Springer protocols feed by Biotechnology 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Mechanical Testing of Cartilage Constructs
A key goal of functional cartilage tissue engineering is to develop constructs with mechanical properties approaching those of the native tissue. Herein we describe a number of tests to characterize the mechanical properties of tissue engineered cartilage. Specifically, methods to determine the equilibrium confined compressive (or aggregate) modulus, the equilibrium unconfined compressive (or Young’s) modulus, and the dynamic modulus of tissue engineered cartilaginous constructs are described. As these measurements are commonly used in both the articular cartilage mechanics literature and the cartilage tissue enginee...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Proteomic Analysis of Engineered Cartilage
Tissue engineering holds promise for the treatment of damaged and diseased tissues, especially for those tissues that do not undergo repair and regeneration readily in situ. Many techniques are available for cell and tissue culturing and differentiation of chondrocytes using a variety of cell types, differentiation methods, and scaffolds. In each case, it is critical to demonstrate the cellular phenotype and tissue composition, with particular attention to the extracellular matrix molecules that play a structural role and that contribute to the mechanical properties of the resulting tissue construct. Mass spectrometry prov...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Transplantation of Tissue-Engineered Cartilage in an Animal Model (Xenograft and Autograft): Construct Validation
Tissue engineering holds great promise for cartilage repair with minimal donor-site morbidity. The in vivo maturation of a tissue-engineered construct can be tested in the subcutaneous tissues of the same species for autografts or of immunocompromised animals for allografts or xenografts. This section describes detailed protocols for the surgical transplantation of a tissue-engineered construct into an animal model to assess construct validity. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Microbioreactors for Cartilage Tissue Engineering
In tissue engineering research, cell-based assays are widely utilized to fundamentally explore cellular responses to extracellular conditions. Nevertheless, the simplified cell culture models available at present have several inherent shortcomings and limitations. To tackle the issues, a wide variety of microbioreactors for cell culture have been actively proposed, especially during the past decade. Among these, micro-scale cell culture devices based on microfluidic biochip technology have particularly attracted considerable attention. In this chapter, we not only discuss the advantageous features of using micro-scale cell...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Shear and Compression Bioreactor for Cartilage Synthesis
Mechanical forces, including hydrodynamic shear, hydrostatic pressure, compression, tension, and friction, can have stimulatory effects on cartilage synthesis in tissue engineering systems. Bioreactors capable of exerting forces on cells and tissue constructs within a controlled culture environment are needed to provide appropriate mechanical stimuli. In this chapter, we describe the construction, assembly, and operation of a mechanobioreactor providing simultaneous dynamic shear and compressive loading on developing cartilage tissues to mimic the rolling and squeezing action of articular joints. The device is suitable for...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Mechanobioreactors for Cartilage Tissue Engineering
Mechanical stimulation is an effective method to increase extracellular matrix synthesis and to improve the mechanical properties of tissue-engineered cartilage constructs. In this chapter, we describe valuable methods of imposing direct mechanical stimuli (compression or shear) to tissue-engineered cartilage constructs as well as some common analytical methods used to quantify the effects of mechanical stimuli after short-term or long-term loading. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Decellularized Extracellular Matrix Scaffolds for Cartilage Regeneration
Decellularized extracellular matrix (ECM) is gaining a lot of attention as a biomaterial for tissue engineering applications. This chapter describes the processing techniques for decellularization of cell-derived ECM and protocols for the fabrication of ECM-based scaffolds in the form of hydrogels or fibrous polymer meshes by electrospinning. It describes the protocols to analyze the morphology and presence of collagen in fabricated scaffolds using scanning electron microscope and Picrosirius Red staining respectively. Methods to evaluate the metabolic activity and proliferation of cells (resazurin-based assay and DNA assa...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Hydrogels with Tunable Properties
This chapter describes the preparation of tissue engineered constructs by immobilizing chondrocytes in hydrogel with independently tunable porosity and mechanical properties. This chapter also presents the methods to characterize these tissue engineered constructs. The resulting tissue engineered constructs can be useful for the generation of cartilage tissue both in vitro and in vivo. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Stratified Scaffolds for Osteochondral Tissue Engineering
