Rheologica Acta

, Volume 58, Issue 8, pp 421–437 | Cite as

Characterizing the dynamic rheology in the pericellular region by human mesenchymal stem cell re-engineering in PEG-peptide hydrogel scaffolds

  • Maryam Daviran
  • Kelly M. SchultzEmail author
Original Contribution


During wound healing, human mesenchymal stem cells (hMSCs) migrate to injuries to regulate inflammation and coordinate tissue regeneration. To enable migration, hMSCs re-engineer the extracellular matrix rheology. Our work determines the correlation between cell-engineered rheology and motility. We encapsulate hMSCs in a cell-degradable peptide-polymeric hydrogel and characterize the change in rheological properties in the pericellular region using multiple particle tracking microrheology. Previous studies determined that pericellular rheology is correlated with motility. Additionally, hMSCs re-engineer their microenvironment by regulating cell-secreted enzyme, matrix metalloproteinases (MMPs), activity by also secreting their inhibitors, tissue inhibitors of metalloproteinases (TIMPs). We independently inhibit TIMPs and measure two different degradation profiles, reaction-diffusion and reverse reaction-diffusion. These profiles are correlated with cell spreading, speed and motility type. We model scaffold degradation using Michaelis-Menten kinetics, finding a decrease in kinetics between joint and independent TIMP inhibition. hMSCs ability to regulate microenvironmental remodeling and motility could be exploited in design of new materials that deliver hMSCs to wounds to enhance healing.


Multiple particle tracking microrheology Cellular degradation Polymeric hydrogel scaffold Michaelis-Menten kinetics Matrix metalloproteinases Tissue inhibitor of metalloproteinases 



We thank Dr. Susan Perry from the Department of Bioengineering at Lehigh University for her useful discussion on Western blot experiments. Research reported in this publication was supported by the National Institute of General Medical Sciences of the National Institutes of Health under award number R15GM119065. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Supplementary material

