title: A comparative analysis of the in vitro effects of pulsed electromagnetic field treatment on osteogenic differentiation of two different mesenchymal cell lineages.
abstract: Human mesenchymal stem cells (MSCs) are a promising candidate cell type for regenerative medicine and tissue engineering applications. Exposure of MSCs to physical stimuli favors early and rapid activation of the tissue repair process. In this study we investigated the in vitro effects of pulsed electromagnetic field (PEMF) treatment on the proliferation and osteogenic differentiation of bone marrow MSCs (BM-MSCs) and adipose-tissue MSCs (ASCs), to assess if both types of MSCs could be indifferently used in combination with PEMF exposure for bone tissue healing. We compared the cell viability, cell matrix distribution, and calcified matrix production in unstimulated and PEMF-stimulated (magnetic field: 2 mT, amplitude: 5 mV) mesenchymal cell lineages. After PEMF exposure, in comparison with ASCs, BM-MSCs showed an increase in cell proliferation (p<0.05) and an enhanced deposition of extracellular matrix components such as decorin, fibronectin, osteocalcin, osteonectin, osteopontin, and type-I and -III collagens (p<0.05). Calcium deposition was 1.5-fold greater in BM-MSC-derived osteoblasts (p<0.05). The immunofluorescence related to the deposition of bone matrix proteins and calcium showed their colocalization to the cell-rich areas for both types of MSC-derived osteoblast. Alkaline phosphatase activity increased nearly 2-fold (p<0.001) and its protein content was 1.2-fold higher in osteoblasts derived from BM-MSCs. The quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis revealed up-regulated transcription specific for bone sialoprotein, osteopontin, osteonectin, and Runx2, but at a higher level for cells differentiated from BM-MSCs. All together these results suggest that PEMF promotion of bone extracellular matrix deposition is more efficient in osteoblasts differentiated from BM-MSCs.
authors.affiliation: Department of Public Health, Neuroscience, and Experimental & Forensic Medicine, University of Pavia , Pavia, Italy . ; Center for Tissue Engineering (C.I.T.), University of Pavia , Pavia, Italy .
authors.forename: Gabriele
authors.initials: G
authors.lastname: Ceccarelli
authors.forename: Nora
authors.initials: N
authors.lastname: Bloise
authors.forename: Melissa
authors.initials: M
authors.lastname: Mantelli
authors.forename: Giulia
authors.initials: G
authors.lastname: Gastaldi
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Fassina
authors.forename: Maria Gabriella Cusella
authors.initials: MG
authors.lastname: De Angelis
authors.forename: Davide
authors.initials: D
authors.lastname: Ferrari
authors.forename: Marcello
authors.initials: M
authors.lastname: Imbriani
authors.forename: Livia
authors.initials: L
authors.lastname: Visai
keywords: human adipose-derived stem cells; human mesenchymal stem cells; osteogenic differentiation; pulsed electromagnetic field
references.citation: Int J Artif Organs. 2011 Mar;34(3):259-70
references.pmid: 21374561
references.citation: J Cell Physiol. 2007 Apr;211(1):121-30
references.pmid: 17187344
references.citation: Biomaterials. 2012 Mar;33(7):2025-31
references.pmid: 22177287
references.citation: Clin Cases Miner Bone Metab. 2009 May;6(2):149-54
references.pmid: 22461165
references.citation: Bioelectromagnetics. 2002 Jul;23(5):398-405
references.pmid: 12111759
references.citation: Cell Biol Int. 1996 Jul;20(7):459-64
references.pmid: 8931312
references.citation: Cell Mol Biol Lett. 2011 Jun;16(2):236-57
references.pmid: 21394447
references.citation: Invest Ophthalmol Vis Sci. 2003 Apr;44(4):1704-15
references.pmid: 12657612
references.citation: Bioelectromagnetics. 2007 Oct;28(7):519-28
references.pmid: 17516509
references.citation: J Cell Physiol. 1990 Jun;143(3):420-30
references.pmid: 1694181
references.citation: J Biol Chem. 2005 Sep 30;280(39):33132-40
references.pmid: 16043491
references.citation: J Bone Miner Res. 1993 Dec;8(12):1499-506
references.pmid: 8304052
references.citation: J Bone Miner Res. 1992 Jun;7(6):683-92
references.pmid: 1414487
references.citation: Clin Chem. 1979 Aug;25(8):1519-20
references.pmid: 455710
references.citation: Clin Orthop Relat Res. 1989 Mar;(240):270-80
