factor Delivery

Authors
Deepti Rana, Vasileios D. Trikalitis (Contributor), Vincent R. Rangel (Contributor), Ajoy Kumar Kandar (Contributor), Nasim Salehi Nik (Contributor), Jeroen Rouwkema* (Contributor)
Abstract
Introduction Spatiotemporally controlled growth factors delivering systems are crucial for tissue engineering. However, most of the current strategies for growth factors delivery often focuses on the immobilization or coupling of growth factors within the engineered matrices (hydrogel) via various linker proteins or peptides. These systems provide passive release rates and growth factor delivery on demand, but fail to adapt their release rates in accordance with the tissue development. To overcome this limitation, the present study employed nucleic acid based aptamers for achieving spatiotemporally controlled growth factor delivery. Aptamers are affinity ligands selected from DNA/RNA libraries to recognize proteins with high affinity and specificity.1 Aptamer based growth factor delivery systems are able to load/release multiple growth factors on demand with high specificity. In the present study, the authors have 3D-bioprinted aptamer-functionalized bio-inks to evaluate their potential for growth factor sequestering, programmable release and for studying their effect on vascular network formation. Methods The aptamer-functionalized hydrogels were prepared via photo-polymerization of gelatin methacryloyl (GelMA) and acrydite functionalized aptamers having sequence specific for binding to vascular endothelial growth factor (VEGF165). Visible light photoinitiator, tris(2,2′-bipyridyl)dichloro-ruthenium(II) hexahydrate with sodium persulfate was used. The 3D-bioprinting experiments were carried out using Rokit Invivo 3D printer. The viscoelastic properties of the bio-inks were evaluated and compared with control GelMA bio-ink. To study the programmable growth factor release efficiency, VEGF antibody immunostaining was used. For studying the effect of triggered growth factor release on vascular network formation, human umbilical vein endothelial cells (HUVECs) and mesenchymal stem cells (MSCs) were encapsulated within the bio-inks. Results & Discussion The results obtained from VEGF antibody immunostainings confirmed the sequestration and triggered release of VEGF in response to complementary sequence addition from the 3D bioprinted construct after 5 days of culture. The bioprinted construct showed high cellular viability. The F-Actin/DAPI staining showed cellular sprouting and vascular network formation within the 3D printing aptamer functionalized bio-ink regions. In addition, the endothelial cells showed variations in cellular organization based on the VEGF bound aptamer availability within the bioprinted construct. These observations altogether confirms the bioactivity of VEGF bound aptamers within the printed constructs. Conclusions The present study shows the vasculogenic potential of 3D bioprinted aptamer-functionalized bio-inks via spatiotemporally controlling VEGF availability within the hydrogel system. Acknowledgements: This work is supported by an ERC Consolidator Grant under grant agreement no 724469. References 1. M.R. Battig, et. al., J. Am. Chem. Soc. 134 (2012) 12410-12413.

April 10, 2020 by admin 0 Comments

Structurally Reinforced Biodegradable Antithrombotic Small-Caliber Vascular Grafts Immobilized with VEGF to Accelerate Endothelialization: When 3D Printing Meets Electrospun Fiber

Authors
Gladys A. Emechebe, Francis O. Obiweluozor, In Seok Jeong, Park June Kyu, Chan Hee Park, Cheol Sang Kim
Abstract
The major challenge of commercially available vascular substitutes come from their limitations in terms of good mechanical strength and host remodeling. To date, tissue-engineered and synthetic grafts have not translated well to clinical trials when looking at small diameters. We conceptualized a cell-free structurally reinforced biodegradable vascular graft recapitulating the anisotropic feature of native blood vessel by using nanofibrous scaffold that will gradually degrade systematically to yield a neo-vessel, facilitated by an immobilized bioactive molecule-vascular endothelial growth factor (VEGF). The nanotopographic cue of the device is capable to directs host cell infiltration. We evaluated the burst pressure, Histology, hemocompatibility, compression test and mechanical analysis of the new graft. Hence, we proposed that future long-term studies of this technology on porcine models due to their similar vasculature regeneration to humans is needed prior to clinical translation. This acellular off-the-shelf approach will mark a paradigm shift from the current dominant focus on cell incorporation in vascular tissue engineering thus strongly influencing regenerative medicine as we move forward in this new decade.

