Glycosaminoglycans enhance osteoblast differentiation of bone marrow derived human mesenchymal stem cells

Abstract

Extracellular matrix plays an important role in regulating cell growth and differentiation. The biomimetic approach of cell‐based tissue engineering is based on mirroring this in vivo micro environment for developing a functional tissue engineered construct. In this study, we treated normal tissue culture plates with selected extracellular matrix components consisting of glycosaminoglycans such as chondroitin‐4‐sulphate, dermatan sulphate, chondroitin‐6‐sulphate, heparin and hyaluronic acid. Mesenchymal stem cells isolated from adult human bone marrow were cultured on the glycosaminoglycan treated culture plates to evaluate their regulatory role in cell growth and osteoblast differentiation. Although no significant improvement on human mesenchymal stem cell adhesion and proliferation was observed on the glycosaminoglycan‐treated tissue culture plates, there was selective osteoblast differentiation, indicating its potential role in differentiation rather than proliferation. Osteoblast differentiation studies showed high osteogenic potential for all tested glycosaminoglycans except chondroitin‐4‐sulphate. Osteoblast differentiation‐associated genes such as osterix, osteocalcin, integrin binding sialoprotein, osteonectin and collagen, type 1, alpha 1 showed significant upregulation. We identified osterix as the key transcription factor responsible for the enhanced bone matrix deposition observed on hyaluronic acid, heparin and chondroitin‐6‐sulphate. Hyaluronic acid provided the most favourable condition for osteoblast differentiation and bone matrix synthesis. Our results confirm and emphasise the significant role of extracellular matrix in regulating cell differentiation. To summarise, glycosaminoglycans of extracellular matrix played a significant role in regulating osteoblast differentiation and could be exploited in the biomimetic approach of fabricating or functionalizing scaffolds for stem cell based bone tissue engineering. Copyright © 2012 John Wiley & Sons, Ltd.

Heparin modification of a biomimetic bone matrix modulates osteogenic and angiogenic cell response in vitro

“…Whereas existing data in the literature primarily show increased ALP expression and activity and mineralisation in response to heparin, the situation regarding the regulation of matrix protein expression is not clear: Hausser and Brenner (2004) described a concentrationdependent upregulation of OC by Saos-2 cells. Mathews et al (2014) found in hBMSC cultured on heparin-treated tissue culture plates an upregulated OC expression too, but no significant upregulation of Col I and BSP II expression. Recently, Simann et al (2015) investigated the effect of heparin on osteogenic lineage commitment of hBMSC and found also no upregulation of OC, but concluded from their observations of ALP stimulation and enhanced mineralisation that heparin supported osteogenic differentiation.…”

“…Benoit et al (2007) for example measured about 30 ng/mL fibronectin and 84 pg/mL bone morphogenetic protein II (BMP II) in cell culture medium containing 10 % FCS. The high affinity of heparin to adhesion molecules like fibronectin and vitronectin facilitates not only a better cell attachment and proliferation (Uygun et al, 2009;Kim et al, 2013) but also promotes the differentiation of pre-osteoblastic cells through integrin mediated signal transduction (Mathews et al, 2012).…”

FOXD1 promotes nephron progenitor differentiation by repressing decorin in the embryonic kidney

“…In this work, we demonstrate that the aberrant ECM is sufficient to reduce differentiation of progenitor cells to epithelial structures. Stem and progenitor cells, in particular, are sensitive to changes in the ECM environment (Ichii et al, 2012;Mathews et al, 2012a;Mathews et al, 2012b;Wang et al, 2010).…”

Osteogenic Potential of Mesenchymal Stromal Cells Contributes to Primary Myelofibrosis

“…In primary myelofibrosis, we demonstrated an increased production of GAGs in vitro by MSCs and in vivo on bone marrow biopsies; this overproduction is higher in evolved patients (fibrosis grade 3). It is reported that enhancement of GAGs promotes osteogenic commitment (34) and that TGFb1 exerts a dual action on osteoblast differentiation. Actually, whereas TGFb1 disruption in mice results in bone defects, its overexpression displays an osteoporosis-like phenotype (35), suggesting that TGFb1 enhances the proliferation and early differentiation of osteoblasts, but impairs their terminal differentiation.…”