title: Osteogenic Differentiation of MSC through Calcium Signaling Activation: Transcriptomics and Functional Analysis.
abstract: The culture of progenitor mesenchymal stem cells (MSC) onto osteoconductive materials to induce a proper osteogenic differentiation and mineralized matrix regeneration represents a promising and widely diffused experimental approach for tissue-engineering (TE) applications in orthopaedics. Among modern biomaterials, calcium phosphates represent the best bone substitutes, due to their chemical features emulating the mineral phase of bone tissue. Although many studies on stem cells differentiation mechanisms have been performed involving calcium-based scaffolds, results often focus on highlighting production of in vitro bone matrix markers and in vivo tissue ingrowth, while information related to the biomolecular mechanisms involved in the early cellular calcium-mediated differentiation is not well elucidated yet. Genetic programs for osteogenesis have been just partially deciphered, and the description of the different molecules and pathways operative in these differentiations is far from complete, as well as the activity of calcium in this process. The present work aims to shed light on the involvement of extracellular calcium in MSC differentiation: a better understanding of the early stage osteogenic differentiation program of MSC seeded on calcium-based biomaterials is required in order to develop optimal strategies to promote osteogenesis through the use of new generation osteoconductive scaffolds. A wide spectrum of analysis has been performed on time-dependent series: gene expression profiles are obtained from samples (MSC seeded on calcium-based scaffolds), together with related microRNAs expression and in vivo functional validation. On this basis, and relying on literature knowledge, hypotheses are made on the biomolecular players activated by the biomaterial calcium-phosphate component. Interestingly, a key role of miR-138 was highlighted, whose inhibition markedly increases osteogenic differentiation in vitro and enhance ectopic bone formation in vivo. Moreover, there is evidence that Ca-P substrate triggers osteogenic differentiation through genes (SMAD and RAS family) that are typically regulated during dexamethasone (DEX) induced differentiation.
authors.affiliation: Institute of Biophysics, National Research Council, Genoa, Italy.|Institute of Biomedical Technologies, National Research Council, Segrate (Mi), Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Viti
authors.affiliation: Institute of Biomedical Technologies, National Research Council, Segrate (Mi), Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Landini
authors.affiliation: Institute of Biomedical Technologies, National Research Council, Segrate (Mi), Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Mezzelani
authors.affiliation: Institute of Biophysics, National Research Council, Genoa, Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Petecchia
authors.affiliation: Institute of Biomedical Technologies, National Research Council, Segrate (Mi), Italy.
authors.forename: Luciano
authors.initials: L
authors.lastname: Milanesi
authors.affiliation: Institute of Electronics, Computer and Telecommunication Engineering, National Research Council, Genoa, Italy.|Advanced Biotechnology Center (CBA), Genoa, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Scaglione
mesh_terms: D000818:Animals; D020013:Calcium Signaling / Q000235:genetics*; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000235:genetics*; D030541:Databases, Genetic; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D006801:Humans; D007107:Immune System / Q000378:metabolism; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D058977:Molecular Sequence Annotation; D010012:Osteogenesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D013194:Staining and Labeling
chemical_list: D035683:MicroRNAs; D012333:RNA, Messenger
references.citation: Biomaterials. 1993 Apr;14(5):341-8
references.pmid: 8389612
references.citation: J Cell Mol Med. 2013 Jul;17(7):823-32
references.pmid: 23672518
references.citation: Genome Biol. 2003;4(10):R70
references.pmid: 14519205
references.citation: Nucleic Acids Res. 2015 Jan;43(Database issue):D1113-6
references.pmid: 25361974
references.citation: J Cell Biochem. 2006 Jan 1;97(1):56-70
references.pmid: 16229015
references.citation: Mol Biol Cell. 2002 Dec;13(12):4279-95
references.pmid: 12475952
references.citation: Nat Biotechnol. 2000 Sep;18(9):959-63
references.pmid: 10973216
references.citation: Biomed Mater Eng. 1999;9(5-6):319-24
references.pmid: 10822487
references.citation: J Mater Sci Mater Med. 2003 Mar;14(3):211-8
references.pmid: 15348466
references.citation: J Cell Biochem. 2002;85(4):737-46
references.pmid: 11968014
references.citation: Drug Discov Today. 2004 Dec 1;9(23):994
references.pmid: 15574312
references.citation: Brain Behav Evol. 2001;58(5):296-305
references.pmid: 11978947
references.citation: Biomaterials. 2006 Jun;27(17):3230-7
references.pmid: 16488007
references.citation: J Bone Miner Res. 1998 Jul;13(7):1101-11
references.pmid: 9661074
