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      Bioinspired nanostructured hydroxyapatite/collagen three-dimensional porous scaffolds for bone tissue engineering

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          Abstract

          A needle punching and bioinspired mineralization strategy has been developed to fabricate a collagen/hydroxyapatite porous scaffold for bone tissue engineering.

          Abstract

          During the biomineralization process of bone minerals, amorphous calcium phosphate (ACP) is converted to apatite crystals by using octacalcium phosphate (OCP) and brushite (DCPD) as transitory precursors, resulting in the formation of hybrid nanostructured collagen/apatite composites. Herein, we report, for the first time, the bioinspired synthesis of a collagen/hydroxyapatite (HA) porous scaffold (CHPS) according to the following stages: (i) fabrication of collagen fibre porous scaffold (CFPS) by a needle-punching process; (ii) deposition of brushite/chitosan (DCPD/CS) on CHPS by a dip-coating method; and (iii) formation of CHPS by in situ conversion of DCPD to HA. The CHPS exhibits three-dimensional (3D) interconnected porous structures with pore sizes of around 60 μm. HA crystals distribute homogeneously on the CHPS, and display wheat-like shapes with a length of approximately 200 nm and a width of approximately 80 nm. The in vitro cell tests by using human bone marrow stromal cells (hBMSCs) indicate that the HA crystals in the CHPS not only promote the cell adhesion and proliferation of the hBMSCs, but also stimulate osteogenic differentiation. The in vivo results reveal that the CHPS exhibits better osteoinductivity than the CFPS because of its similar chemical components, crystallinity and crystallographic texture to natural bone. Moreover, the CHPS can stimulate new bone formation in rat critical-sized calvarial defects within 8 weeks. The CHPS possesses a favourable pore structure, and excellent biocompatibility and osteoinductivity, and thus it has great potential applications for bone tissue engineering.

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          Recent advances in bone tissue engineering scaffolds.

          Bone disorders are of significant concern due to increase in the median age of our population. Traditionally, bone grafts have been used to restore damaged bone. Synthetic biomaterials are now being used as bone graft substitutes. These biomaterials were initially selected for structural restoration based on their biomechanical properties. Later scaffolds were engineered to be bioactive or bioresorbable to enhance tissue growth. Now scaffolds are designed to induce bone formation and vascularization. These scaffolds are often porous, made of biodegradable materials that harbor different growth factors, drugs, genes, or stem cells. In this review, we highlight recent advances in bone scaffolds and discuss aspects that still need to be improved. Copyright © 2012 Elsevier Ltd. All rights reserved.
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            3D printing of composite calcium phosphate and collagen scaffolds for bone regeneration.

            Low temperature 3D printing of calcium phosphate scaffolds holds great promise for fabricating synthetic bone graft substitutes with enhanced performance over traditional techniques. Many design parameters, such as the binder solution properties, have yet to be optimized to ensure maximal biocompatibility and osteoconductivity with sufficient mechanical properties. This study tailored the phosphoric acid-based binder solution concentration to 8.75 wt% to maximize cytocompatibility and mechanical strength, with a supplementation of Tween 80 to improve printing. To further enhance the formulation, collagen was dissolved into the binder solution to fabricate collagen-calcium phosphate composites. Reducing the viscosity and surface tension through a physiologic heat treatment and Tween 80, respectively, enabled reliable thermal inkjet printing of the collagen solutions. Supplementing the binder solution with 1-2 wt% collagen significantly improved maximum flexural strength and cell viability. To assess the bone healing performance, we implanted 3D printed scaffolds into a critically sized murine femoral defect for 9 weeks. The implants were confirmed to be osteoconductive, with new bone growth incorporating the degrading scaffold materials. In conclusion, this study demonstrates optimization of material parameters for 3D printed calcium phosphate scaffolds and enhancement of material properties by volumetric collagen incorporation via inkjet printing. Copyright © 2014 Elsevier Ltd. All rights reserved.
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              The regulation of tendon stem cell differentiation by the alignment of nanofibers.

              Tendon is a specific connective tissue composed of parallel collagen fibers. The effect of this tissue-specific matrix orientation on stem cell differentiation has not been investigated. This study aimed to determine the effects of nanotopography on the differentiation of human tendon stem/progenitor cells (hTSPCs) and develop a biomimetic scaffold for tendon tissue engineering. The immuno-phenotype of fetal hTSPCs was identified by flow cytometry. The multipotency of hTSPCs toward osteogenesis, adipogenesis, and chondrogenesis was confirmed. Then, the hTSPCs were seeded onto aligned or randomly-oriented poly (l-lactic acid) nanofibers. Scanning electron micrographs showed that hTSPCs were spindle-shaped and well orientated on the aligned nanofibers. The expression of tendon-specific genes was significantly higher in hTSPCs growing on aligned nanofibers than those on randomly-oriented nanofibers in both normal and osteogenic media. In addition, alkaline phosphatase activity and alizarin red staining showed that the randomly-oriented fibrous scaffold induced osteogenesis, while the aligned scaffold hindered the process. Moreover, aligned cells expressed significantly higher levels of integrin alpha1, alpha5 and beta1 subunits, and myosin II B. In in vivo experiments, the aligned nanofibers induced the formation of spindle-shaped cells and tendon-like tissue. In conclusion, the aligned electrospun nanofiber structure provides an instructive microenvironment for hTSPC differentiation and may lead to the development of desirable engineered tendons. Copyright (c) 2009 Elsevier Ltd. All rights reserved.
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                Author and article information

                Journal
                RSCACL
                RSC Advances
                RSC Adv.
                Royal Society of Chemistry (RSC)
                2046-2069
                2015
                2015
                : 5
                : 46
                : 36175-36184
                Affiliations
                [1 ]Department of Orthopedics Surgery
                [2 ]Shanghai Jiaotong University Affiliated Sixth People's Hospital
                [3 ]Shanghai 200233
                [4 ]China
                [5 ]The Education Ministry Key Lab of Resource Chemistry
                [6 ]Shanghai Key Laboratory of Rare Earth Functional Materials
                [7 ]Shanghai Normal University
                [8 ]Shanghai 200234
                [9 ]P. R. China
                Article
                10.1039/C5RA01487E
                4b8b1d99-94ea-4c94-b225-e2ea09761c40
                © 2015
                History

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