Early Shifts in Gene Expression during Chondroinduction of Human Dermal Fibroblasts

Yates, Karen E.; Mizuno, Shuichi; Glowacki, Julie

doi:10.1006/excr.2001.5192

Cited by 21 publications

(61 citation statements)

References 48 publications

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“…53 The culture device has also been used to study chondroinduction in other post-natal cell types, such as human gingival fibroblasts from elders and to identify the early gene regulatory events in chondroinduction. 54,55 In Vivo Responses to Collagen/DBP Sponges An early design criteria for a DBP/polymer composite device was that it be osteoinductive in vivo. Evidence showed that the mild inflammation that occurred with the implantation of DBP devices containing intact collagen completely blocked induction, whereas pepsin-digested telopeptide-free collagen-containing devices were inductive.…”

Section: In Vivo Fate Of Chondrocyte/collagen Spongesmentioning

confidence: 99%

Collagen scaffolds for tissue engineering

2007

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There are two major approaches to tissue engineering for regeneration of tissues and organs. One involves cell‐free materials and/or factors and one involves delivering cells to contribute to the regeneraion process. Of the many scaffold materials being investigated, collagen type I, with selective removal of its telopeptides, has been shown to have many advantageous features for both of these approaches. Highly porous collagen lattice sponges have been used to support in vitro growth of many types of tissues. Use of bioreactors to control in vitro perfusion of medium and to apply hydrostatic fluid pressure has been shown to enhance histogenesis in collagen scaffolds. Collagen sponges have also been developed to contain differentiating‐inducing materials like demineralized bone to stimulate differentiation of cartilage tissue both in vitro and in vivo. © 2007 Wiley Periodicals, Inc. Biopolymers 89: 338–344, 2008.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

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Section: In Vivo Fate Of Chondrocyte/collagen Spongesmentioning

confidence: 99%

Collagen scaffolds for tissue engineering

2007

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“…Three days was selected to identify genes that are induced prior to the expression of chondrocyte-specific genes. 17 RNA Isolation and RT-PCR Total RNA was isolated from sponges with Trizol reagent (Invitrogen) after 3 days of treatment. Two micrograms of total RNA was reverse-transcribed into cDNA with SuperScript II in 20 ml reaction volume according to the manufacturer's instructions (Invitrogen).…”

Section: Cell Cultures In Collagen Scaffoldsmentioning

confidence: 99%

Wnt pathway regulation by demineralized bone is approximated by both BMP‐2 and TGF‐β1 signaling

Zhou

Mizuno

Glowacki

2012

Journal Orthopaedic Research

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Allogeneic demineralized bone is used extensively as a clinical graft material because it has osteo/chondroinductive and osteoconductive properties. Demineralized bone powder (DBP) induces chondrogenic differentiation of human dermal fibroblasts (hDFs) in three-dimensional collagen cultures, but the initiating mechanisms have not been fully characterized nor has it been shown that bone morphogenetic proteins (BMPs) recapitulate DBP's effects on target cells. Among the many signaling pathways regulated in hDFs by DBP prior to in vitro chondrogenesis, there are changes in Wnts and their receptors that may contribute to DBP actions. This study tests the hypothesis that DBP modulation of Wnt signaling entails both BMP and TGF-b pathways. We compared the effects of DBP, TGF-b1, or BMP-2 on Wnt signaling components in hDFs by Wnt signaling macroarray, RT-PCR, in situ hybridization, and Western immunoblot analyses. Many effects of DBP on Wnt signaling components were not shared by BMP-2, and likewise DBP effects on Wnt genes and b-catenin only partially required the TGF-b pathway, as shown by selective inhibition of TGF-b/activin receptor-like kinase. The analyses revealed that 64% (16/25) of the Wnt signaling components regulated by DBP were regulated similarly by the sum of effects by BMP-2 and by TGF-b1. In conclusion, signaling mechanisms of inductive DBP in human dermal fibroblasts involve the modulation of multiple Wnt signals through both BMP and TGF-b pathways. ß

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“…Fibroblasts seeded on the sponge surface migrate throughout, contact the DBP and, after 7 days, are induced to express cartilage-specific matrix components such as Type II collagen and Aggrecan (Mizuno and Glowacki, 1996b). Analysis of differential gene expression in the DBP/ collagen sponge system showed that induction of cartilage phenotypic genes is preceded by transient shifts in many other genes (Yates, 2004;Yates et al, 2001;Zhou et al, 2004). Three days after seeding onto sponges, hDFs cultured with DBP show altered expression of cytoskeletal and matrix components, peptide growth factors, and signal transduction proteins.…”

Section: Introductionmentioning

confidence: 99%

“…The DBP-induced gene expression profile suggests that changes in cell shape and interactions with the matrix occur at early stages of chondroinduction in post-natal hDFs. Given that those cellular events are similar to the early stages in chondrocyte differentiation during embryonic development, and that DBP induces extensive changes in gene expression in hDFs by day 3, it is surprising that differential gene expression studies have identified very few transcription factors (although DBP does affect expression of transcriptional regulators that act at the chromatin level) (Yates et al, 2001;Zhou et al, 2004). Moreover, the transcription factors that have been identified (ID2, EGR-1) (Zhou et al, 2004) are not known to be major regulators of chondrogenesis.…”

Section: Introductionmentioning

confidence: 99%

“…Because DBP affects the expression of more than 300 genes (Yates and Glowacki, 2003b;Yates et al, 2001;Zhou et al, 2004), a strategy was devised to identify factor(s) that coordinate DBP-altered gene expression. A group of proteins that are among the most highly upregulated by DBP-TGF-β-Induced/BIGH3 (BIGH3), Type I collagen (COL1A2), and Fibronectin (FN1) (Zhou et al, 2004)-are also known to interact in the matrix (Kim et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

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Identification of cis and trans-acting transcriptional regulators in chondroinduced fibroblasts from the pre-phenotypic gene expression profile

Yates

2006

Gene

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Cell differentiation is regulated via expression of successive sets of genes. In an in vitro model of chondrocyte differentiation, human dermal fibroblasts (hDFs) cultured in collagen sponges are induced to express cartilage matrix genes after 7 days' culture with demineralized bone powder (DBP). A shift in expression of many other genes occurs within 3 days, before chondroblast phenotypic genes are detectable. In this study, the pre-chondrogenic gene expression profile was used as a starting point to derive information on transcriptional regulation of chondrocyte differentiation induced by DBP. Putative cis regulatory elements were identified by comparing promoter regions from three genes that are highly upregulated in chondroinduced hDFs (BIGH3, COL1A2, and FN1) [Zhou, S., Glowacki, J.,

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Early Shifts in Gene Expression during Chondroinduction of Human Dermal Fibroblasts

Cited by 21 publications

References 48 publications

Collagen scaffolds for tissue engineering

Collagen scaffolds for tissue engineering

Wnt pathway regulation by demineralized bone is approximated by both BMP‐2 and TGF‐β1 signaling

Identification of cis and trans-acting transcriptional regulators in chondroinduced fibroblasts from the pre-phenotypic gene expression profile

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