Advertisement

A nanofibrous electrospun patch to maintain human mesenchymal cell stemness

  • L. Pandolfi
  • N. Toledano Furman
  • Xin Wang
  • C. Lupo
  • J. O. Martinez
  • M. Mohamed
  • F. Taraballi
  • E. Tasciotti
Tissue Engineering Constructs and Cell Substrates Original Research
Part of the following topical collections:
  1. Tissue Engineering Constructs and Cell Substrates

Abstract

Mesenchymal stem cells (MSCs) have been extensively investigated in regenerative medicine because of their crucial role in tissue healing. For these properties, they are widely tested in clinical trials, usually injected in cell suspension or in combination with tridimensional scaffolds. However, scaffolds can largely affect the fates of MSCs, inducing a progressive loss of functionality overtime. The ideal scaffold must delay MSCs differentiation until paracrine signals from the host induce their change. Herein, we proposed a nanostructured electrospun gelatin patch as an appropriate environment where human MSCs (hMSCs) can adhere, proliferate, and maintain their stemness. This patch exhibited characteristics of a non-linear elastic material and withstood degradation up to 4 weeks. As compared to culture and expansion in 2D, hMSCs on the patch showed a similar degree of proliferation and better maintained their progenitor properties, as assessed by their superior differentiation capacity towards typical mesenchymal lineages (i.e. osteogenic and chondrogenic). Furthermore, immunohistochemical analysis and longitudinal non-invasive imaging of inflammatory response revealed no sign of foreign body reaction for 3 weeks. In summary, our results demonstrated that our biocompatible patch favored the maintenance of undifferentiated hMSCs for up to 21 days and is an ideal candidate for tridimensional delivery of hMSCs.

Graphical Abstract

Open image in new window

The present work reports a nanostructured patch gelatin-based able to maintain in vitro hMSCs stemness features. Moreover, hMSCs were able to differentiate toward osteo- and chondrogenic lineages once induces by differentiative media, confirming the ability of this patch to support stem cells for a potential in vivo application. These attractive properties together with the low inflammatory response in vivo make this patch a promising platform in regenerative medicine.

Keywords

ThermoFisher Scientific HFIP Multilineage Differentiation Osteogenic Marker Chondrogenic Lineage 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

The authors acknowledge Ms. Nupur Basu for harvesting the cells used in this study and Dr. Junping You and Dr. Armando Torres for helping with the in vivo studies. We also acknowledge Mr. Christopher Candelari and Chih Hao Liu for helping with the mechanical testing. We thank Dr. J. Gu of the HMRI Microscopy-SEM/AFM core, and Dr. David Haviland, Director of the flow cytometry core. We also thank Ms. Megan Livingston for editing this document. The authors gratefully acknowledge funding support from the following sources: Brown Foundation (Project ID, 18130011), the Hearst Foundation (Project ID, 18130017), the Cullen Trust for Health Care Foundation (Project ID, 18130014), and the DoD USAMRMC (Project ID, W81XWH-15-1-0718).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interest.

Supplementary material

10856_2017_5856_MOESM1_ESM.tif (1.5 mb)
Supplementary Figure1
10856_2017_5856_MOESM2_ESM.tif (971 kb)
Supplementary Figure2
10856_2017_5856_MOESM3_ESM.tif (3.9 mb)
Supplementary Figure3
10856_2017_5856_MOESM4_ESM.tif (2.5 mb)
Supplementary Figure4
10856_2017_5856_MOESM5_ESM.tif (2.5 mb)
Supplementary Figure5
10856_2017_5856_MOESM6_ESM.tif (3 mb)
Supplementary Figure6
10856_2017_5856_MOESM7_ESM.docx (14 kb)
Supplementary Information

