Skip to main content

Advertisement

SpringerLink
Log in

Fibronectin/thermo-responsive polymer scaffold as a dynamic ex vivo niche for mesenchymal stem cells

  • Tissue Engineering Constructs and Cell Substrates
  • Original Research
  • Published:
Journal of Materials Science: Materials in Medicine Aims and scope Submit manuscript

Abstract

In this paper, we created a dynamic adhesive environment (DAE) for adipose tissue-derived mesenchymal stem cells (ADMSCs) cultured on smart thermo-responsive substrates, i.e., poly (N-isopropyl acrylamide) (PNIPAM), via introducing periodic changes in the culture temperature. We further explored the particular role of adsorbed fibronectin (FN), an important cell adhesive protein that was recently attributed to the recruitment of stem cells in the niche. The engineered FN/PNIPAM DAE system significantly increased the symmetric renewal of ADMSCs, particularly between passages 7 and 9 (p7–p9), before it dropped down to the level of the control (FN-coated TC polystyrene). This decline in the growth curve was consistent with the increased number of senescent cells, the augmented average cell size and the suppressed FN matrix secretion at late passages (p10–p12), all of them characteristic for stem cells ageing, which equivocally tended to slow down at our DAE system. FN supported also the osteogenic response of ADMSCs (apart from the previous observations with plain PNIPAM substrata) indicated by the significant increase of alkaline phosphatase (ALP) activity at days 7 and 14. The minimal changes in the Ca deposition, however, suggest a restricted effect of DAE on the early osteogenic response of ADMSCs only. Thus, the engineering of niche-like DAE involving FN uncovers a new tissue engineering strategy for gaining larger amounts of functionally active stem cells for clinical application.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Krishnamoorthy N, Tseng YT, Gajendrarao P, Sarathchandra P, McCormack A, Carubelli I, et al. A novel strategy to enhance secretion of ECM components by stem cells: relevance to tissue engineering. Tissue Eng Part A. 2017;24:145–56.

    Google Scholar 

  2. Ferraro F, Lo Celso C. Adult stem cells and their niches. Adv Exp Med Biol. 2010;695:155–68.

    CAS  Google Scholar 

  3. Frese L, Dijkman PE, Hoerstrup SP. Adipose tissue derived stem cells in regenerative medicine. Transfus Med Hemotherapy. 2016;43:268–74.

    Google Scholar 

  4. Caplan AI. Mesenchymal stem cells. J Orthop Res. 1991;9:641–50.

    CAS  Google Scholar 

  5. Caplan AI. Mesenchymal stem cells: cell-based reconstructive therapy in orthopedics. Tissue Eng. 2005;11:1198–211.

    CAS  Google Scholar 

  6. Prockop DJ. Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 1997;276:71–4.

    CAS  Google Scholar 

  7. Baker N, Boyette LB, Tuan RS. Characterization of bone marrow-derived mesenchymal stem cells in ageing. Bone 2015;70:37–47.

    CAS  Google Scholar 

  8. Boyette LB, Tuan RS. Adult stem cells and diseases of ageing. J Clin Med. 2014;3:88–134.

    CAS  Google Scholar 

  9. Sousa-Victor P, Garcia-Prat L, Serrano AL, Perdiguero E, Munoz-Canoves P. Muscle stem cell ageing: regulation and rejuvenation. Trends Endocrinol Metab. 2015;26:287–96.

    CAS  Google Scholar 

  10. Kennedy BK, Berger SL, Brunet A, Campisi J, Cuervo AM, Epel ES, et al. Geroscience: linking ageing to chronic disease. Cell 2014;159:709–13.

    CAS  Google Scholar 

  11. Reitinger S, Schimke M, Klepsch S, de Sneeuw S, Yani SL, Gassner R, et al. Systemic impact molds mesenchymal stromal/stem cell ageing. Transfus Apheresis Sci. 2015;52:285–9.

    Google Scholar 

  12. Scadden DT. The stem-cell niche as an entity of action. Nature 2006;441:1075–9.

    CAS  Google Scholar 

  13. Phadke A, Chang CW, Varghese S. Functional biomaterials for controlling stem cell differentiation. In: Krishnendu R, editor. Biomaterials as stem cells niche. Berlin Heidelberg: Springer-Verlag; 2010.

