Cell‐associated type I collagen in nondegenerate and degenerate human articular cartilage

Chondral knee defects have a limited ability to be repaired. Current surgical interventions have been unable to regenerate articular cartilage with the mechanical properties of native hyaline cartilage. The use of a scaffold-based approach is a potential solution. Scaffolds are often implanted with cells to stimulate cartilage regeneration, but cell-based therapies are associated with additional regulatory restrictions, an additional surgical procedure for cell harvest, time for cell expansion, and the associated costs. To overcome these disadvantages, cell-free scaffolds can be used in isolation allowing native cells to attach over time. This review discusses the optimal properties of scaffolds used for chondral defects, and the evidence for the use of hydrogel scaffolds and hydrogel–synthetic polymer hybrid scaffolds. Preclinical and clinical studies have shown that cell-free scaffolds can support articular cartilage regeneration and have the potential to treat chondral defects. However, there are very few studies in this area and, despite the many biomaterials tested in cell-based scaffolds, most cell-free studies focused on a specific type I collagen scaffold. Future studies on cell-free scaffolds should adopt the modifications made to cell-based scaffolds and replicate them in the clinical setting. More studies are also needed to understand the underlying mechanism of cell-free scaffolds.

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Macromolecular Organization and In Vitro Growth Characteristics of Scaffold-free Neocartilage Grafts

Journal of Histochemistry & Cytochemistry ◽

10.1369/jhc.7a7210.2007 ◽

2007 ◽

Vol 55 (8) ◽

pp. 853-866 ◽

Cited By ~ 63

Author(s):

Anthony J. Hayes ◽

Amanda Hall ◽

Liesbeth Brown ◽

Ross Tubo ◽

Bruce Caterson

Keyword(s):

Articular Cartilage ◽

Type I Collagen ◽

Primary Cultures ◽

Surface Zone ◽

Type I ◽

Focal Lesions ◽

Electron Microscopic ◽

Chondrocyte Phenotype ◽

Macromolecular Organization

Recent advances in tissue engineering offer considerable promise for the repair of focal lesions in articular cartilage. Here we describe (1) the macromolecular organization of tissue-engineered neocartilage grafts at light and electron microscopic levels, (2) their in vitro development, and (3) the effect of chondrocyte dedifferentiation, induced by monolayer expansion, on their resultant structure. We show that grafts produced from primary cultures of chondrocytes are hyaline in appearance with identifiable zonal strata as evidenced by cell morphology, matrix organization, and immunohistochemical composition. Like native articular cartilage, their surface zone contains type I collagen, surface zone proteoglycan, biglycan and decorin with type II collagen, aggrecan, chondroitin sulfate, chondroitin-4-sulfate, and keratan sulfate, becoming more prominent with depth. Assessment of cell viability by Live/Dead staining and cell-cycle analysis with BrDU suggest that the in vitro tissue has a high cellular turnover and develops through both appositional and interstitial growth mechanisms. Meanwhile, cell-tracker studies with CMFDA (5-chloromethyl-fluorescein diacetate) demonstrate that cell sorting in vitro is not involved in their zonal organization. Finally, passage expansion of chondrocytes in monolayer culture causes progressive reductions in graft thickness, loss of zonal architecture, and a more fibrocartilaginous tissue histology, consistent with a dedifferentiating chondrocyte phenotype. (J Histochem Cytochem 55: 853–866, 2007)

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Immunohistochemistry of types I and II collagen in undecalcified skeletal tissues.

Journal of Histochemistry & Cytochemistry ◽

10.1177/31.3.6338106 ◽

1983 ◽

Vol 31 (3) ◽

pp. 417-425 ◽

Cited By ~ 41

Author(s):

W A Horton ◽

C Dwyer ◽

R Goering ◽

D C Dean

Keyword(s):

Articular Cartilage ◽

Growth Plate ◽

Subchondral Bone ◽

Type I Collagen ◽

Sodium Ethoxide ◽

Cartilage Matrix ◽

Antigenic Determinants ◽

Immunoperoxidase Staining ◽

Type I ◽

Intense Staining

Types I and II collagen were demonstrated in semithin sections of undecalcified human endochondral growth plate, articular cartilage, and subchondral bone. The effects of several different methods for fixation, embedding, exposing of antigenic determinants, and immunoperoxidase staining were examined. Fixation in buffered formalin and paraformaldehyde-lysine-periodate solution gave more intense staining for collagens than fixation in paraformaldehyde-gluaraldehyde or Bouin's solution. Specimens embedded in Spurr epoxy resin yielded intense and uniform staining of areas known to contain the particular collagens after the resin had been removed by sodium ethoxide. The staining was enchanced following enzymatic digestion, especially with protease V (Sigma). Staining sensitivity and specificity were comparable with the indirect conjugate and double peroxidase-antiperoxidase (PAP) techniques; the PAP method was less sensitive. Embedment in methacrylate resins proved unsatisfactory because of exaggerated immunostaining of mineralized sites in comparison to unmineralized areas of the same tissues. In the growth plate specimens, type I collagen was identified in the matrices of bone, periosteum, perichondrium, and in the cytoplasm of hypertrophic and degenerative chondrocytes. Type II collagen was found uniformly throughout the cartilage matrix and in spicules of unresorbed cartilage matrix located in subchondral bone. A similar staining pattern was observed for the articular cartilage, except that type I collagen was not detected in chondrocytes.

