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      Macro- and micro-designed chitosan-alginate scaffold architecture by three-dimensional printing and directional freezing

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      Biofabrication

      IOP Publishing

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          Abstract

          While many tissue-engineered constructs aim to treat cartilage defects, most involve chondrocyte or stem cell seeding on scaffolds. The clinical application of cell-based techniques is limited due to the cost of maintaining cellular constructs on the shelf, potential immune response to allogeneic cell lines, and autologous chondrocyte sources requiring biopsy from already diseased or injured, scarce tissue. An acellular scaffold that can induce endogenous influx and homogeneous distribution of native stem cells from bone marrow holds great promise for cartilage regeneration. This study aims to develop such an acellular scaffold using designed, channeled architecture that simultaneously models the native zones of articular cartilage and subchondral bone. Highly porous, hydrophilic chitosan-alginate (Ch-Al) scaffolds were fabricated in three-dimensionally printed (3DP) molds designed to create millimeter scale macro-channels. Different polymer preform casting techniques were employed to produce scaffolds from both negative and positive 3DP molds. Macro-channeled scaffolds improved cell suspension distribution and uptake overly randomly porous scaffolds, with a wicking volumetric flow rate of 445.6 ± 30.3 mm(3) s(-1) for aqueous solutions and 177 ± 16 mm(3) s(-1) for blood. Additionally, directional freezing was applied to Ch-Al scaffolds, resulting in lamellar pores measuring 300 μm and 50 μm on the long and short axes, thus creating micrometer scale micro-channels. After directionally freezing Ch-Al solution cast in 3DP molds, the combined macro- and micro-channeled scaffold architecture enhanced cell suspension uptake beyond either macro- or micro-channels alone, reaching a volumetric flow rate of 1782.1 ± 48 mm(3) s(-1) for aqueous solutions and 440.9 ± 0.5 mm(3) s(-1) for blood. By combining 3DP and directional freezing, we can control the micro- and macro-architecture of Ch-Al to drastically improve cell influx into and distribution within the scaffold, while achieving porous zones that mimic articular cartilage zonal architecture. In future applications, precisely controlled micro- and macro-channels have the potential to assist immediate endogenous bone marrow uptake, stimulate chondrogenesis, and encourage vascularization of bone in an osteochondral scaffold.

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          Most cited references 44

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          Porous chitosan scaffolds for tissue engineering.

          The wide array of tissue engineering applications exacerbates the need for biodegradable materials with broad potential. Chitosan, the partially deacetylated derivative of chitin, may be one such material. In this study, we examined the use of chitosan for formation of porous scaffolds of controlled microstructure in several tissue-relevant geometries. Porous chitosan materials were prepared by controlled freezing and lyophilization of chitosan solutions and gels. The materials were characterized via light and scanning electron microscopy as well as tensile testing. The scaffolds formed included porous membranes, blocks, tubes and beads. Mean pore diameters could be controlled within the range 1-250 microm, by varying the freezing conditions. Freshly lyophilized chitosan scaffolds could be treated with glycosaminoglycans to form ionic complex materials which retained the original pore structure. Chitosan scaffolds could be rehydrated via an ethanol series to avoid the stiffening caused by rehydration in basic solutions. Hydrated porous chitosan membranes were at least twice as extensible as non-porous chitosan membranes, but their elastic moduli and tensile strengths were about tenfold lower than non-porous controls. The methods and structures described here provide a starting point for the design and fabrication of a family of polysaccharide based scaffold materials with potentially broad applicability.
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            Is Open Access

            Freezing as a path to build complex composites

            Materials that are strong, ultra-light weight and tough are in demand for a range of applications, requiring architectures and components carefully designed from the micrometer down to nanometer scales. Nacre-a structure found in many molluscan shells-and bone are frequently used as examples for how nature achieves this through hybrid organic-inorganic composites. Unfortunately, it has proven extremely difficult to transcribe nacre-like clever designs into synthetic materials, partly because their intricate structures need to be replicated at several length scales. We demonstrate how the physics of ice formation can be used to develop sophisticated porous and layered-hybrid materials, including artificial bone, ceramic/metal composites, and porous scaffolds for osseous tissue regeneration with strengths up to four times higher than those currently used for implantation.
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              Making tissue engineering scaffolds work. Review: the application of solid freeform fabrication technology to the production of tissue engineering scaffolds.

              Tissue engineering is a new and exciting technique which has the potential to create tissues and organs de novo. It involves the in vitro seeding and attachment of human cells onto a scaffold. These cells then proliferate, migrate and differentiate into the specific tissue while secreting the extracellular matrix components required to create the tissue. It is evident, therefore, that the choice of scaffold is crucial to enable the cells to behave in the required manner to produce tissues and organs of the desired shape and size. Current scaffolds, made by conventional scaffold fabrication techniques, are generally foams of synthetic polymers. The cells do not necessarily recognise such surfaces, and most importantly cells cannot migrate more than 500 microm from the surface. The lack of oxygen and nutrient supply governs this depth. Solid freeform fabrication (SFF) uses layer-manufacturing strategies to create physical objects directly from computer-generated models. It can improve current scaffold design by controlling scaffold parameters such as pore size, porosity and pore distribution, as well as incorporating an artificial vascular system, thereby increasing the mass transport of oxygen and nutrients into the interior of the scaffold and supporting cellular growth in that region. Several SFF systems have produced tissue engineering scaffolds with this concept in mind which will be the main focus of this review. We are developing scaffolds from collagen and with an internal vascular architecture using SFF. Collagen has major advantages as it provides a favourable surface for cellular attachment. The vascular system allows for the supply of nutrients and oxygen throughout the scaffold. The future of tissue engineering scaffolds is intertwined with SFF technologies.
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                Author and article information

                Journal
                Biofabrication
                Biofabrication
                IOP Publishing
                1758-5090
                March 01 2016
                January 07 2016
                : 8
                : 1
                : 015003
                Article
                10.1088/1758-5090/8/1/015003
                26741113
                © 2016

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