  14. Lukjanenko L, Jung MJ, Hedge N, Perruisseau-Carrier C, Migliavacca E, Rozo M, et al. Loss of fibronectin from the aged stem cell niche affects the regenerative capacity of skeletal muscle in mice. Nat Med. 2016;22:897–905.

    CAS  Google Scholar 

  15. Birmingham E, Niebur GL, McHugh PE, Shaw G, Barry FP, McNamara LM. Osteogenic differentiation of mesenchymal stem cells is regulated by osteocyte and osteoblast cells in a simplified bone niche. Eur Cell Mater. 2012;23:13–27.

    CAS  Google Scholar 

  16. Kuang S, Gillespie MA, Rudnicki MA. Niche regulation of muscle satellite cell self-renewal and differentiation. Cell Stem Cell. 2008;2:22–31.

    CAS  Google Scholar 

  17. Morrison SJ, Kimble J. Asymmetric and symmetric stem-cell divisions in development and cancer. Nature 2006;441:1068–74.

    CAS  Google Scholar 

  18. Teixeira A, Hermanson O, Werner C. Designing and engineering stem cell niches. MRS Bull. 2010;35:591–6.

    CAS  Google Scholar 

  19. Donzelli E, Salvadè A, Mimo P, Viganò M, Morrone M, Papagna R, et al. Mesenchymal stem cells cultured on a collagen scaffold: In vitro osteogenic differentiation. Arch Oral Biol. 2007;52:64–73.

    CAS  Google Scholar 

  20. Salasznyk RM, Williams WA, Boskey A, Batorsky A, Plopper GE. Adhesion to vitronectin and collagen I promotes osteogenic differentiation of human mesenchymal stem cells. J Biomedicine Biotechnol 2004;1:24–34.

    Google Scholar 

  21. Schneider RK, Puellen A, Kramann R, Raupach K, Bornemann J, Knuechel R, et al. Osteogenic differentiation of adult bone marrow and perinatal umbilical mesenchymal stem cells and matrix remodelling in three-dimensional collagen scaffolds. Biomaterials 2010;31:467–80.

    CAS  Google Scholar 

  22. Somaiah C, Kumar A, Mauwrie D, Sharma A, Patil SD, Bhattacharyya J, et al. Collagen promotes higher adhesion, survival and proliferation of mesenchymal stem cells. PLoS ONE. 2015;10:e0145068.

    Google Scholar 

  23. Singh P, Schwarzbauer JE. Fibronectin and stem cell differentiation — lessons from chondrogenesis. J Cell Sci. 2012;125:3703–12.

    CAS  Google Scholar 

  24. Trappmann B, Gautrot JE, Connelly JT, Strange DGT, Li Y, Oyen ML, et al. Extracellular-matrix tethering regulates stem-cell fate. Nat Mater. 2012;11:642–9.

    CAS  Google Scholar 

  25. Gattazzo F, Urciuolo A, Bonardo P. Extracellular matrix: a dynamic microenvironment for stem cell niche. Biochimica Biophysica Acta. 2014;1840:2506–19.

    CAS  Google Scholar 

  26. Bianchi MV, Awaja F, Altankov G. Dynamic adhesive environment alters the differentiation potential of young and ageing mesenchymal stem cells. Mater Sci Eng C 2017;78:467–74.

    CAS  Google Scholar 

  27. Zhang W, Zhang X, Wang S, Xu L, Zhang M, Wang G, et al. Comparison of the use of adipose tissue-derived and bone marrow-derived stem cells for rapid bone regeneration. J Dent Res. 2013;92:1136–41.

    CAS  Google Scholar 

  28. McLaughlin MM, Marra KG. The use of adipose-derived stem cells as sheets for wound healing. Organogenesis 2013;9:79–81.

    Google Scholar 

  29. Cohen S. Emerging applications of stimuli-responsive polymer materials. Nat Mater. 2010;9:101–13.

    Google Scholar 

  30. Kikuchi A, Okano T. Temperature responsive, polymer-modified surfaces for green chromatography. Macromol Symposia. 2004;207:217–27.

    CAS  Google Scholar 

  31. Yamato M, Konno C, Kushida A, Hirose M, Utsumi M, Kikuchi A, et al. Release of adsorbed fibronectin from temperature-responsive culture surfaces requires cellular activity. Biomaterials 2000;21:981–6.

    CAS  Google Scholar 

  32. da Silva RMP, Mano JF, Reis RL. Smart thermoresponsive coatings and surfaces for tissue engineering: switching cell-material boundaries. Trends Biotechnol. 2007;25:577–83.