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Localization of the Expression of Type I, II, III Collagen, and Aggrecan Core Protein Genes in Developing Human Articular Cartilage

Matrix ◽

10.1016/s0934-8832(11)80065-x ◽

1992 ◽

Vol 12 (3) ◽

pp. 221-232 ◽

Cited By ~ 25

Author(s):

Isabelle Treilleux ◽

Frederic Mallein-Gerin ◽

Dominique Le Guellec ◽

Daniel Herbage

Keyword(s):

Articular Cartilage ◽

Core Protein ◽

Type I ◽

Human Articular Cartilage ◽

Aggrecan Core Protein

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Promotion of articular cartilage matrix vesicle mineralization by type I collagen

Arthritis & Rheumatism ◽

10.1002/art.23762 ◽

2008 ◽

Vol 58 (9) ◽

pp. 2809-2817 ◽

Cited By ~ 33

Author(s):

Brian Jubeck ◽

Claudia Gohr ◽

Mark Fahey ◽

Emily Muth ◽

Michele Matthews ◽

...

Keyword(s):

Articular Cartilage ◽

Type I Collagen ◽

Matrix Vesicle ◽

Cartilage Matrix ◽

Type I

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Differential Production of Cartilage ECM in 3D Agarose Constructs by Equine Articular Cartilage Progenitor Cells and Mesenchymal Stromal Cells

International Journal of Molecular Sciences ◽

10.3390/ijms21197071 ◽

2020 ◽

Vol 21 (19) ◽

pp. 7071

Author(s):

Stefanie Schmidt ◽

Florencia Abinzano ◽

Anneloes Mensinga ◽

Jörg Teßmar ◽

Jürgen Groll ◽

...

Keyword(s):

Articular Cartilage ◽

Progenitor Cells ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Type I Collagen ◽

Type Ii Collagen ◽

Cell Types ◽

Type I ◽

Cartilage Engineering ◽

Hypoxic Conditions

Identification of articular cartilage progenitor cells (ACPCs) has opened up new opportunities for cartilage repair. These cells may be used as alternatives for or in combination with mesenchymal stromal cells (MSCs) in cartilage engineering. However, their potential needs to be further investigated, since only a few studies have compared ACPCs and MSCs when cultured in hydrogels. Therefore, in this study, we compared chondrogenic differentiation of equine ACPCs and MSCs in agarose constructs as monocultures and as zonally layered co-cultures under both normoxic and hypoxic conditions. ACPCs and MSCs exhibited distinctly differential production of the cartilaginous extracellular matrix (ECM). For ACPC constructs, markedly higher glycosaminoglycan (GAG) contents were determined by histological and quantitative biochemical evaluation, both in normoxia and hypoxia. Differential GAG production was also reflected in layered co-culture constructs. For both cell types, similar staining for type II collagen was detected. However, distinctly weaker staining for undesired type I collagen was observed in the ACPC constructs. For ACPCs, only very low alkaline phosphatase (ALP) activity, a marker of terminal differentiation, was determined, in stark contrast to what was found for MSCs. This study underscores the potential of ACPCs as a promising cell source for cartilage engineering.

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Expression of cartilage-specific molecules is retained on long-term culture of human articular chondrocytes

Journal of Cell Science ◽

10.1242/jcs.108.5.1991 ◽

1995 ◽

Vol 108 (5) ◽

pp. 1991-1999 ◽

Cited By ~ 3

Author(s):

E. Kolettas ◽

L. Buluwela ◽

M.T. Bayliss ◽

H.I. Muir

Keyword(s):

Type I Collagen ◽

Articular Chondrocytes ◽

Culture Conditions ◽

Human Articular Chondrocytes ◽

Type I ◽

Human Articular Cartilage ◽

Long Term Culture ◽

Chondrocyte Phenotype ◽

Link Protein

Normal human adult articular chondrocytes were used to determine how the chondrocyte phenotype is modulated by culture conditions following long-term culture. We report here for the first time that human articular chondrocytes have a lifespan in the range of 34–37 population doublings. While chondrocytes cultured as monolayers displayed a fibroblastoid morphology and grew faster, those cultured as suspensions over agarose adopted a round morphology and formed clusters of cells reminiscent of chondrocyte differentiation in intact cartilage, with little or no DNA synthesis. These morphologies were independent of the age of the culture. Despite, these morphological differences, however, chondrocytes expressed markers at mRNA and protein levels characteristic of cartilage: namely, types II and IX collagens and the large aggregating proteoglycans, aggrecan, versican and link protein, but not syndecan, under both culture conditions. However, they also expressed type I collagen alpha 1(I) and alpha 2(I) chains. It has been suggested that expression of collagen alpha 1(I) by chondrocytes cultured as monolayers is a marker of the loss of the chondrocyte phenotype. However, we show here, using reverse transcriptase/polymerase chain reaction, that normal fresh intact human articular cartilage expresses collagen alpha 1(I). The data show that following long-term culture human articular chondrocytes retain their differentiated characteristics and that cell shape does not correlate with the expression of the chondrocyte phenotype. It is proposed that loss of the chondrocyte phenotype is marked by the loss of one or more cartilage-specific molecules rather than by the appearance of non-cartilage-specific molecules.

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