Correlating the effects of bone morphogenic protein to secreted soluble factors from fibroblasts and mesenchymal stem cells in regulating regenerative processes in vitro.

The capacity to regenerate complex tissue structures after amputation in humans is limited to the digit tip. In a comparable mouse digit model, which includes both distal regeneration-competent and proximal regeneration-incompetent regions, successful regeneration involves precise orchestration of complex microenvironmental cues, including paracrine signaling via heterogeneous cell-cell interactions. Initial cellular processes, such as proliferation and migration, are critical in the formation of an initial stable cell mass and the ultimate regenerative outcome. Hence, the objective of these in vitro studies was to investigate the effect of soluble factors secreted by fibroblasts and mesenchymal stem cells (MSCs) on the proliferation and migration of cells from the regeneration-competent (P3) and -incompetent (P2) regions of the mouse digit tip. We found that P2 and P3 cells were more responsive to fibroblasts than MSCs and that the effects were mediated by bi-directional communication. To initiate understanding of the specific soluble factors that may be involved in the fibroblast-mediated changes in migration of P2 and P3 cells, bone morphogenic protein 2 (BMP2) was exogenously added to the medium. We found that changes in migration of P3 cells were similar when exposed to BMP2 or co-cultured with fibroblasts, indicating that BMP signaling may be responsible for the migratory response of P3 cells to the presence of fibroblasts. Furthermore, BMP2 expression in fibroblasts was shown to be responsive to tensile strain, as is present during wound closure. Therefore, these in vitro studies indicate that regenerative processes may be regulated by fibroblast-secreted soluble factors, which, in turn, are modulated by both cross-talk between heterogeneous phenotypes and the physical microenvironment of the healing site.