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PubMed2003

The effects of hyaluronic acid on fibronectin fragment mediated cartilage chondrolysis in skeletally mature rabbits.

OBJECTIVE: Intraarticular Na-Hyaluronate (HA) exerts a beneficial effect on adolescent rabbits after fibronectin fragment (Fn-f) mediated cartilage injury. We extended our studies to a population of rabbits which have reached full skeletal maturity. DESIGN: Adult male NZW rabbits received an injury with Fn-f and no further treatment; an injection of HA followed by Fn-f injury, or Fn-f injury followed by a single or weekly intraarticular injection of HA. All animals were sacrificed 38 days after receiving the Fn-f injury. After sacrifice, proteoglycan (PG) content was determined from articular cartilage from the medial femoral condyles and tibial plateaus. The patellae were processed for histology. RESULTS: Cartilage PG contents were significantly reduced after Fn-f injection (P=0.0167) and were only slightly improved with HA pre-treatment. However, post-treatment with HA resulted in significant improvements in cartilage PG content when compared to Fn-f only (single HA, P=0.01; weekly HA,P =0.01). Loss of Safranin-O staining, cell loss, osteophyte formation and inflammation were present in the patellae following Fn-f injection. Pre-treatment with HA reduced these changes. More significant protection of cartilage and restoration of Fn-f injury were noted in animals receiving post-treatment with HA. CONCLUSIONS: These results suggest that 38 days after Fn-f injury the lost PG content induced by Fn-f injection is substantially restored by weekly post-treatment with intraarticular HA in rabbits that have reached full skeletal maturity.

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PubMed2002

Antisense oligonucleotides to the integrin receptor subunit alpha(5) decrease fibronectin fragment mediated cartilage chondrolysis.

OBJECTIVE: To investigate involvement of the integrin alpha(5) subunit of the classical fibronectin receptor in cartilage chondrolytic activities of fibronectin fragments (Fn-f). DESIGN: Bovine chondrocytes and cartilage explants were cultured in the presence of antisense oligonucleotide (ASO), or sense (SO) or scrambled sequence oligonucleotide (SCO) corresponding to the bovine alpha(5) subunit. The effects of the oligonucleotides on mRNA and protein expression of the alpha(5) subunit were analysed by rtPCR and Western blotting, respectively. To test effects on Fn-f activities, three different Fn-f were first added to serum or serum-free cultures, followed by addition of oligonucleotides and the effects on Fn-f mediated proteoglycan (PG) degradation, cartilage PG depletion and PG and general protein synthesis suppression were tested. RESULTS: The ASO decreased alpha(5) mRNA and protein expression to 69% and 55%, respectively, in monolayer cultures and decreased protein expression 67% in cartilage explants, while SO and SCO were ineffective. The ASO partially reversed the ability of the Fn-fs to suppress PG and general protein synthesis in cartilage explant and high density chondrocyte cultures. Concentrations of ASO from 1 nM to 5 microM effectively suppressed Fn-f activities in particular assays and the effects were reversible, while SO and SCO were not significantly effective. ASO also suppressed, in a dose-dependent and reversible fashion, the ability of the Fn-fs to enhance degradation and release of PG from cartilage explants. The ASO were also effective in suppressing the ability of an antibody to the alpha(5) subunit to enhance PG degradation, but were ineffective in blocking endotoxin or IL-1beta enhanced degradation. CONCLUSIONS: These data implicate the alpha(5) integrin subunit in Fn-f mediated activities, consistent with a role for the alpha(5)beta(1) integrin in this pathway.

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