Extracellular matrix recovery by human articular chondrocytes after treatment with hyaluronan hexasaccharides or Streptomyces hyaluronidase

Nishida, Yoshihiro; Knudson, Cheryl B.; Knudson, Warren

doi:10.1007/s101650300009

Cited by 9 publications

(12 citation statements)

References 18 publications

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“…In RT-PCR analysis, the gene expressions of type II collagen and COMP were up-regulated by the treatment with HA 6 . Furthermore, expression of HA synthase 2 mRNA was increased 2.1-fold by HA oligosaccharides, and aggrecan mRNA was increased 1.8-fold by HA oligosaccharides in human articular chondrocytes (47). These results also suggest that anabolic responses, as evidenced by matrix synthesis, take place simultaneously with the catabolic responses induced by small HA oligosaccharides.…”

Section: Discussionmentioning

confidence: 62%

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Ohno¹,

Im²,

Knudson³

et al. 2005

Arthritis & Rheumatism

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Objective. To document the activity profile of transcription factors following chondrocyte stimulation with hyaluronan (HA) hexasaccharides (HA 6 ) and to determine the expression of genes whose transcriptional activation is tightly associated with the transcription factors.Methods. Nuclear extracts from bovine articular chondrocytes treated with HA 6 were subjected to transcription factor protein-DNA array analysis. Electrophoretic mobility shift assay (EMSA) analyses were performed to confirm the results of protein-DNA array. The gene expressions of matrix metalloproteinase 3 (MMP-3), type II collagen, and cartilage oligomeric matrix protein (COMP) were examined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR), and protease activity was assessed by casein zymography.Results. In the protein-DNA array analysis, 12 transcription factors were up-regulated and 2 transcription factors were down-regulated in the chondrocytes treated with HA 6 . The transcription factors retinoic acid receptor (RAR), retinoid X receptor (RXR), and Sp-1 exhibited >2-fold increased activity by HA 6 treatment, as confirmed by EMSA. RT-PCR analysis showed that the expression levels of MMP-3, type II collagen, and COMP messenger RNA, which are tightly associated with the activation of RAR, RXR, or Sp-1, were upregulated by treatment with HA 6 . Addition of high molecular mass HA after HA 6 treatment resulted in abrogation of the MMP-3 induction. Conclusion.These results suggest that HA 6 increase the activity of multiple transcription factors in chondrocytes and signal the enhanced expression of key genes involved in cartilage-matrix remodeling and turnover. The data also demonstrate that high molecular mass HA has a potential to suppress the signaling activated by HA 6 .The glycosaminoglycan hyaluronan (HA) is a critical component of the extracellular matrix of articular cartilage. HA serves as a scaffold for the aggregation of cartilage proteoglycan (1) and, in this manner, forms a continuum of structure throughout the extracellular matrix. In addition, we have shown that HA facilitates the anchorage of these proteoglycan aggregates to the chondrocyte cell surface via its interaction with the membrane HA receptor CD44 (2-4). Thus, it is, in part, the interaction of chondrocytes directly with HA that constitutes the cell-matrix interaction that serves to regulate many aspects of chondrocyte metabolism (5). We have shown previously that one method of interference with the cell-matrix interactions of chondrocytes is through the use of excess small HA oligosaccharides, such as HA hexasaccharides (HA 6 ) (2,4,6). HA 6 compete with high molecular mass HA for CD44 binding; yet, they are too small in size to interfere with HA aggrecan or HA-link protein interactions (1,7,8).The consequences of this loss of cell-matrix interactions can be dramatic. In a previous study, we demonstrated that the addition of small HA oligosaccharides to human or bovine articular cartilage slices resulted in the apparent activation o...

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Section: Discussionmentioning

confidence: 62%

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Ohno¹,

Im²,

Knudson³

et al. 2005

Arthritis & Rheumatism

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“…However, as discussed above, exposure of chondrocytes to IL-1α or IL-1β results in the stimulation of NO, MMP-13, HA synthase-2 (HAS-2) and CD44 but, a dramatic inhibition of aggrecan and type II collagen. HA oligos affect a stimulation of NO, MMP-13 and HAS-2 but, little change in CD44 and a stimulation of aggrecan and type II collagen Nishida et al, 2003;Ohno et al, 2005). In addition, whereas IL-1 mediated signaling terminates in an activation of AP-1 transcription factors, HA oligos affect no activation of AP-1 (Ohno et al, 2005).…”

Section: Discussionmentioning

confidence: 96%

“…Moreover, in vitro studies have demonstrated that HA oligosaccharides alter chondrocyte metabolism and induce chondrocytic chondrolysis. Enhanced MMP activity was observed after HA oligosaccharide treatment of both cartilage explants and chondrocytes in culture balanced by increased synthesis of both HA and aggrecan, but no change in expression of CD44 receptors (Nishida, Knudson, & Knudson, 2003;Ohno, Im, Knudson, & Knudson, 2005). Thus, disruption of HA-chondrocyte interactions by HA oligosaccharides initiates both anabolic and catabolic responses.…”

