Immunolocalization of metalloproteinases and their inhibitor in the rabbit growth plate.

Brown, Cameron C.; Hembry, Rosalind M.; Reynolds, JJ

doi:10.2106/00004623-198971040-00014

Cited by 122 publications

(59 citation statements)

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“…However we are not the only investigators to report the presence of stronielysin in the physis. Brown et al [8] used immunolocalization to detect stromelysin in rabbit physes. Our results are further corroborated by the work of Lee et al [27] who used immunohistochemistry to locate the stromelysin cleavage site in rat growth plates.…”

Section: Resultsmentioning

confidence: 99%

Identification of the metalloproteinase stromelysin in the physis

Barrach

Ehrlich

2002

Journal Orthopaedic Research

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For long bone growth to occur, calcification of the matrix must commence in the lower hypertrophic zone of the growth plate. It is generally accepted that physeal proteoglycans help regulate mineralization, and that at least in vitro, intact proteoglycans can inhibit mineralization. Thus degradation of proteoglycan may be a necessary step prior to calcification. Previous work in o u r laboratory has demonstrated the presence of neutral metallo-proteases in the growth plate with highest levels in the hypertrophic zone, where calcification occurs.Stromelysin (MMP-3) is a connective tissue matrix-degrading enzyme. It was formerly known as proteoglycanase and is generally considered to be one of the major proteoglycan degrading enzymes in cartilage. Stromelysin is implicated in cartilage destruction in osteoarthritis and may also be involved in tissue remodeling in the physis. Our goal was to determine if the neutral protease previously reported by the authors in the physis was stromelysin.In this study we used Western blots and antibodies to stromelysin and to the stromelysin cleavage site in aggrecan, the most common form of proteoglycan, to demonstrate the presence of stromelysin in the bovine physis. When an antibody raised against the stromelysin cleavage site of aggrecan (FVDIPEN) was incubated with a Western blot, which had been run with aggrecan extracted from bovine physes, a positive reaction resulted. This suggests that there is stromelysin degradation in vivo in the physis. Two different polyclonal antibodies to stromelysin gave positive results on Western blots of purified media from growth plate cultures indicating that stromelysin is produced in vitro in culture. These antibodies also reacted with active stromelysin.The presence of stromelysin in the physis implicates it in physeal physiology. The concentration of its activity in the lower hypertrophic zone and zone of provisional calcification suggests that it may be particularly important in mineralization. 0 3002 Orthopaedic Research Society. Published by

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Section: Resultsmentioning

confidence: 99%

Identification of the metalloproteinase stromelysin in the physis

Barrach

Ehrlich

2002

Journal Orthopaedic Research

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“…Increased activities of these metalloproteinases have been reported in cartilages of growth plates (31)(32)(33)(34), OA patients ( 3 3 , and RA patients (36), where cartilage degradation is occurring. In the chick system, we have identified only collagenase and gelatinase associated with collagen degradation and have not been able to identify the enzyme responsible for PG loss.…”

Section: Discussionmentioning

confidence: 99%

Doxycycline disrupts chondrocyte differentiation and inhibits cartilage matrix degradation

Cole

Chubinskaya

Luchene

et al. 1994

Arthritis & Rheumatism

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Objective. The effects of doxycycline were tested in an in vitro system in which the cartilages of embryonic avian tibias are completely degraded.Methods. Tibias were cultured with 5, 20, or 40 pg/ml doxycycline. Control tibias were cultured without doxycycline. Conditioned media and tissue sections were examined for enzyme activity and matrix loss.Results. Cartilages were not resorbed in the presence of doxycycline, whereas control cartilages were completely degraded. Collagen degradation was reduced in association with treatment with doxycycline at all doses studied. Higher concentrations of doxycycline reduced collagenase and gelatinase activity and prevented proteoglycan loss, cell death, and deposition of type X collagen in the cartilage matrix; in addition, treatment with doxycycline at higher concentrations caused increases in the length of the hypertrophic region.Conclusion. The effects of doxycycline extend beyond inhibition of the proteolytic enzymes by stimulating cartilage growth and disrupting the terminal differentiation of chondrocytes.

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“…Matrix metalloprotease activities (Brown et al, 1989) probably are associated with the advancing front of the marrow cavity and might account for some of the loss of immunoreactivity observed. But such activities, even if widely distributed throughout the hypertrophic zone, are unlikely to be affected by both agents we used to inhibit crosslink formation.…”

Section: Immunoreactivity In Pattern Ofmentioning

confidence: 98%

Type II collagen during cartilage and corneal development: Immunohistochemical analysis with an anti‐telopeptide antibody

Chen

Fitch

Gibney

et al. 1993

Developmental Dynamics

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We have examined the pattern of immunoreactivity of a monoclonal antibody, II-5B2, with specificity for an epitope which resides within the NH,-terminal extension peptide (telopeptide) of the avian type I1 collagen molecule. This epitope is available in regions of matrix where de novo synthesis of the molecule is ongoing, but not where synthesis has ceased and maturation and crosslink formation have occurred. Within the cartilaginous growth plate, the epitope disappears from the matrix soon after the chondrocytes become hypertrophic; within the cornea, the epitope disappears subjacent to the epithelium. The 11-5B2 epitope is not made available by a variety of procedures shown to remove potentially masking substances and to disrupt fibrillar organization. It is rendered available, however, when covalent crosslink formation between collagen molecules is blocked through administration of P-aminopropionitrile or penicillamine. In contrast, the epitope of another monoclonal antibody against type I1 collagen, 11-IIGB3, which resides in the triple-helical domain of the molecule, in cartilage is present throughout the growth plate including the hypertrophic zone, and in cornea extends for a considerable distance into the stroma. Thus, it is available for antibody binding regardless of fibril maturation and crosslinking. These data suggest that the 11-5B2 epitope becomes unavailable when the telopeptide becomes crosslinked. By using these two monoclonal antibodies in serial sections, one can establish the crosslinking pattern of type I1 collagen in the tissue. This set of antibodies is a potentially useful tool for analyzing normal and abnormal development, remodeling, and repair processes in the skeletal system and in other tissues where type I1 collagen is involved in organization of the matrix, such as the primary corneal stroma. 0 1993 Wiley-Liss, Inc.

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Immunolocalization of metalloproteinases and their inhibitor in the rabbit growth plate.

Cited by 122 publications

References 0 publications

Identification of the metalloproteinase stromelysin in the physis

Identification of the metalloproteinase stromelysin in the physis

Doxycycline disrupts chondrocyte differentiation and inhibits cartilage matrix degradation

Type II collagen during cartilage and corneal development: Immunohistochemical analysis with an anti‐telopeptide antibody

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