Colony Formation and Matrix Production by Human Anulus Cells

Gruber, Helen E.; Leslie, Kelly; Ingram, Jane A.; Hoelscher, Gretchen L.; Norton, H. James; Hanley, Edward N.

doi:10.1097/01.brs.0000129029.10036.64

Cited by 25 publications

(25 citation statements)

References 25 publications

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“…One recent study showed that human disc annulus fibrosus cells cultured in a 3D pellet system were able to produce a proteoglycan/collagen II matrix, and responded to TGF-b1 by increasing proteoglycan synthesis as compared with monolayer culture. 24 Intriguingly, findings from Gruber et al 25 demonstrated that even annulus cells derived from Figure 7. Real-time RT-PCR results showed there were no statistically significant differences in the gene expression levels between the alginate and alginate/chitosan scaffold group (n ¼ 10 in each group, data showed as Mean 6 SD).…”

Section: Discussionmentioning

confidence: 92%

Developing an alginate/chitosan hybrid fiber scaffold for annulus fibrosus cells

Shao

Hunter

2007

J Biomedical Materials Res

108

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A fibrous scaffold made of alginate or alginate/chitosan was fabricated for annulus fibrosus (AF) cell culture using a wet-spinning and lyophilization technique. The scaffolds were evaluated using several in vitro tests. Scanning electron microscopy showed the scaffold fibers generally aligned in one direction with individual fiber diameters varying between 40-100 microm. The alginate/chitosan hybrid scaffold exhibited a slower degradation rate, while both scaffold types did not display any cyto-toxicity to 3T3 fibroblasts and could maintain canine AF cell growth. The AF cells retained their spherical shape within the fibrous scaffold at the beginning of the culture period and formed into cell clusters at later times. Specific extracellular matrix molecules, including collagen I, collagen II, and aggrecan, could be detected in the AF cell clusters. These results demonstrate the feasibility of using this hybrid alginate/chitosan scaffold for AF cell culture, and the potential application for intervertebral disc tissue engineering.

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Section: Discussionmentioning

confidence: 92%

Developing an alginate/chitosan hybrid fiber scaffold for annulus fibrosus cells

Shao

Hunter

2007

J Biomedical Materials Res

108

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“…Finally, cells were encapsulated in 2% agarose disks (8mm in diameter, 1 mm thick) at a density of 2.4×10 7 cells/ml. Three-dimensional agarose culture has been shown to maintain the phenotype of IVD cells [23][24][25]. After an initial 24-hour culture in the highglucose DMEM containing 10% FBS and 1% antibiotic-antimycotic at 37°C, the agarose disks were separated into four groups which were incubated in the DMEM with glucose concentrations of 1, 2.5, 5, and 25 mM for another 24 hours before the oxygen consumption rate measurement.…”

Section: Methodsmentioning

confidence: 99%

Effects of Low Glucose Concentrations on Oxygen Consumption Rates of Intervertebral Disc Cells

Huang¹,

Yuan²,

Jackson³

et al. 2007

Spine

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Study Design-Investigation of the effects of low glucose concentrations on the oxygen consumption rates of intervertebral disc cells.Objectives-To determine the oxygen consumption rate of porcine annulus fibrosus (AF) cells at different glucose concentrations and to examine the differences in the oxygen consumption rate between AF and nucleus pulposus (NP) cells at different glucose levels.Summary of Background Data-Poor nutrient supply has been suggested as a potential mechanism for degeneration of the intervertebral disc (IVD). Distribution of nutrients in the IVD is strongly dependent on transport properties of the tissue and cellular metabolic rates. Previous studies have shown dependence of oxygen consumption rate of IVD cells on oxygen tension, pH levels, and glucose levels outside the physiological range. However, the oxygen consumption rate of AF cells at in vivo glucose levels has not been investigated.Methods-IVD cells were isolated from the outer AF and NP of 4-5 month-old porcine lumbar discs. The changes in oxygen tension were recorded when cells were cultured in sealed metabolism chamber. The oxygen consumption rate of cells was determined by theoretical curve fitting using the Michaelis-Menten equation.Results-The outer AF cells cultured in high glucose medium (25mM) exhibited the lowest oxygen consumption rate whereas no significant differences in oxygen consumption rates were found among outer AF cells cultured at physiological glucose levels (i.e., 1 mM, 2.5 mM, 5 mM). The oxygen consumption rate of NP cells was significantly greater than that of outer AF cells.Conclusions-Since the oxygen consumption rates determined in this study are comparable to the findings in the literature, this study has developed a new alternative method for determining oxygen consumption rate. The oxygen consumption rates of IVD cells reported in this study will be valuable for theoretically predicting local oxygen concentrations in IVD, which can provide a better understanding of transport of oxygen in the discs.

