The lack of effect of glucosamine sulphate on aggrecan mRNA expression and 35S-sulphate incorporation in bovine primary chondrocytes

Qu, Chengjuan; Karjalainen, Hannu M.; Helminen, Heikki J.

doi:10.1016/j.bbadis.2006.01.005

Cited by 26 publications

(20 citation statements)

References 35 publications

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“…On the other hand, studies with human and MC615 mouse chondrocytes have indicated that glucosamine does not stimulate CS synthesis, and a high concentration of glucosamine even inhibits CS synthesis Silbert 2003, 2004). In our previous studies (Qu et al 2006(Qu et al , 2007, we have been unable to see any positive effect of GS on ECM production or gene expression even at 1 mM concentrations. Since the supplementation period (10 days) was relatively short compared with the recommended minimum used time for GS, we decided to test whether a remarkably longer exposure to GS would show beneficial effects.…”

Section: Discussionmentioning

confidence: 81%

“…Preparation of chondrocyte-seeded culture inserts Isolated primary chondrocytes were obtained from the articular cartilage of bovine (estimated age: 13-22 months) femoral condyles by collagenase (Sigma-Aldrich, St. Louis, Mo., USA) digestion (Qu et al 2006). Subsequently, the isolated chondrocytes were washed twice with PBS, counted, and plated at cell density of 6×10 6 chondrocytes per transwell-COL insert, which was located inside a 24-well plate.…”

Section: Equilibrium Of Inserts Before Cell Seedingmentioning

confidence: 99%

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Five percent oxygen tension is not beneficial for neocartilage formation in scaffold-free cell cultures

Lindeberg

Ylärinne

et al. 2012

Cell Tissue Res

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We have investigated whether 5% oxygen tension (O(2)) is beneficial for neocartilage formation when chondrocytes are cultured in transwell-COL inserts. Six million bovine primary chondrocytes were cultured in an insert with DMEM supplemented with 10% fetal bovine serum and antibiotics, with or without glucosamine sulphate (GS) in a 5% or 20% O(2) environment for 2, 4, or 6 weeks. The samples were collected for the histological staining of proteoglycans (PGs) and type II collagen, quantitative reverse transcription with the polymerase chain reaction (RT-PCR) analyses of the mRNA expression of aggrecan and procollagen α(1)(II), procollagen α(2)(I) and hyaluronan synthase 2, quantitation of PGs, and agarose gel electrophoresis. Neocartilage produced at 20% O(2) appeared larger than that at 5% O(2). Histological staining showed that more PGs and type II collagen and better native cartilage structure were produced at 20% than at 5% O(2). The thickness of neocartilage increased during the culture period. Quantitative RT-PCR showed that the procollagen α(1)(II) mRNA expression level was significantly higher at 20% than at 5% O(2). However, no significant difference in gene expression and PG content was found between control and GS-treated cultures at either 20% or 5% O(2). Thus, in contrast to monolayer cultures, engineered cartilage from scaffold-free cultured chondrocytes at 20% O(2) produced better extracellular matrix (ECM) than that at 5% O(2). PGs were mainly large. Exogenous GS was not beneficial for the ECM in scaffold-free chondrocyte cultures.

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Section: Discussionmentioning

confidence: 81%

Section: Equilibrium Of Inserts Before Cell Seedingmentioning

confidence: 99%

Five percent oxygen tension is not beneficial for neocartilage formation in scaffold-free cell cultures

Lindeberg

Ylärinne

et al. 2012

Cell Tissue Res

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“…Therefore, a question has been raised whether such a low concentration can increase cartilage PG synthesis, as has been suggested for OA chondrocytes (Bassleer et al 1998;Dodge and Jimenez 2003). Previously, we have been unable to confirm that exogenous GS in low-or highglucose DMEM increases aggrecan mRNA expression or GAG synthesis (Qu et al 2006(Qu et al , 2007a.…”

Section: Discussionmentioning

confidence: 94%

“…[ 35 S]-sulphate (5 µCi/ml) was added into each well of the 12-well plates and then the cultures were incubated in humidified 5% or 20% O 2 incubators at 37°C with 5% CO 2 for 24 h. The supernatants were collected and the amounts of incorporated and free [ 35 S]-sulphate were analysed as in our previous study (Qu et al 2006). The volumes used for the analysis were so adjusted that a good separation between the GAG incorporated and the free unincorporated label could be achieved.…”

