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Stepwise morphological changes and cytoskeletal reorganization of human mesenchymal stem cells treated by short-time cyclic uniaxial stretch

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Abstract

This study aimed to investigate stepwise remodeling of human mesenchymal stem cells (hMSCs) in response to cyclic stretch through rearrangement and alignment of cells and cytoskeleton regulation toward smooth muscle cell (SMC) fate in different time spans. Image analysis techniques were utilized to calculate morphological parameters. Cytoskeletal reorganization was observed by investigating F-actin filaments using immunofluorescence staining, and expression level of contractile SMC markers was followed by a quantitative polymerase chain reaction method. Applying cyclic uniaxial stretch on cultured hMSCs, utilizing a costume-made device, led to alteration in fractal dimension (FD) and cytoskeleton structure toward continuous alignment and elongation of cells by elevation of strain duration. Actin filaments became more aligned perpendicular to the axis of mechanical stretch by increasing uniaxial loading duration. At first, FD met a significant decrease in 4 h loading duration then increased significantly by further loading up to 16 h, followed by another decrease up to 1 d of uniaxial stretching. HMSCs subjected to 24 h cyclic uniaxial stretching significantly expressed early and intermediate contractile SM markers. It was hypothesized that the increase in FD after 4 h while cells continuously became more aligned and elongated was due to initiation of change in phenotype that influenced arrangement of cells. At this point, change in cell phenotype started leading to change in morphology while mechanical loading still caused cell alignment and rearrangement. Results can be helpful when optimized engineered cells are needed based on mechanical condition for functional engineered tissue and cell therapy.

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References

  • Alimperti S, You H, George T, Agarwal SK, Andreadis ST (2014) Cadherin-11 regulates both mesenchymal stem cell differentiation into smooth muscle cells and the development of contractile function in vivo. J Cell Sci 127(Pt 12):2627–2638

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bono N, Pezzoli D, Levesque L, Loy C, Candiani G, Fiore GB, Mantovani D (2016) Unraveling the role of mechanical stimulation on smooth muscle cells: a comparative study between 2D and 3D models. Biotechnol Bioeng 113(10):2254–2263

    Article  CAS  PubMed  Google Scholar 

  • Catacchio I, Berardi S, Reale A, De Luisi A, Racanelli V, Vacca A, Ria R (2013) Evidence for bone marrow adult stem cell plasticity: properties, molecular mechanisms, negative aspects, and clinical applications of hematopoietic and mesenchymal stem cells Transdifferentiation. Stem Cells Int 2013:589139

    Article  PubMed  PubMed Central  Google Scholar 

  • Childs PG, Boyle CA, Pemberton GD, Nikukar H, Curtis AS, Henriquez FL, Dalby MJ, Reid S (2016) Use of nanoscale mechanical stimulation for control and manipulation of cell behaviour. Acta Biomater 34:159–168

    Article  PubMed  Google Scholar 

  • Colombo A, Guha S, Mackle JN, Cahill PA, Lally C (2013) Cyclic strain amplitude dictates the growth response of vascular smooth muscle cells in vitro: role in in-stent restenosis and inhibition with a sirolimus drug-eluting stent. Biomech Model Mechanobiol 12(4):671–683

    Article  PubMed  Google Scholar 

  • Dan P, Velot E, Decot V, Menu P (2015) The role of mechanical stimuli in the vascular differentiation of mesenchymal stem cells. J Cell Sci 128(14):2415–2422

    Article  CAS  PubMed  Google Scholar 

  • Ghazanfari S, Tafazzoli-Shadpour M, Ali Shokrgozar MA (2009) Effects of cyclic stretch on proliferation of mesenchymal stem cells and their differentiation to smooth muscle cells. Biochem Biophys Res Commun 388(3):601–605

    Article  CAS  PubMed  Google Scholar 

  • Greiner AM, Chen H, Spatz JP, Kemkemer R (2013) Cyclic tensile strain controls cell shape and directs actin stress fiber formation and focal adhesion alignment in spreading cells. PLoS One 8(10):e77328