References

  1. 1.
    Bianco P, Riminucci M, Gronthos S, Robey PG. Bone marrow stromal stem cells: nature, biology, and potential applications. Stem Cells. 2001;19(3):180–92.CrossRefGoogle Scholar
  2. 2.
    Barry FP, Murphy JM. Mesenchymal stem cells: clinical applications and biological characterization. Int J Biochem Cell Biol. 2004;36(4):568–84.CrossRefGoogle Scholar
  3. 3.
    Corradetti B, Taraballi F, Powell S, Sung D, Minardi S, Ferrari M, et al. Osteoprogenitor cells from bone marrow and cortical bone: understanding how the environment affects their fate. Stem Cells Dev. 2014;24(9):1112–23.CrossRefGoogle Scholar
  4. 4.
    Kalervo Väänänen H. Mesenchymal stem cells. Ann Med. 2005;37(7):469–79.CrossRefGoogle Scholar
  5. 5.
    Rustad KC, Wong VW, Sorkin M, Glotzbach JP, Major MR, Rajadas J, et al. Enhancement of mesenchymal stem cell angiogenic capacity and stemness by a biomimetic hydrogel scaffold. Biomaterials. 2012;33(1):80–90.CrossRefGoogle Scholar
  6. 6.
    Quarto R, Mastrogiacomo M, Cancedda R, Kutepov SM, Mukhachev V, Lavroukov A, et al. Repair of large bone defects with the use of autologous bone marrow stromal cells. New Engl J Med. 2001;344(5):385–6.CrossRefGoogle Scholar
  7. 7.
    Lee CH, Cook JL, Mendelson A, Moioli EK, Yao H, Mao JJ. Regeneration of the articular surface of the rabbit synovial joint by cell homing: a proof of concept study. Lancet. 2010;376(9739):440–8.CrossRefGoogle Scholar
  8. 8.
    O’brien FJ. Biomaterials & scaffolds for tissue engineering. Mater Today. 2011;14(3):88–95.CrossRefGoogle Scholar
  9. 9.
    Geckil H, Xu F, Zhang X, Moon S, Demirci U. Engineering hydrogels as extracellular matrix mimics. Nanomedicine. 2010;5(3):469–84.CrossRefGoogle Scholar
  10. 10.
    Augello A, Kurth TB, De Bari C. Mesenchymal stem cells: a perspective from in vitro cultures to in vivo migration and niches. Eur Cell Mater. 2010;20:121–33.CrossRefGoogle Scholar
  11. 11.
    Peerani R, Rao BM, Bauwens C, Yin T, Wood GA, Nagy A, et al. Niche‐mediated control of human embryonic stem cell self‐renewal and differentiation. EMBO J. 2007;26(22):4744–55.CrossRefGoogle Scholar
  12. 12.
    Tsai C-C, Chen C-L, Liu H-C, Lee Y-T, Wang H-W, Hou L-T, et al. Overexpression of hTERT increases stem-like properties and decreases spontaneous differentiation in human mesenchymal stem cell lines. J Biomed Sci. 2010;17(1):1CrossRefGoogle Scholar
  13. 13.
    Hocking AM, Gibran NS. Mesenchymal stem cells: paracrine signaling and differentiation during cutaneous wound repair. Exp Cell Res. 2010;316(14):2213–9.CrossRefGoogle Scholar
  14. 14.
    Trcin MT, Dekaris I, Mijović B, Bujić M, Zdraveva E, Dolenec T, et al. Synthetic vs natural scaffolds for human limbal stem cells. Croat Med J. 2015;56(3):246CrossRefGoogle Scholar
  15. 15.
    Casper CL, Yang W, Farach-Carson MC, Rabolt JF. Coating electrospun collagen and gelatin fibers with perlecan domain I for increased growth factor binding. Biomacromolecules. 2007;8(4):1116–23.CrossRefGoogle Scholar
  16. 16.
    Matthews JA, Wnek GE, Simpson DG, Bowlin GL. Electrospinning of collagen nanofibers. Biomacromolecules. 2002;3(2):232–8.CrossRefGoogle Scholar
  17. 17.
    Zhang Z, Li G, Shi B. Physicochemical properties of collagen, gelatin and collagen hydrolysate derived from bovine limed split wastes. J Soc Leath Tech Ch. 2006;90(1):23Google Scholar
  18. 18.
    Zhang Y, Venugopal J, Huang ZM, Lim C, Ramakrishna S. Crosslinking of the electrospun gelatin nanofibers. Polymer (Guildf). 2006;47(8):2911–7.CrossRefGoogle Scholar
  19. 19.
    Mano J, Silva G, Azevedo HS, Malafaya P, Sousa R, Silva S, et al. Natural origin biodegradable systems in tissue engineering and regenerative medicine: present status and some moving trends. J R Soc Interface. 2007;4(17):999–1030.CrossRefGoogle Scholar
