title: Boron nitride nanotube-mediated stimulation of cell co-culture on micro-engineered hydrogels.
abstract: In this paper, we describe the effects of the combination of topographical, mechanical, chemical and intracellular electrical stimuli on a co-culture of fibroblasts and skeletal muscle cells. The co-culture was anisotropically grown onto an engineered micro-grooved (10 µm-wide grooves) polyacrylamide substrate, showing a precisely tuned Young's modulus (∼ 14 kPa) and a small thickness (∼ 12 µm). We enhanced the co-culture properties through intracellular stimulation produced by piezoelectric nanostructures (i.e., boron nitride nanotubes) activated by ultrasounds, thus exploiting the ability of boron nitride nanotubes to convert outer mechanical waves (such as ultrasounds) in intracellular electrical stimuli, by exploiting the direct piezoelectric effect. We demonstrated that nanotubes were internalized by muscle cells and localized in both early and late endosomes, while they were not internalized by the underneath fibroblast layer. Muscle cell differentiation benefited from the synergic combination of topographical, mechanical, chemical and nanoparticle-based stimuli, showing good myotube development and alignment towards a preferential direction, as well as high expression of genes encoding key proteins for muscle contraction (i.e., actin and myosin). We also clarified the possible role of fibroblasts in this process, highlighting their response to the above mentioned physical stimuli in terms of gene expression and cytokine production. Finally, calcium imaging-based experiments demonstrated a higher functionality of the stimulated co-cultures.
authors: The BioRobotics Institute, Scuola Superiore Sant'Anna, Pontedera, Pisa, Italy.
authors: Leonardo
authors: L
authors: Ricotti
authors: Toshinori
authors: T
authors: Fujie
authors: Helena
authors: H
authors: Vazão
authors: Gianni
authors: G
authors: Ciofani
authors: Roberto
authors: R
authors: Marotta
authors: Rosaria
authors: R
authors: Brescia
authors: Carlo
authors: C
authors: Filippeschi
authors: Irene
authors: I
authors: Corradini
authors: Michela
authors: M
authors: Matteoli
authors: Virgilio
authors: V
authors: Mattoli
authors: Lino
authors: L
authors: Ferreira
authors: Arianna
authors: A
authors: Menciassi
mesh_terms: D000180:Acrylic Resins / Q000494:pharmacology; D000818:Animals; D001896:Boron Compounds / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D018920:Coculture Techniques / Q000379:methods*; D055119:Elastic Modulus / Q000187:drug effects; D004558:Electric Stimulation; D004705:Endocytosis / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D005353:Fibronectins / Q000494:pharmacology; D006801:Humans; D020100:Hydrogels / Q000494:pharmacology*; D051379:Mice; D055616:Microtechnology / Q000379:methods*; D032446:Myoblasts / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D043942:Nanotubes / Q000737:chemistry*; D000099166:Polyacrylamides
chemical_list: D000180:Acrylic Resins; D001896:Boron Compounds; D002118:Calcium; D005353:Fibronectins; D020100:Hydrogels; D000099166:Polyacrylamides; C017282:boron nitride
references: Biomaterials. 2011 May;32(14):3575-83
references: 21324402
references: Eur J Neurosci. 1997 Apr;9(4):800-8
references: 9153587
references: J Biomech. 2010 May 28;43(8):1514-21
references: 20189177
references: J Biomed Mater Res A. 2004 Jun 1;69(3):373-81
references: 15127383
references: Pathobiology. 2006;73(6):288-94
references: 17374966
references: Am J Physiol Heart Circ Physiol. 2005 Apr;288(4):H1620-6
references: 15550526
references: Science. 2010 Aug 27;329(5995):1078-81
references: 20647425
references: Biotechnol Bioeng. 2008 Nov 1;101(4):850-8
references: 18512259
references: Science. 2007 Sep 7;317(5843):1366-70
references: 17823347
references: Methods Enzymol. 1998;298:489-96
references: 9751904
references: Int J Nanomedicine. 2012;7:19-24
references: 22275819
references: Science. 2005 Nov 18;310(5751):1139-43
references: 16293750
references: Biotechnol Bioeng. 2010 Jun 1;106(2):285-94
references: 20148416
references: ACS Nano. 2010 Oct 26;4(10):6267-77
references: 20925390
references: IEEE Trans Biomed Eng. 2013 Mar;60(3):727-34
references: 23380842
references: J Tissue Eng Regen Med. 2009 Aug;3(6):407-15
references: 19575392
references: Am J Physiol Cell Physiol. 2013 Jan 15;304(2):C128-36
references: 23114963
references: Biotechnol Prog. 2007 Jan-Feb;23(1):265-8
references: 17269697
references: Skelet Muscle. 2011 May 04;1(1):18
references: 21798098
references: Biomed Microdevices. 2011 Apr;13(2):255-66
references: 20981490
references: J Cell Mol Med. 2004 Oct-Dec;8(4):413-22
references: 15601570
references: Clin Plast Surg. 1999 Oct;26(4):647-56, ix-x
references: 10553219
references: Adv Funct Mater. 2008;18(9):1378-1389
references: 18841249
references: Tissue Eng Part A. 2009 Sep;15(9):2447-57
references: 19292666
references: Int J Nanomedicine. 2010 Apr 15;5:285-98
references: 20463944
references: FASEB J. 2001 May;15(7):1169-80
references: 11344085
references: Tissue Eng Part A. 2012 Aug;18(15-16):1664-76
references: 22607460
references: Cell Mol Bioeng. 2008 Sep 1;1(2-3):133-145
references: 19774099
references: FASEB J. 2006 Dec;20(14):2570-2
references: 17060399
references: Biomaterials. 2007 Apr;28(13):2202-10
references: 17267031
references: Am J Physiol Cell Physiol. 2007 May;292(5):C1660-71
references: 17151142
references: Biomed Microdevices. 2010 Oct;12(5):809-19
references: 20552402
references: J Biomed Mater Res A. 2008 Mar 15;84(4):1094-101
references: 17685407
references: Biomaterials. 2012 Jul;33(21):5230-46
references: 22521491
references: Pflugers Arch. 1993 Mar;422(6):591-8
references: 8469610
references: Biomed Mater. 2012 Jun;7(3):035010
references: 22477772
references: Tissue Eng Part A. 2009 Apr;15(4):935-44
references: 18821844
references: Biotechnol Bioeng. 2009 Feb 1;102(2):624-31
references: 18958861
references: J Cell Biol. 2004 Sep 13;166(6):877-87
references: 15364962
references: Biomaterials. 2006 Aug;27(24):4340-7
references: 16650470
references: Cell Motil Cytoskeleton. 2005 Jan;60(1):24-34
references: 15573414
references: Acta Biomater. 2010 Jun;6(6):1948-57
references: 20040385
references: Biomed Microdevices. 2012 Dec;14(6):987-98
references: 22960907
references: Nano Lett. 2006 Mar;6(3):537-42
references: 16522058
references: Cell Motil Cytoskeleton. 2004 Jul;58(3):200-11
references: 15146538
references: Br J Pharmacol. 2005 Oct;146(3):389-96
references: 16041403
references: Arterioscler Thromb Vasc Biol. 2009 May;29(5):725-31
references: 19229069