Section: [ 35 S]-sulphate Incorporation Analysismentioning

confidence: 99%

“…On the other hand, studies with human and MC615 mouse chondrocytes have indicated that GlcN cannot stimulate chondroitin sulphate (CS) synthesis, and that a high concentration of GlcN can even inhibit CS synthesis Silbert 2003, 2004). Our previous studies have shown that GS salt at 20% O 2 tension does not increase aggrecan mRNA expression or GAG synthesis (Qu et al 2006), and the intracellular UDP-N-acetylhexosamines (UDP-HexN) level in bovine primary chondrocytes increases only at a concentration of 1 mM GS (Qu et al 2007a). An increased level of sulphate alone after the administration of GS has also been suggested to mediate its effect, because of the increased sulphate concentration in serum after the oral administration of GS (Hoffer et al 2001).…”

Section: Introductionmentioning

confidence: 97%

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Glucosamine sulphate does not increase extracellular matrix production at low oxygen tension

Pöytäkangas

Jauhiainen

et al. 2009

Cell Tissue Res

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Low oxygen tension may change the dependence of chondrocytes on exogenous carbohydrate sources. In this study, we have investigated whether glucosamine sulphate (GS) stimulates proteoglycan synthesis, the mRNA expression of aggrecan and of type II collagen, and UDP-sugar levels in bovine primary chondrocytes under a low oxygen (O(2)) atmosphere. Chondrocytes from bovine femoral condyles were cultivated with or without GS or sulphate at various concentrations in low- (5.5 mM) or high-glucose (25 mM) DMEM under either a 5% or 20% O(2) atmosphere for 2 or 8 days after isolation. The mRNA expression of aggrecan and type II collagen and the synthesis of glycosaminoglycan (GAG) were determined by quantitative real-time reverse transcription with polymerase chain reaction and a [(35)S]-sulphate incorporation assay, respectively. Aggrecan promoter activity was analysed by a dual-luciferase reporter gene assay. Intracellular UDP-N-acetylhexosamines (UDP-HexN), UDP-glucuronic acid and UDP-hexoses were analysed by reversed-phase high-performance liquid chromatography electrospray ionization mass spectrometry. A low (5%) O(2) atmosphere significantly increased GAG synthesis, mRNA expression of aggrecan and of type II collagen and aggrecan promoter activity in bovine primary chondrocytes. A high (1 mM) concentration of GS was required to increase the level of UDP-HexN. However, GS did not increase GAG synthesis, aggrecan promoter activity or mRNA expression of aggrecan and of type II collagen. Interestingly, a 5% O(2) atmosphere increased the level of UDP-HexN in 8-day cultures without GS treatment. Thus, exogenous GS does not change chondrocyte metabolism, whereas a 5% O(2) atmosphere stimulates extracellular matrix production in bovine primary chondrocytes. The balance of UDP-sugars is changed under a 5% O(2) atmosphere for longer culture periods.

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The effects of glucosamine hydrochloride on subchondral bone changes in an animal model of osteoarthritis

Wang

Laverty

Dumitriu

et al. 2007

Arthritis & Rheumatism

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Objective. To quantify periarticular subchondral bone changes in a rabbit model of experimental osteoarthritis (OA), and to determine the effects of continuous administration of a clinically relevant dose of glucosamine HCl on subchondral bone changes in this model.Methods. Anterior cruciate ligament transection (ACLT) was performed on the left femorotibial joints of 16 rabbits to induce OA. Ten rabbits that did not undergo ACLT served as unoperated controls. Eight rabbits that underwent ACLT and 6 control rabbits were treated with 100 mg of glucosamine daily, and the others were given a placebo. The articular cartilage was evaluated macroscopically and graded at the time of necropsy, 8 weeks after ACLT. Bone mineral density (BMD) was measured using dual-energy x-ray absorptiometry on the dissected distal femur and proximal tibia. Subchondral trabecular bone turnover, architecture, and connectivity, as well as subchondral plate thickness and mineralization were studied on the undecalcified tibia sections from each animal.Results. Eight weeks after ACLT, most of the operated joints had various degrees of cartilage damage and fibrillation. Compared with the control group, the ACLT group had significantly increased subchondral bone turnover and lower BMD, bone volume, connectivity, and bone mineralization. The high bone turnover was significantly reduced in glucosamine-treated animals that underwent ACLT. In fact, there were no significant differences between the ACLT/glucosamine group and the control/glucosamine group in most of the bone parameters studied.Conclusion. This study shows that subchondral bone turnover, structure, and mineralization are significantly altered in the early stages of experimental OA, and that these changes are attenuated by glucosamine treatment.

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The lack of effect of glucosamine sulphate on aggrecan mRNA expression and 35S-sulphate incorporation in bovine primary chondrocytes

Cited by 26 publications

References 35 publications

Five percent oxygen tension is not beneficial for neocartilage formation in scaffold-free cell cultures

Five percent oxygen tension is not beneficial for neocartilage formation in scaffold-free cell cultures

Glucosamine sulphate does not increase extracellular matrix production at low oxygen tension

The effects of glucosamine hydrochloride on subchondral bone changes in an animal model of osteoarthritis

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