  20. 20.
    Schiffman JD, Schauer CL. A review: electrospinning of biopolymer nanofibers and their applications. Polym Rev. 2008;48(2):317–52.CrossRefGoogle Scholar
  21. 21.
    Kriegel C, Arrechi A, Kit K, McClements DJ, Weiss J. Fabrication, functionalization, and application of electrospun biopolymer nanofibers. Crit Rev Food Sci Nutr. 2008;48(8):775–97.CrossRefGoogle Scholar
  22. 22.
    Huang ZM, Zhang YZ, Kotaki M, Ramakrishna S. A review on polymer nanofibers by electrospinning and their applications in nanocomposites. Compos Sci Technol. 2003;63(15):2223–53.CrossRefGoogle Scholar
  23. 23.
    Pham QP, Sharma U, Mikos AG. Electrospun poly (ε-caprolactone) microfiber and multilayer nanofiber/microfiber scaffolds: characterization of scaffolds and measurement of cellular infiltration. Biomacromolecules. 2006;7(10):2796–805.CrossRefGoogle Scholar
  24. 24.
    Yoshimoto H, Shin Y, Terai H, Vacanti J. A biodegradable nanofiber scaffold by electrospinning and its potential for bone tissue engineering. Biomaterials. 2003;24(12):2077–82.CrossRefGoogle Scholar
  25. 25.
    Ghasemi-Mobarakeh L, Morshed M, Karbalaie K, Fesharaki M, Nasr-Esfahani MH, Baharvand H. Electrospun poly (ε-caprolactone) nanofiber mat as extracellular matrix. Yakhteh Med J. 2008;10(3):179–84.Google Scholar
  26. 26.
    Melchels FPW, et al. Effects of the architecture of tissue engineering scaffolds on cell seeding and culturing. Acta Biomater. 2010;6(11):4208–17.Google Scholar
  27. 27.
    Garg K, Bowlin GL. Electrospinning jets and nanofibrous structures. Biomicrofluidics. 2011;5(1):013403CrossRefGoogle Scholar
  28. 28.
    Minardi S, Sandri M, Martinez JO, Yazdi IK, Liu X, Ferrari M, et al. Multiscale patterning of a biomimetic scaffold integrated with composite microspheres. Small. 2014;10(19):3943–53.CrossRefGoogle Scholar
  29. 29.
    Corradetti B, Taraballi F, Powell S, Sung D, Minardi S, Ferrari M, et al. Osteoprogenitor cells from bone marrow and cortical bone: understanding how the environment affects their fate. Stem Cells Dev. 2015;24(9):1112–23.CrossRefGoogle Scholar
  30. 30.
    Taraballi F, Wang S, Li J, Lee FYY, Venkatraman SS, Birch WR, et al. Understanding the nano‐topography changes and cellular influences resulting from the surface adsorption of human hair keratins. Adv Healthc Mater. 2012;1(4):513–9.CrossRefGoogle Scholar
  31. 31.
    Minardi S, Pandolfi L, Taraballi F, De Rosa E, Yazdi IK, Liu X, et al. PLGA-mesoporous silicon microspheres for the in vivo controlled temporospatial delivery of proteins. ACS Appl Mater Inter. 2015;7(30):16364–73.CrossRefGoogle Scholar
  32. 32.
    Torres-Giner S, Gimeno-Alcaniz JV, Ocio MJ, Lagaron JM. Comparative performance of electrospun collagen nanofibers cross-linked by means of different methods. ACS Appl Mater Inter. 2008;1(1):218–23.CrossRefGoogle Scholar
  33. 33.
    Meng L, Arnoult O, Smith M, Wnek GE. Electrospinning of in situ crosslinked collagen nanofibers. J Mater Chem. 2012;22(37):19412–7.CrossRefGoogle Scholar
  34. 34.
    Gross S, Gammon ST, Moss BL, Rauch D, Harding J, Heinecke JW, et al. Bioluminescence imaging of myeloperoxidase activity in vivo. Nat Med. 2009;15(4):455–61.CrossRefGoogle Scholar
  35. 35.
    Li L, Xie T. Stem cell niche: structure and function. Annu Rev Cell Dev Biol. 2005;21:605–31.CrossRefGoogle Scholar
  36. 36.
    Spradling A, Drummond-Barbosa D, Kai T. Stem cells find their niche. Nature. 2001;414(6859):98–104.CrossRefGoogle Scholar
  37. 37.
    Yao W, Lane NE. Targeted delivery of mesenchymal stem cells to the bone. Bone. 2015;70:62–5.CrossRefGoogle Scholar
