title: Ligament-derived matrix stimulates a ligamentous phenotype in human adipose-derived stem cells.
abstract: Human adipose stem cells (hASCs) can differentiate into a variety of phenotypes. Native extracellular matrix (e.g., demineralized bone matrix or small intestinal submucosa) can influence the growth and differentiation of stem cells. The hypothesis of this study was that a novel ligament-derived matrix (LDM) would enhance expression of a ligamentous phenotype in hASCs compared to collagen gel alone. LDM prepared using phosphate-buffered saline or 0.1% peracetic acid was mixed with collagen gel (COL) and was evaluated for its ability to induce proliferation, differentiation, and extracellular matrix synthesis in hASCs over 28 days in culture at different seeding densities (0, 0.25 x 10(6), 1 x 10(6), or 2 x 10(6) hASC/mL). Biochemical and gene expression data were analyzed using analysis of variance. Fisher's least significant difference test was used to determine differences between treatments following analysis of variance. hASCs in either LDM or COL demonstrated changes in gene expression consistent with ligament development. hASCs cultured with LDM demonstrated more dsDNA content, sulfated-glycosaminoglycan accumulation, and type I and III collagen synthesis, and released more sulfated-glycosaminoglycan and collagen into the medium compared to hASCs in COL (p <or= 0.05). Increased seeding density increased DNA content incrementally over 28 days in culture for LDM but not COL constructs (p <or= 0.05). These findings suggest that LDM can stimulate a ligament phenotype by hASCs, and may provide a novel scaffold material for ligament engineering applications.
authors.affiliation: Division of Orthopaedic Surgery, Department of Surgery, Duke University Medical Center, Durham, North Carolina 27710, USA.
authors.forename: Dianne
authors.initials: D
authors.lastname: Little
authors.forename: Farshid
authors.initials: F
authors.lastname: Guilak
authors.forename: David S
authors.initials: DS
authors.lastname: Ruch
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D000818:Animals; D003094:Collagen / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D008022:Ligaments / Q000378:metabolism*; D008875:Middle Aged; D010641:Phenotype; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D034421:Sus scrofa; D013710:Tendons / Q000166:cytology / Q000378:metabolism; D054457:Tissue Scaffolds
chemical_list: D006025:Glycosaminoglycans; C013786:A73025; D003094:Collagen; D004247:DNA
references.citation: Int J Exp Pathol. 2009 Apr;90(2):95-100
references.pmid: 19335547
references.citation: Biol Cell. 1992;75(1):1-9
references.pmid: 1381252
references.citation: Crit Rev Oncol Hematol. 2004 Mar;49(3):199-202
references.pmid: 15036260
references.citation: J Cell Biochem. 2006 Aug 15;98(6):1436-49
references.pmid: 16518859
references.citation: J Orthop Res. 2000 Jul;18(4):557-64
references.pmid: 11052491
references.citation: Methods Mol Biol. 2008;449:69-79
references.pmid: 18370084
references.citation: J Histochem Cytochem. 1990 Oct;38(10):1405-11
references.pmid: 1698203
references.citation: Z Orthop Ihre Grenzgeb. 2004 Mar-Apr;142(2):248-53
references.pmid: 15106072
references.citation: Hum Genet. 2005 Jan;116(1-2):51-61
references.pmid: 15517394
references.citation: Eur J Cell Biol. 1994 Aug;64(2):264-70
references.pmid: 7813514
references.citation: J Orthop Res. 2005 Jan;23(1):164-74
references.pmid: 15607889
references.citation: Biomaterials. 2005 Aug;26(23):4805-16
references.pmid: 15763260
references.citation: Clin Orthop Relat Res. 2002 Apr;(397):414-23
references.pmid: 11953636
references.citation: Biochem Biophys Res Commun. 2002 Jun 7;294(2):371-9
references.pmid: 12051722
references.citation: Am J Sports Med. 2003 Jul-Aug;31(4):555-9
references.pmid: 12860544
references.citation: Anal Biochem. 1996 Dec 1;243(1):189-91
references.pmid: 8954546
references.citation: J Biol Chem. 1994 Jan 14;269(2):931-5
references.pmid: 8288646
references.citation: Genome Biol. 2007;8(5):215
references.pmid: 17543136
