The Structure and Abundance of Cartilage Proteoglycans during Early Development of the Human Fetus

Roughley, Peter J.; White, Robert J.; Glant, Tibor T.

doi:10.1203/00006450-198710000-00009

Cited by 16 publications

(11 citation statements)

References 45 publications

(41 reference statements)

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“…Ion exchange HPLC analyses of fluorotagged chondroitinase digestion products was done under conditions in which all internal ⌬disaccharides and nonreducing terminal monosaccharides and disaccharides are measured (26). The majority of the fluorescent products in the digests of human aggrecan represented the sulfated ⌬disaccharides (Table I), and in agreement with previous reports (3,6,7), the ratio of ⌬Di6S:⌬Di4S was about 0.75 in aggrecan CS from cartilages up to 1 year of age, but increased to a much higher level, ϳ25, in samples from individuals older than 15 years of age.…”

Section: Fluorescent Hplc Analyses Of Chondroitinase Digests Fromsupporting

confidence: 87%

Chemical and Immunological Assay of the Nonreducing Terminal Residues of Chondroitin Sulfate from Human Aggrecan

Plaas¹,

Wong-Palms²,

Roughley³

et al. 1997

Journal of Biological Chemistry

143

138

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Samples of aggrecan chondroitin sulfate, isolated from normal human knee cartilages of individuals from fetal to 72 years of age, were digested with chondroitin lyases. The products were analyzed by fluorescencebased anion exchange high performance liquid chromatography to separate and quantitate nonreducing terminal structures, in addition to internal unsaturated disaccharide products. The predominant terminal structures were the monosaccharides, GalNAc4S and GalNAc4,6S as they were present on 85-90% of all chains. The remaining chains terminated with the disaccharides GlcA␤1,3GalNAc4S and GlcA␤1,3GalNAc6S. Marked changes in the relative abundance of these terminals were identified in the transition from growth cartilage to adult articular cartilage. First, terminal GalNAc residues were almost exclusively 4-sulfated in aggrecan from fetal through 15 years of age, but were ϳ50% 4,6-disulfated in aggrecans from adults (22-72 years of age). Second, the terminal disaccharide GlcA␤1,3GalNAc4S was on ϳ7% of chains on aggrecan from fetal through 15 years of age, but on only ϳ3% of chains on adult aggrecan. In contrast, the proportion of chains terminating in GlcA␤1,3GalNAc6S, ϳ9%, was unchanged from fetal to 72 years of age. This terminal disaccharide is proposed to be recognized by the widely used monoclonal antibody 3B3. However, chemical quantitation of the structure together with solid phase 3B3(؊) immunoassay of fetal and adult aggrecans showed that the content of the terminal disaccharide does not necessarily correlate with immunoreactivity of the proteoglycan, as chain density and presentation on the solid phase are critical factors for recognition of chain terminals by 3B3. The quantitative results obtained from chemical analyses of all nonreducing termini of aggrecan chondroitin sulfate chains revealed important changes in chain termination that occur when cellular activities are altered as adult articular cartilage is formed after removal of growth cartilage. These findings are discussed in relation to specific enzymatic steps that generate the nonreducing termini of chains in the biosynthesis pathway of chondroitin sulfate proteoglycans and their modulation in tissue development and pathology.

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Section: Fluorescent Hplc Analyses Of Chondroitinase Digests Fromsupporting

confidence: 87%

Chemical and Immunological Assay of the Nonreducing Terminal Residues of Chondroitin Sulfate from Human Aggrecan

Plaas¹,

Wong-Palms²,

Roughley³

et al. 1997

Journal of Biological Chemistry

143

138

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“…In adult hyaline cartilage, sulfated GAG content, on a weight basis, is more than an order of magnitude greater than the HA content. During development, however, it has been shown that non-sulfated disaccharide content starts higher than 4-sulfated GAG but decreases significantly with age [39]. Collectively, it suggests that the ECM formed is relatively ‘young’ compared to adult articular cartilage.…”

