scholarly journals Extracellular matrix formation by chondrocytes in monolayer culture.

1981 ◽  
Vol 90 (1) ◽  
pp. 78-83 ◽  
Author(s):  
W Dessau ◽  
B M Vertel ◽  
H von der Mark ◽  
K von der Mark
Keyword(s):  
Extracellular Space ◽  
Monolayer Culture ◽  
Spherical Shape ◽  
Type Ii Collagen ◽  
Cartilage Matrix ◽  
Pericellular Matrix ◽  
Type Ii ◽  
Culture Dish ◽  
Immunofluorescence Labeling ◽  
Matrix Characteristic

In previous studies were have reported on the secretion and extracellular deposition of type II collagen and fibronectin (Dessau et al., 1978, J. Cell Biol., 79:342-355) and chondroitin sulfate proteoglycan (CSPG) (Vertel and Dorfman, 1979, Proc. Natl. Acad. Sci. U. S. A. 76:1261-1264) in chondrocyte cultures. This study describes a combined effort to compare sequence and pattern of secretion and deposition of all three macromolecules in the same chondrocyte culture experiment. By immunofluorescence labeling experiments, we demonstrate that type II collagen, fibronectin, and CSPG reappear on the cell surface after enzymatic release of chondrocytes from embryonic chick cartilage but develop different patterns in the pericellular matrix. When chondrocytes spread on the culture dish, CSPG is deposited in the extracellular space as an amorphous mass and fibronectin forms fine, intercellular strands, whereas type II collagen disappears from the chondrocyte surface and remains absent from the extracellular space in early cultures. Only after cells in the center of chondrocyte colonies shape reassume spherical shape does the immunofluorescence reveal type II collagen in the refractile matrix characteristic of differentiated cartilage. By immunofluorescence double staining of the newly formed cartilage matrix, we demonstrate that CSPG spreads farther out into the extracellular space that type II collagen. Fibronectin finally disappears from the cartilage matrix.

scholarly journals Extracellular Nucleotides Selectively Induce Migration of Chondrocytes and Expression of Type II Collagen

2020 ◽  
Vol 21 (15) ◽  
pp. 5227
Author(s):  
Marcin Szustak ◽  
Edyta Gendaszewska-Darmach
Keyword(s):  
Migration Rate ◽  
Monolayer Culture ◽  
Type Ii Collagen ◽  
Extracellular Nucleotides ◽  
Nonbridging Oxygen ◽  
Nucleotide Receptors ◽  
Type Ii ◽  
Mrna Transcripts ◽  
Chondrogenic Phenotype

The migration of chondrocytes from healthy to injured tissues is one of the most important challenges during cartilage repair. Additionally, maintenance of the chondrogenic phenotype remains another limitation, especially during monolayer culture in vitro. Using both the differentiated and undifferentiated chondrogenic ATDC5 cell line, we showed that extracellular nucleotides are able to increase the migration rate of chondrocytes without affecting their chondrogenic phenotype. We checked the potency of natural nucleotides (ATP, ADP, UTP, and UDP) as well as their stable phosphorothioate analogs, containing a sulfur atom in the place of one nonbridging oxygen atom in a phosphate group. We also detected P2y1, P2y2, P2y4, P2y6, P2y12, P2y13, and P2y14 mRNA transcripts for nucleotide receptors, demonstrating that P2y1 and P2y13 are highly upregulated in differentiated ATDC5 cells. We showed that ADPβS, UDPβS, and ADP are the best stimulators of migration of differentiated chondrocytes. Additionally, ADP and ADPβS positively affected the expression of type II collagen, a structural component of the cartilage matrix.

scholarly journals Turnover of type II collagen and aggrecan in cartilage matrix at the onset of inflammatory arthritis in humans: Relationship to mediators of systemic and local inflammation

2003 ◽  
Vol 48 (11) ◽  
pp. 3085-3095 ◽  
Author(s):  
Alexander Fraser ◽  
Ursula Fearon ◽  
R. Clark Billinghurst ◽  
Mirela Ionescu ◽  
Richard Reece ◽  
...  
Keyword(s):  
Inflammatory Arthritis ◽  
Type Ii Collagen ◽  
Cartilage Matrix ◽  
Type Ii ◽  
Local Inflammation

Achondrogensis type II: A heterozygous point mutation prevents deposition of type II collagen into the cartilage matrix

