scholarly journals Bioprinting small-diameter vascular vessel with endothelium and smooth muscle by the approach of two-step crosslinking process

Author(s):  
Qianheng Jin ◽  
Guangzhe Jin ◽  
Jihui Ju ◽  
Lei Xu ◽  
Linfeng Tang ◽  
...  

Three-dimensional (3D) bioprinting shows great potential for autologous vascular grafts due to its simplicity, accuracy, and flexibility. 6mm diameter vascular grafts are used in clinic. However, producing small-diameter vascular grafts are still an enormous challenge. Normally, sacrificial hydrogels are used as temporary lumen support to mold tubular structure which will affect the structure’s stability. In this study, we develop a new bioprinting approach to fabricating small-diameter vessel using two-step crosslinking process. ¼ lumen wall of bioprinted gelatin mechacrylate (GelMA) flat structure is exposed to ultraviolet (UV) light briefly for having certain strength, while ¾ lumen wall shows as concave structure remained uncrosslinked. Pre-crosslinked flat structure is merged towards the uncrosslinked concave structure. Two individual structures will be combined tightly into an intact tubular structure by receiving more UV exposure time. Complicated tubular structures are constructed by these method. Notably, the GelMA-based bioink loaded with smooth muscle cells (SMCs) are bioprinted as the outer layer and human umbilical vein endothelial cells (HUVECs) are seeded onto the inner surface. A bionic vascular vessel with dual layers is fabricated successfully and keeps good viability, and functionality. This study may provide a novel idea for fabricating biomimetic vascular network or other more complicated organs.

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Manabu Itoh ◽  
Koichi Nakayama ◽  
Ryo Noguchi ◽  
Keiji Kamohara ◽  
Kojirou Furukawa ◽  
...  

Introduction: Small caliber synthetic vascular grafts are not clinically available. We developed a novel method to create scaffold-free tubular tissue from multicellular spheroids (MCS) using a “Bio-3D printer”-based system, which enables the creation of various three-dimensional structures pre-designed using a computer system. With this system, we created a tubular structure (Fig. 1), and studied its biological features. Methods: We made 1.5 mm in diameter scaffold-free tubular tissues from MCS (1.25 x 10[[Unable to Display Character: ⁷]] cells) composed of human umbilical vein endothelial cells (40%), human aortic smooth muscle cells (10%) and normal human dermal fibroblasts (50%) using a Bio-3D printer. The vessels were cultured in a perfusion system. We implanted grafts into the abdominal aortas of F344 nude rats, and assessed the flow by ultrasonography and performed histological examinations on the second (N=5) and fifth (N=5) days after implantation. Results: All grafts were patent. Remodeling of the vessel (enlargement of the lumen area and thinning of the wall) was observed (Fig. 2). A layer of endothelial cells was developed after implantation of the graft (Fig. 3). Conclusions: The scaffold-free vascular grafts made of MCS using a Bio-3D printer showed biological features comparable to native vessels. Further studies are warranted toward the clinical application of this novel technology.


2021 ◽  
Author(s):  
Qianheng Jin ◽  
Yi Fu ◽  
Guangliang Zhang ◽  
Lei Xu ◽  
Guangzhe Jin ◽  
...  

Abstract Background: Cardiovascular disease is responsible for a large number of deaths each year. Autologous vascular transplantation is still the best surgical intervention option, but the success rate is affected by many factors. Tissue engineering is a growing research area of great interest because it can produce bionic grafts to replace autologous tissue. Although many molding strategies have been tried, precellularization of small-diameter vascular grafts remains a research challenge. Here, a novel approach for fabricating bionic small-diameter vascular vessels with endothelial and smooth muscle cells is developed through combining nanofiber electrospinning and a specially-designed rotary bioprinter.Results: Combining and utilizing the advantages of nanofiber electrospinning and rotary printing, a tissue-engineered vascular tissue more suitable for biological transplantation is fabricated. Electrospun poly(ε-caprolactone) (PCL) provides good elasticity, and the electrospinning modification is beneficial for adhesion and functionalization of endothelial cells. A flat monolayer on the surface of PCL is formed after 7 days cultivation. Modification of the traditional three-dimensional (3D) bioprinter to increase rotation of the central axis used dual motors rotating clockwise and anticlockwise at the same speed increase stability during the printing process. This allowed a uniform dense methacrylated gelatin (GelMA) structure containing smooth muscle cells to be bioprinted with the cells are arranged linearly along the horizontal axis of rotation. Perfused with umbilical vein endothelial cells, a monolayer endothelial structure is formed. The two type cells maintain viability and proliferation in the structure during the process of cultivation. In addition, the bionic structureis superior to the natural blood vessel in anti-burst pressure and suture retention strength.Conclusion: By combining nanofiber electrospinning and modified rotary bioprinter, we successfully formed a small-diameter bionic vascular vessel with smooth muscle cells and endothelial cells. This method takes advantages of two advanced technologies and provides a new strategy for the development of bionic blood vascular tissue.


Polymers ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1217
Author(s):  
Jang Ho Ha ◽  
Jae Hyun Lim ◽  
Ji Woon Kim ◽  
Hyeon-Yeol Cho ◽  
Seok Geun Jo ◽  
...  

