Chick Cartilage Fibronectin Differs in Structure from the Fibronectin in Limb Mesenchyme

White, D.; Hall, Jeremy L.; Brandli, David W.; Gehris, Amy L.; Bennett, Vickie D.

doi:10.1006/excr.1996.0149

Cited by 11 publications

(7 citation statements)

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“…However, previous studies have also revealed that fibronectin is present throughout chondrogenesis and remains in differentiated cartilage. [21,22] This spatial expression of fibronectin has been found in accordance with the trend of cell viability analysis in this study, which shows that most hMSCs remained viable initially in the presence of fibronectin derived RGD peptide, followed by a significant decrease after 7 d of in vitro chondrogenic culture. This is because the expression of integrins a5b1 on cell surfaces correlates with that of fibronectin during the development of chondrocytes.…”

Section: Immunohistological Analysissupporting

confidence: 91%

Injectable Biodegradable Poly(ethylene glycol)/RGD Peptide Hybrid Hydrogels for in vitro Chondrogenesis of Human Mesenchymal Stem Cells

Liu

Tian

Wang

et al. 2010

Macromol. Rapid Commun.

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In this study, an injectable and biodegradable poly(ethylene glycol) (PEG)/arginine-glycine-aspartic (RGD) peptide hybrid hydrogel has been synthesized and used as a biomimetic scaffold for encapsulation of human mesenchymal stem cells (hMSCs). Tetrahydroxyl PEG was functionalized with acrylate, and then reacted with thiol-containing peptide (RGD). Gelation occurred within 30 min with the addition of cells and PEG-dithiol via Michael addition. The hydrogels synthesized with a peptide concentration of 1.0-5.0 mM achieved significantly greater cell viability when compared to the hydrogels without the RGD peptide. However, the effect of RGD on chondrogenesis was found to be dose-dependent. Immunohistology studies demonstrated that hMSCs encapsulated in the hydrogel matrix with 1.0 mM RGD and TGF-β3 showed enhanced positive staining for aggrecan and type II collagen as compared to that with 5.0 mM RGD and unmodified PEG hydrogels. RT-PCR results further revealed that the cells in hydrogels with 1 mM RGD expressed significantly higher levels of type II collagen than those in PEG hydogels without RGD peptide. These findings have demonstrated that the PEG-RGD hydrogels can be a promising scaffold to deliver hMSCs for cartilage repair.

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Section: Immunohistological Analysissupporting

confidence: 91%

Injectable Biodegradable Poly(ethylene glycol)/RGD Peptide Hybrid Hydrogels for in vitro Chondrogenesis of Human Mesenchymal Stem Cells

Liu

Tian

Wang

et al. 2010

Macromol. Rapid Commun.

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“…Thus, in the IVD tissues, there are as many as 32 splice forms. Although the functions of these diverse forms have not been thoroughly elucidated yet, 7-9 their potential roles in IVD development early in life and repair in adults should not be overlooked.…”

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confidence: 99%

Fibronectin Fragments and the Cleaving Enzyme ADAM-8 in the Degenerative Human Intervertebral Disc

Ruel

Markova

Adams

et al. 2014

Spine

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Study Design The presence fibronectin fragments (FN-fs) and the cleaving enzyme, A disintegrin and metalloproteinase domain-containing protein (ADAM)-8 were examined in human intervertebral disc (IVD) tissue in vitro. Objective To investigate the presence and pathophysiological concentration of FN-fs and their cleaving enzyme, ADAM-8, in the human IVD tissue. Summary of Background Data The 29kDa FN-f has been shown to result in extracellular matrix loss in rabbit IVDs. However, the concentration of this biologically active fragment in the degenerative human IVD tissue has previously not been determined. Further, it is critical to identify the enzyme(s) responsible for FN cleavage in the IVD. Methods Human degenerative IVD tissues were removed during spinal surgery. A normal appearing young adult and an infant human cadaveric sample were obtained as controls. Soluble proteins were extracted, and analyzed by Western blotting utilizing antibodies specific for the human FN neoepitope VRAA271. A purified 29 kDa FN-f was used to allow estimation of the concentration of FN-fs in the tissues. ADAM-8, a FN-cleaving enzyme, was analyzed by Western blotting and immunostaining. Results All adult IVD tissues contain many FN-f species, but these species were absent from the infant disc tissue. Moderately degenerative discs contained the highest amount of FN-fs; the concentration was estimated to be in the nanomolar range per gram of tissue. ADAM-8, known to cleave FN resulting in the VRAA271 neoepitope, was present in the human disc. ADAM-8 primarily localized in the pericellular matrix of the nucleus pulposus (NP) tissue, as determined by immunostaining. Conclusion This is the first report that N-terminal FN-fs are consistently present in IVD tissues from adult subjects. The pathophysiological concentration of these fragments is estimated to be at nanomolar range per gram of IVD tissue. Further, ADAM-8, known to cleave FN, is present at the pericellular matrix of disc cells.

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“…The rationale for using four distinct peptides as immunogens and developing six distinct ELISA systems is as follows. The quantitation of secreted proteins and cytokines by ELISA can be dependent on the nature of the antibody used [White et al, 1996;Krakauer, 1998] and OPN is susceptible to proteolytic fragmentation by thrombin and this process may have physiologic importance Senger et al, 1994]. Therefore a system to detect thrombin-cleaved forms of OPN is needed.…”

Section: Nature Of Antibodiesmentioning

confidence: 99%

Antibodies to different peptides in osteopontin reveal complexities in the various secreted forms

et al. 2000

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We have generated synthetic peptides corresponding to various portions of human osteopontin (OPN) and have immunized rabbits and mice with these peptides to generate polyclonal and monoclonal antibodies specific to human OPN. We then generated six distinct sandwich enzyme-linked immunoabsorbent assay (ELISA) systems by using different pairs of polyclonal and monoclonal antibodies against human OPN. These systems allowed us to detect not only various isoforms and truncated forms of recombinant OPN, but also the glycosylated form of native urinary OPN. Most importantly, tumor-derived OPN was differentially detected by the six ELISA systems. The ELISA systems that we have developed will be useful for clarifying the functional roles for OPN in vivo in various physiologic and pathologic conditions.

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Chick Cartilage Fibronectin Differs in Structure from the Fibronectin in Limb Mesenchyme

Cited by 11 publications

References 0 publications

Injectable Biodegradable Poly(ethylene glycol)/RGD Peptide Hybrid Hydrogels for in vitro Chondrogenesis of Human Mesenchymal Stem Cells

Injectable Biodegradable Poly(ethylene glycol)/RGD Peptide Hybrid Hydrogels for in vitro Chondrogenesis of Human Mesenchymal Stem Cells

Fibronectin Fragments and the Cleaving Enzyme ADAM-8 in the Degenerative Human Intervertebral Disc

Antibodies to different peptides in osteopontin reveal complexities in the various secreted forms

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