title: Compressive force promotes sox9, type II collagen and aggrecan and inhibits IL-1beta expression resulting in chondrogenesis in mouse embryonic limb bud mesenchymal cells.
abstract: The initial modeling and subsequent development of the skeleton is controlled by complex gene-environment interactions. Biomechanical forces may be one of the major epigenetic factors that determine the form and differentiation of skeletal tissues. In order to test the hypothesis that static compressive forces are transduced into molecular signals during early chondrogenesis, we have developed a unique three-dimensional collagen gel cell culture system which is permissive for the proliferation and differentiation of chondrocytes. Mouse embryonic day 10 (E10) limb buds were microdissected and dissociated into cells which were then cultured within a collagen gel matrix and maintained for up to 10 days. Static compressive forces were exerted onto these cultures. The time course for expression pattern and level for cartilage specific markers, type II collagen and aggrecan, and regulators of chondrogenesis, Sox9 and IL-1beta, were analyzed and compared with non-compressed control cultures. Under compressive conditions, histological evaluation showed an apparent acceleration in the rate and extent of chondrogenesis. Quantitatively, there was a significant 2- to 3-fold increase in type II collagen and aggrecan expression beginning at day 5 of culture and the difference was maintained through 10 days of cultures. Compressive force also causes an elevated level of Sox9, a transcriptional activator of type II collagen. In contrast, the expression and accumulation of IL-1beta, a transcriptional repressor of type II collagen was down-regulated. We conclude that static compressive forces promote chondrogenesis in embryonic limb bud mesenchyme, and propose that the signal transduction from a biomechanical stimuli can be mediated by a combination of positive and negative effectors of cartilage specific extracellular matrix macromolecules.
authors.affiliation: Craniofacial Development Section, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: I
authors.initials: I
authors.lastname: Takahashi
authors.forename: G H
authors.initials: GH
authors.lastname: Nuckolls
authors.forename: K
authors.initials: K
authors.lastname: Takahashi
authors.forename: O
authors.initials: O
authors.lastname: Tanaka
authors.forename: I
authors.initials: I
authors.lastname: Semba
authors.forename: R
authors.initials: R
authors.lastname: Dashner
authors.forename: L
authors.initials: L
authors.lastname: Shum
authors.forename: H C
authors.initials: HC
authors.lastname: Slavkin
mesh_terms: D053674:Aggrecans; D000818:Animals; D015415:Biomarkers; D002356:Cartilage / Q000166:cytology / Q000196:embryology*; D002478:Cells, Cultured; D011508:Chondroitin Sulfate Proteoglycans / Q000096:biosynthesis; D003094:Collagen / Q000096:biosynthesis*; D004622:Embryo, Mammalian; D005314:Embryonic and Fetal Development; D016326:Extracellular Matrix Proteins; D018507:Gene Expression Regulation, Developmental; D005865:Gestational Age; D006609:High Mobility Group Proteins / Q000096:biosynthesis*; D007375:Interleukin-1 / Q000096:biosynthesis; D007700:Kinetics; D037181:Lectins, C-Type; D018878:Limb Buds; D008648:Mesoderm / Q000166:cytology / Q000502:physiology*; D051379:Mice; D016133:Polymerase Chain Reaction; D011509:Proteoglycans / Q000096:biosynthesis*; D055758:SOX9 Transcription Factor; D012733:Sex Differentiation; D013314:Stress, Mechanical; D013997:Time Factors; D014157:Transcription Factors / Q000096:biosynthesis*
chemical_list: C508320:Acan protein, mouse; D053674:Aggrecans; D015415:Biomarkers; D011508:Chondroitin Sulfate Proteoglycans; D016326:Extracellular Matrix Proteins; D006609:High Mobility Group Proteins; D007375:Interleukin-1; D037181:Lectins, C-Type; D011509:Proteoglycans; D055758:SOX9 Transcription Factor; C527933:Sox9 protein, mouse; D014157:Transcription Factors; D003094:Collagen