Perfusable System Using Porous Collagen Gel Scaffold Actively Provides Fresh Culture Media to a Cultured 3D Tissue
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Culturing three-dimensional (3D) tissues with an appropriate microenvironment is a critical and fundamental technology in broad areas of cutting-edge bioengineering research. In addition, many technologies have engineered tissue functions. However, an effective system for transporting nutrients, waste, or oxygen to affect the functions of cell tissues has not been reported. In this study, we introduce a novel system that employs diffusion and convection to enhance transportation. To demonstrate the concept of the proposed system, three layers of normal human dermal fibroblast cell sheets are used as a model tissue, which is cultured on a general dish or porous collagen scaffold with perfusable channels for three days with and without the perfusion of culture media in the scaffold. The results show that the viability of the cell tissue was improved by the developed system. Furthermore, glucose consumption, lactate production, and oxygen transport to the tissues were increased, which might improve the viability of tissues. However, mechanical stress in the proposed system did not cause damage or unintentional functional changes in the cultured tissue. We believe that the introduced culturing system potentially suggests a novel standard for 3D cell cultures.Keywords:
Viability assay
Abstract Tissue engineering has attracted attention worldwide because of its application in regenerative medicine, drug screening, and cultured meat. Numerous biofabrication techniques for producing tissues have been developed, including various scaffold and printing methods. Here, we have proposed a novel tissue engineering method using a net metal mould without the use of a scaffold. Briefly, normal human dermal fibroblasts seeded on a dimple plate were subjected to static culture technique for several days to form spheroids. Spheroids of diameter ⩾200 μ m were poured into a net-shaped mould of gap ⩽100 μ m and subjected to shake-cultivation for several weeks, facilitating their fusion to form a three-dimensional (3D) tissue. Through this study, we successfully constructed a scaffold-free 3D tissue having strength that can be easily manipulated, which was difficult to construct using conventional tissue engineering methods. We also investigated the viability of the 3D tissue and found that the condition of the tissues was completely different depending on the culture media used. Collectively, this method allows scaffold-free culture of 3D tissues of unprecedented thickness, and may contribute largely to next-generation tissue engineering products.
Biofabrication
Regenerative Medicine
Spheroid
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The scaffold materials of skin tissue engineering play a key role in construction of artificial skins,which is an important research content in skin tissue engineering by providing the seed cells with an environment of adherence,growth,migration,proliferation,differentiation as well as metabolization.This paper expounds on the progress of tissue engineering skin scaffold materials and preparation methods in recent years,with analysis on developing trend of skin tissue engineering scaffolds in the future.
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FUNCTIONS OF CELLS SCAFFOLD IN THE TISSUE ENGINEERINGCell, cells scaffold and the construction of tissue and organ are three main factors for the Tissue Engineering. A main function of cells scaffold in tissue engineering is to provide an environment for cells propagation.