1994 ◽  
Vol 14 (5) ◽  
pp. 389
Author(s):  
Danny Chan ◽  
C.W. Chow ◽  
William G. Cole ◽  
John F. Bateman
Keyword(s):  
Point Mutation ◽  
Type Ii Collagen ◽  
Cartilage Matrix ◽  
Type Ii

Disproportionate micromelia (Dmm): an incomplete dominant mouse dwarfism with abnormal cartilage matrix

Development ◽  
1981 ◽  
Vol 62 (1) ◽  
pp. 165-182
Author(s):  
Kenneth S. Brown ◽  
Robert E. Cranley ◽  
Robert Greene ◽  
Hynda K. Kleinman ◽  
John P. Pennypacker
Keyword(s):  
Type Ii Collagen ◽  
Cartilage Matrix ◽  
Type Ii ◽  
Cartilage Growth ◽  
Normal Cartilage ◽  
The Matrix ◽  
Collagen Antibodies ◽  
Biochemical Analyses ◽  
Cartilage Proteoglycans ◽  
Cartilage Growth Plate

This paper describes a new autosomal incomplete dominant dwarfism, disproportionate micromelia, which has been characterized genetically and phenotypically, and the cartilage of homozygotes, and heterozygotes has been examined by histochemical, immunofluorescence and biochemical methods. Homozygotes, which die at birth, are disproportionately short and have cleft palates. The heterozygotes appear normal at birth but beginning at 1 week of age dwarfism is apparent and increases during growth. Histochemical and biochemical analyses of the cartilage rudiments of homozygotes at day 18 of gestation demonstrate that the cartilage growth plate is disorganized and that the matrix components, collagen and proteoglycan, are altered. Total collagen synthesis is reduced by approximately 30% and the amount of type II collagen is greatly reduced. By immunofluorescence staining with collagen antibodies, it appears that type II collagen is located primarily near the cell surface of chondrocytes but is poorly distributed throughout the remainder of the matrix. The amount of proteoglycan in the cartilage matrix is reduced by approximately 70% as determined by chemical analysis of hexosamines and by [35S]sulfate incorporation. Although the proteoglycans synthesized by the mutant are normal in size and in glycosaminoglycan composition, they were more easily extractable from the matrix than were normal cartilage proteoglycans. Heterozygotes had reduced cartilage matrix proteoglycan by histochemical methods, but the organization of the epiphyseal cartilage was not abnormal. These data suggest that a reduced or abnormal cartilage matrix is the cause of the dwarfism.

Type II collagen Pro-α-chains containing a Gly574Ser mutation are not incorporated into the cartilage matrix of transgenic mice

1997 ◽  
Vol 16 (3) ◽  
pp. 93-103 ◽  
Author(s):  
B.Kerry Maddox ◽  
Silvio Garofalo ◽  
Douglas R. Keene ◽  
Chad Smith ◽  
William A. Horton
Keyword(s):  
Transgenic Mice ◽  
Type Ii Collagen ◽  
Cartilage Matrix ◽  
Type Ii

scholarly journals The calcification of cartilage matrix in chondrocyte culture: studies of the C-propeptide of type II collagen (chondrocalcin).

1987 ◽  
Vol 104 (5) ◽  
pp. 1435-1441 ◽  
Author(s):  
A Hinek ◽  
A Reiner ◽  
A R Poole
Keyword(s):  
Growth Plate ◽  
Type Ii Collagen ◽  
Close Correlation ◽  
Cartilage Matrix ◽  
Type Ii ◽  
Growth Plate Chondrocytes ◽  
Culture Studies ◽  
Chondrocyte Culture ◽  
Intracellular Content ◽  
Bovine Fetuses

We have shown that when chondrocytes are isolated by collagenase digestion of hyaline cartilage from growth plate, nasal, and epiphyseal cartilages of bovine fetuses they rapidly elaborate an extracellular matrix in culture. Only growth plate chondrocytes can calcify this matrix as ascertained by incorporation of 45Ca2+, detection of mineral with von Kossa's stain and electron microscopy. There is an extremely close direct correlation between 45Ca2+ incorporation in the first 24 h of culture and the content of the C-propeptide of type II collagen, measured by radioimmunoassay, at the time of isolation and during culture. Moreover, growth plate cells have an increased intracellular content of the C-propeptide per deoxyribonucleic acid and, during culture, per hydroxyproline (as a measure of helical collagen) compared with nasal and epiphyseal chondrocytes. In growth plate chondrocytes 24,25-dihydroxycholecalciferol (24,25-[OH]2D3), but not 1,25-dihydroxycholecalciferol alone, stimulates the net synthesis of the C-propeptide and calcification; proteoglycan net synthesis is unaffected. Together, these metabolites of vitamin D further stimulate C-propeptide net synthesis but do not further increase calcification stimulated by 24,25-(OH)2D3. These observations further demonstrate the close correlation between the C-propeptide of type II collagen and the calcification of cartilage matrix.