397_2019_1142_MOESM1_ESM.pdf (8.6 mb)
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  1. abcam (2018) buffer and stock solutions for western blot. https://www.abcamcom/protocols/buffer-and-stock-solutions-for-western-blot
  2. Adolf D, Martin JE (1990) Time-cure superposition during crosslinking. Macromolecules 23:3700–3704CrossRefGoogle Scholar
  3. Aimetti AA, Machen AJ, Anseth KS (2009) Poly(ethylene glycol) hydrogels formed by thiol-ene photopolymerization for enzyme-responsive protein delivery. Biomaterials 30:6048–6054CrossRefGoogle Scholar
  4. Anderson SB, Lin CC, Kuntzler DV, Anseth KS (2011) The performance of human mesenchymal stem cells encapsulated in cell-degradable polymer-peptide hydrogels. Biomaterials 32:3564–3574CrossRefGoogle Scholar
  5. Bassi EJ, Candido de ALmeida D, Moraes-Vieira PMM, Camara NOS (2012) Exploring the role of soluble factors associated with immune regulatory properties of mesenchymal stem cells. Stem Cell Rev and Rep 8:329–342CrossRefGoogle Scholar
  6. Bear JE, Haugh JM (2014) Directed migration of mesenchymal cells: where signaling and the cytoskeleton meet. Curr Opin in Cell Biology 30:78–82CrossRefGoogle Scholar
  7. Benton JA, Fairbanks BD, Anseth KS (2009) Characterization of valvular interstitial cell function in three dimensional matrix metalloproteinase degradable PEG hydrogels. Biomaterials 30: 6593–6603CrossRefGoogle Scholar
  8. Bloom RJ, George JP, Celedon A, Sun SX, Wirtz D (2008) Mapping local matrix remodeling induced by a migrating tumor cell using three-dimensional multiple-particle tracking. Biophys J 95:4077–4088CrossRefGoogle Scholar
  9. Brew K, Nagase H (2010) The tissue inhibitors of metalloproteinases (TIMPs): an ancient family with structural and functional diversity. Biochim Biophys Acta 1(1803):55–71CrossRefGoogle Scholar
  10. Brew K, Dinakarpandian D, Nagase H (2000) Tissue inhibitors of metalloproteinases: evolution, structure and function. Biochemica et Biophysica Acta 1477:267–283CrossRefGoogle Scholar
  11. Bryant SJ, Anseth KS (2003) Controlling the spatial distribution of ECM components in degradable PEG hydrogels for tissue engineering cartilage. Biomed Mater Res 64A:70–79CrossRefGoogle Scholar
  12. Buxboim A, Ivanovska IL, Discher DE (2010) Matrix elasticity, cytoskeletal forces and physics of the nucleus: how deeply do cells ‘feel’ outside and in? Cell Science 123:297–308CrossRefGoogle Scholar
  13. Caplan AI (2009) Why are MSCs therapeutic? New data: new insight. J Pathol 217:318–324CrossRefGoogle Scholar
  14. Chambon F, Winter HH (1987) Linear viscoelasticity at the gel point of a crosslinking PDMS with imbalanced stoichiometry. J Rheol 31:683–697CrossRefGoogle Scholar
  15. Cheng Y, Prudhomme RK (2000) Enzymatic degradation of guar and substituted guar galactomannans. Biomacromolecules 1:782–788CrossRefGoogle Scholar
  16. Corrigan AM, Donald AM (2009) Passive microrheology of solvent-induced fibrillar protein networks. Langmuir 25:8599–8605CrossRefGoogle Scholar
  17. Cox TR, Erler JT (2009) Remodeling and homeostasis of the extracellular matrix: implications for fibrotic diseases and cancer. Disease Models and Mechanisms 4:165–178CrossRefGoogle Scholar
  18. Crocker JC, Grier DG (1996) Methods of digital video microscopy for colloidal studies. J Colloid Interface Sci 179:298–310CrossRefGoogle Scholar
  19. Crocker JC, Weeks ER (2011) Particle tracking using idl. http://www.physicsemoryedu/faculty/weeks//idl/
  20. Daviran M, Caram HS, Schultz KM (2018a) Role of cell-mediated enzymatic degradation and cytoskeletal tension on dynamic changes in the rheology of the pericellular region prior to human mesenchymal stem cell motility. ACS Biomater Sci Eng 4:468–472CrossRefGoogle Scholar
  21. Daviran M, Longwill SM, Casella JF, Schultz KM (2018b) Rheological characterization of dynamic remodeling of the pericellular region by human mesenchymal stem cell-secreted enzymes in well-defined synthetic hydrogel scaffolds. Soft Matter 14:3078–3089CrossRefGoogle Scholar
  22. Eggenhofer E, Luk F, Dahlke MH, Hoogduijn MJ (2014) The life and fate of mesenchymal stem cells. Front Immunol 5:148–1–148-6CrossRefGoogle Scholar