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Current therapies of organ failure or a wide range of tissue defect are often not ideal. Transplantation is the only effective way for long time survival. But it is hard to meet huge patients demands because of donor shortage, immune rejection and other problems. Tissue engineering could be a potential option. Choosing a suitable scaffold material is an essential part of it. According to different sources, tissue engineering scaffold materials could be divided into three types which are natural and its modified materials, artificial and composite ones. The purpose of tissue engineering scaffold is to repair the tissues or organs damage, so could reach the ideal recovery in its function and structure aspect. Therefore, tissue engineering scaffold should even be as close as much to the original tissue or organs in function and structure. We call it "organic scaffold" and this strategy might be the drastic perfect substitute for the tissues or organs in concern. Optimized organization with each kind scaffold materials could make up for biomimetic structure and function of the tissue or organs. Scaffold material surface modification, optimized preparation procedure and cytosine sustained-release microsphere addition should be considered together. This strategy is expected to open new perspectives for tissue engineering. Multidisciplinary approach including material science, molecular biology, and engineering might find the most ideal tissue engineering scaffold. Using the strategy of drawing on each other strength and optimized organization with each kind scaffold material to prepare a multifunctional biomimetic tissue engineering scaffold might be a good method for choosing tissue engineering scaffold materials. Our research group had differentiated bone marrow mesenchymal stem cells into bile canaliculi like cells. We prepared poly(L-lactic acid)/poly(ε-caprolactone) biliary stent. The scaffold's internal played a part in the long-term release of cytokines which mixed with sustained-release nano-microsphere containing growth factors. What's more, the stent internal surface coated with glue/collagen matrix mixing layer containing bFGF and EGF so could supplying the early release of the two cytokines. Finally, combining the poly(L-lactic acid)/poly(ε-caprolactone) biliary stent with the induced cells was the last step for preparing tissue-engineered bile duct. This literature reviewed a variety of the existing tissue engineering scaffold materials and briefly introduced the impact factors on the characteristics of tissue engineering scaffold materials such as preparation procedure, surface modification of scaffold, and so on. We explored the choosing strategy of desired tissue engineering scaffold materials.
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Cell scaffold material is the framework of regeneration in tissue engineering.The property of cell scaffold material is the direct effect on the adhesion,proliferation and metabolism functions.Therefore,cell scaffold material is one of the critical factors in tissue engineering.Currently,the scaffold material of tissue engineering is categorized as the synthesized absorbable macromolecule polymer,natural material extracts and extracellular matrix.Based on the pertinent iterature in recent years,the research progress of scaffold material in tissue engineering was summarized in this paper.
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Scaffold plays a critical role in tissue engineering where it provides necessary structural support for the cells to accommodate and to guide their growth in the three dimensional space into a specific tissue. Therefore, engineering scaffolds favorable for cell/tissue growth is of great importance and a pre-requisite for scaffold-based tissue engineering. Electrospinning is a versatile method that has been recently adapted in engineering nano-fibrous scaffolds that mimic the structural features of biological extracellular matrix (ECM). It offers many advantages over conventional scaffold methodologies, for example, capable of producing ultra-fine fibers with high porosity, high spatial orientation, high aspect ratio, and high surface area, which are highly required for the initial cell attachment, tissue formation, and continued function. Considering these astonishing merits, this article emphasis on nano-fibrous scaffold engineering by electrospinning.
Electrospinning
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Matrix (chemical analysis)
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Purpose The 3 D bioprinting technology is used to prepare the tissue engineering scaffold with precise structures for the cell proliferation and differentiation. Design/methodology/approach According to the characteristics of the ideal tissue engineering scaffold, the microstructural design of the tissue engineering scaffold is carried out. The bioprinter is used to fabricate the tissue engineering scaffold with different structures and spacing sizes. Finally, the scaffold with good connectivity is achieved and used to cell PC12 culture. Findings The results show that the pore structure with the line spacing of 1 mm was the best for cell culture, and the survival rate of the inoculated cells PC12 is as high as 90%. The influence of the pore shape on the cell survival is not evidence. Originality/value This study shows that tissue engineering scaffolds prepared by 3 D bioprinting have graded structure for three-dimensional cell culture, which lays the foundation for the later detection of drug resistance.
3D cell culture
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Natural polymers
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Tissue engineeringによって肝組織を再構築するためには, 細胞, 細胞の足場となる基質 (scaffold), 細胞を接着させる環境を適当に機能させることが必要である。このうち細胞選択のための研究はかなり進歩してきているが, 特に, 細胞の接着性, 動態をどのように制御するかというようなscaffoldと細胞との環境を整備する研究が遅れている。肝組織再構築のためにはscaffold内において肝細胞がどのような動態を示すかを解明し, 自由に接着させる方法を開発することが必要である。さらに, 近未来には, tissue engineeringの構造物として, 生物学的シグナルも組み込んだ, 生態系や生細胞と共存できるscaffold作製が期待される。
Liver tissue
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