    Google Scholar 

  33. Ward MA, Georgiou TK. Thermoresponsive polymers for biomedical applications. Polymers 2011;3:1215–42.

    CAS  Google Scholar 

  34. Okano T, Yamada N, Okuhara M, Sakai H, Sakurai Y. Mechanism of cell detachment from temperature-modulated, hydrophilic-hydrophobic polymer surfaces. Biomaterials 1995;16:297–303.

    CAS  Google Scholar 

  35. Groth T, Altankov G, Kostadinova A, Krasteva N, Albrecht W, Paul D. Altered vitronectin receptor (alpha-v integrin) function in fibroblasts adhering on hydrophobic glass. J Biomed Mater Res. 1998;44:341–51.

    Google Scholar 

  36. Toromanov G, Gugutkov D, Gustavsson J, Planell J, Salmeron-Sanchez M, Altankov G. Dynamic behavior of vitronectin at the cell-material interface. ACS Biomater Sci Eng. 2015;1:927–34.

    CAS  Google Scholar 

  37. Greenwood SK, Hill RB, Sun JT, Armstrong MJ, Johnson TE, Gara JP, et al. Population doubling: a simple and more accurate estimation of cell growth suppression in the in vitro assay for chromosomal aberrations that reduces irrelevant positive results. Environ Mol. Mutagenesis 2004;43:36–44.

    CAS  Google Scholar 

  38. Yamada N, Okano T, Sakai H, Karikusa F, Sawasaki Y, Sakurai Y. Thermo‐responsive polymeric surfaces; control of attachment and detachment of cultured cells. Macromol Rapid Commun. 1990;11:571–6.

    CAS  Google Scholar 

  39. Shimizu K, Fujita H, Nagamori E. Oxygen plasma‐treated thermoresponsive polymer surfaces for cell sheet engineering. Biotechnol Bioeng. 2010;106:303–10.

    CAS  Google Scholar 

  40. Muniandy MKR, Ruzanna AK, Intan Zarina ZA, Rohaya MAW, Sahidan S, Majlis BY, et al. Proliferation rate and cell size analyses of human peripheral blood suspension stem cells from three culturing terms populations. Malays Appl Biol. 2013;42:59–63.

    Google Scholar 

  41. Raven PH How cells divide. In: Biology 10th, Raven PH, Johnson G, Mason K, editors. New York: McGraw-Hill; 2011.

  42. Holstein TW. Cell cycle of stem cells in hydra. Developmental Biol. 1990;142:392–400.

    CAS  Google Scholar 

  43. Geissler S, Textor M, Kuehnisch J, Koennig D, Klein O, Ode A, et al. Functional comparison of chronological and in vitro ageing: differential role of the cytoskeleton and mitochondria in mesenchymal stromal cells. PLoS ONE. 2012;7:e52700.

    CAS  Google Scholar 

  44. Legzdina D, Romanauska A, Nikulshin S, Kozlovska T, Berzins U. Characterization of senescence of culture-expanded human adipose-derived mesenchymal stem cells. Int J Stem Cells. 2016;9:124–36.

    CAS  Google Scholar 

  45. Lee BY, Han JA, Im JS, Morrone A, Johung K, Goodwin EC, et al. Senescence‐associated β‐galactosidase is lysosomal β‐galactosidase. Ageing Cell. 2006;5:187–95.

    CAS  Google Scholar 

  46. Gordon J, Hassan MQ, Koss M, Montecino M, Selleri L, van Vijnen AL, et al. Epigenetic regulation of early osteogenesis and mineralized tissue formation by a HOXA10-PBX1-associated complex. Cells Tissues Organs. 2011;194:146–50.

    CAS  Google Scholar 

  47. Shi D, Ma D, Dong F, Zong C, Liu L, Shen D, et al. Proliferation and multi-differentiation potentials of human mesenchymal stem cells on thermoresponsive PDMS surfaces grafted with PNIPAM. Biosci Rep. 2009;30:149–58.

    Google Scholar 

  48. Dedhar S. Signal transduction via the b1 integrins is a required intermediate in interleukin-1 b induction of alkaline phosphatase activity in human osteosarcoma cells. Exp Cell Res. 1989;183:207–14.