  23. Engler AJ, Sen S, Lee Sweeney H, Discher DE (2006) Matrix elasticity directs stem cell lineage specification. Cell 126:677–689CrossRefGoogle Scholar
  24. Escobar F, Anseth KS, Schultz KM (2017) Dynamic changes in material properties and degradation of poly(ethylene glycol)hydrazone gels as a function of pH. Macromolecules 50:7351–7360CrossRefGoogle Scholar
  25. Fairbanks BD, Schwartz MP, Bowman CN, Anseth KS (2009) Photoinitiated polymerization of PEG-diacrylate with lithium phenyl-2,4,6-trimethylbenzoylphosphinate: polymerization rate and cytocompatibility. Biomaterials 30:6702–6707CrossRefGoogle Scholar
  26. Ferreira LS, Gerecht S, Fuller J, Shieh HF, Vunjak-Novakovic G, Langer R (2007) Bioactive hydrogel scaffolds for controllable vascular differentiation of human embryonic stem cells bioactive hydrogel scaffolds for controllable vascular differentiation of human embryonic stem cells. Biomaterials 28:2706–2717CrossRefGoogle Scholar
  27. Ferry JD (1980) Viscoelastic properties of polymers. Wiley, New YorkGoogle Scholar
  28. Friedl P (2004) Prespecification and plasticity: shifting mechanisms of cell migration. Curr Opin Cell Biol 16:14–23CrossRefGoogle Scholar
  29. Furst EM, Squires TM (2017) Microrheology, 1st edn. Oxford University Press, OxfordGoogle Scholar
  30. Grim JC, Marozas IA, Anseth KS (2015) Thiol-ene and photo-cleavage chemistry for controlled presentation of biomolecules in hydrogels. J Controlled Release 219:95–106CrossRefGoogle Scholar
  31. Guvendiren M, Burdick JA (2013) Engineering synthetic hydrogel microenvironments to instruct stem cells. Curr Opin in Biotech 24:841–846CrossRefGoogle Scholar
  32. Jackson WM, Nesti LJ, Tuan RS (2012) Concise review: Clinical translation of wound healing therapies based on mesenchymal stem cells. Stem Cells Transl Med 1:44–50CrossRefGoogle Scholar
  33. Kawada H, Fujita J, Kinjo K, Matsuzaki Y, Tsuma M, Miyatake H, Muguruma Y, Tsuboi K, Itabashi Y, Ikeda Y, Ogawa S, Okano H, Hotta T, Ando K, Fukuda K (2004) Nonhematopoietic mesenchymal stem cells can be mobilized and differentiate into cardiomyocytes after myocardial infarction. Blood 104:3581–3587CrossRefGoogle Scholar
  34. Kloxin AM, Kloxin CJ, Bowman CN, Anseth KS (2010) Mechanical properties of cellularly responsive hydrogels and their experimental determination. Biomaterials 22:3484–3494Google Scholar
  35. Kyburz KA, Anseth KS (2013) Three-dimensional hMSC motility within peptide-functionalized PEG-based hydrogels of varying adhesivity and crosslinking density. Acta Biomater 9:6381–6392CrossRefGoogle Scholar
  36. Larsen T, Schultz K, Furst EM (2008) Hydrogel microrheology near the liquid-solid transition. Korea-Australia Rheology Journal 20:165–173Google Scholar
  37. Larsen TH, Furst EM (2008) Microrheology of the liquid-solid transition during gelation. Phys Rev Lett 100:146001–4CrossRefGoogle Scholar
  38. Latifi-Pupovci H, Kuçi Z, Wehner S, Bönig H, Lieberz R, Klingebiel T, Bader P, Kuçi S (2015) In vitro migration and proliferation (“wound healing”) potential of mesenchymal open access stromal cells generated from human cd271 bone marrow mononuclear cells. J Transl Med 13:315–323CrossRefGoogle Scholar
  39. Lauffenburger DA, Horwitz AF (1996) Cell migration: a physically integrated molecular process. Cell 84:359–369CrossRefGoogle Scholar
  40. Legant WR, Miller JS, Blakely BL, Cohen DM, Genin GM, Chen CS (2010) Measurement of mechanical tractions exerted by cells in three-dimensional matrices. Nat Methods 7:969–973CrossRefGoogle Scholar
  41. Longhurst CM, Jennings LK (1998) Integrin-mediated signal transduction. Cell Mol Life Sci 54:514–526CrossRefGoogle Scholar
  42. Lozito TP, Tuan RS (2011) Endothelial cell microparticles act as centers of matrix metalloproteinsase-2 (MMP-2) activation and vascular matrix remodeling. Cell Physiology 227:534–549CrossRefGoogle Scholar
  43. Lozito TP, Jackson WM, Nesti LJ, Tuan RS (2014) Human mesenchymal stem cells generate a distinct pericellular zone of MMP activities via binding of MMPs and secretion of high levels of TIMPs. Matrix Biol 34:132–143CrossRefGoogle Scholar
