title: A hexadecylamide derivative of hyaluronan (HYMOVIS®) has superior beneficial effects on human osteoarthritic chondrocytes and synoviocytes than unmodified hyaluronan.
abstract: BACKGROUND
  Intra-articular hyaluronan (HA) injection provides symptomatic benefit in the treatment of osteoarthritis (OA). Previously we found superior beneficial effects in a large animal OA model of a hexadecylamide derivative compared with unmodified HA of the same initial molecular weight. The current study sought to define possible molecular mechanisms whereby this enhanced relief of symptoms was occurring.
  
  
  METHODS
  Chondrocytes and synovial fibroblasts were isolated from tissues of patients undergoing arthroplasty for knee OA. Monolayer cultures of cells were treated with 0, 0.5, 1.0 or 1.5 mg/mL of unmodified HA (500-730 kDa) or a hexadecylamide derivative of HA of the same initial molecular weight (HYADD4®-G; HYMOVIS®) simultaneously or 1 hour before incubation with interleukin (IL)-1beta (2 ng/mL). Cultures were terminated 15 or 30 minutes later (chondrocytes and synovial fibroblasts, respectively) for quantitation of phosphorylated-(p)-JNK, p-NFkappaB, p-p38, or at 24 hours for quantitation of gene expression (MMP1 &13, ADAMTS4 &5, TIMP1 &3, CD44, COL1A1 &2A1, ACAN, PTGS2, IL6, TNF) and matrix metalloproteinase (MMP)-13 activity.
  
  
  RESULTS
  The hexadecylamide derivative of HA had significantly better amelioration of IL-1beta-induced gene expression of key matrix degrading enzymes (MMP1, MMP13, ADAMTS5), and inflammatory mediators (IL6, PTGS2) by human OA chondrocytes and synovial fibroblasts. Pre-incubation of cells with the derivatized HA for 1 hour prior to IL-1beta exposure significantly augmented the inhibition of MMP1, MMP13, ADAMTS4 and IL6 expression by chondrocytes. The reduction in MMP13 mRNA by the amide derivative of HA was mirrored in reduced MMP-13 protein and enzyme activity in IL-1beta-stimulated chondrocytes. This was associated in part with a greater inhibition of phosphorylation of the cell signalling molecules JNK, p38 and NF-kappaB.
  
