Mechanical and failure properties of extracellular matrix sheets as a function of structural protein composition.

The goal of this study was to determine how alterations in protein composition of the extracellular matrix (ECM) affect its functional properties. To achieve this, we investigated the changes in the mechanical and failure properties of ECM sheets generated by neonatal rat aortic smooth muscle cells engineered to contain varying amounts of collagen and elastin. Samples underwent static and dynamic mechanical measurements before, during, and after 30 min of elastase digestion followed by a failure test. Microscopic imaging was used to measure thickness at two strain levels to estimate the true stress and moduli in the ECM sheets. We found that adding collagen to the ECM increased the stiffness. However, further increasing collagen content altered matrix organization with a subsequent decrease in the failure strain. We also introduced collagen-related percolation in a nonlinear elastic network model to interpret these results. Additionally, linear elastic moduli correlated with failure stress which may allow the in vivo estimation of the stress tolerance of ECM. We conclude that, in engineered replacement tissues, there is a tradeoff between improved mechanical properties and decreased extensibility, which can impact their effectiveness and how well they match the mechanical properties of native tissue.

[1]  J. Paul Robinson,et al.  Tensile mechanical properties of three-dimensional type I collagen extracellular matrices with varied microstructure. , 2002, Journal of biomechanical engineering.

[2]  K. Burridge,et al.  Rho-mediated Contractility Exposes a Cryptic Site in Fibronectin and Induces Fibronectin Matrix Assembly , 1998, The Journal of cell biology.

[3]  P. Stone,et al.  Repair of elastase-digested elastic fibers in acellular matrices by replating with neonatal rat-lung lipid interstitial fibroblasts or other elastogenic cell types. , 1997, American journal of respiratory cell and molecular biology.

[4]  Ann E Rundell,et al.  Biaxial strength of multilaminated extracellular matrix scaffolds. , 2004, Biomaterials.

[5]  E. Dempsey,et al.  The structure and chemical characterization of elastic fibers as reveled by elastase and by electron microscopy , 1952, The Anatomical record.

[6]  B Suki,et al.  Roles of mechanical forces and collagen failure in the development of elastase-induced emphysema. , 2001, American journal of respiratory and critical care medicine.

[7]  M. Nugent,et al.  Elastase-mediated Release of Heparan Sulfate Proteoglycans from Pulmonary Fibroblast Cultures , 1999, The Journal of Biological Chemistry.

[8]  Elliot L Elson,et al.  The relationship between cell and tissue strain in three-dimensional bio-artificial tissues. , 2005, Biophysical journal.

[9]  H. Rosenfeldt,et al.  Fibroblast Quiescence and the Disruption of ERK Signaling in Mechanically Unloaded Collagen Matrices* , 2000, The Journal of Biological Chemistry.

[10]  S. Shapiro,et al.  Proteinases in chronic obstructive pulmonary disease. , 2001, Biochemical Society transactions.

[11]  J. Fredberg,et al.  Force heterogeneity in a two-dimensional network model of lung tissue elasticity. , 1998, Journal of applied physiology.

[12]  K. Lutchen,et al.  Pseudorandom signals to estimate apparent transfer and coherence functions of nonlinear systems: applications to respiratory mechanics , 1992, IEEE Transactions on Biomedical Engineering.

[13]  K. Kokini,et al.  Biomechanics and Mechanotransduction in Cells and Tissues Extracellular matrix ( ECM ) microstructural composition regulates local cell-ECM biomechanics and fundamental fibroblast behavior : a multidimensional perspective , 2005 .

[14]  C. Franzblau,et al.  Elastin in a neonatal rat smooth muscle cell culture has greatly decreased susceptibility to proteolysis by human neutrophil elastase. An in vitro model of elastolytic injury , 1987, In Vitro Cellular & Developmental Biology.

[15]  D. Stamenović,et al.  Dynamic moduli of rabbit lung tissue and pigeon ligamentum propatagiale undergoing uniaxial cyclic loading. , 1994, Journal of applied physiology.

[16]  C. Schmidt,et al.  Acellular vascular tissues: natural biomaterials for tissue repair and tissue engineering. , 2000, Biomaterials.

[17]  Arnab Majumdar,et al.  Mechanical interactions between collagen and proteoglycans: implications for the stability of lung tissue. , 2005, Journal of applied physiology.

[18]  C. S. Chen,et al.  Geometric control of cell life and death. , 1997, Science.

[19]  G. Genin,et al.  Cellular and Matrix Contributions to Tissue Construct Stiffness Increase with Cellular Concentration , 2006, Annals of Biomedical Engineering.

[20]  Arbabi,et al.  Elastic properties of three-dimensional percolation networks with stretching and bond-bending forces. , 1988, Physical review. B, Condensed matter.

[21]  D. Bader,et al.  The influence of swelling and matrix degradation on the microstructural integrity of tendon. , 2006, Acta biomaterialia.

