Chondroitin sulfate based niches for chondrogenic differentiation of mesenchymal stem cells.

Bone marrow-derived mesenchymal stem cells (MSCs) have strong potential in regeneration of musculoskeletal tissues including cartilage and bone. The microenvironment, comprising of scaffold and soluble factors, plays a pivotal role in determining the efficacy of cartilage tissue regeneration from MSCs. In this study, we investigated the effect of a three-dimensional synthetic-biological composite hydrogel scaffold comprised of poly (ethylene glycol) (PEG) and chondroitin sulfate (CS) on chondrogenesis of MSCs. The cells in CS-based bioactive hydrogels aggregated in a fashion which mimicked the mesenchymal condensation and produced cartilaginous tissues with characteristic morphology and basophilic extracellular matrix production. The aggregation of cells resulted in an enhancement of both chondrogenic gene expressions and cartilage specific matrix production compared to control PEG hydrogels containing no CS-moieties. Moreover, a significant down-regulation of type X collagen expression was observed in PEG/CS hydrogels, indicating that CS inhibits the further differentiation of MSCs into hypertrophic chondrocytes. Overall, this study demonstrates the morphogenetic role of bioactive scaffold-mediated microenvironment on temporal pattern of cartilage specific gene expressions and subsequent matrix production during MSC chondrogenesis.

[1]  Gordana Vunjak-Novakovic,et al.  Tissue Engineering of Cartilage , 1999 .

[2]  Freddie H. Fu,et al.  GAG-augmented polysaccharide hydrogel: a novel biocompatible and biodegradable material to support chondrogenesis. , 2000, Journal of biomedical materials research.

[3]  Nathaniel S. Hwang,et al.  Chondrogenic differentiation of human embryonic stem cell-derived cells in arginine-glycine-aspartate-modified hydrogels. , 2006, Tissue engineering.

[4]  F. Flamigni,et al.  Chondrocyte hypertrophy and apoptosis induced by GROα require three‐dimensional interaction with the extracellular matrix and a co‐receptor role of chondroitin sulfate and are associated with the mitochondrial splicing variant of cathepsin B , 2007, Journal of cellular physiology.

[5]  R. Tuan Stemming cartilage degeneration: adult mesenchymal stem cells as a cell source for articular cartilage tissue engineering. , 2006, Arthritis and rheumatism.

[6]  D. Huang EFFECT OF EXTRACELLULAR CHONDROITIN SULFATE ON CULTURED CHONDROCYTES , 1974, The Journal of cell biology.

[7]  Christopher G Williams,et al.  In vitro chondrogenesis of bone marrow-derived mesenchymal stem cells in a photopolymerizing hydrogel. , 2003, Tissue engineering.

[8]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[9]  A. Grodzinsky,et al.  Fluorometric assay of DNA in cartilage explants using Hoechst 33258. , 1988, Analytical biochemistry.

[10]  Yang Luo,et al.  N‐cadherin is not essential for limb mesenchymal chondrogenesis , 2005, Developmental dynamics : an official publication of the American Association of Anatomists.

[11]  Dong-An Wang,et al.  Heterogeneous-Phase Reaction of Glycidyl Methacrylate and Chondroitin Sulfate: Mechanism of Ring-Opening−Transesterification Competition , 2003 .

[12]  R. Tuan,et al.  Expression and functional involvement of N-cadherin in embryonic limb chondrogenesis. , 1994, Development.

[13]  P. Prendergast,et al.  A collagen-glycosaminoglycan scaffold supports adult rat mesenchymal stem cell differentiation along osteogenic and chondrogenic routes. , 2006, Tissue engineering.

[14]  Darwin J. Prockop,et al.  In vitro cartilage formation by human adult stem cells from bone marrow stroma defines the sequence of cellular and molecular events during chondrogenesis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Nagarajan Vaidehi,et al.  Sulfation patterns of glycosaminoglycans encode molecular recognition and activity , 2006, Nature chemical biology.

[16]  Yumiko Saga,et al.  Cell‐Cell Interaction Mediated by Cadherin‐11 Directly Regulates the Differentiation of Mesenchymal Cells Into the Cells of the Osteo‐Lineage and the Chondro‐Lineage , 2004, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[17]  J. F. Woessner,et al.  The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. , 1961, Archives of biochemistry and biophysics.

[18]  F. Barry,et al.  Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. , 2001, Experimental cell research.

[19]  D. Huang Extracellular Matrix‐Cell Interactions and Chondrogenesis , 1977, Clinical orthopaedics and related research.

[20]  Joseph D. Bronzino,et al.  The Biomedical Engineering Handbook , 1995 .

[21]  Jun Wang,et al.  Photocrosslinkable polysaccharides based on chondroitin sulfate. , 2004, Journal of biomedical materials research. Part A.

[22]  S. Varghese,et al.  Designing new thermoreversible gels by molecular tailoring of hydrophilic-hydrophobic interactions , 2000 .

[23]  R. Tuan,et al.  Analysis of N‐cadherin function in limb mesenchymal chondrogenesis in vitro , 2002, Developmental dynamics : an official publication of the American Association of Anatomists.

[24]  J. Elisseeff,et al.  Hydrogels for musculoskeletal tissue engineering , 2006 .

[25]  D. Buttle,et al.  Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. , 1986, Biochimica et biophysica acta.

[26]  V. Pipitone Chondroprotection with chondroitin sulfate. , 1991, Drugs under experimental and clinical research.

[27]  V. Lee,et al.  Versican modulates embryonic chondrocyte morphology via the epidermal growth factor-like motifs in G3. , 2001, Experimental cell research.

[28]  K. Shimizu,et al.  Versican/PG-M Regulates Chondrogenesis as an Extracellular Matrix Molecule Crucial for Mesenchymal Condensation* , 2006, Journal of Biological Chemistry.

[29]  F. Ronca,et al.  Anti-inflammatory activity of chondroitin sulfate. , 1998, Osteoarthritis and cartilage.

[30]  Jeffrey A Hubbell,et al.  Materials as morphogenetic guides in tissue engineering. , 2003, Current opinion in biotechnology.