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“…These include a stimulation of synthesis of proteoglycan and collagen type II; stimulation of cell proliferation and enhancement of cell survival (Gruber et al 1997(Gruber et al , 2004(Gruber et al , 2010Masuda and An 2004;Zhang et al 2006;Risbud et al 2006). Of relevance, Gruber et al (2010) have recently shown that adult human annulus cells grown under micromass conditions in the presence of TGF-increase their synthesis of ECM and sulphated PG.…”

Section: Typementioning

confidence: 97%

“…Furthermore, during ageing there is a decreased expression of TGF-s, and their receptors, within the IVD (Matsunaga et al 2003), which appears to correlate with degenerative changes in the disc. The TGF-s have also been shown to variously modulate IVD cell behaviour in vitro; promoting cell proliferation and extracellular matrix (ECM) synthesis-stimulating secretion and accumulation of proteoglycan and collagen type II-and enhancing cell viability (Gruber et al 1997(Gruber et al , 2004(Gruber et al , 2010Masuda and An 2004;Zhang et al 2006;Risbud et al 2006).…”

Section: Introductionmentioning

confidence: 98%

“…Osada et al 1996;Gruber et al 2000Gruber et al , 2004Gruber et al , 2008Masuda and An 2004;Zhang et al 2006;Pratsinis and Kletsas 2008). When used in combination with PDGF, they have an antiapoptotic eVect on human annulus cells (Gruber et al 2000) and, in concert with TGF-, they enhance their colony forming eYciency and ECM production (Gruber et al 2004).…”

Section: Introductionmentioning

confidence: 99%

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The response of foetal annulus fibrosus cells to growth factors: modulation of matrix synthesis by TGF-β1 and IGF-1

Hayes

Ralphs

2011

Histochem Cell Biol

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The annulus fibrosus of the intervertebral disc is a complex radial-ply tissue that derives initially from segmental condensations of axial mesenchyme surrounding the notochord. These mesenchymal condensations differentiate into the early annulus fibrosus during foetal development-their outer part becoming fibrous, containing collagen type I; and their inner part cartilaginous, containing type II collagen and aggrecan. With post-natal growth and ageing, there is a switch from type I to type II collagen and an increase in proteoglycan synthesis in the outer annulus. This fibrocartilaginous metaplasia appears to occur in response to compressive loading of the tissue as occurs in tendons that wrap around bony pulleys, and driven by growth factors, such as TGF-β. In this study, using high-density micromass cultures, we have assessed the response of foetal outer annulus cells to growth factor stimulation with TGF-β1 and IGF-1, growth factors known to occur within the developing disc. We qualitatively and quantitatively describe the stimulatory effects of these growth factors, both alone and in combination, on the synthesis of sulphated glycosaminoglycan, and collagen types I and II by annulus cells. We show a potential role for TGF-β1 in pushing cells towards a fibrocartilaginous phenotype, with possible complementary effects of IGF-1.

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Colony Formation and Matrix Production by Human Anulus Cells

Cited by 25 publications

References 25 publications

Developing an alginate/chitosan hybrid fiber scaffold for annulus fibrosus cells

Developing an alginate/chitosan hybrid fiber scaffold for annulus fibrosus cells

Effects of Low Glucose Concentrations on Oxygen Consumption Rates of Intervertebral Disc Cells

The response of foetal annulus fibrosus cells to growth factors: modulation of matrix synthesis by TGF-β1 and IGF-1

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