Hydrostatic Pressure Enhances Chondrogenic Differentiation of Human Bone Marrow Stromal Cells in Osteochondrogenic Medium

Abstract

This study demonstrated the chondrogenic effect of hydrostatic pressure on human bone marrow stromal cells (MSCs) cultured in a mixed medium containing osteogenic and chondrogenic factors. MSCs seeded in type I collagen sponges were exposed to 1 MPa of intermittent hydrostatic pressure at a frequency of 1 Hz for 4 h per day for 10 days, or remained in identical culture conditions but without exposure to pressure. Afterwards, we compared the proteoglycan content of loaded and control cell/scaffold constructs with Alcian blue staining. We also used real-time PCR to evaluate the change in mRNA expression of selected genes associated with chondrogenic and osteogenic differentiation (aggrecan, type I collagen, type II collagen, Runx2 (Cbfa-1), Sox9, and TGF-beta1). With the hydrostatic pressure loading regime, proteoglycan staining increased markedly. Correspondingly, the mRNA expression of chondrogenic genes such as aggrecan, type II collagen, and Sox9 increased significantly. We also saw a significant increase in the mRNA expression of type I collagen, but no change in the expression of Runx2 or TGF-beta1 mRNA. This study demonstrated that hydrostatic pressure enhanced differentiation of MSCs in the presence of multipotent differentiation factors in vitro, and suggests the critical role that this loading regime may play during cartilage development and regeneration in vivo.

The Role of Environmental Factors in Regulating the Development of Cartilaginous Grafts Engineered Using Osteoarthritic Human Infrapatellar Fat Pad–Derived Stem Cells

“…This is in contrast to the results of previous studies demonstrating that cyclic HP can enhance chondrogenesis of bone marrow 42,[80][81][82][83][84][85] and adipose-derived 86 stem cells. These FIG.…”

The role of mechanical signals in regulating chondrogenesis and osteogenesis of mesenchymal stem cells

“…This is in contrast to previously described increases observed in MSC aggregates , which may be due to greater cell-cell proximity in these pellets compared to agarose hydrogels (Finger et al, 2007). In bone marrow derived MSCs embedded in type I collagen sponges and subjected to hydrostatic pressure in a media containing osteogenic and chondrogenic factors, mRNA expression of chondrogenic genes such as aggrecan, type II collagen and Sox9 increased, with no change in the expression of the osteogenic marker Runx2 (Wagner et al, 2008). Significantly lower magnitudes of dynamic pressure have also been shown to promote chondrogenesis of MSCs embedded in alginate beads, and this response was at least partially regulated via activated phosphorylation of p38 MAPK ).…”

Cyclic hydrostatic pressure promotes a stable cartilage phenotype and enhances the functional development of cartilaginous grafts engineered using multipotent stromal cells isolated from bone marrow and infrapatellar fat pad

“…Articular cartilage is a multiphasic tissue, the solid phase of which is composed predominately of a collagen (mainly type II) fibrillar network (around 10-22% by wet weight) enmeshing proteoglycan macromolecules (2-7% by wet weight) (Maroudas et al, 1969;Muir et al, 1970;Bayliss and Ali, 1978;Inerot et al, 1978;Maroudas et al, 1980;Zirn et al, 1984;Mow et al, 1992;Rieppo et al, 2009;Gannon et al, 2012). Previous studies have demonstrated that the application of dynamic compression (Huang et al, 2010;Bian et al, 2012;Thorpe et al, 2013) or hydrostatic pressure (Miyanishi et al, 2006;Wagner et al, 2008;Ogawa et al, 2009;Huang et al, 2010;Meyer et al, 2011;Correia et al, 2012;Liu et al, 2012;Steward et al, 2012;Vinardell et al, 2012a;Liu et al, 2013) can enhance the sGAG and collagen content as well as the mechanical functionality of cartilage constructs engineered using MSCs. However, these studies often fail to produce cartilage grafts with mechanical functionality or ECM content approaching that of native cartilage.…”