February 14, 2020 by admin 0 Comments

Photocurable chitosan as bioink for cellularized therapies towards personalized scaffold architecture

Authors
Chiara Tonda-Turo (a) (b), Irene Carmagnola (a) (b), Annalisa Chiappone (b) (c), Zhaoxuan Feng (d), Gianluca Ciardelli (a) (b), Minna Hakkarainen (d), Marco Sangermano (b) (c)
Abstract
Recent progresses in tissue engineering are directed towards the development of technologies able to provide personalized scaffolds recreating the defect shape in a patient specific manner. To achieve this ambitious goal, 3D bioprinting can be combined with a suitable bioink, able to create a physiological milieu for cell growth. In this work, a novel chitosan-based hydrogel was developed combining photocrosslinking and thermo-sensitive properties. Commercial chitosan (CS) was first methacrylated and then mixed with β-glycerol phosphate salt (β-GP) to impart a thermally induced phase transition. The absence of cytotoxic degradation products and the excellent biocompatibility of the developed hydrogel was confirmed through in vitro tests using different cell lines (NIH/3T3, Saos-2, SH-SY5Y). Cellularized 3D structures were obtained though 3D bioprinting technologies confirming the processability of the developed hydrogels and its unique biological properties.

August 30, 2019 by admin 0 Comments

Bioprinting Technologies in Tissue Engineering

Authors
Bengi Yilmaz, Aydin Tahmasebifar, Erkan Türker Baran
Abstract
Bioprinting technology is a strong tool in producing living functional tissues and organs from cells, biomaterial-based bioinks, and growth factors in computer-controlled platform. The aim of this chapter is to present recent progresses in bioprinting of nerve, skin, cardiac, bone, cartilage, skeletal muscle, and other soft tissues and highlight the challenges in these applications. Various composite bioinks with bioactive ceramic-based scaffolds having patient-specific design and controlled micro-architectures were used at clinical and preclinical applications successfully for regeneration of bone. In nerve tissue engineering, bioprinting of alginate- and gelatin-based gel bioinks by extrusion presented a controllable 3D microstructures and showed satisfactory cytocompatibility and axonal regeneration. Bioprinting of cardiac progenitors in biopolymers resulted in limited success, while the use of bioinks from extracellular matrix induced satisfactory results in cardiac regeneration. Osteochondral scaffold bioprinting is challenging due to the complex hierarchical structure and limited chondral regeneration. Therefore, current approaches focused on osteochondral scaffold with vascular network and mimicking hierarchical structures. The applications of bioprinting in other types of tissues were also studied, and results showed significant potentials in regeneration of tissues such as cornea, liver, and urinary bladder.

June 1, 2018 by admin 0 Comments

The Arrival of Commercial Bioprinters – Towards 3D Bioprinting Revolution!

Authors
Deepak Choudhury*, Shivesh Anand, May Win Naing*
Abstract
The dawn of commercial bioprinting is rapidly advancing the tissue engineering field. In the past few years, new bioprinting approaches as well as novel bioinks formulations have emerged, enabling biological research groups to demonstrate the use of such technology to fabricate functional and relevant tissue models. In recent years, several companies have launched bioprinters pushing for early adoption and democratisation of bioprinting. This article reviews the progress in commercial bioprinting since the inception, with a particular focus on the comparison of different available printing technologies and important features of the individual technologies as well as various existing applications. Various challenges and potential design considerations for next generations of bioprinters are also discussed.

January 1, 2018 by admin 0 Comments

Design, fabrication and evaluation of a hybrid biomanufacturing system for tissue engineering

Authors
Fengyuan Liu
Abstract
Plasma-assisted Bio-extrusion System (PBS System) is an innovative hybrid bio-manufacturing system to produce complex multi-material and functionally graded scaffolds combining multiple pressure-assisted and screw-assisted printing heads and plasma jets. This approach, which represents a step forward regarding the current state of the art technology in the field of biomanufacturing, enables to design and fabricate more effective scaffolds matching the mechanical and surface characteristics of the surrounding tissue, enabling the incorporation of high number of cells uniformly distributed and the introduction of multiple cell types with positional specificity. The system requires complex control software to manipulate different materials, scaffold designs and processing parameters. This software, developed using MATLAB GUI, is detailed in this paper. It provides high freedom of design allowing the users to create single or multi-material constructs with uniform pore size or pores size gradients by changing the operation parameters, such as geometric parameters, lay-down pattern, filament distance, feed rate and layer thickness. Functionally graded scaffolds can also be designed considering different layer-by-layer coating/surface modification strategies using the multi-jet plasma system. Based on the user definition, G programming codes are generated enabling fully integration and synchronization with the hardware of the PBS system. Examples will be provided describing the design of single, multi-material and functionally graded scaffolds.