  44. Lutolf MP, Lauer-Fields JL, Schoekel HG, Metters AT, Weber FE, Fields GB, Hubbell JA (2003) Synthetic matrix metalloproteinase-sensitive hydrogels for the conduction of tissue regeneration: engineering cell-invasion characteristics. PNAS 100:5413–5418CrossRefGoogle Scholar
  45. Mackenzie TC, Flake AW (2001) Human mesenchymal stem cells persist, demonstrate site-specific multipotential differentiation, and are present in sites of wound healing and tissue regeneration after transplantation into fetal sheep. Blood Cell Mol Dis 27:601–604CrossRefGoogle Scholar
  46. Maskarine SA, Franck C, Tirrell DA, Ravichandran G (2009) Quantifying cellular traction forces in three dimensions. PNAS 106:22108–22113CrossRefGoogle Scholar
  47. Mason TG, Weitz DA (1995) Optical measurements of frequency-dependent linear viscoelastic moduli of complex fluids. Phys Rev Lett 74(7):1250–1253CrossRefGoogle Scholar
  48. Mason TG, Ganesan K, van Zanten JH, Wirtz D, Kuo SC (1997) Particle tracking microrheology of complex fluids. Phys Rev Lett 79:3282–3285CrossRefGoogle Scholar
  49. Mazzeo MS, Chai T, Daviran M, Schultz KM (2019) Characterization of the kinetics and mechanism of degradation of human mesenchymal stem cell-laden poly(ethylene glycol) hydrogels. ACS Appl Bio Mater 2:81–92CrossRefGoogle Scholar
  50. Metzger S, Blache U, Lienemann PS, Karlsson M, Weber FE, Weber W, Ehrbar M (2016) Cell-mediated proteolytic release of growth factors from poly(ethylene glycol) matrices. Macromol Biosci 16:1703–1713CrossRefGoogle Scholar
  51. Miller JS, Shen CJ, Legant WR, Baranski JD, Blakely BL, Chen CS (2010) Bioactive hydrogels made from step-growth derived PEG-peptide macromers. Biomaterials 31:3736–3743CrossRefGoogle Scholar
  52. Munevar S, Wang Y, Dembo M (2001) Traction force microscopy of migrating normal and h-ras transformed 3t3 fibroblasts. Biophys J 80:1744–1757CrossRefGoogle Scholar
  53. Muthukumar M, Winter HH (1986) Fractal dimension of a crosslinking polymer at the gel point. Macromolecules 19:1284–1285CrossRefGoogle Scholar
  54. Nagase H, Visse R, Murphy G (2006) Structure and function of matrix metalloproteinases and timps. Cardiovasc Res 69:562–573CrossRefGoogle Scholar
  55. Olson MW, Gervasi DC, Mobashery S, Fridman R (1997) Kinetic analysis of the binding of human matrix metalloproteinase-2 and -9 to tissue inhibitor of metalloproteinase TIMP-1 and TIMP-2. Biol Chem 272 (47):29975–29983CrossRefGoogle Scholar
  56. Palmer A, Xu J, Wirtz D (1998) High-frequency viscoelasticity of crosslinked actin filament networks measured by diffusing wave spectroscopy. Rheol Acta 33:97–106CrossRefGoogle Scholar
  57. Patterson J, Hubbell JA (2010) Enhanced proteolytic degradation of molecularly engineered PEG hydrogels in response to MMP-1 and MMP-2. Biomaterials 31:7836–7845CrossRefGoogle Scholar
  58. Peppas NA, Hilt JZ, Khademhosseini A, Langer R (2006) Hydrogels in biology and medicine: From molecular principles to bionanotechnology. Adv Mater 18:1345–1360CrossRefGoogle Scholar
  59. Peyton SR, Raub CB, Keschrumrus VP, Putnam AJ (2006) The use of poly(ethylene glycol) hydrogels to investigate the impact of ECM chemistry and mechanics on smooth muscle cells. Biomaterials 27:4881–4893CrossRefGoogle Scholar
  60. Piperigkou Z, Götte M, Theocharis AD, Karamanos NK (2017) Insights into the key roles of epigenetics in matrix macromolecules-associated wound healing. Adv Drug Deliv Rev 129:16–36CrossRefGoogle Scholar
  61. Ponte AL, Marais E, Gallay N, Langonne A, Delorme B, Herault O, Charbord P, Domenech J (2007) The in vitro migration capacity of human bone marrow mesenchymal stem cells: comparison of chemokine and growth factor chemotactic activities. Stem Cells 25:1737–1745CrossRefGoogle Scholar
  62. Raeber GP, Lutolf MP, Hubbell JA (2007) Mechanisms of 3-d migration and matrix remodeling of fibroblasts within artificial ECMs. Acta Biomater 3:615–629CrossRefGoogle Scholar
  63. Rice MA, Sanchez-Adams J, Anseth KS (2006) Exogenously triggered, enzymatic degradation of photopolymerized hydrogels with polycaprolactone subunits: experimental observation and modeling of mass loss behavior. Biomacromolecules 7:1968–1975CrossRefGoogle Scholar
