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      Bone Tissue Engineering with Adipose-Derived Stem Cells in Bioactive Composites of Laser-Sintered Porous Polycaprolactone Scaffolds and Platelet-Rich Plasma

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          Abstract

          Three-dimensional porous polycaprolactone (PCL) scaffolds with consistent inter-pore channels, 83% porosity and 300–400 μm pore size were fabricated via selective laser sintering. The PCL scaffold was combined with platelet-rich plasma (PRP) to form a bioactive composite and studied for potential application in bone tissue engineering using porcine adipose-derived stem cells (PASCs). The PCL/PRP/PASCs construct showed enhanced cell seeding efficiency and synergistically increased the differentiation capability of PASCs in osteogenic medium toward the osteoblast lineage, judging from elevated alkaline phosphatase activity and up-regulated osteogenic genes expression. For in vivo study, a 3 cm × 3 cm mandible defect was created in pigs and reconstructed by implanting acellular PCL scaffolds or PCL/PRP/PASCs constructs. Both groups showed new bone formation, however, the new bone volume was 5.1 times higher for PCL/PRP/PASCs 6 months post-operation. The bone density was less and loose in the acellular PCL group and the Young’s modulus was only 29% of normal bone. In contrast, continued and compact bone formation was found in PCL/PRP/PASCs and the Young’s modulus was 81% that of normal bone. Masson’s trichrome stain, immunohistochemical analysis of osteocalcin and collagen type I also confirmed new bone formation.

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          Most cited references28

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          Platelet-rich plasma: evidence to support its use.

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            Platelet-rich plasma: Growth factor enhancement for bone grafts.

            Platelet-rich plasma is an autologous source of platelet-derived growth factor and transforming growth factor beta that is obtained by sequestering and concentrating platelets by gradient density centrifugation. This technique produced a concentration of human platelets of 338% and identified platelet-derived growth factor and transforming growth factor beta within them. Monoclonal antibody assessment of cancellous cellular marrow grafts demonstrated cells that were capable of responding to the growth factors by bearing cell membrane receptors. The additional amounts of these growth factors obtained by adding platelet-rich plasma to grafts evidenced a radiographic maturation rate 1.62 to 2.16 times that of grafts without platelet-rich plasma. As assessed by histomorphometry, there was also a greater bone density in grafts in which platelet-rich plasma was added (74.0% +/- 11%) than in grafts in which platelet-rich plasma was not added (55.1% +/- 8%; p = 0.005).
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              Bone tissue engineering using polycaprolactone scaffolds fabricated via selective laser sintering.

              Polycaprolactone (PCL) is a bioresorbable polymer with potential applications for bone and cartilage repair. In this work, porous PCL scaffolds were computationally designed and then fabricated via selective laser sintering (SLS), a rapid prototyping technique. The microstructure and mechanical properties of the fabricated scaffolds were assessed and compared to the designed porous architectures and computationally predicted properties. Scaffolds were then seeded with bone morphogenetic protein-7 (BMP-7) transduced fibroblasts and implanted subcutaneously to evaluate biological properties and to demonstrate tissue in-growth. The work done illustrates the ability to design and fabricate PCL scaffolds with porous architecture that have sufficient mechanical properties for bone tissue engineering applications using SLS. Compressive modulus and yield strength values ranged from 52 to 67 MPa and 2.0 to 3.2 Mpa, respectively, lying within the lower range of properties reported for human trabecular bone. Finite element analysis (FEA) results showed that mechanical properties of scaffold designs and of fabricated scaffolds can be computationally predicted. Histological evaluation and micro-computed tomography (microCT) analysis of implanted scaffolds showed that bone can be generated in vivo. Finally, to demonstrate the clinical application of this technology, we designed and fabricated a prototype mandibular condyle scaffold based on an actual pig condyle. The integration of scaffold computational design and free-form fabrication techniques presented here could prove highly useful for the construction of scaffolds that have anatomy specific exterior architecture derived from patient CT or MRI data and an interior porous architecture derived from computational design optimization.
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                Author and article information

                Journal
                Materials (Basel)
                Materials (Basel)
                materials
                Materials
                MDPI
                1996-1944
                25 October 2013
                November 2013
                : 6
                : 11
                : 4911-4929
                Affiliations
                [1 ]Department of Chemical and Materials Engineering, Chang Gung University, Kweishan, Taoyuan 333, Taiwan; E-Mail: lia01211@ 123456gmail.com
                [2 ]Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital, Craniofacial Research Center, College of Medicine, Chang Gung University, Kweishan, Taoyuan 333, Taiwan
                [3 ]Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Kweishan, Taoyuan 333, Taiwan
                [4 ]Department of Mechanical Engineering, Chang Gung University, Kweishan, Taoyuan 333, Taiwan; E-Mail: leemiy@ 123456mail.cgu.edu.tw
                Author notes
                [* ]Author to whom correspondence should be addressed; E-Mail: jpchen@ 123456mail.cgu.edu.tw ; Tel.: +886-3-211-8800; Fax: +886-3-211-8668.
                Article
                materials-06-04911
                10.3390/ma6114911
                5452768
                a17cb800-62fc-4cdf-a4b9-0d210e70594f
                © 2013 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/3.0/).

                History
                : 07 August 2013
                : 09 October 2013
                : 18 October 2013
                Categories
                Article

                bone tissue engineering,selective laser sintering,polycaprolactone,platelet-rich plasma,adipose-derived stem cells

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