[22]  C. Franzblau,et al.  Isolation of hydroxylysyl pyridinoline, a mature collagen crosslink from neonatal rat aorta smooth muscle cell cultures. , 1992, Matrix.

[23]  A Giudiceandrea,et al.  The Mechanical Behavior of Vascular Grafts: A Review , 2001, Journal of biomaterials applications.

[24]  K J Halbhuber,et al.  Impact of decellularization of xenogeneic tissue on extracellular matrix integrity for tissue engineering of heart valves. , 2003, Journal of structural biology.

[25]  Y. Fung,et al.  Biomechanics: Mechanical Properties of Living Tissues , 1981 .

[26]  J. Fredberg,et al.  Input impedance and peripheral inhomogeneity of dog lungs. , 1992, Journal of applied physiology.

[27]  Yoshiki Sawa,et al.  Novel method of preparing acellular cardiovascular grafts by decellularization with poly(ethylene glycol). , 2003, Journal of biomedical materials research. Part A.

[28]  Chrysanthi Williams,et al.  Cell sourcing and culture conditions for fibrin-based valve constructs. , 2006, Tissue engineering.

[29]  J. Bates,et al.  A distributed nonlinear model of lung tissue elasticity. , 1997, Journal of applied physiology.

[30]  Elliot L Elson,et al.  Thin bio-artificial tissues in plane stress: the relationship between cell and tissue strain, and an improved constitutive model. , 2005, Biophysical journal.

[31]  Yu-Ting Tsai,et al.  Process development of an acellular dermal matrix (ADM) for biomedical applications. , 2004, Biomaterials.

[32]  Sangeeta N Bhatia,et al.  Assessing porcine liver-derived biomatrix for hepatic tissue engineering. , 2004, Tissue engineering.

[33]  S. Redner,et al.  Introduction To Percolation Theory , 2018 .

[34]  Qijin Lu,et al.  Novel porous aortic elastin and collagen scaffolds for tissue engineering. , 2004, Biomaterials.

[35]  D. Ingber,et al.  Altering the cellular mechanical force balance results in integrated changes in cell, cytoskeletal and nuclear shape. , 1992, Journal of cell science.

[36]  P. Bell,et al.  The association between venous structural alterations and biomechanical weakness in patients with abdominal aortic aneurysms. , 2002, Journal of vascular surgery.

[37]  A Haverich,et al.  Tissue engineering of heart valves--human endothelial cell seeding of detergent acellularized porcine valves. , 1998, European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery.

[38]  M. DeRuiter,et al.  Histological evaluation of decellularised porcine aortic valves: matrix changes due to different decellularisation methods. , 2005, European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery.

[39]  Arnab Majumdar,et al.  Lung and alveolar wall elastic and hysteretic behavior in rats: effects of in vivo elastase treatment. , 2003, Journal of applied physiology.

[40]  S. Badylak,et al.  The use of xenogeneic small intestinal submucosa as a biomaterial for Achilles tendon repair in a dog model. , 1995, Journal of biomedical materials research.

[41]  B. Suki,et al.  Effects of elastase on the mechanical and failure properties of engineered elastin-rich matrices. , 2005, Journal of applied physiology.

[42]  S. Chien,et al.  Role of integrins in cellular responses to mechanical stress and adhesion. , 1997, Current opinion in cell biology.

[43]  M. Sacks,et al.  Biaxial mechanical properties of muscle-derived cell seeded small intestinal submucosa for bladder wall reconstitution. , 2005, Biomaterials.

[44]  T. M. Parker,et al.  Elastin: a representative ideal protein elastomer. , 2002, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[45]  P. Stone,et al.  Immunocytochemical study of the degradation of elastic fibers in a living extracellular matrix. , 1995, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[46]  J. Scott,et al.  Tendon response to tensile stress: an ultrastructural investigation of collagen:proteoglycan interactions in stressed tendon. , 1995, Journal of anatomy.

[47]  R T Tranquillo,et al.  A fibrin-based arterial media equivalent. , 2003, Journal of biomedical materials research. Part A.

[48]  Stephen F Badylak,et al.  Production and characterization of ECM powder: implications for tissue engineering applications. , 2005, Biomaterials.

[49]  John Fisher,et al.  Development and characterisation of a full-thickness acellular porcine bladder matrix for tissue engineering. , 2007, Biomaterials.

[50]  Y. Wang,et al.  Cell locomotion and focal adhesions are regulated by substrate flexibility. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[51]  F H Silver,et al.  Viscoelasticity of the vessel wall: the role of collagen and elastic fibers. , 2001, Critical reviews in biomedical engineering.

[52]  H. C. Robinson,et al.  Cleavage of proteoglycan aggregate by leucocyte elastase. , 1992, Archives of biochemistry and biophysics.

[53]  F. Grinnell,et al.  Microtubule function in fibroblast spreading is modulated according to the tension state of cell–matrix interactions , 2007, Proceedings of the National Academy of Sciences.