references.citation: J Mater Sci Mater Med. 2008 Apr;19(4):1565-73
references.pmid: 18299963
references.citation: Biomaterials. 2012 Apr;33(11):3205-15
references.pmid: 22285104
references.citation: Nucleic Acids Res. 2005 Jan 1;33(Database issue):D54-8
references.pmid: 15608257
references.citation: Nat Genet. 2001 Dec;29(4):365-71
references.pmid: 11726920
references.citation: Nucleic Acids Res. 2004 Jan 1;32(Database issue):D258-61
references.pmid: 14681407
references.citation: Tissue Eng Part A. 2011 Apr;17(7-8):1083-97
references.pmid: 21091326
references.citation: Vision Res. 2003 Apr;43(8):927-36
references.pmid: 12668062
references.citation: Biomed Eng Online. 2011;10:9
references.pmid: 21269490
references.citation: Res Vet Sci. 2012 Oct;93(2):1021-5
references.pmid: 22280550
references.citation: J Endocrinol. 1999 Sep;162(3):371-9
references.pmid: 10467228
references.citation: PLoS One. 2011;6(10):e26211
references.pmid: 22022571
references.citation: Bioinformatics. 2003 Jan 22;19(2):185-93
references.pmid: 12538238
references.citation: J Cell Mol Med. 2010 Jun;14(6B):1845-56
references.pmid: 19538476
references.citation: Biochem Biophys Res Commun. 2004 Apr 9;316(3):651-8
references.pmid: 15033449
references.citation: Genomics. 2012 Sep;100(3):141-8
references.pmid: 22735742
references.citation: Cell Res. 2009 Jan;19(1):128-39
references.pmid: 19114990
references.citation: J Biomed Mater Res A. 2009 Aug;90(2):533-42
references.pmid: 18563821
references.citation: Plast Reconstr Surg. 2005 Sep 15;116(4):1044-52
references.pmid: 16163093
references.citation: Bone. 1999 Aug;25(2 Suppl):51S-54S
references.pmid: 10458275
references.citation: J Surg Res. 2013 Nov;185(1):278-85
references.pmid: 23972621
references.citation: Sci Rep. 2012;2:274
references.pmid: 22355786
references.citation: Proc Natl Acad Sci U S A. 2011 Apr 12;108(15):6139-44
references.pmid: 21444814
references.citation: J Biomed Mater Res A. 2010 May;93(2):567-75
references.pmid: 19591232
references.citation: Clin Orthop Relat Res. 1991 Jan;(262):298-311
references.pmid: 1984928
references.citation: Exp Cell Res. 1988 Apr;175(2):266-76
references.pmid: 3360056
references.citation: Nucleic Acids Res. 2003 Jan 1;31(1):258-61
references.pmid: 12519996
references.citation: J Biomed Mater Res. 2000 Mar 5;49(3):328-37
references.pmid: 10602065
references.citation: Bone. 1992;13(1):81-8
references.pmid: 1581112
references.citation: J Biomed Mater Res B Appl Biomater. 2014 Jul;102(5):922-32
references.pmid: 24259264
references.citation: PLoS One. 2010;5(3):e9690
references.pmid: 20300619
references.citation: Bone. 1998 May;22(5 Suppl):131S-134S
references.pmid: 9600769
references.citation: J Mater Sci Mater Med. 2010 May;21(5):1649-54
references.pmid: 20162336
references.citation: Acta Physiol (Oxf). 2014 Jun;211(2):285-96
references.pmid: 24702694
references.citation: Vitam Horm. 2011;87:417-80
references.pmid: 22127254
references.citation: Biomaterials. 1990 Sep;11(7):513-7
references.pmid: 2242401
references.citation: J Biol Chem. 1998 Feb 6;273(6):3129-31
references.pmid: 9452419
references.citation: PLoS One. 2014;9(4):e94353
references.pmid: 24728045
references.citation: Nucleic Acids Res. 2011 Jul;39(Web Server issue):W307-15
references.pmid: 21646343
references.citation: Biomed Res Int. 2013;2013:154919
references.pmid: 23862134
references.citation: J Nippon Med Sch. 2003 Aug;70(4):300-6
references.pmid: 12928709
references.citation: J Orthop Res. 2000 Nov;18(6):933-44
references.pmid: 11192254
references.citation: Cytotherapy. 2011 Jul;13(6):661-74
references.pmid: 21250865
references.citation: N Engl J Med. 2001 Feb 1;344(5):385-6
references.pmid: 11195802
references.citation: Plast Reconstr Surg. 2013 Sep;132(3):580-9
references.pmid: 23985633
references.citation: J Clin Invest. 2000 Jun;105(12):1663-8
references.pmid: 10862779
references.citation: Acta Biomater. 2011 Oct;7(10):3573-85
references.pmid: 21723966
references.citation: Med Sci (Paris). 2004 Mar;20(3):265-7
references.pmid: 15067567
references.citation: J Hematother Stem Cell Res. 2000 Dec;9(6):841-8
references.pmid: 11177595
references.citation: Nature. 2001 Nov 1;414(6859):118-21
references.pmid: 11689957
references.citation: FASEB J. 2007 May;21(7):1472-80
references.pmid: 17264165
references.citation: Biomaterials. 1998 Jan-Feb;19(1-3):133-9
references.pmid: 9678860
references.citation: MedGenMed. 2001 Feb 5;:E1
references.pmid: 11320340
references.citation: Biomaterials. 2005 Sep;26(27):5474-91
references.pmid: 15860204
references.citation: Stem Cell Res. 2014 Jan;12(1):153-65
references.pmid: 24239963
references.citation: Ann Transplant. 2004;9(4):34-6
references.pmid: 15884434
references.citation: Nature. 1996 Aug 1;382(6590):448-52
references.pmid: 8684484
references.citation: J Biomed Inform. 2011 Oct;44(5):839-47
references.pmid: 21605702
references.citation: Biomaterials. 2004 Aug;25(19):4639-45
references.pmid: 15120510
references.citation: Genome Biol. 2007;8(9):R183
references.pmid: 17784955
references.citation: Curr Pharm Biotechnol. 2011 Feb 1;12(2):151-9
references.pmid: 21044012
references.citation: Nucleic Acids Res. 2011 Jan;39(Database issue):D163-9
references.pmid: 21071411