Incorporation of Structurally Defective Type II Collagen into Cartilage Matrix in Kniest Chondrodysplasia

1998 ◽  
Vol 355 (2) ◽  
pp. 282-290 ◽  
Author(s):  
Russell J. Fernandes ◽  
Douglas J. Wilkin ◽  
Mary Ann Weis ◽  
William R. Wilcox ◽  
Daniel H. Cohn ◽  
...  
Keyword(s):  
Type Ii Collagen ◽  
Cartilage Matrix ◽  
Type Ii

scholarly journals Chondrocalcin is identical with the C-propeptide of type II procollagen

1986 ◽  
Vol 237 (3) ◽  
pp. 923-925 ◽  
Author(s):  
M Van der Rest ◽  
L C Rosenberg ◽  
B R Olsen ◽  
A R Poole
Keyword(s):  
Primary Structure ◽  
Triple Helix ◽  
Type Ii Collagen ◽  
Cartilage Matrix ◽  
Type Ii ◽  
Sequence Analyses ◽  
Tryptic Peptides ◽  
Cartilage Calcification ◽  
Matrix Molecule

The primary structure of the cartilage matrix molecule chondrocalcin has been found to be identical with that of the C-propeptide of type II procollagen by comparing sequence analyses of the N-terminal regions and of tryptic peptides derived from chondrocalcin. This implies that in type II collagen the C-propeptide of type II collagen is employed not only in the assembly of the triple helix of type II collagen, as demonstrated previously, but in calcifying cartilage it may also be involved in those events leading to cartilage calcification, as earlier indicated.

scholarly journals Role of anchorin CII, a 31,000-mol-wt membrane protein, in the interaction of chondrocytes with type II collagen.

1984 ◽  
Vol 98 (4) ◽  
pp. 1572-1579 ◽  
Author(s):  
J Mollenhauer ◽  
J A Bee ◽  
M A Lizarbe ◽  
K von der Mark
Keyword(s):  
Membrane Protein ◽  
Monolayer Culture ◽  
Collagen Matrix ◽  
Type Ii Collagen ◽  
Type Ii ◽  
Fab Fragments ◽  
Cell Processes ◽  
Matrix Removal ◽  
Binding Glycoprotein

We have previously reported the isolation of a hydrophobic, type-II collagen-binding glycoprotein of molecular weight 31,000 (31,000-mol-wt protein) from chick chondrocyte membranes (Mollenhauer, J., and K. von der Mark, EMBO Eur. Mol. Biol. Organ. J., 2:45-50). The function of this protein in anchoring pericellular type II collagen to the chondrocyte surface was inferred from its ability to bind native type-II collagen either when detergent solubilized or when inserted into liposomes. In the present study we have used specific antibodies to localize this protein, which we now call anchorin CII, to the surface of chondrocytes in both cartilage sections, and in cell culture. In immunofluorescence studies of isolated chondrocytes we observed a dense, punctate distribution of anchorin CII on the cell surface when chondrocytes were enclosed by a pericellular type II collagen matrix. Removal of the pericellular matrix with trypsin also removed anchorin CII. The membrane protein character of anchorin CII was indicated by the demonstration of antibody-induced patching and capping on the chondrocyte surface at 22 degrees C and 37 degrees C, respectively. In monolayer culture, the amount of anchorin CII appeared reduced on flattened chondrocytes lacking a pericellular type II collagen matrix but was prominent upon intercellular cell processes. Fab' fragments prepared from either anchorin CII antiserum or an antiserum directed against the entire chondrocyte membrane inhibited the attachment of chondrocytes to a type II collagen substrate. In each case, the inhibition of attachment was neutralized by preincubation of Fab' fragments with purified anchorin CII.

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