Global MicroRNA Profiling in Human Bone Marrow Skeletal—Stromal or Mesenchymal–Stem Cells Identified Candidates for Bone Regeneration

Abstract

Bone remodeling and regeneration are highly regulated multistep processes involving posttranscriptional regulation by microRNAs (miRNAs). Here, we performed a global profiling of differentially expressed miRNAs in bone-marrow-derived skeletal cells (BMSCs; also known as stromal or mesenchymal stem cells) during in vitro osteoblast differentiation. We functionally validated the regulatory effects of several miRNAs on osteoblast differentiation and identified 15 miRNAs, most significantly miR-222 and miR-423, as regulators of osteoblastogenesis. In addition, we tested the possible targeting of miRNAs for enhancing bone tissue regeneration. Scaffolds functionalized with miRNA nano-carriers enhanced osteoblastogenesis in 3D culture and retained this ability at least 2 weeks after storage. Additionally, anti-miR-222 enhanced in vivo ectopic bone formation through targeting the cell-cycle inhibitor CDKN1B (cyclin-dependent kinase inhibitor 1B). A number of additional miRNAs exerted additive osteoinductive effects on BMSC differentiation, suggesting that pools of miRNAs delivered locally from an implanted scaffold can provide a promising approach for enhanced bone regeneration.

MiR-33a Controls hMSCS Osteoblast Commitment Modulating the Yap/Taz Expression Through EGFR Signaling Regulation

“…In our previous studies, we identified two miRNAs involved in bone regeneration: (i) miRNA-675-5p as a modulator of HIF-1α and Wnt/β-catenin signalings in human mesenchymal stromal cells (hMSCs) [6]; and (ii) miRNA-31-5p, a mechanosensitive miRNA involved in hMSCs hypoxia response [7]. Microarray analysis revealed that hMSCs and osteoblast cells showed strong differences in terms of miRNAs expression and relative signaling activation [8,9], such as epithelial-to-mesenchymal transition (EMT) pathway [9] and the epidermal growth factor (EGF) signaling [10], which are strongly correlated to hypoxia cell response during osteoblast differentiation [6].…”

“…In this study, we wanted to understand the role of the miR-33a family during hMSCs osteoblast commitment and their relative targets using hMSCs as a model of stromal cells, hMSCs maintained in osteogenic medium as a model of pre-osteoblast cells, and Nh-Ost as a model of osteoblast cells [8,9]. Starting from the recent evidence that displayed a different miRNAs expression profiling between hMSCs and Nh-Ost as well as from our previous studies in which we demonstrated the roles of miR-675-5p and miR-31-5p in hypoxia and cytoskeletal modulation during hMSCs osteoblast differentiation [6,7], we decided to investigate another important signaling involved in hMSCs osteoblast commitment, such as EMT, for which it has been demonstrated that miR-33a shows a different mRNAs interaction.…”

Single-cell high-content imaging parameters predict functional phenotype of cultured human bone marrow stromal stem cells

“…1,5,6 These markers are sensitive but not predictive of the differentiation capacity of the cells. 9,14 Several studies have identified a molecular signature for bone-forming hBM-MSCs by applying global analysis of gene expression, 9,15,16 miRNA expression, 17,18 or proteome analysis. 19,20 Although these approaches contribute significantly to understanding the biology and functions of hBM-MSCs, they are labor-intensive, use a large number of cells, and may not be easy to implement in a clinical setting.…”

Regional difference in microRNA regulation in the skull vault

“…We previously identified unrecognized differences in a potential gene regulatory network in mouse frontal bone and parietal bone . Recently, micro ribonucleic acids (miRNAs) have been reported to play regulatory roles in the skeletal homeostasis . MiRNA‐378 can fuel osteogenesis‐angiogenesis for bone regeneration, and miR‐335‐5p can down‐regulate DKK1to boost osteogenic differentiation, and the modification of miR‐335‐5p was sufficient to enhance calvarial bone regeneration and minimize bone defects .…”

“…6 Recently, micro ribonucleic acids (miRNAs) have been reported to play regulatory roles in the skeletal homeostasis. [13][14][15] MiRNA-378 can fuel osteogenesis-angiogenesis for bone regeneration, 16 and miR-335-5p can down-regulate DKK1to boost osteogenic differentiation, and the modification of miR-335-5p was sufficient to enhance calvarial bone regeneration and minimize bone defects. 17 Therefore, miRNAs can be potential therapeutic targets to treat the skeletal disorders, but the potential miRNAs in the frontal bone and parietal bone are still illusive.…”