  64. Ries C, Egea V, Karow M, Kolb H, Jochum M, Neth P (2007) MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: differential regulation by inflammatory cytokines. Blood 109(9):4055–4063CrossRefGoogle Scholar
  65. Savin T, Doyle PS (2005) Static and dynamic errors in particle tracking microrheology. Biophys J 88:623–638CrossRefGoogle Scholar
  66. Schultz KM, Anseth KS (2013) Monitoring degradation of matrix metalloproteinases cleavable PEG hydrogels via multiple particle tracking microrheology. Soft Matter 9:1570–1579CrossRefGoogle Scholar
  67. Schultz KM, Furst EM (2012) Microrheology of biomaterial hydrogelators. Soft Matter 8:6198–6205CrossRefGoogle Scholar
  68. Schultz KM, Bayles AV, Baldwin AD, Kiick KL, Furst EM (2011) Rapid, high resolution screening of biomaterial hydrogelators by μ 2rheology. Biomacromolecules 12:4178–4182CrossRefGoogle Scholar
  69. Schultz KM, Baldwin AD, Kiick KL, Furst EM (2012) Measuring the modulus and reverse percolation transition of a degrading hydrogel. ACS Macro Lett 1:706–708CrossRefGoogle Scholar
  70. Schultz KM, Kyburz KA, Anseth KS (2015) Measuring dynamic cell-material interactions and remodeling during 3d human mesenchymal stem cell migration in hydrogels. PNAS 112(29):E3757–E3764CrossRefGoogle Scholar
  71. Schwartz MP, Fairbanks BD, Rogers RE, Rangarajan R, Zaman MH, Anseth KS (2010) A synthetic strategy for mimicking the extracellular matrix provides insight about tumor cell migration. Integr Biol 2:32–40CrossRefGoogle Scholar
  72. Singer AJ, Clark RA (1999) Cutaneous wound healing. N Engl J Med 341:738–746CrossRefGoogle Scholar
  73. Soiné JRD, Brand CA, Stricker J, Oakes PW, Gardel ML, Schwarz US (2015) Model-based traction force microscopy reveals differential tension in cellular actin bundles. PLOS Comput Biol 11:e1004076CrossRefGoogle Scholar
  74. Squires TM, Mason TG (2010) Fluid mechanics of microrheology. Annu Rev Fluid Mech 42:413–438CrossRefGoogle Scholar
  75. Stauffer D, Coniglio A, Adam M (1982) Gelation and critical phenomena. Adv Polym Sci 44:103–158CrossRefGoogle Scholar
  76. Suga K, Dedem GV, Moo-Young M (1975) Enzymatic breakdown of water insoluble substrates. Biotechnol Bioeng 17:185–201CrossRefGoogle Scholar
  77. Tse JR, Engler AJ (2011) Stiffness gradients mimicking in vivo tissue variation regulate mesenchymal stem cell fate. PLoS one 6:e15978CrossRefGoogle Scholar
  78. Vincent LG, Choi YS, Alonso-Latorre B, del Álamo JC, Engler AJ (2013) Mesenchymal stem cell durotaxis depends on substrate stiffness gradient strength. Biotechnol J 8:472–484CrossRefGoogle Scholar
  79. Vu TH, Werb Z (2000) Matrix metalloproteinases: effectors of development and normal physiology. Genes Dev 14:2123–2133CrossRefGoogle Scholar
  80. Waigh TA (2005) Microrheology of complex fluids. Rep Prog Phys 68:685–742CrossRefGoogle Scholar
  81. Wehrman MD, Lindberg S, Schultz KM (2016) Quantifying the dynamic transition of hydrogenated castor oil gels measured via multiple particle tracking microrheology. Soft Matter 12:6463–6472CrossRefGoogle Scholar
  82. Wehrman MD, Lindberg S, Schultz KM (2018) Impact of shear on the structure and rheological properties of a hydrogenated castor oil colloidal gel during dynamic phase transitions. J Rheol 62:437–446CrossRefGoogle Scholar
  83. West JL, Hubbell JA (1999) Polymeric biomaterials with degradation sites for proteases involved in cell migration. Macromolecules 32:241–244CrossRefGoogle Scholar
  84. Winter HH (1987) Can the gel point of a cross-linking polymer be detected by the G’-G” crossover? Polym Eng Sci 27:1698–1702CrossRefGoogle Scholar
  85. Wolf K, Te Lindert M, Krause M, Alexander S, Te Riet J, Willis AL, Hoffman RM, Figdor CG, Weiss SJ, Friedl P (2013) Physical limits of cell migration: control by ECM space and nuclear deformation and tuning by proteolysis and traction force. Cell Biol 7(201):1069–1084CrossRefGoogle Scholar

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© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Chemical and Biomolecular EngineeringLehigh UniversityBethlehemUSA

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