references.pmid: 2645077
references.citation: Bull Exp Biol Med. 2005 Jul;140(1):138-43
references.pmid: 16254640
references.citation: Keio J Med. 2005 Sep;54(3):132-41
references.pmid: 16237275
references.citation: Biotechnol Bioeng. 2012 Jan;109(1):252-61
references.pmid: 21965169
references.citation: Curr Stem Cell Res Ther. 2012 Mar;7(2):127-33
references.pmid: 22023633
references.citation: J Orthop Res. 2005 Sep;23(5):1035-46
references.pmid: 15936919
references.citation: Clin Orthop Relat Res. 1998 Oct;(355 Suppl):S90-104
references.pmid: 9917630
references.citation: J Cell Biochem. 1997 Feb;64(2):278-94
references.pmid: 9027588
references.citation: J Orthop Res. 2007 Sep;25(9):1213-20
references.pmid: 17503520
references.citation: Calcif Tissue Int. 1994 Nov;55(5):376-80
references.pmid: 7866919
references.citation: Methods. 2008 Jun;45(2):115-20
references.pmid: 18593609
references.citation: J Bone Joint Surg Br. 1980 Nov;62-B(4):465-70
references.pmid: 6968752
references.citation: Clin Cases Miner Bone Metab. 2012 Jan;9(1):24-7
references.pmid: 22783331
references.citation: Bioelectromagnetics. 1999;20(3):177-82
references.pmid: 10194560
references.citation: J Athl Train. 2007 Oct-Dec;42(4):530-5
references.pmid: 18174942
references.citation: Clin Orthop Relat Res. 1981 Nov-Dec;(161):58-66
references.pmid: 6975692
references.citation: Exp Biol Med (Maywood). 2009 Jan;234(1):1-9
references.pmid: 19109553
references.citation: N Engl J Med. 2003 Aug 7;349(6):570-82
references.pmid: 12904523
references.citation: Stem Cells Dev. 2012 Sep 20;21(14):2724-52
references.pmid: 22468918
references.citation: J Bone Joint Surg Am. 1966 Jul;48(5):915-23
references.pmid: 5942807
references.citation: Arch Immunol Ther Exp (Warsz). 2011 Oct;59(5):369-78
references.pmid: 21789625
references.citation: Orthopedics. 1998 Mar;21(3):297-302
references.pmid: 9547814
references.citation: Science. 1999 Apr 2;284(5411):143-7
references.pmid: 10102814
references.citation: J Biochem. 2004 Sep;136(3):377-86
references.pmid: 15598896
references.citation: Cells Tissues Organs. 2003;174(3):101-9
references.pmid: 12835573
references.citation: Chin Med J (Engl). 2008 Oct 20;121(20):2095-9
references.pmid: 19080282
references.citation: FASEB J. 1993 Dec;7(15):1475-82
references.pmid: 8262332
references.citation: J Bone Miner Res. 1994 Apr;9(4):487-96
references.pmid: 7518179
references.citation: J Cell Biochem. 2003 May 1;89(1):9-18
references.pmid: 12682904
references.citation: J Cell Biochem. 2006 Dec 1;99(5):1233-9
references.pmid: 16795049
references.citation: J Struct Biol. 2008 Jun;162(3):468-79
references.pmid: 18424074
references.citation: Bioelectromagnetics. 2010 Apr;31(3):209-19
references.pmid: 19866474
references.citation: J Musculoskelet Neuronal Interact. 2001 Sep;2(1):71-6
references.pmid: 15758478
references.citation: J Cell Biochem. 2006 Dec 1;99(5):1285-97
references.pmid: 16795045
references.citation: Tissue Eng Part A. 2010 Mar;16(3):995-1008
references.pmid: 19839719
references.citation: J Cell Biochem. 1993 May;52(1):37-41
references.pmid: 8320273
references.citation: Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2087-92
references.pmid: 10051599
references.citation: Arch Med Res. 2012 May;43(4):274-82
references.pmid: 22704852
references.citation: Differentiation. 1990 Sep;44(3):207-15
references.pmid: 2272412
references.citation: Cancer Res. 2007 Oct 1;67(19):9142-9
references.pmid: 17909019
references.citation: Exp Cell Res. 1995 Jan;216(1):35-45
references.pmid: 7813631
references.citation: Int J Immunopathol Pharmacol. 2011 Jan-Mar;24(1 Suppl 2):1-6
references.pmid: 21669129
references.citation: Eur Cell Mater. 2011 Jan 14;21:59-72; discussion 72
references.pmid: 21240845
references.citation: J Cell Biochem. 2003 Jan 1;88(1):104-12
references.pmid: 12461779
references.citation: J Biomech Eng. 2011 Sep;133(9):091005
references.pmid: 22010740
references.citation: Biomaterials. 2012 May;33(13):3515-28
references.pmid: 22333987
references.citation: Cytometry. 1998 Jan 1;31(1):1-9
references.pmid: 9450519
references.citation: J Biomed Mater Res A. 2008 Dec 1;87(3):750-9
references.pmid: 18200542
references.citation: J Tissue Eng Regen Med. 2015 Oct;9(10):1161-71
references.pmid: 23281198