Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro.

Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro. Research Abstract Details 

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Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro. Abstract Text:

Ronald R Gomes,Mary C Farach Carson,Daniel D Carson,

The heparan sulfate proteoglycan, perlecan, is localized to hypertrophic chondrocytes in the growth plates of long bones. Mice mutants for perlecan display severe cartilage and skeletal defects. Previously, we demonstrated that C3H10T1/2 fibroblasts cultured on perlecan stimulated extensive formation of dense nodules reminiscent of embryonic cartilaginous condensations. These nodules stain intensely with Alcian blue, and antibodies specific for collagen type II and aggrecan; however, nodules do not express collagen type X, a marker of chondrogenic maturation. In this investigation, we tested the hypothesis that addition of rhBMP-2 to perlecan-induced nodules would promote chondrogenic maturation in vitro. C3H10T1/2 fibroblasts were seeded in Lab-Tek chambered "Permanox" slides uncoated or coated with perlecan (B&D, 5 microg/well), at a density of 2 x 10(5) cells/well. The cells were maintained in CMRL-1066 media supplemented with ascorbic acid, citrate, and pyruvate (50 ng/ml). C3H10T1/2 fibroblasts seeded on perlecan-coated wells began to condense and form cell aggregates within 15 min. On the third day postplating, the media was replaced and supplemented with or without rhBMP-2 (50 ng/ml, Genetics Institute). On day 6 of culture, microscopy revealed that rhBMP-2-treated cultures had significantly proliferated; however, untreated cultures had not. By day 12 of culture, confocal microscopy revealed that perlecan-stimulated nodules treated with rhBMP-2 express a late stage marker of chondrogenesis (collagen type X). Morphologically, cells expressing collagen type X in rhBMP-2-treated nodules appear larger in diameter, relative to cells not expressing collagen type X. Cells cultured on plastic and treated with rhBMP-2 did not form nodules, but attached and spread, yielding a high-density monolayer. In response to rhBMP-2 treatment, these cells also express collagen type X. However, the appearance of collagen type X occurs at a later time point relative to the appearance of collagen type X in perlecan-stimulated nodules. Thus, perlecan-stimulated nodules do mature at a faster rate when treated with rhBMP-2 relative to monolayer cells.

Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro. Publishing Authors By Initials

RR Gomes,MC Farach Carson,DD Carson,

For similar peptides: intercellular signaling peptides and proteins: cytokines: transforming growth factor beta research abstracts see: peptides: intercellular signaling peptides and proteins: cytokines: transforming growth factor beta research

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Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Connective tissue research

VOLUME: 44 Suppl 1

Page Numbers: 196-201

Journal Abbreviation: Connect. Tissue Res.

ISSN: 0300-8207

DAY: 19

MONTH: 11

YEAR: 2003

Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 365263

Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro. Keywords Mesh Terms:

KEYWORDS: Transforming Growth Factor beta

MESH TERMS: pharmacology

Chemical & Substance for Abstract: Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro. Information

Substance Name: perlecan

Registry Number: 143972-95-6

Grant and Affiliation Information for Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro.

AFFILIATION: Department of Biological Sciences, University of Delaware, Newark, Delaware 19716, USA.

Country: United States

AGENCY: United States NIDCR

GRANT: R01-DE13542

ACRONYM: DE

MEDLINETA: Connect Tissue Res

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