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

Qu, Chengjuan; Pöytäkangas, Teemu; Jauhiainen, Marjo; Auriola, Seppo

doi:10.1007/s00441-009-0797-7

Cited by 7 publications

(7 citation statements)

References 49 publications

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“…Our previous study has shown that 5% O 2 significantly increases GAG synthesis and the expression of aggrecan and procollagen α 1 (II) mRNA when bovine primary chondrocytes are cultured in monolayers (Qu et al 2009). Thus, our hypothesis has been that a positive effect would be obtained when normal mature chondrocytes are cultured under hypoxic conditions.…”

Section: Discussionmentioning

confidence: 98%

“…The quantitative real-time RT-PCR analyses of these genes were performed by using the Mx3000P Real-Time PCR System (Stratagene, La Jolla, Calif., USA; Qu et al 2007). The primer sequences of GAPDH (Qu et al 2009), aggrecan (Bosnakovski et al 2006), procollagen α 1 (II) (Galois et al 2006), procollagen α 2 (I) (Galois et al 2006) and HAS2 (Schoenfelder and Einspanier 2003) used for quantitative RT-PCR in this study are available in the cited references.…”

Section: Pg Separation With Agarose Gel Electrophoresismentioning

confidence: 99%

“…Low (5%) O 2 might actually favour the production of PGs (Rajpurohit et al 1996;Saini and Wick 2004). Our previous study has indicated that 5% O 2 increases PG biosynthesis and the expression of mRNA for aggrecan and procollagen α 1 (II) in monolayer cultures of bovine primary chondrocytes (Qu et al 2009). Moreover, 5% O 2 enhances cartilage ECM production when human meniscus cells are cultured in three-dimensional cellaggregate cultures with fibroblast growth factor-2 (FGF-2; Adesida et al 2006).…”

Section: Introductionmentioning

confidence: 96%

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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: 98%

Section: Pg Separation With Agarose Gel Electrophoresismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 96%

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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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“…Renilla luciferase activity was used to normalize the transfection efficiency. The transfection efficiency was previously shown to be 27 % (Qu et al 2009). The experiment was repeated three times.…”

Section: Taurine Transporter Promoter Activity In Hypoand Hypertonic mentioning

confidence: 95%

“…Dual-luciferase ® reporter gene assay (Promega, Madison, WI, USA) was performed as previously described (Qu et al 2009), using taurine transporter firefly luciferase reporter plasmid, which contains −124 to +46 region of rat taurine transporter promoter (Han et al 2000;Ito et al 2004), and Renilla luciferase plasmid at 10:1 ratio. We also used a taurine transporter reporter plasmid, which has a mutated tonicity-responsive element (TonE) (Ito et al 2004) to verify that the induction of the reporter gene by hypertonicity was being regulated by this region.…”

Section: Taurine Transporter Promoter Activity In Hypoand Hypertonic mentioning

confidence: 99%

Chondrocytic cells express the taurine transporter on their plasma membrane and regulate its expression under anisotonic conditions

Karjalainen

Leskelä

et al. 2014

Amino Acids

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Taurine is a small organic osmolyte which participates in cell volume regulation. Chondrocytes have been shown to accumulate and release taurine; in bone, taurine participates in bone metabolism. However, its role in skeletal cells is poorly understood, especially in chondrocytes. This study investigated the regulation of taurine transporter in chondrocytic cells. We examined the transcriptional regulation of the taurine transporter under anisotonia by reporter gene and real-time RT-PCR assays. The effect of providing supplementary taurine on cell viability was evaluated with the lactate dehydrogenase release assay. The localization of the taurine transporter in human chondrosarcoma cells was studied by overexpressing a taurine transporter-enhanced green fluorescent protein. We observed that the transcription of the taurine transporter gene was up-regulated in hypertonic conditions. Hyperosmolarity-related cell death could be partly abolished by taurine supplementation in the medium. As expected, the fluorescently labeled taurine transporter localized at the plasma membrane. In polarized epithelial MDCK cells, the strongest fluorescence signal was located in the lateral cell membrane area. We also observed that the taurine transporter gene was expressed in several human tissues and malignant cell lines. This is the first study to present information on the transcriptional regulation of taurine transporter gene and the localization of the taurine transporter protein in chondrocytic cells.

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Hypertonic conditions enhance cartilage formation in scaffold-free primary chondrocyte cultures

Ylärinne

2014

Cell Tissue Res

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The potential of hypertonic conditions at in vivo levels to promote cartilage extracellular matrix accumulation in scaffold-free primary chondrocyte cultures was investigated. Six million bovine primary chondrocytes were cultured in transwell inserts in low glucose (LG), high glucose (HG), or hypertonic high glucose (HHG) DMEM supplemented with fetal bovine serum, antibiotics, and ascorbate under 5 % or 20 % O2 tension with and without transforming growth factor (TGF)-β3 for 6 weeks. Samples were collected for histological staining of proteoglycans (PGs) and type II collagen, analysis by quantitative reverse transcription plus the polymerase chain reaction (RT-PCR) of mRNA expression of aggrecan and procollagen α1 (II) and of Sox9 and procollagen α2 (I), and quantitation of PGs and PG separation in agarose gels. Cartilage tissues produced at 20 % O2 tension were larger than those formed at 5 % O2 tension. Compared with LG, the tissues grew to larger sizes in HG or HHG medium. Histological staining showed the strongest PG and type II collagen staining in cartilage generated in HG or HHG medium at 20 % O2 tension. Quantitative RT-PCR results indicated significantly higher expression of procollagen α1 (II) mRNA in cartilage generated in HHG medium at 20 % O2 tension compared with that in the other samples. TGF-β3 supplements in the culture medium provided no advantage for cartilage formation. Thus, HHG medium used at 20 % O2 tension is the most beneficial combination of the tested culture conditions for scaffold-free cartilage production in vitro and should improve cell culture for research into cartilage repair or tissue engineering.

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

Cited by 7 publications

References 49 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

Chondrocytic cells express the taurine transporter on their plasma membrane and regulate its expression under anisotonic conditions

Hypertonic conditions enhance cartilage formation in scaffold-free primary chondrocyte cultures

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