Jens Tuischer

Mesenchymal Stem Cells Regulate Angiogenesis According to Their Mechanical Environment

In fracture and bone defect healing, MSCs largely drive tissue regeneration. MSCs have been shown to promote angiogenesis both in vivo and in vitro. Angiogenesis is a prerequisite to large tissue reconstitution. The present study investigated how mechanical loading of MSCs influences their proangiogenic capacity. The results show a significant enhancement of angiogenesis by conditioned media from mechanically stimulated compared with unstimulated MSCs in two‐dimensional tube formation and three‐dimensional spheroid sprouting assays. In particular, proliferation but not migration or adhesion of endothelial cells was elevated. Promotion of angiogenesis was dependent upon fibroblast growth factor receptor 1 (FGFR1) signaling. Moreover, stimulation of tube formation was inhibited by vascular endothelial growth factor receptor (VEGFR) tyrosine kinase blocking. Screening for the expression levels of different soluble regulators of angiogenesis revealed an enrichment of matrix metalloprotease 2, transforming growth factor β1, and basic fibroblast growth factor but not of vascular endothelial growth factor in response to mechanical stimulation. In conclusion, mechanical loading of MSCs seems to result in a paracrine stimulation of angiogenesis, most likely by the regulation of a network of several angiogenic molecules. The underlying mechanism appears to be dependent on the FGFR and VEGFR signaling cascades and might be mediated by an additional cross‐talk with other pathways.

Matrix Metalloprotease Activity Is an Essential Link Between Mechanical Stimulus and Mesenchymal Stem Cell Behavior

Progenitor cells are involved in the regeneration of the musculoskeletal system, which is known to be influenced by mechanical boundary conditions. Furthermore, matrix metalloproteases (MMPs) and tissue‐specific inhibitors of metalloproteases (TIMPs) are crucial for matrix remodelling processes that occur during regeneration of bone and other tissues. This study has therefore investigated whether MMP activity affects mesenchymal stem cell (MSC) behavior and how MMP activity is influenced by the mechanical stimulation of these cells. Broad spectrum inhibition of MMPs altered the migration, proliferation, and osteogenic differentiation of MSCs. Expression analysis detected MMP‐2, ‐3, ‐10, ‐11, ‐13, and ‐14, as well as TIMP‐2, in MSCs at the mRNA and protein levels. Mechanical stimulation of MSCs led to an upregulation of their extracellular gelatinolytic activity, which was consistent with the increased protein levels seen for MMP‐2, ‐3, ‐13, and TIMP‐2. However, mRNA expression levels of MMPs/TIMPs showed no changes in response to mechanical stimulation, indicating an involvement of post‐transcriptional regulatory processes such as alterations in MMP secretion or activation. One potential regulatory molecule might be the furin protease. Specific inhibition of MMP‐2, ‐3, and ‐13 showed MMP‐13 to be involved in osteogenic differentiation. The results of this study suggest that MSC function is controlled by MMP activity, which in turn is regulated by mechanical stimulation of cells. Thus, MMP/TIMP balance seems to play an essential role in transferring mechanical signals into MSC function.

Local application of a collagen type I/hyaluronate matrix and growth and differentiation factor 5 influences the closure of osteochondral defects in a minipig model by enchondral ossification.

This pilot study evaluated the effect of growth and differentiation factor-5 (rhGDF-5) combined with a collagen type I/hyaluronate matrix (c/h) on osteochondral defect repair in a minipig model. Defects created in both medial femoral condyles of 20 minipigs were treated with c/h (n = 10), c/h + rhGDF-5 (n = 10) or were left empty. After 3 and 12 months, five animals of each group were sacrificed. Evaluation included macroscopic and histological scoring and quantitative histomorphometry of synthesized bone. C/h and c/h + rhGDF-5 treatment increased trabecular bone formation in the upper third of the defect compared to empty controls, showing significance for c/h + rhGDF-5 (p = 0.05) but not between c/h and c/h + rhGDF-5 treatment. Cartilage regeneration and macroscopic outcome were not improved by c/h or c/h + rhGDF-5 treatment. Since c/h remnants were seen even one year postoperatively in the defect, possibly inhibiting further bone and cartilage healing, other matrices in combination with rhGDF-5 may provide further improvement in osteochondral defect treatment.

Effects of tacrolimus, cyclosporin A and sirolimus on MG63 cells

The reduction in bone mineral density after organ transplantation results in increased morbidity (post‐transplantation bone disease) and remains an unsolved problem. A connection with the long‐term application of nonglucocorticoidal immunosuppressants is the subject of controversial discussion. We hypothesized that such substances have an influence on the skeletal system on the cellular level by modulating osteoblast differentiation. Therefore, we investigated the effects of tacrolimus, cyclosporin A and sirolimus as representative substances of nonglucocorticoidal immunosuppressants on cell proliferation and expression of bone tissue‐specific genes of human osteoblasts (MG63). None of the examined substances affected cell proliferation, but all influenced the gene expression pattern towards change in cell differentiation. In detail, collagen III and XII, matrix metalloproteinase 2, SMAD2, epithelial growth factor receptor, annexin V and osteonectin expression were increased by all of the examined substances. Tacrolimus, cyclosporin A and sirolimus influence intracellular signalling pathways, transmembranous receptors and bone‐specific matrix synthesis. They do not have antiproliferative or toxic effects. We postulate that the shown changes of osteoblast differentiation cause post‐transplantation disease.