  38. 38.
    Simpson D, Liu H, Fan THM, Nerem R, Dudley SC. A tissue engineering approach to progenitor cell delivery results in significant cell engraftment and improved myocardial remodeling. Stem Cells. 2007;25(9):2350–7.CrossRefGoogle Scholar
  39. 39.
    Kouris NA, Squirrell JM, Jung JP, Pehlke CA, Hacker T, Eliceiri KW, et al. A nondenatured, noncrosslinked collagen matrix to deliver stem cells to the heart. Regen Med. 2011;6(5):569–82.CrossRefGoogle Scholar
  40. 40.
    Li M, Yu J, Li Y, Li D, Yan D, Ruan Q. CXCR4+ progenitors derived from bone mesenchymal stem cells differentiate into endothelial cells capable of vascular repair after arterial injury. Cell Reprogram. 2010;12(4):405–15.CrossRefGoogle Scholar
  41. 41.
    Silva GV, Litovsky S, Assad JA, Sousa AL, Martin BJ, Vela D, et al. Mesenchymal stem cells differentiate into an endothelial phenotype, enhance vascular density, and improve heart function in a canine chronic ischemia model. Circulation. 2005;111(2):150–6.CrossRefGoogle Scholar
  42. 42.
    Galperin A, Long TJ, Ratner BD. Degradable, thermo-sensitive poly (N-isopropyl acrylamide)-based scaffolds with controlled porosity for tissue engineering applications. Biomacromolecules. 2010;11(10):2583–92.CrossRefGoogle Scholar
  43. 43.
    Discher DE, Janmey P, Wang YL. Tissue cells feel and respond to the stiffness of their substrate. Science. 2005;310(5751):1139–43.CrossRefGoogle Scholar
  44. 44.
    Wang H-B, Dembo M, Wang YL. Substrate flexibility regulates growth and apoptosis of normal but not transformed cells. Am J Physiol-Cell Ph. 2000;279(5):C1345–C50.Google Scholar
  45. 45.
    Nam J, Huang Y, Agarwal S, Lannutti J. Improved cellular infiltration in electrospun fiber via engineered porosity. Tiss Eng. 2007;13(9):2249–57.CrossRefGoogle Scholar
  46. 46.
    Manning C, Schwartz A, Liu W, Xie J, Havlioglu N, Sakiyama-Elbert S, et al. Controlled delivery of mesenchymal stem cells and growth factors using a nanofiber scaffold for tendon repair. Acta Biomater. 2013;9(6):6905–14.CrossRefGoogle Scholar
  47. 47.
    Kuhn NZ, Tuan RS. Regulation of stemness and stem cell niche of mesenchymal stem cells: implications in tumorigenesis and metastasis. J Cell Physiol. 2010;222(2):268–77.CrossRefGoogle Scholar
  48. 48.
    Edmondson R, Broglie JJ, Adcock AF, Yang L. Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay Drug Dev Technol. 2014;12(4):207–18.CrossRefGoogle Scholar
  49. 49.
    Westhrin M, Xie M, Olderøy MØ, Sikorski P, Strand BL, Standal T. Osteogenic differentiation of human mesenchymal stem cells in mineralized alginate matrices. PloS One. 2015;10(3):2583–92CrossRefGoogle Scholar
  50. 50.
    Page H, Flood P, Reynaud EG. Three-dimensional tissue cultures: current trends and beyond. Cell Tissue Res. 2013;352(1):123–31.CrossRefGoogle Scholar
  51. 51.
    Celiz AD, Smith JG, Langer R, Anderson DG, Winkler DA, Barrett DA, et al. Materials for stem cell factories of the future. Nat Mater. 2014;13(6):570–9.CrossRefGoogle Scholar
  52. 52.
    Kastellorizios M, Tipnis N, Burgess DJ. Foreign body reaction to subcutaneous implants. In: John DL, Kristina NE, Daniel R, Bo N, editors. Immune responses to biosurfaces. Springer;2015. pp. 93–108.Google Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Regenerative MedicineHouston Methodist Research InstituteHoustonUSA
  2. 2.College of Materials Science and EngineeringUniversity of Chinese Academy of ScienceBeijingChina
  3. 3.Department of Biomedical EngineeringUniversity of HoustonHoustonUSA
  4. 4.Department of Orthopedics and Sports MedicineHouston Methodist HospitalHoustonUSA

Personalised recommendations