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
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.
Similar content being viewed by others
Explore related subjects
Discover the latest articles, news and stories from top researchers in related subjects.References
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.
Barry FP, Murphy JM. Mesenchymal stem cells: clinical applications and biological characterization. Int J Biochem Cell Biol. 2004;36(4):568–84.
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.
Kalervo Väänänen H. Mesenchymal stem cells. Ann Med. 2005;37(7):469–79.
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.
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.
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.
O’brien FJ. Biomaterials & scaffolds for tissue engineering. Mater Today. 2011;14(3):88–95.
Geckil H, Xu F, Zhang X, Moon S, Demirci U. Engineering hydrogels as extracellular matrix mimics. Nanomedicine. 2010;5(3):469–84.
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.
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.
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):1
Hocking AM, Gibran NS. Mesenchymal stem cells: paracrine signaling and differentiation during cutaneous wound repair. Exp Cell Res. 2010;316(14):2213–9.
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):246
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.
Matthews JA, Wnek GE, Simpson DG, Bowlin GL. Electrospinning of collagen nanofibers. Biomacromolecules. 2002;3(2):232–8.
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):23
Zhang Y, Venugopal J, Huang ZM, Lim C, Ramakrishna S. Crosslinking of the electrospun gelatin nanofibers. Polymer (Guildf). 2006;47(8):2911–7.
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.
Schiffman JD, Schauer CL. A review: electrospinning of biopolymer nanofibers and their applications. Polym Rev. 2008;48(2):317–52.
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.
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.
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.
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.
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.
Melchels FPW, et al. Effects of the architecture of tissue engineering scaffolds on cell seeding and culturing. Acta Biomater. 2010;6(11):4208–17.
Garg K, Bowlin GL. Electrospinning jets and nanofibrous structures. Biomicrofluidics. 2011;5(1):013403
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.
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.
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.
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.
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.
Meng L, Arnoult O, Smith M, Wnek GE. Electrospinning of in situ crosslinked collagen nanofibers. J Mater Chem. 2012;22(37):19412–7.
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.
Li L, Xie T. Stem cell niche: structure and function. Annu Rev Cell Dev Biol. 2005;21:605–31.
Spradling A, Drummond-Barbosa D, Kai T. Stem cells find their niche. Nature. 2001;414(6859):98–104.
Yao W, Lane NE. Targeted delivery of mesenchymal stem cells to the bone. Bone. 2015;70:62–5.
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.
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.
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.
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.
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.
Discher DE, Janmey P, Wang YL. Tissue cells feel and respond to the stiffness of their substrate. Science. 2005;310(5751):1139–43.
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.
Nam J, Huang Y, Agarwal S, Lannutti J. Improved cellular infiltration in electrospun fiber via engineered porosity. Tiss Eng. 2007;13(9):2249–57.
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.
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.
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.
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):2583–92
Page H, Flood P, Reynaud EG. Three-dimensional tissue cultures: current trends and beyond. Cell Tissue Res. 2013;352(1):123–31.
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.
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.
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).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no competing interest.
Rights and permissions
About this article
Cite this article
Pandolfi, L., Furman, N.T., Wang, X. et al. A nanofibrous electrospun patch to maintain human mesenchymal cell stemness. J Mater Sci: Mater Med 28, 44 (2017). https://doi.org/10.1007/s10856-017-5856-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10856-017-5856-0