title: Applied osmotic loading for promoting development of engineered cartilage.
abstract: This study investigated the potential use of static osmotic loading as a cartilage tissue engineering strategy for growing clinically relevant grafts from either synovium-derived stem cells (SDSCs) or chondrocytes. Bovine SDSCs and chondrocytes were individually encapsulated in 2% w/v agarose and divided into chondrogenic media of osmolarities 300 (hypotonic), 330 (isotonic), and 400 (hypertonic, physiologic) mOsM for up to 7 weeks. The application of hypertonic media to constructs comprised of SDSCs or chondrocytes led to increased mechanical properties as compared to hypotonic (300mOsM) or isotonic (330mOsM) media (p<0.05). Constant exposure of SDSC-seeded constructs to 400mOsM media from day 0 to day 49 yielded a Young's modulus of 513±89kPa and GAG content of 7.39±0.52%ww on day 49, well within the range of values of native, immature bovine cartilage. Primary chondrocyte-seeded constructs achieved almost as high a Young's modulus, reaching 487±187kPa and 6.77±0.54%ww (GAG) for the 400mOsM condition (day 42). These findings suggest hypertonic loading as a straightforward strategy for 3D cultivation with significant benefits for cartilage tissue engineering strategies. In an effort to understand potential mechanisms responsible for the observed response, cell volume measurements in response to varying osmotic conditions were evaluated in relation to the Boyle-van't Hoff (BVH) law. Results confirmed that chondrocytes behave as perfect osmometers; however SDSCs deviated from the BVH relation.
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY, USA.
authors.forename: Sonal R
authors.initials: SR
authors.lastname: Sampat
authors.forename: Matthew V
authors.initials: MV
authors.lastname: Dermksian
authors.forename: Sevan R
authors.initials: SR
authors.lastname: Oungoulian
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Winchester
authors.forename: J Chloë
authors.initials: JC
authors.lastname: Bulinski
authors.forename: Gerard A
authors.initials: GA
authors.lastname: Ateshian
authors.forename: Clark T
authors.initials: CT
authors.lastname: Hung
mesh_terms: D000818:Animals; D002356:Cartilage / Q000166:cytology / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D055119:Elastic Modulus; D009997:Osmotic Pressure / Q000502:physiology*; D013997:Time Factors; D023822:Tissue Engineering
keywords: Cartilage; Chondrocytes; Static osmotic loading; Synovium-derived stem cells; Tissue engineering
references.citation: Tissue Eng Part A. 2009 Aug;15(8):2065-72
references.pmid: 19196151
references.citation: Biomed Mater. 2010 Oct;5(5):055005
references.pmid: 20826911
references.citation: Arch Histol Cytol. 2003 May;66(2):145-53
references.pmid: 12846554
references.citation: Tissue Eng Part C Methods. 2010 Oct;16(5):957-63
references.pmid: 20001269
references.citation: Arthritis Rheum. 2009 Oct;60(10):3028-37
references.pmid: 19790068
references.citation: Tissue Eng Part A. 2011 Sep;17(17-18):2259-65
references.pmid: 21542714
references.citation: J Tissue Eng Regen Med. 2007 Nov-Dec;1(6):436-42
references.pmid: 18265417
references.citation: J Anat. 1994 Jun;184 ( Pt 3):493-501
references.pmid: 7928638
references.citation: Tissue Eng Part C Methods. 2011 Nov;17(11):1041-9
references.pmid: 21797756
references.citation: J Orthop Res. 2010 Oct;28(10):1283-91
references.pmid: 20225285
references.citation: Tissue Eng Part A. 2011 Feb;17(3-4):371-80
references.pmid: 20799905
references.citation: Arthritis Rheum. 2008 May;58(5):1389-98
references.pmid: 18438859
references.citation: Cryobiology. 2011 Jun;62(3):232-41
references.pmid: 21376029
references.citation: Biorheology. 2003;40(1-3):61-72
references.pmid: 12454388
references.citation: Ann Biomed Eng. 2002 Sep;30(8):1046-56
references.pmid: 12449765
references.citation: Birth Defects Res C Embryo Today. 2003 May;69(2):144-55
references.pmid: 12955858
references.citation: Ann Anat. 2010 Apr 20;192(2):116-21
references.pmid: 20149606
references.citation: Acta Biomater. 2013 Mar;9(3):5554-61
references.pmid: 23128162
references.citation: J Biomech. 2008 Nov 14;41(15):3253-9
references.pmid: 18823628
references.citation: Biomed Mater Eng. 2010;20(3):145-58
references.pmid: 20930322
references.citation: J Cell Physiol. 2006 Nov;209(2):481-92
references.pmid: 16897756
references.citation: J Orthop Res. 2001 Sep;19(5):768-78
references.pmid: 11562120
references.citation: Arthritis Rheum. 2003 May;48(5):1315-25
references.pmid: 12746904
references.citation: Tissue Eng Part A. 2010 Sep;16(9):2699-708
references.pmid: 20367254
references.citation: Cell Mol Bioeng. 2008 Dec 1;1(4):339-348
references.pmid: 20011231
references.citation: Clin Orthop Relat Res. 2008 Aug;466(8):1880-9
references.pmid: 18512111
references.citation: J Cell Physiol. 1993 Feb;154(2):262-70
references.pmid: 8425907
references.citation: Arthritis Rheum. 2001 Aug;44(8):1928-42
references.pmid: 11508446
references.citation: Am J Physiol Cell Physiol. 2006 Oct;291(4):C718-25
references.pmid: 16928775
references.citation: Open Rheumatol J. 2011;5:100-6
references.pmid: 22279508
references.citation: Tissue Eng. 2007 Jun;13(6):1227-34
references.pmid: 17518725
references.citation: Differentiation. 2008 Dec;76(10):1044-56
references.pmid: 18637024
references.citation: Clin Orthop Relat Res. 1988 Oct;(235):289-95
references.pmid: 3416536
references.citation: Clin Chim Acta. 1967 Nov;18(2):267-73
references.pmid: 4864804
references.citation: Osteoarthritis Cartilage. 2001 Nov;9(8):761-70
references.pmid: 11795996
references.citation: Cell Physiol Biochem. 2011;28(6):1111-22
references.pmid: 22179000
references.citation: Tissue Eng. 2007 Oct;13(10):2465-73
references.pmid: 17665999
references.citation: Br J Rheumatol. 1994 Oct;33(10):901-8
references.pmid: 7921748
references.citation: Tissue Eng. 2003 Aug;9(4):597-611
references.pmid: 13678439
references.citation: Macromol Biosci. 2010 Nov 10;10(11):1355-64
references.pmid: 20857388
references.citation: Osteoarthritis Cartilage. 2007 Sep;15(9):1025-33
references.pmid: 17498976