title: Engineered cartilage via self-assembled hMSC sheets with incorporated biodegradable gelatin microspheres releasing transforming growth factor-β1.
abstract: Self-assembling cell sheets have shown great potential for use in cartilage tissue engineering applications, as they provide an advantageous environment for the chondrogenic induction of human mesenchymal stem cells (hMSCs). We have engineered a system of self-assembled, microsphere-incorporated hMSC sheets capable of forming cartilage in the presence of exogenous transforming growth factor β1 (TGF-β1) or with TGF-β1 released from incorporated microspheres. Gelatin microspheres with two different degrees of crosslinking were used to enable different cell-mediated microsphere degradation rates. Biochemical assays, histological and immunohistochemical analyses, and biomechanical testing were performed to determine biochemical composition, structure, and equilibrium modulus in unconfined compression after 3 weeks of culture. The inclusion of microspheres with or without loaded TGF-β1 significantly increased sheet thickness and compressive equilibrium modulus, and enabled more uniform matrix deposition by comparison to control sheets without microspheres. Sheets incorporated with fast-degrading microspheres containing TGF-β1 produced significantly more GAG and GAG per DNA than all other groups tested and stained more intensely for type II collagen. These findings demonstrate improved cartilage formation in microsphere-incorporated cell sheets, and describe a tailorable system for the chondrogenic induction of hMSCs without necessitating culture in growth factor-containing medium.
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH, USA.
authors.forename: Loran D
authors.initials: LD
authors.lastname: Solorio
authors.forename: Eran L
authors.initials: EL
authors.lastname: Vieregge
authors.forename: Chirag D
authors.initials: CD
authors.lastname: Dhami
authors.forename: Phuong N
authors.initials: PN
authors.lastname: Dang
authors.forename: Eben
authors.initials: E
authors.lastname: Alsberg
mesh_terms: D002356:Cartilage / Q000166:cytology / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D005780:Gelatin / Q000008:administration & dosage*; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D008863:Microspheres; D023822:Tissue Engineering / Q000379:methods*; D053773:Transforming Growth Factor beta1 / Q000008:administration & dosage*
chemical_list: D006025:Glycosaminoglycans; D053773:Transforming Growth Factor beta1; D005780:Gelatin; D004247:DNA
references.citation: J Cell Physiol. 2005 May;203(2):398-409
references.pmid: 15521064
references.citation: Biomacromolecules. 2005 Mar-Apr;6(2):975-87
references.pmid: 15762668
references.citation: J Bone Joint Surg Am. 1998 Dec;80(12):1745-57
references.pmid: 9875932
references.citation: Biorheology. 2000;37(1-2):141-7
references.pmid: 10912186
references.citation: Tissue Eng Part A. 2010 Feb;16(2):513-21
references.pmid: 19715388
references.citation: Stem Cells. 2007 Nov;25(11):2786-96
references.pmid: 17656642
references.citation: Biotechnol Lett. 2010 Sep;32(9):1339-46
references.pmid: 20464452
references.citation: J Control Release. 2004 Jan 8;94(1):101-14
references.pmid: 14684275
references.citation: J Tissue Eng Regen Med. 2010 Oct;4(7):514-23
references.pmid: 20872738
references.citation: Tissue Eng. 1998 Winter;4(4):415-28
references.pmid: 9916173
references.citation: Acta Biomater. 2011 Jul;7(7):2797-803
references.pmid: 21549223
references.citation: Am J Sports Med. 2011 Apr;39(4):753-63
references.pmid: 21257846
references.citation: Exp Cell Res. 1998 Jan 10;238(1):265-72
references.pmid: 9457080
references.citation: Tissue Eng Part A. 2008 Dec;14(12):2041-9
references.pmid: 18636944
references.citation: Bone. 1992;13(1):81-8
references.pmid: 1581112
references.citation: Biomaterials. 2005 Dec;26(34):7095-103
references.pmid: 16023196
references.citation: J Biomech Eng. 1995 May;117(2):179-92
references.pmid: 7666655
references.citation: Expert Opin Biol Ther. 2006 Sep;6(9):847-66
references.pmid: 16918253
references.citation: J Biomed Mater Res A. 2010 Mar 1;92(3):1139-44
references.pmid: 19322820
references.citation: Tissue Eng. 2006 Dec;12(12):3307-39
references.pmid: 17518671
references.citation: Adv Drug Deliv Rev. 2007 May 30;59(4-5):263-73
references.pmid: 17507111
references.citation: Stem Cells. 2007 Dec;25(12):3244-51
references.pmid: 17872501
references.citation: J Mater Sci Mater Med. 2007 Feb;18(2):273-81
references.pmid: 17323158
references.citation: Tissue Eng Part A. 2009 Oct;15(10):3025-36
references.pmid: 19335060
references.citation: Osteoarthritis Cartilage. 2002 Jul;10(7):580-7
references.pmid: 12127839
references.citation: Biotechnol Bioeng. 2011 Jun;108(6):1421-9
references.pmid: 21274847
references.citation: Mol Pharm. 2011 Aug 1;8(4):994-1001
references.pmid: 21500855
references.citation: Knee. 2007 Jun;14(3):177-82
references.pmid: 17428666
references.citation: J Biomater Sci Polym Ed. 1999;10(1):63-78
references.pmid: 10091923
references.citation: Adv Drug Deliv Rev. 1998 May 4;31(3):287-301
references.pmid: 10837630
references.citation: J Biomater Sci Polym Ed. 2001;12(1):77-88
references.pmid: 11334191
references.citation: Arthritis Rheum. 2008 Jan;58(1):26-35
references.pmid: 18163497
references.citation: Biochim Biophys Acta. 1986 Sep 4;883(2):173-7
references.pmid: 3091074
references.citation: Tissue Eng. 2006 Apr;12(4):969-79
references.pmid: 16674308
references.citation: Tissue Eng. 2007 Apr;13(4):843-53
references.pmid: 17394383
references.citation: J Control Release. 2005 Dec 5;109(1-3):256-74
references.pmid: 16266768
references.citation: Tissue Eng. 2006 May;12(5):1337-44
references.pmid: 16771646
references.citation: Methods Mol Med. 2004;100:53-68
references.pmid: 15280587
references.citation: Tissue Eng. 2004 Nov-Dec;10(11-12):1695-706
references.pmid: 15684678
references.citation: Annu Rev Biomed Eng. 2002;4:175-209
references.pmid: 12117756
references.citation: Osteoarthritis Cartilage. 2007 Sep;15(9):1025-33
references.pmid: 17498976