Stratified scaffolds are promising devices finding application in the field of osteochondral tissue engineering. In this scaffold type, different biomaterials are chosen to fulfill specific features required to mimic the complex osteochondral tissue interface, including cartilage, interlayer tissue, and subchondral bone. Here, the biomaterials and fabrication methods currently used to manufacture stratified multilayered scaffolds as well as cell seeding techniques for their characterization are presented. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Nanostructured Capsules for Cartilage Tissue Engineering
Polymeric multilayered capsules (PMCs) have found great applicability in bioencapsulation, an evolving branch of tissue engineering and regenerative medicine. Here, we describe the production of hierarchical PMCs composed by an external multilayered membrane by layer-by-layer assembly of poly(l-lysine), alginate, and chitosan. The core of the PMCs is liquified and encapsulates human adipose stem cells and surface-functionalized collagen II-TGF-β3 poly(l-lactic acid) microparticles for cartilage tissue engineering. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Scaffolds for Controlled Release of Cartilage Growth Factors
In recent years, cell-based therapies using adult stem cells have attracted considerable interest in regenerative medicine. A tissue-engineered construct for cartilage repair should provide a support for the cell and allow sustained in situ delivery of bioactive factors capable of inducing cell differentiation into chondrocytes. Pharmacologically active microcarriers (PAMs), made of biodegradable and biocompatible poly (d,l-lactide-co-glycolide acid) (PLGA), are a unique system which combines these properties in an adaptable and simple microdevice. This device relies on nanoprecipitation of proteins encapsulated in polymer...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Bioprinted Scaffolds for Cartilage Tissue Engineering
Researchers are focusing on bioprinting technology as a viable option to overcome current difficulties in cartilage tissue engineering. Bioprinting enables a three-dimensional (3-D), free-form, computer-designed structure using biomaterials, biomolecules, and/or cells. The inner and outer shape of a scaffold can be controlled by this technology with great precision. Here, we introduce a hybrid bioprinting technology that is a co-printing process of multiple materials including high-strength synthetic polymer and cell-laden hydrogel. The synthetic polymer provides mechanical support for shape maintenance and load bearing, w...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Use of Interim Scaffolding and Neotissue Development to Produce a Scaffold-Free Living Hyaline Cartilage Graft
The fabrication of three-dimensional (3D) constructs relies heavily on the use of biomaterial-based scaffolds. These are required as mechanical supports as well as to translate two-dimensional cultures to 3D cultures for clinical applications. Regardless of the choice of scaffold, timely degradation of scaffolds is difficult to achieve and undegraded scaffold material can lead to interference in further tissue development or morphogenesis. In cartilage tissue engineering, hydrogel is the highly preferred scaffold material as it shares many similar characteristics with native cartilaginous matrix. Hence, we employed gelatin...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Gene Transfer and Gene Silencing in Stem Cells to Promote Chondrogenesis
In stem cell-based chondrogenesis for articular cartilage regeneration, TGF-β3 is dosed to the stem cells to drive differentiation into chondrocytic cells. Meanwhile, type I collagen, which is endogenously expressed in some stem cells (e.g., synovium-derived mesenchymal stem cells) and upregulated by TGF-β3, poses a threat to chondrogenesis, as type I collagen may alter the components and stiffness of articular cartilage. Therefore, a wiser strategy would be to feed the cells with TGF-β3 while at the same time silencing the expression of type I collagen. In this chapter, methods for construction of adenovira...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
Differentiation of Human Induced Pluripotent Stem Cells to Chondrocytes
Human induced pluripotent stem (iPS) cells are relevant tools for modeling human skeletal development and disease, and represent a promising source of patient-specific cells for the regeneration of skeletal tissue, such as articular cartilage. Devising efficient and reproducible strategies, which closely mimic the physiological chondrogenic differentiation process, will be necessary to generate functional chondrocytes from human iPS cells. Our previous study demonstrated the generation of chondrogenically committed human iPS cells via the enrichment of a mesenchymal-like progenitor population, application of appropriate hi...
Source: Springer protocols feed by Biotechnology - September 14, 2015 Category: Biotechnology Source Type: news
