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      Plasma-Derived Fibronectin Stimulates Chondrogenic Differentiation of Human Subchondral Cortico-Spongious Progenitor Cells in Late-Stage Osteoarthritis

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          Abstract

          Migration and chondrogenesis of human subchondral cortico-spongious progenitor cells (SPCs) are the key steps in the repair of microfracture-induced articular cartilage defects. The aim of this study was to evaluate the effect of human plasma-derived fibronectin (Fn) on the chondrogenic differentiation of SPCs, which was isolated from subchondrol cortico-spongious bone of late-stage osteoarthritis (OA) patients. SPCs were isolated and cultured for three passages. Stem cell surface antigens of SPCs were analyzed by flow cytometry. The osteogenic, chondrogenic and adipogenic differentiation potential were detected by histological staining. The chondrogenesis potential of SPCs with or without stimulation of either Fn or BMP-2 were studied by immunochemical staining and gene expression analysis. Cells isolated from subchondral bone presented to be positive for CD44, CD73, CD90, and CD166, and showed high capacity of osteogenic, adipogenic and chondrogenic differentiation, which suggested this cell population to be MSC-like cells. Stimulating with Fn increased the expression of SOX-9, aggrecan, collagen II while decreased the formation of collagen I by immunochemical staining. Gene expression analysis showed similar results. These results suggest that plasma-derived Fn can increase the chondrogenic differentiation of SPCs isolated from late-stage OA and improve cartilage repair after microfracture.

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          Most cited references21

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          The regulation of differentiation in mesenchymal stem cells.

          Mesenchymal stromal/stem cells (MSCs) are a population of stromal cells present in the bone marrow and most connective tissues, capable of differentiation into mesenchymal tissues such as bone and cartilage. MSCs are attractive candidates for biological cell-based tissue repair approaches because of their extensive proliferative ability in culture while retaining their mesenchymal multilineage differentiation potential. In addition to its undoubted scientific interest, the prospect of monitoring and controlling MSC differentiation is a crucial regulatory and clinical requirement. Hence, the molecular regulation of MSC differentiation has been extensively studied. Most of the studies are in vitro, because the identity of MSCs in their tissues of origin in vivo remains undefined. This review addresses the current knowledge of the molecular basis of differentiation of cultured MSCs, with a particular focus on chondrogenesis and osteogenesis. Building on the information coming from developmental biology studies of embryonic skeletogenesis, several signaling pathways and transcription factors have been investigated and shown to play critical roles in MSC differentiation. In particular, the Wnt and transforming growth factor-β/bone morphogenetic protein signaling pathways are well known to modulate in MSCs the molecular differentiation into cartilage and bone. Relevant to the emerging concept of stem cell niches is the demonstration that physical factors can also participate in the regulation of MSC differentiation. Knowledge of the regulation of MSC differentiation will be critical in the design of three-dimensional culture systems and bioreactors for automated bioprocessing through mathematical models applied to systems biology and network science.
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            Osteochondral lesions of the knee: a new one-step repair technique with bone-marrow-derived cells.

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              Cartilage healing after microfracture in osteoarthritic knees.

              To evaluate the clinical and radiologic results, second-look arthroscopic findings, histologic evaluation, and results of immunohistochemical staining and the Western blotting test for type II collagen after microfracture for full-thickness chondral defects in patients with osteoarthritic knee. Between October 1997 and December 1998, 49 knees in 46 patients who had moderate osteoarthritic changes underwent microfracture; 44 patients (47 cases) had second-look arthroscopy and biopsy performed 1 year after surgery. The clinical outcomes were assessed by use of Baumgaertner's 9-point scale and joint space changes were measured radiographically; 18 knees underwent immunohistochemical study and 21 knees underwent Western blotting tests to identify formation of type II collagen. Significant improvements were noted for the parameters of daily living activity and pain (P < .05). The joint spaces were widened by 1.06 mm on standing anteroposterior and by 1.37 mm on standing lateral radiographs, which showed statistical significance (P < .05). Individual defects were filled with white tissue resembling cartilage on second-look arthroscopy, and the cartilage healing was found by histologic evaluation. Type II collagen formation was identified qualitatively by immunohistochemical staining. Quantitative collagen formation by Western blotting showed 44% growth compared with the normal control. This healed tissue is a combination, or hybrid, of fibrocartilage and hyaline-like cartilage, and it is shown to contain type II collagen on immunohistochemical staining and the Western blotting test. Patients with full-thickness chondral defects in the osteoarthritic knee can have improved function and see an increase in joint space after microfracture. Cartilaginous tissue containing type II collagen is formed after the microfracture procedure in the osteoarthritic knee. Level IV, retrospective case series.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                18 August 2015
                August 2015
                : 16
                : 8
                : 19477-19489
                Affiliations
                Department of Orthopedic Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China; E-Mails: jiangchao2189@ 123456sina.com (C.J.); dr_mapei@ 123456163.com (P.M.); mbp350579121@ 123456163.com (B.M.); wuzhihong139@ 123456139.com (Z.W.); qiugx@ 123456medmail.com.cn (G.Q.); suxinlin1988@ 123456163.com (X.S.); tienan523523@ 123456163.com (Z.X.); yezx06@ 123456mails.tsinghua.edu.cn (Z.Y.)
                Author notes
                [†]

                These authors contributed equally to this work.

                [* ]Author to whom correspondence should be addressed; E-Mail: wangyipengjc@ 123456sina.com ; Tel./Fax: +86-10-6915-2809.
                Article
                ijms-16-19477
                10.3390/ijms160819477
                4581308
                26295224
                7c3b17b7-730f-4abd-9a81-8b0204a91dfc
                © 2015 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/4.0/).

                History
                : 27 June 2015
                : 07 August 2015
                Categories
                Article

                Molecular biology
                plasma-derived fibronectin,cartilage regeneration,chondrogenesis,subchondral progenitor cells

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