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      Human adipose derived stem cells are superior to human osteoblasts (HOB) in bone tissue engineering on a collagen-fibroin-ELR blend

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          Abstract

          The ultrastructure of the bone provides a unique mechanical strength against compressive, torsional and tensional stresses. An elastin-like recombinamer (ELR) with a nucleation sequence for hydroxyapatite was incorporated into films prepared from a collagen – silk fibroin blend carrying microchannel patterns to stimulate anisotropic osteogenesis. SEM and fluorescence microscopy showed the alignment of adipose-derived stem cells (ADSCs) and the human osteoblasts (HOBs) on the ridges and in the grooves of microchannel patterned collagen-fibroin-ELR blend films. The Young's modulus and the ultimate tensile strength (UTS) of untreated films were 0.58 ± 0.13 MPa and 0.18 ± 0.05 MPa, respectively. After 28 days of cell culture, ADSC seeded film had a Young's modulus of 1.21 ± 0.42 MPa and UTS of 0.32 ± 0.15 MPa which were about 3 fold higher than HOB seeded films. The difference in Young's modulus was statistically significant (p: 0.02). ADSCs attached, proliferated and mineralized better than the HOBs. In the light of these results, ADSCs served as a better cell source than HOBs for bone tissue engineering of collagen-fibroin-ELR based constructs used in this study. We have thus shown the enhancement in the tensile mechanical properties of the bone tissue engineered scaffolds by using ADSCs.

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          Highlights

          • ADSCs and HOBs aligned along the microchannel patterns which is important in the mimicking of the bone tissue ultrastructure.

          • ADSCs deposited statistically significant amounts of calcium phosphate.

          • Deposited calcium phosphate by ADSCs led to an increase in the Young’s modulus and UTS.

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          Most cited references42

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          Bone substitutes: an update.

          Autograft is considered ideal for grafting procedures, providing osteoinductive growth factors, osteogenic cells, and an osteoconductive scaffold. Limitations, however, exist regarding donor site morbidity and graft availability. Allograft on the other hand, posses the risk of disease transmission. Synthetic graft substitutes lack osteoinductive or osteogenic properties. Composite grafts combine scaffolding properties with biological elements to stimulate cell proliferation and differentiation and eventually osteogenesis. We present here an overview of bone grafts and graft substitutes available for clinical applications.
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            Bioprintable, cell-laden silk fibroin-gelatin hydrogel supporting multilineage differentiation of stem cells for fabrication of three-dimensional tissue constructs.

            Bioprinting has exciting prospects for printing three-dimensional (3-D) tissue constructs by delivering living cells with appropriate matrix materials. However, progress in this field is currently extremely slow due to limited choices of bioink for cell encapsulation and cytocompatible gelation mechanisms. Here we report the development of clinically relevant sized tissue analogs by 3-D bioprinting, delivering human nasal inferior turbinate tissue-derived mesenchymal progenitor cells encapsulated in silk fibroin-gelatin (SF-G) bioink. Gelation in this bioink was induced via in situ cytocompatible gelation mechanisms, namely enzymatic crosslinking by mushroom tyrosinase and physical crosslinking via sonication. Mechanistically, tyrosinases oxidize the accessible tyrosine residues of silk and/or gelatin into reactive o-quinone moieties that can either condense with each other or undergo nonenzymatic reactions with available amines of both silk and gelatin. Sonication alters the hydrophobic interaction and accelerates self-assembly of silk fibroin macromolecules to form β-sheet crystals, which physically crosslink the hydrogel. However, sonication has no effect on the conformation of gelatin. The effect of optimized rheology, secondary conformations of silk-gelatin bioink, temporally controllable gelation strategies and printing parameters were assessed to achieve maximum cell viability and multilineage differentiation of the encapsulated human nasal inferior turbinate tissue-derived mesenchymal progenitor cells. This strategy offers a unique path forward in the direction of direct printing of spatially customized anatomical architecture in a patient-specific manner.
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              Effect of fiber diameter on spreading, proliferation, and differentiation of osteoblastic cells on electrospun poly(lactic acid) substrates.

              Electrospinning is a promising method to construct fused-fiber biomaterial scaffolds for tissue engineering applications, but the efficacy of this approach depends on how substrate topography affects cell function. Previously, it has been shown that linear, parallel raised features with length scales of 0.5-2 microm direct cell orientation through the phenomenon of contact guidance, and enhance phenotypic markers of osteoblastic differentiation. To determine how the linear, random raised features produced by electrospinning affect proliferation and differentiation of osteoprogenitor cells, poly(lactic acid) and poly(ethylene glycol)-poly(lactic acid) diblock copolymers were electrospun with mean fiber diameters of 0.14-2.1 microm onto rigid supports. MC3T3-E1 osteoprogenitor cells cultured on fiber surfaces in the absence of osteogenic factors exhibited a lower cell density after 7 and 14 days of culture than cells cultured on spin-coated surfaces, but cell density increased with fiber diameter. However, in the presence of osteogenic factors (2 mM beta-glycerophosphate, 0.13 mM L-ascorbate-2-phosphate), cell density after 7 and 14 days of culture on fiber surfaces was comparable to or exceeded spin-coated controls, and alkaline phosphatase activity after 14 days was comparable. Examination of cell morphology revealed that cells grown on fibers had smaller projected areas than those on planar surfaces. However, cells attached to electrospun substrates of 2.1 microm diameter fibers exhibited a higher cell aspect ratio than cells on smooth surfaces. These studies show that topographical factors designed into biomaterial scaffolds can regulate spreading, orientation, and proliferation of osteoblastic cells.
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                Author and article information

                Contributors
                Journal
                Bioact Mater
                Bioact Mater
                Bioactive Materials
                KeAi Publishing
                2452-199X
                27 April 2017
                June 2017
                27 April 2017
                : 2
                : 2
                : 71-81
                Affiliations
                [a ]METU, Department of Biotechnology, Ankara, Turkey
                [b ]BIOMATEN, METU Center of Excellence in Biomaterials and Tissue Engineering, Dumlupinar Blvd No: 1, 06800 Ankara, Turkey
                [c ]University of Liverpool, School of Engineering, L69 3GH Liverpool, UK
                [d ]BIOFORGE, CIBER-BBN, Campus “Miguel Delibes” Edificio LUCIA, Universidad de Valladolid, Paseo Belén 19, 47011 Valladolid, Spain
                [e ]METU, Department of Biological Sciences, Ankara, 06800, Turkey
                Author notes
                []Corresponding author. METU, Department of Biological Sciences, Ankara, Turkey. Tel.: +90 312 2105180. vhasirci@ 123456metu.edu.tr
                Article
                S2452-199X(17)30003-8
                10.1016/j.bioactmat.2017.04.001
                5935045
                29744414
                52d9e46d-bbfb-4d15-9c4b-d07ae197be4a
                © 2017 The Authors

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 19 January 2017
                : 10 April 2017
                : 12 April 2017
                Categories
                Bioactive Polymer

                adipose-derived stem cells,human osteoblasts,tissue engineering,bone,mechanical properties

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