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guilak F, Butler DL, Goldstein SA, Baaijens FP (2014) Biomechanics and mechanobiology in functional tissue engineering. J Biomech 47(9):1933–1940

    Article  PubMed  PubMed Central  Google Scholar 

  • Khani MM, Tafazzoli-Shadpour M, Rostami M, Peirovi H, Janmaleki M (2014) Evaluation of mechanical properties of human mesenchymal stem cells during differentiation to smooth muscle cells. Ann Biomed Eng 42(7):1373–1380

    Article  PubMed  Google Scholar 

  • Khani MM, Tafazzoli-Shadpour M, Goli-Malekabadi Z, Haghighipour N (2015) Mechanical characterization of human mesenchymal stem cells subjected to cyclic uniaxial strain and TGF-β1. J Mech Behav Biomed Mater 43:18–25

    Article  CAS  PubMed  Google Scholar 

  • Kurpinski K, Park G, Thakar RG, Li S (2006) Regulation of vascular smooth muscle cells and mesenchymal stem cells by mechanical strain. Mol Cell Biomech 3(1):21–34

    PubMed  Google Scholar 

  • Lei Z, Kahn CJ, Chen HQ, Tran N, Wang X (2008) Effect of uniaxial stretching on rat bone mesenchymal stem cell: orientation and expressions of collagen types I and III and tenascin-C. Cell Biol Int 32(3):344–352

    Article  Google Scholar 

  • Li SC, Wang L, Jiang H, Acevedo J, Chang AC, Loudon WG (2009) Stem cell engineering for treatment of heart diseases: potentials and challenges. Cell Biol Int 33(3):255–267

    Article  CAS  PubMed  Google Scholar 

  • Liu Q, Cheng G, Wang Z, Zhan S, Xiong B, Zhao X (2015) Bone marrow-derived mesenchymal stem cells differentiate into nerve-like cells in vitro after transfection with brain-derived neurotrophic factor gene. In Vitro Cell Dev Biol Anim 51(3):319–327

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mammoto T, Mammoto A, Ingber DE (2013) Mechanobiology and developmental control. Annu Rev Cell Dev Biol 29:27–61

    Article  CAS  PubMed  Google Scholar 

  • McGarry JP, Murphy BP, McHugh PE (2005) Computational mechanics modelling of cell–substrate contact during cyclic substrate deformation. J Mech Phys Solids 53(12):2597–2637

    Article  Google Scholar 

  • Nekouzadeh A, Pryse KM, Elson EL, Genin GM (2008) Stretch-activated force shedding, force recovery, and cytoskeletal remodeling in contractile fibroblasts. J Biomech 41(14):2964–2971

    Article  PubMed  PubMed Central  Google Scholar 

  • Obbink-Huizer C, Oomens CW, Loerakker S, Foolen J, Bouten CV, Baaijens FP (2014) Computational model predicts cell orientation in response to a range of mechanical stimuli. Biomech Model Mechanobio 13(1):227–236

    Article  Google Scholar 

  • Owatverot TB, Oswald SJ, Chen Y, Wille JJ, Yin FC (2005) Effect of combined cyclic stretch and fluid shear stress on endothelial cell morphological responses. J Biomech Eng 127(3):374–382

    Article  PubMed  Google Scholar 

  • Park JS, Chu JS, Cheng C, Chen F, Chen D, Li S (2004) Differential effects of equiaxial and uniaxial strain on mesenchymal stem cells. Biotechnol Bioeng 88(3):359–368

    Article  CAS  PubMed  Google Scholar 

  • Sanina C, Joshua MH (2015) Mesenchymal stem cells as a biological drug for heart disease: where are we with cardiac cell-based therapy? Circ Res 117(3):229–233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sawyer AA, Hennessy KM, Bellis SL (2005) Regulation of mesenchymal stem cell attachment and spreading on hydroxyapatite by RGD peptides and adsorbed serum proteins. Biomaterials 26(13):1467–1475

    Article  CAS  PubMed  Google Scholar 

  • Schürmann S, Wagner S, Herlitze S, Fischer C, Gumbrecht S, Wirth-Hücking A, Prölß G, Lautscham LA, Fabry B, Goldmann WH, Nikolova-Krstevski V, Martinac B, Friedrich O (2016) The IsoStretcher: an isotropic cell stretch device to study mechanical biosensor pathways in living cells. Biosens Bioelectron 81:363–372

