Biological Responses and Mechanisms of Human Bone Marrow Mesenchymal Stem Cells to Zn and Mg Biomaterials

Zn biomaterials attract strong attentions recently for load-bearing medical implants because of their mechanical properties similar to bone, biocompatibility, and degradability at a more matched rate to tissue healing. It has been shown previously that Zn alloys are beneficial for bone regeneration, but the supporting mechanisms have not been explored in detail. Here, we studied the biological responses of human bone marrow mesenchymal stem cells (hMSC) to Zn and the underlying cellular signaling mechanisms. Typical Mg material AZ31 was used as a comparative benchmark control. Direct culture of cells on the materials revealed that cell adhesion, proliferation, and motility were higher on Zn than on AZ31. Significant cytoskeletal reorganizations induced by Zn or AZ31 were also observed. Mineralization of extracellular matrix (ECM) and hMSC osteogenic differentiation, measured by Alizarin red and ALP staining and activities, were significantly enhanced when cells were cultured with Zn or AZ31. Quantitative PCR also showed the increased expression of bone-related genes including ALP, collagen I, and osteopontin. Using small RNA interference to knockdown related key molecules, we illustrated the mechanisms of Zn-induced cellular signaling. TRPM7 and GPR39 appear to be the major cellular receptors facilitating Zn²⁺-entry into hMSC. The intracellular Zn²⁺ then activates the cAMP-PKA pathway and triggers intracellular Ca²⁺ responses, leading to activation of MAPK. In addition, Zn²⁺ activates the Gαq-PLC-AKT pathway as well. Eventually, all of this signaling would lead to enhanced differential regulation of genes, cell survival/growth and differentiation, ECM mineralization, osteogenesis, and other cellular activities.