tRNA‐derived small RNA changes in bone marrow stem cells under hypoxia and osteogenic conduction

Abstract

BackgroundTissue engineering using bone mesenchymal stem cells (BMSCs) transplantation is a promising therapeutic for bone regeneration. However, the effect of bone regeneration remains unsatisfactory due to the BMSCs' functional abnormality influenced by hypoxia. In this study, we attempt to explore the mechanism of osteogenic differentiation of BMSCs under hypoxic conditions from the perspective of non‐coding RNA regulation.MethodsThe study employed BMSCs obtained from healthy donors and simulated hypoxia using CoCl2 stimulation. High‐throughput sequencing technique was used to identify differential expression profiles of tRNA‐derived small RNA (tsRNA) in three experimental groups: BMSCs‐0d, BMSCs‐7d and BMSCs‐0d‐CoCl2. TargetScan and miRanda algorithms were used to determine tsRNA target genes, while Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis were employed for the prediction of biological functions. Real‐time reverse transcriptase‐polymerase chain reaction (Real‐time RT‐PCR) was carried out on four selected differentially expressed tsRNAs.ResultsAfter the osteogenic induction and CoCl2 stimulated separately, there were 19 tsRNAs differentially expressed in BMSCs, including 14 upregulated and five downregulated. According to the analysis of biological information, these tsRNAs may regulate 311 potential target genes and mainly enrich the pathways such as metabolic pathways, Wnt signalling pathway, osteoclast differentiation, cellular senescence and mTOR signalling pathway. The results of Real‐time RT‐PCR for 3′tiRNA‐41‐GlnTTG‐6, 3′tiRNA‐42‐LysTTT‐8, 5′tiRNA‐35‐CysACA‐1 and tRF3a‐AsnGTT‐9 were consistent with small RNA sequencing data.ConclusionWe discovered the tsRNA that changes the process of osteogenesis and hypoxia, which provides new targets for promoting survival and regeneration functions after BMSCs transplantation.

The Exosome‐Mediated Bone Regeneration: An Advanced Horizon Toward the Isolation, Engineering, Carrying Modalities, and Mechanisms

“…Oxygen tension, a pivotal part of the culture process, has been dramatically discussed in terms of cell behaviors over a long time. When the BMSCs are treated with an oxygen tension more similar to 1–2% of the physiological condition of bone marrow, [ 132,133 ] better proliferation, [ 134,135 ] and differentiation [ 136,137 ] performance of cells facilitated by exosomes were found. [ 138 ] However, the discrepancies of different treating times, various hypoxia concentrations, and diverse treatment modes in many articles put this vital consideration to a status of lacking consensus.…”