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      3D uniaxial mechanical stimulation induces tenogenic differentiation of tendon‐derived stem cells through a PI3K/AKT signaling pathway

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          Most cited references 42

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            Mechanical regulation of signaling pathways in bone.

            A wide range of cell types depend on mechanically induced signals to enable appropriate physiological responses. The skeleton is particularly dependent on mechanical information to guide the resident cell population towards adaptation, maintenance and repair. Research at the organ, tissue, cell and molecular levels has improved our understanding of how the skeleton can recognize the functional environment, and how these challenges are translated into cellular information that can site-specifically alter phenotype. This review first considers those cells within the skeleton that are responsive to mechanical signals, including osteoblasts, osteoclasts, osteocytes and osteoprogenitors. This is discussed in light of a range of experimental approaches that can vary parameters such as strain, fluid shear stress, and pressure. The identity of mechanoreceptor candidates is approached, with consideration of integrins, pericellular tethers, focal adhesions, ion channels, cadherins, connexins, and the plasma membrane including caveolar and non-caveolar lipid rafts and their influence on integral signaling protein interactions. Several mechanically regulated intracellular signaling cascades are detailed including activation of kinases (Akt, MAPK, FAK), β-catenin, GTPases, and calcium signaling events. While the interaction of bone cells with their mechanical environment is complex, an understanding of mechanical regulation of bone signaling is crucial to understanding bone physiology, the etiology of diseases such as osteoporosis, and to the development of interventions to improve bone strength. Copyright © 2012 Elsevier B.V. All rights reserved.
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              Mechanoactive tenogenic differentiation of human mesenchymal stem cells.

              A mesenchymal stem cell (MSC)-seeded collagen gel under static or dynamic tension is a well-established model to study the potential of MSCs in regenerating a tendon- or ligament-like tissue. Using this model, upregulation of fibrillar collagen mRNA expression and protein production has been demonstrated in response to cyclic tensile mechanical stimulation. However, the mechanisms driving MSC tenogenesis (differentiation into tendon or ligament fibroblasts) have not been elucidated. This study investigated the mechanisms of tenogenesis of human bone marrow-derived MSCs in a dynamic, three-dimensional (3D) tissue-engineering model by investigating the effects of cyclic stretching on matrix production and gene expression of candidate tendon and ligament markers. The 3D MSC tenogenesis culture system upregulated scleraxis, but cyclic stretching was required to maintain expression of this putative tendon marker over time. Enhanced tendinous neo-tissue development demonstrated with extracellular matrix staining was largely due to changes in matrix deposition and remodeling activity under dynamic loading conditions, as evidenced by differential regulation of matrix metalloproteinases at a transcriptional level with minimal changes in collagen mRNA levels. Regulation of Wnt gene expression with cyclic stimulation suggested a similar role for Wnt4 versus Wnt5a in tenogenesis as in cartilage development. This first report of the potential involvement of matrix remodeling and Wnt signaling during tenogenesis of human MSCs in a dynamic, 3D tissue-engineering model provides insights into the mechanisms of tenogenesis in a mechanoactive environment and supports the therapeutic potential of adult stem cells.
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                Author and article information

                Journal
                The FASEB Journal
                FASEB j.
                Wiley
                0892-6638
                1530-6860
                August 20 2018
                September 2018
                March 29 2018
                September 2018
                : 32
                : 9
                : 4804-4814
                Affiliations
                [1 ]Division of Orthopaedic SurgeryDepartment of SurgeryGuangdong General HospitalGuangdong Academy of Medical SciencesGuangzhouChina
                [2 ]Centre for Orthopaedic Translational ResearchSchool of Biomedical SciencesUniversity of Western AustraliaNedlandsWestern AustraliaAustralia
                [3 ]Department of OrthopaedicsThe General Hospital of Chinese People's Liberation ArmyBeijingChina
                [4 ]Sir Charles Gairdner HospitalPerthWestern AustraliaAustralia
                [5 ]Department of Sports Medicine and Adult ReconstructionDrum Tower HospitalMedical School of Nanjing UniversityNanjingChina
                [6 ]Department of Orthopaedic SurgeryCenter for Cellular and Molecular EngineeringUniversity of Pittsburgh School of MedicinePittsburghPennsylvaniaUSA
                [7 ]Institute for Tissue Engineering and Regenerative MedicineThe Chinese University of Hong KongHong KongChina
                Article
                10.1096/fj.201701384R
                © 2018

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