Section: Introductionmentioning

confidence: 96%

Hyaluronan fragments activate nitric oxide synthase and the production of nitric oxide by articular chondrocytes

Iacob

Knudson

2006

The International Journal of Biochemistry & Cell Biology

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Chondrocyte CD44 receptors anchor hyaluronan to the cell surface, enabling the assembly and retention of proteoglycan aggregates in the pericellular matrix. Hyaluronan-CD44 interactions also provide signaling important for maintaining cartilage homeostasis. Disruption of chondrocyte-hyaluronan contact alters CD44 occupancy, initiating alternative signaling cascades. Treatment with hyaluronan oligosaccharides is one approach to uncouple CD44 receptors from its native ligand, hyaluronan. In bovine articular chondrocytes, treatment with hyaluronan oligosaccharides or purified hyaluronan hexasaccharides induced the production of nitric oxide that mirrored nitric oxide production following interleukin-1 treatment. In contrast, 120 and 1260 kDa hyaluronan did not induce production of nitric oxide. Human chondrocytes responded similarly to treatment with hyaluronan or hyaluronan oligosaccharides. Nitric oxide production from chondrocytes was mediated by activation of the inducible nitric oxide synthase, as confirmed by mRNA expression and inhibition of nitric oxide production by diphenyleneiodonium. Cotreatment of chondrocytes with hyaluronan oligosaccharides and interleukin-1 did not demonstrate additive effects. Blocking interleukin-1 receptors with an antagonist did not abolish the production of nitric oxide induced by treatment with hyaluronan oligosaccharides. Moreover, only COS-7 following transfection with a pCD44, not the CD44-null parental cells, responded to treatment with hyaluronan oligosaccharides by releasing nitric oxide. This study demonstrates a novel signaling potential by hyaluronan fragments, in lieu of endogenous hyaluronan-chondrocyte interactions, resulting in the activation of inducible nitric oxide synthase.

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“…After removal of HA8, the cell-associated matrices of MG-63 and articular chondrocytes were recovered. 32 Recovery of LM-8 cell matrices following washout were also observed (data not Mouse LM8 osteosarcoma cells were transplanted subcutaneously into C3H/He mice. From day 14 to day 23, 250 g/ml HA4, HA8, HMWHA, or control medium was injected daily, directly into the tumor mass.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, investigators have used small oligosaccharides of HA to deplete HA-rich matrices from cells. 31,32 The proposed mechanism is that these small oligosaccharides compete with the binding of high-molecular-mass HA with cell surface receptors such as CD44. Given their size, purity, and ease of permeability into tissues, the use of such small oligosaccharides may have suitable clinical applicability.…”

mentioning

confidence: 99%

Hyaluronan Oligosaccharides Inhibit Tumorigenicity of Osteosarcoma Cell Lines MG-63 and LM-8 in Vitro and in Vivo via Perturbation of Hyaluronan-Rich Pericellular Matrix of the Cells

Hosono

Nishida

Knudson

et al. 2007

The American Journal of Pathology

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Numerous studies have demonstrated a correlation between hyaluronan expression and the malignant properties of various kinds of cancer, and inhibition of hyaluronan production causes decreased tumor growth. Hyaluronan oligosaccharides have been shown to inhibit several tumor cell types via disruption of receptor-hyaluronan interaction. However, few studies have addressed hyaluronan with respect to osteosarcoma. In this study, we examined the effects of exogenously added hyaluronan oligosaccharides on tumorigenicity of murine osteosarcoma cells, LM-8, and human osteoblastic osteosarcoma cells, MG-63. Moreover, the critical size of oligomers needed to inhibit malignant properties was defined. Fluorescent hyaluronan oligosaccharides accumulated both on the surface of cells and in the cytoplasm, and this retention was blocked by pretreatment with an anti-CD44 monoclonal antibody. Hyaluronan octasaccharides significantly inhibited cell viability and induced apoptosis as defined by cell proliferation and terminal deoxynucleotidyl transferase dUTP nick-end labeling assays, respectively. Octasaccharides also abrogated functional cell-associated matrices and significantly reduced the retention of endogenous hyaluronan. Further, octasaccharide treatment affected an inhibition of cell motility as well as cell invasiveness. Pretreatment of the cells with anti-CD44 antibody reduced the antitumor effect of the octasaccharides. In vivo, intratumoral injection of hyaluronan octasaccharides reduced the hyaluronan accumulation in local tumors, resulting in significant suppression of the formation of distant lung metastasis. Together these data suggest that hyaluronan oligosaccharides have potent antitumor effects functioning in part by the abrogation of hyaluronan-rich cell-associated matrices.

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Extracellular matrix recovery by human articular chondrocytes after treatment with hyaluronan hexasaccharides or Streptomyces hyaluronidase

Cited by 9 publications

References 18 publications

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Hyaluronan fragments activate nitric oxide synthase and the production of nitric oxide by articular chondrocytes

Hyaluronan Oligosaccharides Inhibit Tumorigenicity of Osteosarcoma Cell Lines MG-63 and LM-8 in Vitro and in Vivo via Perturbation of Hyaluronan-Rich Pericellular Matrix of the Cells

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