Chondrogenic Differentiation on Perlecan Domain I, Collagen II, and Bone Morphogenetic Protein-2-Based Matrices

Yang, Weidong; Gomes, Ronald R.; Brown, Anissa J.; Burdett, Ashley R.; Alicknavitch, Michael; Farach‐Carson, Mary C.; Carson, Daniel D.

doi:10.1089/ten.2006.12.ft-105

Cited by 12 publications

(18 citation statements)

References 56 publications

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“…To measure the chondrogenic differentiation potential of BMP-2 loaded HGPs (HGP-B 2 and HGP-P 1 -B 2 ), micromass of murine mesenchymal stem cells (MSCs, C3H10T1/2 cell line, ATCC, Manassas, VA) was cultured following previously reported procedures [25]. Particles (3 mg) suspended in 200 µL of media were subsequently added directly to the micromass.…”

Section: Methodsmentioning

confidence: 99%

“…After 9 days of incubation, micromass cultures were washed with PBS, fixed with 10% (v/v) buffered formalin acetate containing 0.5% (w/v) cetylpyridinium chloride for 10 min at room temperature, briefly rinsed with 3% (v/v) glacial acetic acid (pH 1.0) and incubated in 1 mL of 0.5% (w/v) Alcian blue in 3% (v/v) glacial acetic acid (pH 1.0) overnight at room temperature [25]. The stained micromass culture was photographed with a digital camera attached to a Nikon microscope (Coolpix 990, Nikon, Kanagawa, Japan).…”

Section: Methodsmentioning

confidence: 99%

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Perlecan domain I-conjugated, hyaluronic acid-based hydrogel particles for enhanced chondrogenic differentiation via BMP-2 release

et al. 2009

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We have developed a biomimetic growth factor delivery system that effectively stimulates the chondrogenic differentiation of the cultured mesenchymal stem cells via the controlled presentation of bone morphogenetic protein 2 (BMP-2). Hyaluronic acid (HA)-based, microscopic hydrogel particles (HGPs) with inherent nanopores and defined functional groups were synthesized by an inverse emulsion polymerization technique. Recombinantly produced, heparan sulfate (HS)-bearing perlecan domain I (PlnDI) was covalently immobilized to HA HGPs (HGP-P1) via a flexible poly(ethylene glycol) (PEG) linker through the lysine amines in the core protein of PlnDI employing reductive amination. Compared to HGP without PlnDI, HGP-P1 exhibited significantly (p<0.05) higher BMP-2 binding capacity and distinctly different BMP-2 release kinetics. Heparitinase treatment increased the amount of BMP-2 released from HGP-P1, confirming the HS-dependent BMP-2 binding. While BMP-2 was released from HGPs with a distinct burst release followed by a minimal cumulative release, its release from HGP-P1 exhibited a minimal burst release followed by linear release kinetics over 15 days. The bioactivity of the hydrogel particles was evaluated using micromass culture of multipotent mesenchymal stem cells (MSCs), and the chondrogenic differentiation was assessed by the production of glycosaminoglycan, aggrecan and collagen type II. Our results revealed that BMP-2 loaded HGP-P1 stimulates more robust cartilage specific ECM production as compared to BMP-2 loaded HGP, due to the ability of HGP-P1 to potentiate BMP-2 and modulate its release with a near zero-order release kinetics. The PlnDI conjugated, HA HGPs provide an improved BMP-2 delivery system for stimulating chondrogenic differentiation in vitro, with potential therapeutic application for cartilage repair and regeneration.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Perlecan domain I-conjugated, hyaluronic acid-based hydrogel particles for enhanced chondrogenic differentiation via BMP-2 release

et al. 2009

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“…(12) Type II collagen was reported to promote chondrogenesis and maintain chondrocyte morphology and phenotype. (13)(14)(15) Therefore, cell surface collagen receptors play critical roles in collagen control of osteoblast and chondrocyte differentiation. Two collagen receptor subfamilies have been identified, integrins and discoidin domain receptors (DDRs).…”

