Construction of autonomous osteogenic microtissues based on human dental pulp derived mesenchymal stem cells

Bone regeneration is currently a major clinical challenge. Adult human mesenchymal stem cells (MSCs) possess a unique advantage in organ tissue repair and physiological function recovery. Human dental pulp derived MSCs (hDPSCs) have the dual advantages of proliferation and osteogenic differentiation abilities. They can be induced to secrete bone matrix proteins to form osteogenic microtissues by constructing an appropriate osteogenic three-dimensional microenvironment. The biological characteristics of hDPSCs from microtissues and two-dimensional cultured cells are compared using RNA sequencing. According to RNA sequencing and subsequent functional verification, the osteogenic activations of hDPSCs might be related to cell adhesion related signaling pathways. In this process, cells could sense the tension from the environment to activate related signaling pathways, such as the FAK/RhoA/Rock signaling pathway. Further, the downstream osteogenic transcription factors could be activited into the nucleus, thereby promoting the expression of osteogenic proteins. To construct human dental pulp derived mesenchymal stem cells (hDPSCs) based osteogenic microtissues (hDOMTs) and evaluate its bioactivity and osteogenic potential. We performed gene expression profiling analysis using data obtained from RNA-seq of hDPSCs. We compared the differential expression genes between the RNA-seq data of two-dimensional cultured hDPSCs and hDPSCs from the microtissues, and their osteogenic induced patterns, respectively.