    Article  PubMed  Google Scholar 

  • Simmons CA, Matlis S, Thornton AJ, Chen S, Wang CY, Mooney DJ (2003) Cyclic strain enhances matrix mineralization by adult human mesenchymal stem cells via the extracellular signal-regulated kinase (ERK1/2) signaling pathway. J Biomech 36(8):1087–1096

    Article  PubMed  Google Scholar 

  • Smith TG Jr, Lange GD, Marks WB (1996) Fractal methods and results in cellular morphology--dimensions, lacunarity and multifractals. J Neurosci Methods 69(2):123–136

    Article  PubMed  Google Scholar 

  • Sonowal H, Kumar A, Bhattacharyya J, Gogoi PK, Jaganathan BG (2013) Inhibition of actin polymerization decreases osteogeneic differentiation of mesenchymal stem cells through p38 MAPK pathway. J Biomed Sci 20(1):71

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sun K, Liu F, Wang J, Guo Z, Ji Z, Yao M (2016a) The effect of mechanical stretch stress on the differentiation and apoptosis of human growth plate chondrocytes. In Vitro Cell Dev Biol Anim. doi:10.1007/s11626-016-0090-5

  • Sun K, Liu F, Wang J, Guo Z, Ji Z, Yao M (2016b) The effect of mechanical stretch stress on the differentiation and apoptosis of human growth plate chondrocytes. In Vitro Cell Dev Biol Anim . doi:10.1007/s11626-016-0090-5September

    Google Scholar 

  • Tréguer K, Naye F, Thiébaud P, Fédou S, Soulet F, Thézé N, Faucheux C (2009) Smooth muscle cell differentiation from human bone marrow: variations in cell type specific markers and id gene expression in a new model of cell culture. Cell Biol Int 33(6):621–631

    Article  PubMed  Google Scholar 

  • Ugolini GS, Rasponi M, Pavesi A, Santoro R, Kamm R, Fiore GB, Pesce M, Soncini M (2016) On-chip assessment of human primary cardiac fibroblasts proliferative responses to uniaxial cyclic mechanical strain. Biotechnol Bioeng 113(4):859–869

    Article  CAS  PubMed  Google Scholar 

  • Wang JH, Goldschmidt-Clermont P, Wille J, Yin FC (2001) Specificity of endothelial cell reorientation in response to cyclic mechanical stretching. J Biomech 34(12):1563–1572

    Article  CAS  PubMed  Google Scholar 

  • Yao R, Wong JY (2015) The effects of mechanical stimulation on controlling and maintaining marrow stromal cell differentiation into vascular smooth muscle cells. J Biomech Eng 137(2):020907

    Article  PubMed  Google Scholar 

  • Yourek G, Hussain MA, Mao JJ (2007) Cytoskeletal changes of mesenchymal stem cells during differentiation. ASAIO J 53(2):219–228

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhang L, Cyril K, Huai-Qing C, Nguyen T, Xiong W (2008) Effect of uniaxial stretching on rat bone mesenchymal stem cell: rientation and expressions of collagen types I and III and tenascin-C. Cell Biol Int 32 (3): 344–52. doi:10.1016/j.cellbi.2007.12.018

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Acknowledgements

This study was supported by Regenerative Medicine and Stem Cell Research Network of Shahid Beheshti University of Medical Sciences with grant number 7691.

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Correspondence to Mohammad Tafazzoli-Shadpour or Mohammad-Mehdi Khani.

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Editor: Tetsuji Okamoto

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Parandakh, A., Tafazzoli-Shadpour, M. & Khani, MM. Stepwise morphological changes and cytoskeletal reorganization of human mesenchymal stem cells treated by short-time cyclic uniaxial stretch. In Vitro Cell.Dev.Biol.-Animal 53, 547–553 (2017). https://doi.org/10.1007/s11626-017-0131-8

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  • DOI: https://doi.org/10.1007/s11626-017-0131-8

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