Osteogenic differentiation of human mesenchymal stromal cells and fibroblasts differs depending on tissue origin and replicative senescence

Abstract

The need for an autologous cell source for bone tissue engineering and medical applications has led researchers to explore multipotent mesenchymal stromal cells (MSC), which show stem cell plasticity, in various human tissues. However, MSC with different tissue origins vary in their biological properties and their capability for osteogenic differentiation. Furthermore, MSC-based therapies require large-scale ex vivo expansion, accompanied by cell type-specific replicative senescence, which affects osteogenic differentiation. To elucidate cell type-specific differences in the osteogenic differentiation potential and replicative senescence, we analysed the impact of BMP and TGF-β signaling in adipose-derived stromal cells (ASC), fibroblasts (FB), and dental pulp stromal cells (DSC). We used inhibitors of BMP and TGF-β signaling, such as SB431542, dorsomorphin and/or a supplemental addition of BMP-2. The expression of high-affinity binding receptors for BMP-2 and calcium deposition with alizarin red S were evaluated to assess osteogenic differentiation potential. Our study demonstrated that TGF-β signaling inhibits osteogenic differentiation of ASC, DSC and FB in the early cell culture passages. Moreover, DSC had the best osteogenic differentiation potential and an activation of BMP signaling with BMP-2 could further enhance this capacity. This phenomenon is likely due to an increased expression of activin receptor-like kinase-3 and -6. However, in DSC with replicative senescence (in cell culture passage 10), osteogenic differentiation sharply decreased, and the simultaneous use of BMP-2 and SB431542 did not result in further improvement of this process. In comparison, ASC retain a similar osteogenic differentiation potential regardless of whether they were in the early (cell culture passage 3) or later (cell culture passage 10) stages. Our study elucidated that ASC, DSC, and FB vary functionally in their osteogenic differentiation, depending on their tissue origin and replicative senescence. Therefore, our study provides important insights for cell-based therapies to optimize prospective bone tissue engineering strategies.

The Role of MSCs and Cell Fusion in Tissue Regeneration

“…By the use of an IL-1R1/MyD88 inhibitor, the regenerative potential of MSCs could be improved. In cell culture studies, TGFβ, an anti-inflammatory cytokine, has inhibited the osteogenic differentiation of MSCs [49]. Similar has been reported for muscle cell differentiation [52].…”

“…There seem to be various factors being involved in the regulation of MSCs and their regenerative capacity. In addition to cultivation, the origin of the MSCs is another important factor, because the osteogenic capacity can vary between cells due to the usage of different signaling cascades [49]. Nevertheless, MSCs are the most interesting agents in bone remodeling.…”

Probing Notch1-Dll4 Signaling in Regulating Osteogenic Differentiation of Human Mesenchymal Stem Cells using Single Cell Nanobiosensor

“…These results indicate that Dll4 mRNA expression dynamics indicate two different stages during osteogenic differentiation: early differentiation stage (day 1day 5) and calcium mineralization stage (after day 5). It has been reported that the process of hMSCs osteogenic differentiation includes early lineage progression (proliferation), early differentiation, and later stage differentiation (calcium mineralization) [28,[30][31][32]. Our results indicate that Dll4 mRNA expression could potentially be the molecular signature of early osteogenic differentiation.…”

Heterogeneity of In Vitro Expanded Mesenchymal Stromal Cells and Strategies to Improve Their Therapeutic Actions

“…A comparative analysis demonstrated that MSCs obtained from bone marrow present a higher osteogenic differentiation potential than MSCs derived from adipose tissue, which are more adipogenic [ 15 ]. Similarly, dental pulp stromal cells displayed a greater capacity to differentiate into osteocytes than Ad-MSCs [ 58 ]. Another study found that umbilical cord blood MSCs possess a reduced capacity to differentiate into adipocytes, in contrast to BM-MSCs and Ad-MSCs [ 33 , 59 ].…”