references.citation: J Am Acad Orthop Surg. 2008 Apr;16(4):177-87
references.pmid: 18390480
references.citation: Acta Biomater. 2007 Sep;3(5):651-61
references.pmid: 17513181
references.citation: Knee Surg Sports Traumatol Arthrosc. 2008 Sep;16(9):834-42
references.pmid: 18516592
references.citation: J Shoulder Elbow Surg. 2008 Jan-Feb;17(1 Suppl):35S-39S
references.pmid: 18201655
references.citation: Int Orthop. 2007 Dec;31(6):783-9
references.pmid: 17583812
references.citation: Am J Vet Res. 2008 Jan;69(1):148-56
references.pmid: 18167101
references.citation: Hum Mol Genet. 1993 Jul;2(7):961-8
references.pmid: 8364578
references.citation: Proc Natl Acad Sci U S A. 1977 Oct;74(10):4453-7
references.pmid: 270689
references.citation: Arthroscopy. 2006 Jul;22(7):700-9
references.pmid: 16843804
references.citation: J Biomed Mater Res. 2000 Aug;51(2):233-40
references.pmid: 10825223
references.citation: J Biomed Mater Res A. 2008 Feb;84(2):500-7
references.pmid: 17618495
references.citation: Biochem Biophys Res Commun. 2006 May 5;343(2):644-52
references.pmid: 16554022
references.citation: J Orthop Res. 2006 Apr;24(4):811-9
references.pmid: 16514641
references.citation: Tissue Eng Part A. 2008 Oct;14(10):1615-27
references.pmid: 18759661
references.citation: Biomed Mater. 2008 Sep;3(3):034002
references.pmid: 18689924
references.citation: J Orthop Res. 2003 Mar;21(2):238-44
references.pmid: 12568954
references.citation: Tissue Eng. 2006 Oct;12(10):2905-12
references.pmid: 17518658
references.citation: Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3049-54
references.pmid: 17360607
references.citation: J Orthop Res. 2009 Jul;27(7):964-71
references.pmid: 19030174
references.citation: J Cell Biol. 2001 Jul 23;154(2):459-68
references.pmid: 11470832
references.citation: Foot Ankle Spec. 2009 Oct;2(5):235-9
references.pmid: 19825780
references.citation: J Orthop Res. 2009 Jul;27(7):937-42
references.pmid: 19105224
references.citation: Clin Orthop Relat Res. 1999 Oct;(367 Suppl):S333-43
references.pmid: 10546657
references.citation: J Tissue Eng Regen Med. 2009 Jan;3(1):43-53
references.pmid: 19039798
references.citation: J Biochem. 1979 Jul;86(1):111-9
references.pmid: 573261
references.citation: Tissue Eng Part A. 2009 Sep;15(9):2435-45
references.pmid: 19292650
references.citation: Biomaterials. 2003 Sep;24(21):3805-13
references.pmid: 12818553
references.citation: J Biomech Eng. 2005 Feb;127(1):181-5
references.pmid: 15868800
references.citation: J Periodontol. 2000 Jun;71(6):974-80
references.pmid: 10914801
references.citation: J Hand Surg Am. 2007 May-Jun;32(5):597-605
references.pmid: 17481995
references.citation: Biomaterials. 2005 Dec;26(36):7530-6
references.pmid: 16045982
references.citation: Arthroscopy. 2006 May;22(5):534-8
references.pmid: 16651164
references.citation: Tissue Eng. 2006 Oct;12(10):2853-62
references.pmid: 17518654
references.citation: Plast Reconstr Surg. 2003 Nov;112(6):1613-9
references.pmid: 14578792
references.citation: J Orthop Res. 2003 May;21(3):420-31
references.pmid: 12706014
references.citation: Cell Differ Dev. 1989 Jul;27(2):103-20
references.pmid: 2475230
references.citation: Amino Acids. 2006 Jun;30(4):445-51
references.pmid: 16583319
references.citation: Matrix Biol. 1994 Jan;14(1):87-99
references.pmid: 8061923
references.citation: Am J Sports Med. 2002 Mar-Apr;30(2):287-305
references.pmid: 11912103
references.citation: Tissue Eng. 2006 Apr;12(4):681-9
references.pmid: 16674283
references.citation: Tissue Eng Part B Rev. 2009 Jun;15(2):171-93
references.pmid: 19207040
references.citation: Wound Repair Regen. 2008 May-Jun;16(3):370-8
references.pmid: 18471255
references.citation: J Neurol. 2009 Jan;256(1):13-27
references.pmid: 19221853
references.citation: Ann Biomed Eng. 1993 May-Jun;21(3):289-305
references.pmid: 8328728
references.citation: Plast Reconstr Surg. 2005 Oct;116(5):1393-404; discussion 1405-6
references.pmid: 16217485
references.citation: J Biomed Mater Res B Appl Biomater. 2005 Aug;74(2):789-91
references.pmid: 15981174