Blended hydrogels play an important role in enhancing the properties (e.g., mechanical properties and conductivity) of hydrogels. In this study, we generated a conductive blended hydrogel, which was achieved by mixing gelatin methacrylate (GelMA) with collagen, and silver nanowire (AgNW). The ratio of GelMA, collagen and AgNW was optimized and was subsequently gelated by ultraviolet light (UV) and heat. The scanning electron microscope (SEM) image of the conductive blended hydrogels showed that collagen and AgNW were present in the GelMA hydrogel. Additionally, rheological analysis indicated that the mechanical properties of the conductive GelMA–collagen–AgNW blended hydrogels improved. Biocompatibility analysis confirmed that the human umbilical vein endothelial cells (HUVECs) encapsulated within the three-dimensional (3D), conductive blended hydrogels were highly viable. Furthermore, we confirmed that the molecule in the conductive blended hydrogel was released by electrical stimuli-mediated structural deformation. Therefore, this conductive GelMA–collagen–AgNW blended hydrogel could be potentially used as a smart actuator for drug delivery applications.


Micromachines ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 3
Author(s):  
Kyo-in Koo ◽  
Andreas Lenshof ◽  
Le Thi Huong ◽  
Thomas Laurell

In the field of engineered organ and drug development, three-dimensional network-structured tissue has been a long-sought goal. This paper presents a direct hydrogel extrusion process exposed to an ultrasound standing wave that aligns fibroblast cells to form a network structure. The frequency-shifted (2 MHz to 4 MHz) ultrasound actuation of a 400-micrometer square-shaped glass capillary that was continuously perfused by fibroblast cells suspended in sodium alginate generated a hydrogel string, with the fibroblasts aligned in single or quadruple streams. In the transition from the one-cell stream to the four-cell streams, the aligned fibroblast cells were continuously interconnected in the form of a branch and a junction. The ultrasound-exposed fibroblast cells displayed over 95% viability up to day 10 in culture medium without any significant difference from the unexposed fibroblast cells. This acoustofluidic method will be further applied to create a vascularized network by replacing fibroblast cells with human umbilical vein endothelial cells.


2017 ◽  
Vol 33 (9) ◽  
pp. 592-599 ◽  
Author(s):  
Francesca Felice ◽  
Ester Belardinelli ◽  
Alessandro Frullini ◽  
Tatiana Santoni ◽  
Egidio Imbalzano ◽  
...  

Objectives Aminaphtone, a naphtohydrochinone used in the treatment of capillary disorders, may affect oedema in chronic venous insufficiency. Aim of study is to investigate the effect of aminaphtone on vascular endothelial permeability in vitro and its effects on three-dimensional capillary-like structures formed by human umbilical vein endothelial cells. Method Human umbilical vein endothelial cells were treated with 50 ng/ml VEGF for 2 h and aminaphtone for 6 h. Permeability assay, VE-cadherin expression and Matrigel assay were performed. Results VEGF-induced permeability was significantly decreased by aminaphtone in a range concentration of 1–20 µg/ml. Aminaphtone restored VE-cadherin expression. Finally, 6 h pre-treatment with aminaphtone significantly preserved capillary-like structures formed by human umbilical vein endothelial cells on Matrigel up to 48 h compared to untreated cells. Conclusions Aminaphtone significantly protects endothelium permeability and stabilises endothelial cells organised in capillary-like structures, modulating VE-cadherin expression. These data might explain the clinical benefit of aminaphtone on chronic venous insufficiency.


2002 ◽  
Vol 11 (4) ◽  
pp. 369-377 ◽  
Author(s):  
Makarand V. Risbud ◽  
Erdal Karamuk ◽  
René Moser ◽  
Joerg Mayer

Three-dimensional (3-D) scaffolds offer an exciting possibility to develop cocultures of various cell types. Here we report chitosan–collagen hydrogel-coated fabric scaffolds with defined mesh size and fiber diameter for 3-D culture of human umbilical vein endothelial cells (HUVECs). These scaffolds did not require pre-coating with fibronectin and they supported proper HUVEC attachment and growth. Scaffolds preserved endothelial cell-specific cobblestone morphology and cells were growing in compartments defined by the textile mesh. HUVECs on the scaffold maintained the property of contact inhibition and did not exhibit overgrowth until the end of in vitro culture (day 6). MTT assay showed that cells had preserved mitochondrial functionality. It was also noted that cell number on the chitosan-coated scaffold was lower than that of collagen-coated scaffolds. Calcein AM and ethidium homodimer (EtD-1) dual staining demonstrated presence of viable and metabolically active cells, indicating growth supportive properties of the scaffolds. Actin labeling revealed absence of actin stress fibers and uniform distribution of F-actin in the cells, indicating their proper attachment to the scaffold matrix. Confocal microscopic studies showed that HUVECs growing on the scaffold had preserved functionality as seen by expression of von Willebrand (vW) factor. Observations also revealed that functional HUVECs were growing at various depths in the hydrogel matrix, thus demonstrating the potential of these scaffolds to support 3-D growth of cells. We foresee the application of this scaffold system in the design of liver bioreactors wherein hepatocytes could be cocultured in parallel with endothelial cells to enhance and preserve liver-specific functions.


Sign in / Sign up

Export Citation Format

Share Document