    CAS  Google Scholar 

  49. Zhao M, Harris SE, Horn D, Geng Z, Nishimura R, Mundy GR, et al. Bone morphogenetic protein receptor signaling is necessary for normal murine postnatal bone formation. J Cell Biol. 2002;157:1049–60.

    CAS  Google Scholar 

  50. An J, Leeuwenburgh S, Wolke J, Jansen J Mineralization processes in hard tissues: Bone. In: Aparicio C, Ginebra MP, editors. Biomineralization and biomaterials fundamentals and application. Woodhead Publishing - Elsevier, Cambridge, United Kingdom. 2016. (pp. 129–46).

  51. Fakhry M, Hamade E, Badran B, Buchet R, Magne D. Molecular mechanisms of mesenchymal stem cell differentiation towards osteoblasts. World J Stem Cells. 2013;5:136–48.

    Google Scholar 

  52. Viti F, Landini M, Mezzelani A, Petecchia L, Milanesi L, Scaglione S. Osteogenic differentiation of MSC through calcium signaling activation: transcriptomics and functional analysis. PLoS ONE. 2016;11:e0148173.

    Google Scholar 

  53. Murshed M, Harmey D, Millan JL, McKee MD, Karsenty G. Unique coexpression in osteoblasts of broadly expressed genes accounts for the spatial restriction of ECM mineralization to bone. Genes Dev. 2005;19:1093–104.

    CAS  Google Scholar 

  54. Keselowsky BG, Wang L, Schwartz Z, Garcia AJ, Boyan BD. Integrin α5 controls osteoblastic proliferation and differentiation responses to titanium substrates presenting different roughness characteristics in a roughness independent manner. J Biomed Mater Res Part A. 2007;80:700–10.

    CAS  Google Scholar 

  55. Hristova-Panusheva K, Keremidarska-Markova M, Altankov G, Krasteva N. Age-related changes in adhesive phenotype of bone marrow-derived mesenchymal stem cells on extracellular matrix proteins. J N. Results Sci. 2017;6:11–9.

    Google Scholar 

  56. Nedjari S, Awaja F, Altankov G. Three dimensional honeycomb patterned fibrinogen based nanofibers induce substantial osteogenic response of mesenchymal stem cells. Sci Rep. 2017;7:15947.

    Google Scholar 

  57. Cosgrove BD, Sacco A, Gilbert PM, Blau HM. A home away from home, challenges and opportunities in engineering in vitro muscle satellite cell niches. Differentiation 2010;78:185–94.

    Google Scholar 

Download references

Acknowledgements

This work was mainly supported by the European Commission through FP7 Industry-Academia Partnerships and Pathways (IAPP) project FIBROGELNET. The valuable contribution of project MAT 2015–69315–C3 MYOHEAL with the Spanish Ministry of Science and Innovation and CIBER-BBN are also acknowledged. FA acknowledges the partial support of FWF under Lise Meitner program (M-1777) and grant No 713690 provided by European Commission within the Horizon 2020 Marie Skłodowska-Curie program.

Author information

Author notes

  1. Firas Awaja

    Present address: Regenerative Medicine Institute (REMEDI) and Centre for Research in Medical Devices (CÚRAM) at National University of Ireland, Galway, Ireland

  2. George Altankov

    Present address: Associate Member Institute for Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Sofia, Bulgaria

Authors and Affiliations

  1. ICREA, Barcelona, Spain

    Laura Ramalho, Salima Nedjari, Dencho Gugutkov & George Altankov

  2. Institute of Biophysics and Biomedical Engineering, Faculty of Sciences, University of Lisbon, Lisbon, Portugal

    Laura Ramalho

  3. École Polytechnique Fédérale de Lausanne (EPFL), Swiss Plasma Center (SPC), CH-5232, Villigen PSI, Switzerland

    Roberto Guarino

  4. Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain

    Firas Awaja & George Altankov

  5. Engmat Ltd., Clybaun Road, Galway, Ireland

    Firas Awaja

Authors
  1. Laura Ramalho
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Salima Nedjari
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Roberto Guarino
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Firas Awaja
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dencho Gugutkov
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. George Altankov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to George Altankov.

Ethics declarations

Conflict of interests

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ramalho, L., Nedjari, S., Guarino, R. et al. Fibronectin/thermo-responsive polymer scaffold as a dynamic ex vivo niche for mesenchymal stem cells. J Mater Sci: Mater Med 31, 129 (2020). https://doi.org/10.1007/s10856-020-06461-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s10856-020-06461-y

Search

Navigation