references.citation: J Orthop Res. 2008 Mar;26(3):364-73
references.pmid: 17972327
references.citation: J Biomech Eng. 2000 Dec;122(6):570-5
references.pmid: 11192376
references.citation: J Orthop Res. 2008 Jan;26(1):1-9
references.pmid: 17676628
references.citation: Biochem Biophys Res Commun. 2002 Jan 18;290(2):763-9
references.pmid: 11785965
references.citation: Biomaterials. 2006 Jul;27(19):3675-83
references.pmid: 16519932
references.citation: Science. 1991 Jun 21;252(5013):1705-8
references.pmid: 1646484
references.citation: Tissue Eng. 2005 Nov-Dec;11(11-12):1887-97
references.pmid: 16411835
references.citation: Tissue Eng. 2006 Jun;12(6):1405-18
references.pmid: 16846339
references.citation: Annu Rev Biomed Eng. 2004;6:131-56
references.pmid: 15255765
references.citation: Nucleic Acids Res. 2001 May 1;29(9):e45
references.pmid: 11328886
references.citation: Acta Orthop. 2005 Jun;76(3):421-7
references.pmid: 16156473
references.citation: J Biol Chem. 1986 Mar 25;261(9):4337-45
references.pmid: 3456347
references.citation: J Orthop Res. 2003 Jul;21(4):617-21
references.pmid: 12798060
references.citation: Nat Med. 2007 Oct;13(10):1219-27
references.pmid: 17828274
references.citation: Eur J Cell Biol. 2009 May;88(5):273-83
references.pmid: 19167776
references.citation: J Biol Chem. 1998 Dec 4;273(49):33054-63
references.pmid: 9830060
references.citation: FEBS Lett. 1995 Jul 17;368(2):307-10
references.pmid: 7628627
references.citation: Birth Defects Res C Embryo Today. 2008 Sep;84(3):228-44
references.pmid: 18773462
references.citation: J Orthop Res. 2004 Jan;22(1):214-20
references.pmid: 14656683
references.citation: Front Biosci. 2004 May 01;9:1422-32
references.pmid: 14977557
references.citation: Cytotherapy. 2005;7(5):447-55
references.pmid: 16236634
references.citation: J Bone Joint Surg Am. 2007 Mar;89(3):621-30
references.pmid: 17332112
references.citation: Orthop Rev. 1989 Aug;18(8):857-63
references.pmid: 2671872
references.citation: J Hand Surg Am. 2006 Oct;31(8):1279-87
references.pmid: 17027787
references.citation: Nature. 1990 Jul 19;346(6281):281-4
references.pmid: 2374594
references.citation: Tissue Eng Part A. 2008 Apr;14(4):559-69
references.pmid: 18377199
references.citation: Tissue Eng Part A. 2008 Sep;14(9):1489-95
references.pmid: 18694325
references.citation: J Shoulder Elbow Surg. 2007 Sep-Oct;16(5 Suppl):S171-8
references.pmid: 17560804
references.citation: Cell Tissue Bank. 2004;5(1):23-36
references.pmid: 15256837
references.citation: Dev Biol. 2006 Oct 1;298(1):234-47
references.pmid: 16876153
references.citation: Acta Biomater. 2009 Jan;5(1):1-13
references.pmid: 18938117
references.citation: Tissue Cell. 2009 Apr;41(2):85-9
references.pmid: 18789468
references.citation: Anat Rec. 2002 Dec 1;268(4):430-40
references.pmid: 12420291
references.citation: J Orthop Res. 1995 Mar;13(2):184-90
references.pmid: 7722755
references.citation: Tissue Eng Part A. 2008 Dec;14(12):2089-104
references.pmid: 19093294
references.citation: J Orthop Res. 2006 Jun;24(6):1299-309
references.pmid: 16649228
references.citation: J Orthop Res. 2002 Nov;20(6):1170-4
references.pmid: 12472225
references.citation: Tissue Eng Part A. 2009 Feb;15(2):231-41
references.pmid: 18950290
references.citation: J Orthop Res. 1997 Jan;15(1):18-23
references.pmid: 9066522
references.citation: J Mater Sci Mater Med. 2007 Apr;18(4):537-43
references.pmid: 17546412
references.citation: Biomaterials. 2002 Sep;23(17):3717-31
references.pmid: 12109697
references.citation: Science. 1965 Nov 12;150(3698):893-9
references.pmid: 5319761
references.citation: J Orthop Res. 2008 Feb;26(2):255-63
references.pmid: 17763435
references.citation: J Cell Biochem. 1999 Jun 15;73(4):445-57
references.pmid: 10733339
references.citation: Int J Obes Relat Metab Disord. 1994 Aug;18(8):532-6
references.pmid: 7951472
references.citation: Kidney Int. 1990 Feb;37(2):689-95
references.pmid: 2407884
references.citation: Biomaterials. 2008 May;29(15):2378-87
references.pmid: 18313139
references.citation: J Biomech. 2007;40(8):1686-1693
references.pmid: 17056048