Section: Discussionmentioning

confidence: 99%

Micrometer scale guidance of mesenchymal stem cells to form structurally oriented large-scale tissue engineered cartilage

et al. 2017

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Current clinical methods to treat articular cartilage lesions provide temporary relief of the symptoms but fail to permanently restore the damaged tissue. Tissue engineering, using mesenchymal stem cells (MSCs) combined with scaffolds and bioactive factors, is viewed as a promising method for repairing cartilage injuries. However, current tissue engineered constructs display inferior mechanical properties compared to native articular cartilage, which could be attributed to the lack of structural organization of the extracellular matrix (ECM) of these engineered constructs in comparison to the highly oriented structure of articular cartilage ECM. We previously showed that we can guide MSCs undergoing chondrogenesis to align using microscale guidance channels on the surface of a two-dimensional (2-D) collagen scaffold, which resulted in the deposition of aligned ECM within the channels and enhanced mechanical properties of the constructs. In this study, we developed a technique to roll 2-D collagen scaffolds containing MSCs within guidance channels in order to produce a large-scale, three-dimensional (3-D) tissue engineered cartilage constructs with enhanced mechanical properties compared to current constructs. After rolling the MSC-scaffold constructs into a 3-D cylindrical structure, the constructs were cultured for 21 days under chondrogenic culture conditions. The microstructure architecture and mechanical properties of the constructs were evaluated using imaging and compressive testing. Histology and immunohistochemistry of the constructs showed extensive glycosaminoglycan (GAG) and collagen type II deposition. Second harmonic generation imaging and Picrosirius red staining indicated alignment of neo-collagen fibers within the guidance channels of the constructs. Mechanical testing indicated that constructs containing the guidance channels displayed enhanced compressive properties compared to control constructs without these channels. In conclusion, using a novel roll-up method, we have developed large scale MSC based tissue-engineered cartilage that show microscale structural organization and enhanced compressive properties compared to current tissue engineered constructs.

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“…33,39 In the fetus, much of the N-acetylgalactosamine may be nonsulfated. 42 In contrast to chondroitin sulfate, keratan sulfate chain length increases during juvenile development. 5,46 Although the aggrecan core protein shows structural variation attributable to alternative splicing, this does not seem to be age-related.…”

Section: Number 391s October 2001mentioning

confidence: 98%

Age-Associated Changes in Cartilage Matrix

Roughley

2001

Clinical Orthopaedics and Related Research

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S153The structure of the extracellular matrix of articular cartilage varies considerably with age. These changes are attributable to variations in molecular abundance and structure, and they can affect all the matrix components, but none more so than the proteoglycans. Some of these changes are attributable to variations in synthesis whereas others are attributable to variations in degradation, some of the changes occur during juvenile development whereas others occur throughout life, and some of the changes are beneficial to cartilage function whereas others are detrimental. These variations result in a cartilage that not only changes in its phenotype with age, but also in one whose functional properties are changing continuously throughout life. In a similar manner, the cartilage formed during repair also may show considerable variation in structure and function, depending on whether tissue is being replaced or regenerated and whether mature or immature cells are being used. Because all cartilage is not ceated equal, different repair techniques may not be equally efficacious.The repair of articular cartilage lesions involves the substitution of the old, damaged cartilage by a new, intact tissue. This can be achieved by either replacement via cartilage grafting or regeneration via chondrocyte implantation, and various techniques exist for both approaches. 7,32,51 In either case, it is necessary that the new tissue has and maintains a cartilage phenotype, requiring the absence of blood vessels and nerves, the absence of mineralization, List of Abbreviations Used BMPbone morphogenetic protein COMP cartilage oligomeric protein EGF epidermal growth factor GAG glycosaminoglycan MMP matrix metalloproteinase PRELP proline and arginine-rich end leucine-rich repeat protein GlossaryLP3 ϭ a component of link protein that is derived by proteolytic cleavage in the amino terminal region of the intact molecule. The intact link proteins may possess two or one N-linked oligosaccharides in this amino terminal region and are referred to as LP1 or LP2, respectively. Proteolytic cleavage of either LP1 or LP2 generates the same LP3 component.

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The Structure and Abundance of Cartilage Proteoglycans during Early Development of the Human Fetus

Cited by 16 publications

References 45 publications

Chemical and Immunological Assay of the Nonreducing Terminal Residues of Chondroitin Sulfate from Human Aggrecan

Chemical and Immunological Assay of the Nonreducing Terminal Residues of Chondroitin Sulfate from Human Aggrecan

Micrometer scale guidance of mesenchymal stem cells to form structurally oriented large-scale tissue engineered cartilage

Age-Associated Changes in Cartilage Matrix

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