Modulation of Matrix Metalloprotease-2 Levels by Mechanical Loading of Three-Dimensional Mesenchymal Stem Cell Constructs: Impact onIn VitroTube Formation

Abstract

Angiogenesis is essential to tissue reconstitution, is sensitive to mechanical stresses, and currently represents one of the major challenges in tissue engineering. The pro-angiogenic matrix metalloprotease-2 (MMP-2) is upregulated in mechanically loaded mesenchymal stem cells (MSCs). Therefore, MMP-2 may provide a regulating link between angiogenesis and the surrounding mechanical conditions. This study aimed to modulate MMP-2 levels by mechanical loading of MSCs embedded in a three-dimensional matrix as well as to investigate the mechanism of MMP-2 regulation along with its contribution to angiogenesis stimulation. MMP-2-inducing conditions (30% compression, 1 Hz, 72 h) were defined after varying loading parameters. Addition of the Golgi-disturbing agent Brefeldin A suppressed this mechanical upregulation of MMP-2. Analysis of enzymatic activities demonstrated an enhancement of pro-MMP-2, mature MMP-2, and tissue inhibitor of metalloproteases-2. Further, mechano-regulation of MMP-14 and mature MMP-2 was dependent upon the activity of furin, a proprotein processing endoprotease. Angiogenesis was stimulated by conditioned media from MSCs loaded at inducing conditions. This augmentation of angiogenesis was hindered by inhibition of pro-MMP-2 and mature MMP-2. In conclusion, mechanical stimulation of MSCs in a three-dimensional matrix induces pro-MMP-2 secretion and MMP-2 activation, potentially via the activation complex consisting of MMP-2/-14/tissue inhibitor of metalloproteases-2. Mechano-regulated pro-MMP-2 and mature MMP-2 seem to contribute to angiogenesis stimulation. Thus, an application of these loading parameters could augment vascularization of tissue-engineered constructs based on the described MMP-2-dependent mechanism.

Paracrine Factors Secreted by Umbilical Cord-Derived Mesenchymal Stem Cells Induce Angiogenesis In Vitro by a VEGF-Independent Pathway

“…Similarly, the ability of a higher concentration of GM6001 to block CM-induced capillary network formation as compared to the control media suggests that MMP-2 present in the CM plays an important role in CM-induced angiogenesis. The role of MMP-2 in inducing endothelial network formation reported in this study is supported by Glaeser et al who had reported that pro-MMP-2 secreted by MSCs is further activated by endothelial cells and the total MMP-2 plays a pivotal role in endothelial network formation [74].…”

Effects of matrix metalloproteinases on the fate of mesenchymal stem cells

“…MMPs that are expressed by MSCs can contribute to that role during the interaction of MSCs and ECs [4]. Glaeser and colleagues [59] demonstrated that the inhibition of MMP-2 attenuated the tube-like formation. Under mechanical stimulation by MSCs, the secretion of proMMP-2 and the activity of MMP-2 were induced and, as a result, this induction contributes to angiogenesis stimulation.…”

In serum veritas—in serum sanitas? Cell non-autonomous aging compromises differentiation and survival of mesenchymal stromal cells via the oxidative stress pathway

“…1, 2 This specific regeneration potential arises not only from the MSC ability to differentiate into various mesenchymal cell types to replace cells in damaged tissues but also from paracrine effects of the cells modulating injury or immune responses. 3, 4, 5, 6 However, healing capacity of mesenchymal tissues, especially of bone and muscle, has been shown to decline with increasing age. 7, 8, 9 In particular, our previous in vivo study showed an age-related delay in the course of bone healing, resulting in an altered microstructure and in reduced mechanical properties of the regenerated tissue.…”