Section: Introductionmentioning

confidence: 99%

An essential role of discoidin domain receptor 2 (DDR2) in osteoblast differentiation and chondrocyte maturation via modulation of Runx2 activation

Zhang

et al. 2010

Journal of Bone and Mineral Research

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Discoidin domain receptor 2 (DDR2) belongs to receptor tyrosine kinase (RTK) family and is activated by collagen binding. Although the bone defects in Ddr2 null mice have been reported for a decade, the molecular mechanism remains unclear. This study sought to investigate the function and detailed mechanism of DDR2 in osteogenic and chondrogenic differentiation. Herein we found that in preosteoblastic cells, DDR2 activation was enhanced by osteogenic induction but was not paralleled with the alteration of DDR2 expression. Under differentiated condition, downregulation of endogenous DDR2 through specific shRNA dramatically repressed osteoblastic marker gene expression and osteogenic differentiation. Enforced expression of constitutively activated DDR2 increased the expression of bone markers in both undifferentiated and differentiated osteoblasts. Importantly, molecular evidence showed that DDR2 regulated the transactivity of Runx2, a master transcription factor involved in skeletal development, by modulating its phosphorylation. Analysis of candidate protein kinases indicated that extracellular signal-regulated kinase (ERK) activation is responsive to DDR2 signaling and involved in DDR2 regulation of Runx2 phosphorylation and transcriptional activity. Notably, a gain-of-function mutant of Runx2 with enhanced ERK-independent phosphorylation rescued the impaired osteogenic phenotypes observed in Ddr2-silenced cells, whereas a Runx2 mutant devoid of phosphorylation regulation by ERK inhibited DDR2 induction of osteogenesis. In addition, DDR2 facilitated Runx2 transactivation and type X collagen expression in hypertrophic chondrocytes. Thus this study reveals for the first time that DDR2 plays an essential role in osteoblast and chondrocyte differentiation. The mechanism disclosure may provide therapeutic targets for human genetic disorders caused by DDR2 deficiency. ß

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“…29 Heparin has been shown to be necessary for FGF signaling, 31 and there have been a number of models proposed to explain this phenomenon, all of which involve the presence of the heparin/HS polysaccharide, growth factor and growth factor receptor that result in growth factor and receptor dimerization and signaling. 32 -35 Recently, HS containing fragments of mouse perlecan have been shown to control chondrocyte phenotype in vitro, 36 bind FGF2 in combination with collagen-based scaffolds, 37 be biologically active 38 and shown to be manufactured in a way that is suitable for tissue engineering. 39 Endothelial cell adhesion to the recombinant material was analysed and compared to full-length perlecan (Fig.…”

Section: Biological Activity Of Recombinant Materialsmentioning

confidence: 99%

Recombinant heparan sulfate for use in tissue engineering applications

Whitelock

Leo

Davies

et al. 2008

J of Chemical Tech & Biotech

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BACKGROUND: Heparan sulfate (HS) is an important component of many extracellular matrices that interacts with mitogens and morphogens to guide and control tissue and organ development. These interactions are controlled by its structure, which varies when produced by different cell types and different species. The major aim of the studies reported here was to isolate and characterize the HS expressed on the N-terminal domain of human perlecan when it is expressed in human cells.

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Chondrogenic Differentiation on Perlecan Domain I, Collagen II, and Bone Morphogenetic Protein-2-Based Matrices

Cited by 12 publications

References 56 publications

Perlecan domain I-conjugated, hyaluronic acid-based hydrogel particles for enhanced chondrogenic differentiation via BMP-2 release

Perlecan domain I-conjugated, hyaluronic acid-based hydrogel particles for enhanced chondrogenic differentiation via BMP-2 release

An essential role of discoidin domain receptor 2 (DDR2) in osteoblast differentiation and chondrocyte maturation via modulation of Runx2 activation

Recombinant heparan sulfate for use in tissue engineering applications

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