  
  CONCLUSIONS
  The present studies have demonstrated several potential key mechanisms whereby the intra-articular injection of a hexadecylamide derivative of HA may be acting in joints with OA.
authors.affiliation: Raymond Purves Bone and Joint Research Laboratories; Kolling Institute of Medical Research, Institute of Bone and Joint Research, University of Sydney at Royal North Shore Hospital, St Leonards, NSW, 2065, Australia.
authors.forename: Margaret M
authors.initials: MM
authors.lastname: Smith
authors.affiliation: Raymond Purves Bone and Joint Research Laboratories; Kolling Institute of Medical Research, Institute of Bone and Joint Research, University of Sydney at Royal North Shore Hospital, St Leonards, NSW, 2065, Australia.
authors.forename: Amy K
authors.initials: AK
authors.lastname: Russell
authors.affiliation: Fidia Farmaceutici SpA, Abano Terme, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Schiavinato
authors.affiliation: Raymond Purves Bone and Joint Research Laboratories; Kolling Institute of Medical Research, Institute of Bone and Joint Research, University of Sydney at Royal North Shore Hospital, St Leonards, NSW, 2065, Australia.
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Little
keywords: Chondrocyte; Hyaluronan; Osteoarthritis; Synovial fibroblasts
references.citation: Arthritis Res Ther. 2009;11(1):203
references.pmid: 19232065
references.citation: Osteoarthritis Cartilage. 2008 Aug;16(8):947-55
references.pmid: 18258458
references.citation: Arthritis Rheum. 1990 Nov;33(11):1695-701
references.pmid: 2242066
references.citation: Biochem Biophys Res Commun. 2010 Dec 10;403(2):184-9
references.pmid: 21059338
references.citation: J Orthop Res. 2010 Jun;28(6):726-31
references.pmid: 20058278
references.citation: J Rheumatol. 2003 Jun;30(6):1164-72
references.pmid: 12784385
references.citation: J Rheumatol. 2006 May;33(5):946-50
references.pmid: 16652425
references.citation: Connect Tissue Res. 2013;54(1):49-54
references.pmid: 23020698
references.citation: Rheumatology (Oxford). 2008 Aug;47(8):1172-8
references.pmid: 18565987
references.citation: Osteoarthritis Cartilage. 2011 Jun;19(6):611-9
references.pmid: 21443958
references.citation: J Orthop Res. 2011 Feb;29(2):258-64
references.pmid: 20706991
references.citation: J Biol Chem. 2000 Sep 1;275(35):26967-75
references.pmid: 10871609
references.citation: J Mol Endocrinol. 2000 Oct;25(2):169-93
references.pmid: 11013345
references.citation: J Clin Immunol. 1995 Nov;15(6):300-11
references.pmid: 8576316
references.citation: J Biol Chem. 2003 Oct 10;278(41):39762-72
references.pmid: 12888570
references.citation: J Pathol. 2009 Jun;218(2):241-7
references.pmid: 19267386
references.citation: Arthritis Rheum. 2008 Apr;58(4):1055-66
references.pmid: 18383380
references.citation: J Trauma. 2010 Jan;68(1):146-52
references.pmid: 20065770
references.citation: Osteoarthritis Cartilage. 2005 Mar;13(3):216-24
references.pmid: 15727888
references.citation: J Clin Invest. 1996 Nov 15;98(10):2425-30
references.pmid: 8941662
references.citation: Biochim Biophys Acta. 1990 May 22;1052(3):366-78
references.pmid: 2162214
references.citation: J Rheumatol. 2000 Apr;27(4):997-1004
references.pmid: 10782829
references.citation: Arthritis Res Ther. 2005;7(4):R852-61
references.pmid: 15987487
references.citation: Osteoarthritis Cartilage. 2006 Dec;14(12):1237-47
references.pmid: 16806998
references.citation: Arthritis Rheum. 2011 Sep;63(9):2732-43
references.pmid: 21590680
references.citation: Agents Actions. 1992 Sep;37(1-2):155-6
references.pmid: 1456177
references.citation: Inflamm Res. 2007 Jun;56(6):246-53
references.pmid: 17607549
references.citation: Rheumatol Int. 2006 Jan;26(3):185-90
references.pmid: 15580348
references.citation: J Biomed Mater Res B Appl Biomater. 2012 Nov;100(8):2073-81
references.pmid: 22865740
references.citation: Trends Immunol. 2005 Dec;26(12):637-43
references.pmid: 16214414
references.citation: Semin Arthritis Rheum. 1993 Jun;22(6 Suppl 1):9-17
references.pmid: 8342053
references.citation: Arthritis Rheum. 2000 Apr;43(4):801-11
references.pmid: 10765924
references.citation: Osteoarthritis Cartilage. 1996 Mar;4(1):77-84
references.pmid: 8731398
references.citation: Osteoarthritis Cartilage. 1999 Mar;7(2):182-90
references.pmid: 10222217
references.citation: Knee Surg Relat Res. 2012 Sep;24(3):164-72
references.pmid: 22977794
references.citation: J Orthop Res. 2008 Jul;26(7):1032-7
references.pmid: 18302264
references.citation: Cell. 1990 Jun 29;61(7):1303-13
references.pmid: 1694723
references.citation: Arthritis Rheum. 2004 Feb;50(2):516-25
references.pmid: 14872494
references.citation: Rheumatol Int. 1987;7(3):113-22
references.pmid: 3671989
references.citation: Eur Spine J. 2007 Dec;16(12):2193-205
references.pmid: 17899219
references.citation: Tohoku J Exp Med. 2010 Mar;220(3):229-35
references.pmid: 20208419
references.citation: J Orthop Res. 2008 Sep;26(9):1230-7
references.pmid: 18404652
references.citation: Anal Biochem. 1985 Oct;150(1):76-85
references.pmid: 3843705
references.citation: Ann Rheum Dis. 2009 Jun;68(6):1051-8
references.pmid: 18662930
references.citation: J Rheumatol. 1998 Sep;25(9):1813-9
references.pmid: 9733465
references.citation: Osteoarthritis Cartilage. 2003 Dec;11(12):839-44
references.pmid: 14629959
references.citation: Mod Rheumatol. 2008;18(3):263-70
references.pmid: 18427719
references.citation: Arthritis Rheum. 1993 Feb;36(2):247-53
references.pmid: 8431214
references.citation: J Orthop Sci. 2008 Nov;13(6):514-23
references.pmid: 19089539
references.citation: J Immunol. 1999 Nov 1;163(9):5049-55
references.pmid: 10528210
references.citation: Eur Spine J. 2003 Feb;12(1):57-65
references.pmid: 12592548
references.citation: Mod Rheumatol. 2013 Nov;23(6):1116-23
references.pmid: 23224053
references.citation: J Pharmacol Exp Ther. 1996 Jun;277(3):1672-5
references.pmid: 8667237
references.citation: Biorheology. 2002;39(1-2):237-46
references.pmid: 12082286
references.citation: Biorheology. 2011;48(5):263-75
references.pmid: 22433568
references.citation: Clin Rheumatol. 2001;20(2):98-103
references.pmid: 11346239
references.citation: Cochrane Database Syst Rev. 2006 Apr 19;(2):CD005321
references.pmid: 16625635
references.citation: J Mol Endocrinol. 2002 Aug;29(1):23-39
references.pmid: 12200227
references.citation: Arthritis Res Ther. 2003;5(2):54-67
references.pmid: 12718745
references.citation: Nat Rev Rheumatol. 2011 Jan;7(1):33-42
references.pmid: 21119608
references.citation: J Rheumatol Suppl. 1995 Feb;43:109-14
references.pmid: 7752112
references.citation: Res Vet Sci. 1999 Oct;67(2):183-90
references.pmid: 10502490